WorldWideScience

Sample records for monolithic stationary phase

  1. Thiol-yne Click Adamantane Monolithic Stationary Phase for Capillary Electrochromatography

    Institute of Scientific and Technical Information of China (English)

    Dao, Thi Thu Hien; Guerrouache, Mohamed; Carbonnier, Benjamin

    2012-01-01

    A porous crosslinked organic polymer based on N-acryloxysuccinimide (NAS) and ethylene dimethacrylate (EDMA) was prepared inside 75 μm i.d. fused silica capillary as functionalizable monolithic stationary phase for electrochromatographic applications. Succinimide groups on the monolith surface provide reactive sites able to re- act readily through standard electrophile-nucleophile chemistry. Propargylamine was used to prepare alkyne func- tionalized poly(NAS-co-EDMA). Onto this thiol-reactive polymer surface was grafted adamantane units via a photochemically-driven addition reaction. Chemical characterization was performed in situ after each synthetic step by means of Raman spectroscopy and grafting kinetics was investigated to ensure quantitative grafting of 1-adamantanethiol. The as-designed monolithic stationary phase exhibited typical reversed-phase separation mechanism as evidenced by the linear increase of the logarithm of retention factor of neutral aromatic solutes with the increase of the aqueous buffer content in the mobile phase.

  2. Silica-based polypeptide-monolithic stationary phase for hydrophilic chromatography and chiral separation.

    Science.gov (United States)

    Zhao, Licong; Yang, Limin; Wang, Qiuquan

    2016-05-13

    Glutathione (GSH)-, somatostatin acetate (ST)- and ovomucoid (OV)-functionalized silica-monolithic stationary phases were designed and synthesized for HILIC and chiral separation using capillary electrochromatography (CEC). GSH, ST and OV were covalently incorporated into the silica skeleton via the epoxy ring-opening reaction between their amino groups and the glycidyl moiety in γ-glycidoxypropyltrimethoxysilane (GPTMS) together with polycondensation and copolymerization of tetramethyloxysilane and GPTMS. Not only could the direction and electroosmotic flow magnitude on the prepared GSH-, ST- and OV-silica hybrid monolithic stationary phases be controlled by the pH of the mobile phase, but also a typical HILIC behavior was observed so that the nucleotides and HPLC peptide standard mixture could be baseline separated using an aqueous mobile phase without any acetonitrile during CEC. Moreover, the prepared monolithic columns had a chiral separation ability to separate dl-amino acids. The OV-silica hybrid monolithic column was most effective in chiral separation and could separate dl-glutamic acid (Glu) (the resolution R=1.07), dl-tyrosine (Tyr) (1.57) and dl-histidine (His) (1.06). Importantly, the chiral separation ability of the GSH-silica hybrid monolithic column could be remarkably enhanced when using gold nanoparticles (AuNPs) to fabricate an AuNP-mediated GSH-AuNP-GSH-silica hybrid monolithic column. The R of dl-Glu, dl-Tyr and dl-His reached 1.19, 1.60 and 2.03. This monolithic column was thus applied to separate drug enantiomers, and quantitative separation of all four R/S drug enantiomers were achieved with R ranging from 4.36 to 5.64. These peptide- and protein-silica monolithic stationary phases with typical HILIC separation behavior and chiral separation ability implied their promise for the analysis of not only the future metabolic studies, but also drug enantiomers recognition.

  3. Convective Interaction Media (CIM)--short layer monolithic chromatographic stationary phases.

    Science.gov (United States)

    Podgornik, Ales; Strancar, Ales

    2005-01-01

    Modern downstream processing requires fast and highly effective methods to obtain large quantities of highly pure substances. Commonly applied method for this purpose is chromatography. However, its main drawback is its throughput since purification, especially of large molecules, requires long process time. To overcome this problem several new stationary phases were introduced, among which short layer monoliths show superior properties for many applications. The purpose of this review is to give an overview about short methacrylate monolithic columns commercialised under the trademark Convective Interaction Media (CIM). Their unique properties are described from different perspectives, explaining reasons for their application on various areas. Approaches to prepare large volume methacrylate monolithic column are discussed and optimal solutions are given. Different examples of CIM monolithic column implementation are summarised in the last part of the article to give the reader an idea about their advantages.

  4. Preparation and evaluation of monolithic molecularly imprinted stationary phase for S-naproxen

    Institute of Scientific and Technical Information of China (English)

    De-Miao Chen; Qiang Fu; Wei Du; Si-Juan Sun; Ping Huang; Chun Chang

    2011-01-01

    An S-naproxen(S-NAP)molecularly imprinted monolithic stationary phase(MIMSP)with specific recognition for S-NAP and naproxen(NAP)was prepared by in situ technique,utilizing 4-vinylpridine(4-VP)as a function monomer,ethylene glycol dimethacrylate(EDMA)as a

  5. A weak cation-exchange monolith as stationary phase for the separation of peptide diastereomers by CEC.

    Science.gov (United States)

    Ludewig, Ronny; Nietzsche, Sandor; Scriba, Gerhard K E

    2011-01-01

    A CEC weak cation-exchange monolith has been prepared by in situ polymerization of acrylamide, methylenebisacrylamide and 4-acrylamidobutyric acid in a decanol-dimethylsulfoxide mixture as porogen. The columns were evaluated by SEM and characterized with regard to the separation of diastereomers and α/β-isomers of aspartyl peptides. Column preparation was reproducible as evidenced by comparison of the analyte retention times of several columns prepared simultaneously. Analyte separation was achieved using mobile phases consisting of acidic phosphate buffer and ACN. Under these conditions the peptides migrated due to their electrophoretic mobility but the EOF also contributed as driving force as a function of the pH of the mobile phase due to increasing dissociation of the carboxyl groups of the polymer. Raising the pH of the mobile phase also resulted in deprotonation of the peptides reducing analyte mobility. Due to these mechanisms each pair of diastereomeric peptides displayed the highest resolution at a different pH of the buffer component of the mobile phase. Comparing the weak-cation exchange monolith to an RP monolith and a strong cation-exchange monolith different elution order of some peptide diastereomers was observed, clearly illustrating that interactions with the stationary phase contribute to the CEC separations.

  6. Microchip free flow planar reversed phase electrochromatography with monolithic stationary phase.

    Science.gov (United States)

    Wang, Pingli; Tao, Dingyin; Zhang, Lihua; Liang, Zhen; Zhang, Yukui

    2009-08-01

    In this study, microchip free flow planar RP electrochromatography (microFF-PRPEC) was developed by in situ polymerization of monolithic materials in microchamber, and successfully applied for the separation of dyes and proteins. Poly(butyle methyacrylate-co-ethylene dimethacrylate) was prepared by UV-initiated polymerization in a glass microchamber (42 mm long, 23 mm wide, and 28 microm deep). A mixture of 1-propanol, 1,4-butanediol, and water was chosen as porogens, and 1.2% (wt%) 2-acrylamide-2-methyl-propanesulfonic acid (AMPS) was added into the polymerization solution to generate EOF. With 30% v/v ACN-15 mM Tris-HCl as the mobile phase, rhodamine B and methyl green were separated from each other with 400 V transverse voltage applied, and resolution as high as 4.6 was obtained, much higher than that obtained by microFFE under optimal conditions. Furthermore, microFF-PRPEC was also successfully applied into the separation of lysozyme and ribonuclease B, and resolution as high as 9.4 was obtained. All these results demonstrate that microFF-RPPEC might have great potential in the microscale continuous preparation of samples with improved resolution compared to microFFE.

  7. Monolithic stationary phases with incorporated fumed silica nanoparticles. Part I. Polymethacrylate-based monolithic column with incorporated bare fumed silica nanoparticles for hydrophilic interaction liquid chromatography.

    Science.gov (United States)

    Aydoğan, Cemil; El Rassi, Ziad

    2016-05-01

    Fumed silica nanoparticles (FSNPs), were incorporated for the first time into a polymethacrylate monolithic column containing glyceryl monomethacrylate (GMM) and ethylene dimethacrylate (EDMA) in order to develop a new monolithic column for hydrophilic interaction high performance liquid chromatography (HILIC). When compared to poly(GMM-EDMA) monolithic column without FSNPs, the same monolithic column with incorporated FSNPs yielded important effects on HILIC separations. The effects of monomers and FSNPs content of the polymerization mixture on the performance of the monolithic column were examined in details, and the optimized stationary phase was investigated over a wide range of mobile phase composition with polar acidic, weakly basic and neutral analytes including hydroxy benzoic acids, nucleotides, nucleosides, dimethylformamide, formamide and thiourea. The retention of these analytes was mainly controlled by hydrophilic interactions with the FSNPs and electrostatic repulsion from the negatively charged silica surface in the case of hydroxy benzoic acids and nucleotides. The electrostatic repulsion was minimized by decreasing the pH of the aqueous component of the mobile phase, which in turn enhanced the retention of acidic solutes. Nucleotides were best separated using step gradient elution at decreasing pH as well as ACN concentration in the mobile phase. Improved peak shape and faster analysis of nucleosides were attained by a fast linear gradient elution with a shallow decrease in the ACN content of the ACN-rich mobile phase. The run-to-run and column-to-column reproducibility were satisfactory. The percent relative standard deviations (%RSDs) for the retention times of tested solutes were lower than 2.5% under isocratic conditions and lower than 3.5 under gradient conditions.

  8. A new anion-exchange/hydrophobic monolith as stationary phase for nano liquid chromatography of small organic molecules and inorganic anions.

    Science.gov (United States)

    Aydoğan, Cemil

    2015-05-01

    In this study, an anion-exchange/hydrophobic polymethacrylate-based stationary phase was prepared for nano-liquid chromatography of small organic molecules and inorganic anions. The stationary phase was synthesized by in situ polymerization of 3-chloro-2-hydroxypropylmethacrylate and ethylene dimethacrylate inside silanized 100 μm i.d. fused silica capillary. The porogen mixture consisted of toluene and dodecanol. The pore size distrubution profiles of the resulting monolith were determined by mercury intrusion porosimetry and the morphology of the prepared monolith was investigated by scanning electron microscope. Good permeability, stability and column efficiency were observed on the monolithic column with nano flow. The produced monolithic column, which contains reactive chloro groups, was then modified by reaction with N,N-dimethyl-N-dodecylamine to obtain an anion-exchange/hydrophobic monolithic stationary phase. The functionalized monolith contained ionizable amine groups and hydrophobic groups that are useful of anion-exchange/hydrophobic mixed-mode chromatography. The final monolithic column performance with respect to anion-exchange and hydrophobic interactions was assesed by the separation of alkylbenzene derivatives, phenolic compounds and inorganic anions, respectively. Theoretical plate numbers up to 23,000 plates/m were successfully achieved in the separation of inorganic anions.

  9. Monolithic stationary phases with incorporated fumed silica nanoparticles. Part II. Polymethacrylate-based monolithic column with "covalently" incorporated modified octadecyl fumed silica nanoparticles for reversed-phase chromatography.

    Science.gov (United States)

    Aydoğan, Cemil; El Rassi, Ziad

    2016-05-01

    This study is concerned with the incorporation of surface modified fumed silica nanoparticles (FSNPs) into polymethacrylate based monolithic columns for use in reversed phase chromatography (RPC) of small solutes and proteins. First, FSNPs were modified with 3-(trimethoxysilyl)propylmethacrylate (TMSPM) to yield the "hybrid" methacryloyl fumed silica nanoparticle (MFSNP) monomer. The resulting MFSNP was then mixed with glyceryl monomethacrylate (GMM) and ethylene dimethacrylate (EDMA) in a binary porogenic solvent composed of cyclohexanol and dodecanol, and the in situ copolymerization of MFSNP, GMM and EDMA was performed in a stainless steel column of 4.6 mm i.d. The silanol groups of the hybrid monolith thus obtained were grafted with octadecyl ligands by perfusing the hybrid monolithic column with a solution of 4% w/v of dimethyloctadecylchlorosilane (DODCS) in toluene while the column was maintained at 110°C for 6h (in a heated HPLC oven). One of the originalities of this study was to demonstrate MFSNP as a novel derivatized "hybrid monomer" in making RPC monolithic columns with surface bound octadecyl ligands. In this respect, the RPC behavior of the monolithic column with "covalently" incorporated FNSPs having surface grafted octadecyl ligands was evaluated with alkylbenzenes, aniline derivatives and phenolic compounds. The results showed that the hybrid poly(GMA-EDMA-MFSNP) having surface bound octadecyl ligands exhibited hydrophobic interactions under reversed phase elution conditions. Furthermore, six standard proteins were baseline separated on the column using a 10min linear gradient elution at increasing ACN concentration in the mobile phase at a flow rate of 1.0mL/min using a 10 cm×4.6mm i.d. column. The relative standard deviations (RSDs) for the retention times of the tested solutes were lower than 2.1% and 2.4% under isocratic elution and gradient elution conditions, respectively.

  10. Porous Monoliths: Stationary Phases of Choice for High Performance Liquid Chromatography in Various Formats%用于高效液相色谱的不同形式的多孔整体固定相

    Institute of Scientific and Technical Information of China (English)

    SVEC Frantisek

    2005-01-01

    Modern porous monoliths have been conceived as a new class of stationary phases for high performance liquid chromatography (HPLC) in classical columns in the early 1990s and later extended to the capillary format. These monolithic materials are prepared using simple processes carried out in an external mold (inorganic monoliths) or within the confines of the column (organic monoliths and all capillary columns). These methods afford macroporous materials with large through-pores that enable applications in a rapid flow-through mode. Since all the mobile phase must flow through the monolith, the convection considerably accelerates mass transport within the monolithic separation medium and improves the separations. As a result, the monolithic columns perform well even at very high flow rates. The applications of monolithic capillary columns are demonstrated on numerous separations in the HPLC mode.

  11. Preparation and characterization of silica monolith modified with bovine serum albumin-gold nanoparticles conjugates and its use as chiral stationary phases for capillary electrochromatography.

    Science.gov (United States)

    Lu, Junyu; Ye, Fanggui; Zhang, Aizhu; Wei, Zong; Peng, Yan; Zhao, Shulin

    2011-08-01

    This paper describes the development of silica monolith modified with bovine serum albumin-gold nanoparticles (BSA-GNPs) conjugates as chiral stationary phases for capillary electrochromatography (CEC). The bare monolithic silica column was prepared by a sol-gel process and has been modified chemically with 3-mercaptopropyltrimethoxysilane to provide thiol groups, followed by immobilization of gold nanoparticles via the formation of an Au-S bond and modification with BSA as the chiral selector via the nitrogen lone pair of electrons. It has been demonstrated that the monolithic chiral stationary phases can be used for the enantioseparation of a number of phenylthiocarbamyl amino acids (PTC-D/L-AAs) by CEC. Ten pairs of tested amino acids enantiomers were successfully resolved within 18 min under optimized conditions, and the resolution values were in the range of 1.486-2.083. With PTC-D/L-tryptophan used as the probe solute, the influences of applied voltage, organic modifier and buffer pH in mobile phase on apparent retention factor, enantioselectivity and resolution factor were also investigated.

  12. Penicillin analyses by capillary electrochromatography-mass spectrometry with different charged poly(stearyl methacrylate-divinylbenzene) monoliths as stationary phases.

    Science.gov (United States)

    Liu, Wan-Ling; Wu, Ching-Yi; Li, Yi-Ting; Huang, Hsi-Ya

    2012-11-15

    This study describes the ability of an on-line concentration capillary electrochromatography (CEC) coupled with mass spectrometry (MS) for the determination of eight common penicillin antibiotics. Poly(stearyl methacrylate-divinylbenzene) (poly(SMA-DVB)) based monolithic columns prepared under the same conditions but differing only in the charged monomer were used as separation columns. Vinylbenzyl trimethylammonium chloride (VBTA) and vinylbenzenesulfonate (VBSA) were employed as the positively charged monolith and negatively charged monolith, respectively. Results indicated that poly(SMA-DVB-VBTA) monolithic column provided reproducible performance for penicillin separation through ion-exchange interaction, while the negatively charged poly(SMA-DVB-VBSA) column produced unstable separation due to the electrostatic repulsion between the electrophilic analytes and the negatively charged stationary phase. On-line concentration steps of step-gradient elution combined with anion selective injection (ASEI) were used to enhance the detection sensitivity of the CEC-MS method and all penicillin detection sensitivities were further improved (reduction in the limits of detection from 1.9-31 μg/L (normal injection mode) to 0.05-0.2 μg/L (on-line concentration mode)). Finally, this optimal on-line concentration CEC-MS method was applied to trace penicillin analyses in milk samples.

  13. Preparation of low flow-resistant methacrylate-based monolithic stationary phases of different hydrophobicity and the application to rapid reversed-phase liquid chromatographic separation of alkylbenzenes at high flow rate and elevated temperature.

    Science.gov (United States)

    Ueki, Yuji; Umemura, Tomonari; Iwashita, Yoshikazu; Odake, Tamao; Haraguchi, Hiroki; Tsunoda, Kin-ichi

    2006-02-17

    Low flow-resistant alkyl methacrylate-based monolithic stationary phases of different hydrophobicity were constructed for reversed-phase capillary liquid chromatography by thermally initiated radical polymerization of respective methacrylate ester monomer with different alkyl chain (C2, C4, C6, C12, C18) and ethylene glycol dimethacrylate (EDMA) in a 250 microm i.d. fused silica capillary. The hydrophobicity was basically controlled by changing the length and/or the density of the alkyl-chain, while the composition and the ratio of porogenic solvent were adjusted to obtain highly permeable rigid monoliths with adequate column efficiency. Among the prepared monolithic stationary phases, C18-methacrylate monoliths polymerized from a binary porogenic solvent of isoamyl alcohol and 1,4-buthandiol exhibited the most promising performance in terms of hydraulic resistance and column efficiency. The pressure drops of 20-cm long monolithic columns were below approximately 0.4 MPa at a normal linear velocity of 1mm/s (a flow rate of 3 microL/min), and the numbers of theoretical plates for alkylbenzenes mostly exceeded 3000 plates/20 cm. The produced monolithic columns had good mechanical strength for high pressure and temperature, and could be properly operated even at a temperature of 80 degrees C and at a pressure of at least 33 MPa. At 80 degrees C, the theoretical plate numbers reached 6000 plates/20 cm because of the enhanced mass transfer. Due to the novel hydraulic resistance and mechanical strength, the separation time could be reduced 120-fold simply by raising the flow rate and column temperature.

  14. Monolithic poly (SPE-co-BVPE) capillary columns as a novel hydrophilic interaction liquid chromatography stationary phase for the separation of polar analytes.

    Science.gov (United States)

    Foo, Hsiao Ching; Heaton, James; Smith, Norman W; Stanley, Shawn

    2012-10-15

    A novel hydrophilic interaction liquid chromatography (HILIC) stationary phase was prepared by the co-polymerisation of zwitterionic N,N'-dimethyl-N-methacryloxyethyl-N-(3-sulfopropyl) ammonium betaine (SPE) and the crosslinker 1,2-bis(p-vinylphenyl) ethane (BVPE) in the presence of the porogens, toluene and methanol. Monolithic columns were produced by carrying out the α,α'-azoisobutyronitrile (AIBN) initiated reaction for 1, 2, 4, 8 and 12 h inside a 200 μm i.d. fused silica capillary at 75°C (water bath). The optimum polymerisation time was shown to be 2 h, as this resulted in good porosity, due to enlarged flow-channels and the presence of a higher proportion of mesopores provided a relatively larger surface area than the other columns. The chromatographic properties of the optimised poly (SPE-co-BVPE) monolithic column were evaluated with test mixtures containing both basic and neutral compounds in the HILIC gradient separation mode. This produced relatively sharp peaks (average peak width at half height=0.1 min) with average asymmetry factors of 1.4 and baseline resolution was obtained for all the compounds. Using the isocratic separation of the test mixture, the number of theoretical plates (N) per metre calculated was between 26,888 and 35,930 by using average values obtained for triplicate injections of the compounds thiourea, toluene and acrylamide.

  15. Development and validation of a rapid stability indicating HPLC-method using monolithic stationary phase and two spectrophotometric methods for determination of antihistaminic acrivastine in capsules

    Science.gov (United States)

    Gouda, Ayman A.; Hashem, Hisham; Jira, Thomas

    2014-09-01

    Simple, rapid and accurate high performance liquid chromatographic (HPLC) and spectrophotometric methods are described for determination of antihistaminic acrivastine in capsules. The first method (method A) is based on accurate, sensitive and stability indicating chromatographic separation method. Chromolith® Performance RP-18e column, a relatively new packing material consisting of monolithic rods of highly porous silica, was used as stationary phase applying isocratic binary mobile phase of ACN and 25 mM NaH2PO4 pH 4.0 in the ratio of 22.5:77.5 at flow rate of 5.0 mL/min and 40 °C. A diode array detector was used at 254 nm for detection. The elution time of acrivastine was found to be 2.080 ± 0.032. The second and third methods (methods B and C) are based on the oxidation of acrivastine with excess N-bromosuccinimide (NBS) and determination of the unconsumed NBS with, metol-sulphanilic acid (λmax: 520 nm) or amaranth dye (λmax: 530 nm). The reacted oxidant corresponds to the drug content. Beer’s law is obeyed over the concentration range 1.563-50, 2.0-20 and 1.0-10 μg mL-1 for methods A, B and C, respectively. The limits of detection and quantitation were 0.40, 0.292 and 0.113 μg mL-1 and 0.782, 0.973 and 0.376 μg mL-1 for methods A, B and C, respectively. The HPLC method was validated for system suitability, linearity, precision, limits of detection and quantitation, specificity, stability and robustness. Stability tests were done through exposure of the analyte solution for four different stress conditions and the results indicate no interference of degradants with HPLC-method. The proposed methods was favorably applied for determination of acrivastine in capsules formulation. Statistical comparison of the obtained results from the analysis of the studied drug to those of the reported method using t- and F-tests showed no significant difference between them.

  16. Automated pre-column derivatization of thiolic fruit-antibrowning agents by sequential injection coupled to high-performance liquid chromatography using a monolithic stationary phase and an in-loop stopped-flow approach.

    Science.gov (United States)

    Karakosta, Theano D; Tzanavaras, Paraskevas D; Themelis, Demetrius G

    2011-08-01

    The present study reports the very first application of ethyl propiolate (EP) as an advantageous pre-column derivatization reagent for the determination of thiols by liquid chromatography (LC). Cysteine (CYS), glutathione (GSH) and N-acetylcysteine (NAC) were derivatized online under stopped-flow conditions in a sequential injection (SI) system coupled to HPLC. The formed derivatives were separated isocratically with a monolithic stationary phase (100×4.6 mm id) and UV detected at 285 nm. Critical parameters that affected the online pre-column derivatization reaction (e.g. the reaction time and the amount concentration of EP) and the separation (e.g. pH and the composition of the mobile phase) were investigated. The developed analytical scheme offers a total analysis time of less than 10 min, limits of detection in the range of 0.24-0.35 μmol/L and satisfactory linearity up to 200 μmol/L for all analytes. The proposed method was applied to the analysis of the selected thiols--that are often employed as antibrowning agents--in fresh fruit samples.

  17. Separation of intact proteins on γ-ray-induced polymethacrylate monolithic columns: A highly permeable stationary phase with high peak capacity for capillary high-performance liquid chromatography with high-resolution mass spectrometry.

    Science.gov (United States)

    Simone, Patrizia; Pierri, Giuseppe; Foglia, Patrizia; Gasparrini, Francesca; Mazzoccanti, Giulia; Capriotti, Anna Laura; Ursini, Ornella; Ciogli, Alessia; Laganà, Aldo

    2016-01-01

    Polymethacrylate-based monolithic capillary columns, prepared by γ-radiation-induced polymerization, were used to optimize the experimental conditions (nature of the organic modifiers, the content of trifluoroacetic acid and the column temperature) in the separation of nine standard proteins with different hydrophobicities and a wide range of molecular weights. Because of the excellent permeability of the monolithic columns, an ion-pair reversed-phase capillary liquid chromatography with high-resolution mass spectrometry method has been developed by coupling the column directly to the mass spectrometer without a flow-split and using a standard electrospray interface. Additionally, the high working flow and concomitant high efficiency of these columns allowed us to employ a longer column (up to 50 cm) and achieve a peak capacity value superior to 1000. This work is motivated by the need to develop new materials for high-resolution chromatographic separation that combine chemical stability at elevated temperatures (up to 75°C) and a broad pH range, with a high peak capacity value. The advantage of the γ-ray-induced monolithic column lies in the batch-to-batch reproducibility and long-term high-temperature stability. Their proven high loading capacity, recovery, good selectivity and high permeability, moreover, compared well with that of a commercially available poly(styrene-divinylbenzene) monolithic column, which confirms that such monolithic supports might facilitate analysis in proteomics.

  18. Construção de câmara de luz ultravioleta para fotopolimerização de fases estacionárias monolíticas Coinstruction of ultraviolet-light chamber for monolithic stationary phases photo-polymerization

    Directory of Open Access Journals (Sweden)

    Fernando Antonio Simas Vaz

    2008-01-01

    Full Text Available The assembly of a photochemical reactor with six fluorescent lamps, used for photopolymerizations is described. This chamber presents a mobile support, allowing the placement of samples at different heights and a safety lock that interrupts the radiation, if it is opened during operation. The mirrored internal walls avoid the dispersion and non-uniform distribution of light. There is no high heating because the own character of the used lamps. All parts could be purchased in commerce with less than U$ 150,00. This reactor was successfully used for monolithic stationary phase photopolymerization.

  19. Stationary phase deposition based on onium salts

    Science.gov (United States)

    Wheeler, David R.; Lewis, Patrick R.; Dirk, Shawn M.; Trudell, Daniel E.

    2008-01-01

    Onium salt chemistry can be used to deposit very uniform thickness stationary phases on the wall of a gas chromatography column. In particular, the stationary phase can be bonded to non-silicon based columns, especially microfabricated metal columns. Non-silicon microfabricated columns may be manufactured and processed at a fraction of the cost of silicon-based columns. In addition, the method can be used to phase-coat conventional capillary columns or silicon-based microfabricated columns.

  20. Monolithic aerogels with nanoporous crystalline phases

    Science.gov (United States)

    Daniel, Christophe; Guerra, Gaetano

    2015-05-01

    High porosity monolithic aerogels with nanoporous crystalline phases can be obtained from syndiotactic polystyrene and poly(2,6-dimethyl-1,4-phenylene)oxide thermoreversible gels by removing the solvent with supercritical CO2. The presence of crystalline nanopores in the aerogels based on these polymers allows a high uptake associated with a high selectivity of volatile organic compounds from vapor phase or aqueous solutions even at very low activities. The sorption and the fast kinetics make these materials particularly suitable as sorption medium to remove traces of pollutants from water and moist air.

  1. Incorporation of ionic liquid into porous polymer monoliths to enhance the separation of small molecules in reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Wang, Jiafei; Bai, Ligai; Wei, Zhen; Qin, Junxiao; Ma, Yamin; Liu, Haiyan

    2015-06-01

    An ionic liquid was incorporated into the porous polymer monoliths to afford stationary phases with enhanced chromatographic performance for small molecules in reversed-phase high-performance liquid chromatography. The effect of the ionic liquid in the polymerization mixture on the performance of the monoliths was studied in detail. While monoliths without ionic liquid exhibited poor resolution and low efficiency, the addition of ionic liquid to the polymerization mixture provides highly increased resolution and high efficiency. The chromatographic performances of the monoliths were demonstrated by the separations of various small molecules including aromatic hydrocarbons, isomers, and homologues using a binary polar mobile phase. The present column efficiency reached 27 000 plates/m, which showed that the ionic liquid monoliths are alternative stationary phases in the separation of small molecules by high-performance liquid chromatography.

  2. Carbon nanotube stationary phases for microchip electrochromatography

    DEFF Research Database (Denmark)

    Mogensen, Klaus Bo; Bøggild, Peter; Kutter, Jörg Peter

    nanotubes are very interesting for integration in especially microfluidic devices, because they can readily be grown on planar substrates by means of chemical vapour deposition. In this way the cumbersome process of packing of the stationary phase in the finished microfluidic channels is avoided and the CNT......, microfluidic devices with microfabricated carbon nanotube columns for electrochromatographic separations will be presented. The electrically conductive carbon nanotube layer has been patterned into hexoganol micropillars in order to support electroosmotic flow without forming gas bubbles from electrolysis...

  3. Stationary phases for superheated water chromatography

    CERN Document Server

    Saha, S

    2002-01-01

    This project focused on the comparison of conventional liquid chromatography and superheated water chromatography. It examined the differences in efficiency and retention of a range of different stationary phases. Alkyl aryl ketones and eight aromatic compounds were separated on PBD-zirconia, Xterra RP 18, Luna C sub 1 sub 8 (2) and Oasis HLB columns using conventional LC and superheated water chromatography system. The retention indices were determined in the different eluents. On changing the organic component of the eluent from methanol to acetonitrile to superheated water considerable improvements were found in the peak shapes and column efficiencies on the PBD-zirconia and Oasis HLB columns. PS-DVB, PBD-zirconia and Xterra RP 18 columns have been used in efficiency studies. It was found that simply elevating the column temperature did not increase the efficiency of a separation in superheated water chromatography. The efficiency depended on flow rate, injection volume and also mobile phase preheating sys...

  4. Stationary phase optimized selectivity supercritical fluid chromatography (SOS-SFC)

    OpenAIRE

    Delahaye, Sander; Lynen, Frederic

    2013-01-01

    In stationary phase optimized selectivity liquid chromatography (SOS-LC) the stationary phase becomes a tunable parameter by connecting column segments with variable lengths of different stationary phases. An optimization procedure and algorithm based on the PRISMA model for optimization of the mobile phase in LC was developed to apply this strategy for isocratic and gradient separations. An optimized column segment combination, giving the highest separation selectivity for all compounds in a...

  5. Fast, high-efficiency peptide separations on a 50-mu m reversed-phase silica monolith in a nanoLC-MS set-up

    NARCIS (Netherlands)

    Rieux, L; Lubda, D; Niederlander, HAG; Verpoorte, E; Bischoff, R

    2006-01-01

    Proteomic studies have stimulated the development of novel stationary phases in miniaturised chromatographic columns that permit high linear flow velocities and exhibit high resolving power. In this work, a 50-mu m reversed-phase silica-based monolith was chromatographically characterised for its us

  6. A Ferrite LTCC-Based Monolithic SIW Phased Antenna Array

    KAUST Repository

    Nafe, Ahmed

    2016-11-17

    In this work, we present a novel configuration for realizing monolithic SIW-based phased antenna arrays using Ferrite LTCC technology. Unlike the current common schemes for realizing SIW phased arrays that rely on surface-mount component (p-i-n diodes, etc) for controlling the phase of the individual antenna elements, here the phase is tuned by biasing of the ferrite filling of the SIW. This approach eliminates the need for mounting of any additional RF components and enables seamless monolithic integration of phase shifters and antennas in SIW technology. As a proof of concept, a two-element slotted SIW-based phased array is designed, fabricated and measured. The prototype exhibits a gain of 4.9 dBi at 13.2 GHz and a maximum E-plane beam-scanning of 28 degrees using external windings for biasing the phase shifters. Moreover, the array can achieve a maximum beam-scanning of 19 degrees when biased with small windings that are embedded in the package. This demonstration marks the first time a fully monolithic SIW-based phased array is realized in Ferrite LTCC technology and paves the way for future larger-size implementations.

  7. Polymethacrylate monolithic and hybrid particle-monolithic columns for reversed-phase and hydrophilic interaction capillary liquid chromatography.

    Science.gov (United States)

    Jandera, Pavel; Urban, Jirí; Skeríková, Veronika; Langmaier, Pavel; Kubícková, Romana; Planeta, Josef

    2010-01-01

    We prepared hybrid particle-monolithic polymethacrylate columns for micro-HPLC by in situ polymerization in fused silica capillaries pre-packed with 3-5microm C(18) and aminopropyl silica bonded particles, using polymerization mixtures based on laurylmethacrylate-ethylene dimethacrylate (co)polymers for the reversed-phase (RP) mode and [2-(methacryloyloxy)ethyl]-dimethyl-(3-sulfopropyl) zwitterionic (co)polymers for the hydrophilic interaction (HILIC) mode. The hybrid particle-monolithic columns showed reduced porosity and hold-up volumes, approximately 2-2.5 times lower in comparison to the pure monolithic columns prepared in the whole volume of empty capillaries. The elution volumes of sample compounds are also generally lower in comparison to packed or pure monolithic columns. The efficiency and permeability of the hybrid columns are intermediate in between the properties of the reference pure monolithic and particle-packed columns. The chemistries of the embedded solid particles and of the interparticle monolithic moiety in the hybrid capillary columns contribute to the retention to various degrees, affecting the selectivity of separation. Some hybrid columns provided improved separations of proteins in comparison to the reference particle-packed columns in the reversed-phase mode. Zwitterionic hybrid particle-monolithic columns show dual mode retention HILIC/RP behaviour depending on the composition of the mobile phase and allow separations of polar compounds such as phenolic acids in the HILIC mode at lower concentrations of acetonitrile and, often in shorter analysis time in comparison to particle-packed and full-volume monolithic columns.

  8. Retention of Halogenated Solutes on Stationary Phases Containing Heavy Atoms

    Directory of Open Access Journals (Sweden)

    Toshio Miwa

    2013-05-01

    Full Text Available To examine the effects of weak intermolecular interactions on solid-phase extraction (SPE and chromatographic separation, we synthesized some novel stationary phases with a heavy atom effect layer by immobilizing halogenated aromatic rings and hydroxyl groups onto the surface of a hydrophilic base polymer. Using SPE cartridges packed with the functionalized materials, we found that the heavy atom stationary phases could selectively retain halophenols in organic solvents, such as 1-propanol which blocks the hydrogen bonding, or acetonitrile which blocks the p-p interaction. The extraction efficiency of the materials toward the halophenols depended on the dipole moments of phenoxy groups present as functional groups. On the other hand, the extraction efficiency of solutes toward the functional group depended on their molar refractions, i.e., induced dipole moments. The retention of the solutes to the stationary phase ultimately depended on not only strong intermolecular interactions, but also the effects of weak interactions such as the dispersion force.

  9. Effects of pressure and frictional heating on protein separation using monolithic columns in reversed-phase chromatography.

    Science.gov (United States)

    Mann, Benjamin F; Makarov, Alexey A; Wang, Heather; Welch, Christopher J

    2017-03-17

    Pressure is not typically controlled or adjusted independently of flow rate during method development in reversed-phase LC (RPLC). However, it has been shown that pressure has an effect on analyte molecular molar volume, and the magnitude of this effect is greater for proteins and ionizable compounds than neutral small molecules. This phenomenon has received attention recently in the context of porous sub-2-micron particle packed columns. The present study surveys the effect of pressure and frictional heating on RPLC separations using commercially-available monolithic columns at constant flow rate and with controlled external temperature. Because the current monoliths cannot be operated at high pressures, all experiments were conducted with pressures at or below 200bar. Nonetheless, substantial changes in retention were still observed; for example, an increase in pressure of 75bar shifted the retention factor for bovine insulin from 1.27 to 1.78, a 40% increase, while a similar experiment with the neutral small molecule, toluene, showed no change in retention. Results are presented from investigations of model peptides and proteins ranging in size from 1kDa to 30kDa, as well as experiments performed with a silica-based C18 monolith and a polystyrene divinylbenzene monolith functionalized with a phenyl stationary phase. This work indicates that protein separations in monoliths are highly pressure sensitive, and pressure should therefore be considered as an additional parameter in method development for optimizing retention and selectivity. Given these findings, and the ever-increasing importance of chromatographic separations of proteins in both industrial and academic laboratories, improved instrumentation and mechanisms for directly controlling system backpressure could be of great practical value. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Luciferase detection during stationary phase in Lactococcus lactis

    NARCIS (Netherlands)

    Bachmann, H.; Santos, dos F.; Kleerebezem, M.; Hylckama Vlieg, van J.E.T.

    2007-01-01

    The luminescence signal of luxAB-encoded bacterial luciferase strongly depends on the metabolic state of the host cell, which restricts the use of this reporter system to metabolically active bacteria. Here we show that in stationary-phase cells of Lactococcus lactis, detection of luciferase is sign

  11. Ultrarapid quantitation of maize proteins by perfusion and monolithic reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Rodríguez-Nogales, J M; del Alamo, M; García, M C; Cifuentes, A; Marina, M L

    2009-04-22

    The main objective of this study was to develop a new methodology alternative to the classical Kjeldahl analysis for determining maize proteins in maize products and seeds. For that purpose, two different chromatographic methodologies using perfusion and monolithic stationary phases, both enabling rapid separations of maize proteins, were investigated. Due to the difficulty to find suitable standards for this type of analysis, three different maize products were initially tested as proteins standards: zein F4000, corn gluten meal, and maize flour. Different figures of merit (i.e., linearity, correlation coefficient, precision, limits of detection and quantitation), as well as the presence of matrix inferences, were investigated. The results obtained for the different chromatographic stationary phases and protein standards were compared in order to select the most suitable analytical conditions. Despite both perfusion and monolithic methodologies resulting, in general, as appropriate for the quantitation of maize proteins, the highest reduction of analysis time and lowest detection and determination limits provided by perfusion methodology enabled to select this one as the method of choice for the quantitation of maize proteins. Regarding the different protein standards studied in this work, in general the best results were obtained using the zein standard. Compared to Kjeldahl methodology, perfusion chromatography yields total protein contents in shorter analysis time while enabling the separation of the different kinds of proteins. Due to the high diversity and complexity of industrial maize products, the proposed chromatographic method could be a very useful tool for their routine analysis.

  12. Gas chromatography on wall-coated open-tubular columns with ionic liquid stationary phases.

    Science.gov (United States)

    Poole, Colin F; Lenca, Nicole

    2014-08-29

    Ionic liquids have moved from novel to practical stationary phases for gas chromatography with an increasing portfolio of applications. Ionic liquids complement conventional stationary phases because of a combination of thermophysical and solvation properties that only exist for ionic solvents. Their high thermal stability and low vapor pressure makes them suitable as polar stationary phases for separations requiring high temperatures. Ionic liquids are good solvents and can be used to expand the chemical space for separations. They are the only stationary phases with significant hydrogen-bond acidity in common use; they extend the hydrogen-bond basicity of conventional stationary phases; they are as dipolar/polarizable as the most polar conventional stationary phases; and some ionic liquids are significantly less cohesive than conventional polar stationary phases. Problems in column coating techniques and related low column performance, column activity, and stationary phase reactivity require further exploration as the reasons for these features are poorly understood at present.

  13. Nonequilibrium stationary states and phase transitions in directed Ising models

    Science.gov (United States)

    Godrèche, Claude; Bray, Alan J.

    2009-12-01

    We study the nonequilibrium properties of directed Ising models with non-conserved dynamics, in which each spin is influenced by only a subset of its nearest neighbours. We treat the following models: (i) the one-dimensional chain; (ii) the two-dimensional square lattice; (iii) the two-dimensional triangular lattice and (iv) the three-dimensional cubic lattice. We raise and answer the question: (a) under what conditions is the stationary state described by the equilibrium Boltzmann-Gibbs distribution? We show that, for models (i), (ii) and (iii), in which each spin 'sees' only half of its neighbours, there is a unique set of transition rates, namely with exponential dependence in the local field, for which this is the case. For model (iv), we find that any rates satisfying the constraints required for the stationary measure to be Gibbsian should satisfy detailed balance, ruling out the possibility of directed dynamics. We finally show that directed models on lattices of coordination number z>=8 with exponential rates cannot accommodate a Gibbsian stationary state. We conjecture that this property extends to any form of the rates. We are thus led to the conclusion that directed models with Gibbsian stationary states only exist in dimensions one and two. We then raise the question: (b) do directed Ising models, augmented by Glauber dynamics, exhibit a phase transition to a ferromagnetic state? For the models considered above, the answers are open problems, with the exception of the simple cases (i) and (ii). For Cayley trees, where each spin sees only the spins further from the root, we show that there is a phase transition provided the branching ratio, q, satisfies q>=3.

  14. Stationary phase optimized selectivity liquid chromatography: Basic possibilities of serially connected columns using the "PRISMA" principle.

    Science.gov (United States)

    Nyiredy, Sz; Szucs, Zoltán; Szepesy, L

    2007-07-20

    A new procedure (stationary phase optimized selectivity liquid chromatography: SOS-LC) is described for the optimization of the HPLC stationary phase, using serially connected columns and the principle of the "PRISMA" model. The retention factors (k) of the analytes were determined on three different stationary phases. By use of these data the k values were predicted applying theoretically combined stationary phases. These predictions resulted in numerous intermediate theoretical separations from among which only the optimal one was assembled and tested. The overall selectivity of this separation was better than that of any individual base stationary phase. SOS-LC is independent of the mechanism and the scale of separation.

  15. Microstructure-based analysis and simulation of flow and mass transfer in chromatographic stationary phases

    Science.gov (United States)

    Koku, Harun

    Limitations of mass transfer in chromatographic bioseparations employing traditional packed particles have fuelled the inception and development of alternative stationary phases with improved performance characteristics. This work investigates case studies in two categories of these alternative media, namely polymer-modified packed particles and continuous monolithic phases, for insight into their enhanced properties. Specifically, high-resolution microscopy techniques and image-based analysis algorithms were implemented to extract morphology information for these materials, in an attempt to elucidate the relation between microstructure and performance. For the monolith, mesoscopic simulation methods were also employed for a more rigorous analysis of the flow and dispersion behavior. Scanning and transmission electron microscopy images of the commercial polymer-modified, agarose-based particle Sepharose XL were compared to those for its unmodified counterpart, Sepharose FF. Local regions in the composite dextran-agarose Sepharose XL particles were noted to exhibit a denser network of fibers and smaller pore sizes overall, compared to those in the traditional Sepharose FF particles. Images of particles equilibrated with high concentrations of protein revealed a significant difference in protein localization patterns, with the stained protein in XL occupying a markedly higher area fraction of the images. This suggests a higher volume available for adsorption and provides visual clues into how the consistently higher static capacity of these polymer-modified particles is manifested. Treatment of the XL particles with dextranase, an enzyme that breaks down dextran, resulted in a reduction of protein coverage, providing evidence that it is indeed the dextran that is responsible for the improved static capacity in this polymer-modified stationary phase. Imaging and image analysis techniques were also used to analyze the commercial CIM(TM) disk monolith. Two- and three

  16. Monolithically integrated heterodyne optical phase-lock loop with RF XOR phase detector.

    Science.gov (United States)

    Steed, Robert J; Pozzi, Francesca; Fice, Martyn J; Renaud, Cyril C; Rogers, David C; Lealman, Ian F; Moodie, David G; Cannard, Paul J; Lynch, Colm; Johnston, Lilianne; Robertson, Michael J; Cronin, Richard; Pavlovic, Leon; Naglic, Luka; Vidmar, Matjaz; Seeds, Alwyn J

    2011-10-10

    We present results for an heterodyne optical phase-lock loop (OPLL), monolithically integrated on InP with external phase detector and loop filter, which phase locks the integrated laser to an external source, for offset frequencies tuneable between 0.6 GHz and 6.1 GHz. The integrated semiconductor laser emits at 1553 nm with 1.1 MHz linewidth, while the external laser has a linewidth less than 150 kHz. To achieve high quality phase locking with lasers of these linewidths, the loop delay has been made less than 1.8 ns. Monolithic integration reduces the optical path delay between the laser and photodiode to less than 20 ps. The electronic part of the OPLL was implemented using a custom-designed feedback circuit with a propagation delay of ~1 ns and an open-loop bandwidth greater than 1 GHz. The heterodyne signal between the locked slave laser and master laser has phase noise below -90 dBc/Hz for frequency offsets greater than 20 kHz and a phase error variance in 10 GHz bandwidth of 0.04 rad2.

  17. Stationary Optical Concentrator Designs and Wafer Scale Monolithic Integration of Semiconductor Devices for Next Generation Photovoltaic Panels

    Science.gov (United States)

    Kim, Jung Min

    A major barrier in utilizing solar energy for large scale deployment is the cost of the photovoltaic (PV) systems. Several approaches have been used for the cost reduction such as by modifying PV system designs in addition to enhancing the efficiency of solar cells. Due to the high cost of materials, minimizing the use of solar cells such as in concentrator type systems is highly attractive for reducing the cost of the PV modules by focusing the incident light onto the PV cell. However concentrator PV systems (CPV) require constant tracking of the sun and hence are complex in design and expensive to operate, except in limited situations such as large scale PV power plants. It is desirable to design new concentrator type systems that do not require continuous tracking of the sun. These systems could ultimately reduce the PV system cost to a minimum while maximizing the power conversion efficiency. In this thesis we propose a simple design for a stationary concentrator photovoltaic (SCPV) system that could significantly reduce the cost of generating electricity using PV devices. Using optical ray tracing simulations, we have been able to design SCPV systems that could reduce the PV module cost by 2--10 times without compromising on the power conversion efficiency of the system. Another alternative approach for sustainable high efficiency PV system design is to develop low cost PV cells for terrestrial applications. To meet the demands of low cost and large scale production, larger and thinner (or flexible) substrates are required. We demonstrated the feasibility of fabricating monolithic interconnected PV devices at the wafer scale (2 inch wafers). In this study, GaSb PV cells grown on semi-insulating GaAs were used as the model material. Crucial device fabrication steps such as a selective etching process have been developed that is necessary for isolating individual devices on the wafer and interconnecting them with sub-micron scale accuracy. Selective etching of

  18. Retention mechanism for polycyclic aromatic hydrocarbons in reversed-phase liquid chromatography with monomeric stationary phases.

    Science.gov (United States)

    Rafferty, Jake L; Siepmann, J Ilja; Schure, Mark R

    2011-12-23

    Reversed-phase liquid chromatography (RPLC) is the foremost technique for the separation of analytes that have very similar chemical functionalities, but differ only in their molecular shape. This ability is crucial in the analysis of various mixtures with environmental and biological importance including polycyclic aromatic hydrocarbons (PAHs) and steroids. A large amount of effort has been devoted to studying this phenomenon experimentally, but a detailed molecular-level description remains lacking. To provide some insight on the mechanism of shape selectivity in RPLC, particle-based simulations were carried out for stationary phases and chromatographic parameters that closely mimic those in an experimental study by Sentell and Dorsey [J. Chromatogr. 461 (1989) 193]. The retention of aromatic hydrocarbons ranging in size from benzene to the isomeric PAHs of the formula C(18)H(12) was examined for model RPLC systems consisting of monomeric dimethyl octadecylsilane (ODS) stationary phases with surface coverages ranging from 1.6 to 4.2 μmol/m(2) (i.e., stationary phases yielding low to intermediate shape selectivity) in contact with a 67/33 mol% acetonitrile/water mobile phase. The simulations show that the stationary phase acts as a very heterogeneous environment where analytes with different shapes prefer different spatial regions with specific local bonding environments of the ODS chains. However, these favorable retentive regions cannot be described as pre-existing cavities because the chain conformation in these local stationary phase regions adapts to accommodate the analytes.

  19. Facile synthesis of gradient mesoporous carbon monolith based on polymerization-induced phase separation

    Science.gov (United States)

    Xu, Shunjian; Luo, Yufeng; Zhong, Wei; Xiao, Zonghu; Luo, Yongping; Ou, Hui; Zhao, Xing-Zhong

    2014-06-01

    In this paper, a gradient mesoporous carbon (GMC) monolith derived from the mixtures of phenolic resin (PF) and ethylene glycol (EG) was prepared by a facile route based on polymerization-induced phase separation under temperature gradient (TG). A graded biphasic structure of PF-rich and EG-rich phases was first formed in preform under a TG, and then the preform was pyrolyzed to obtain the GMC monolith. The TG is mainly induced by the thermal resistance of the preferential phase separation layer at high temperature region. The pore structure of the monolith changes gradually along the TG direction. When the TG varies from 58°C to 29°C, the pore size, apparent porosity and specific surface area of the monolith range respectively from 18 nm to 83 nm, from 32% to 39% and from 140.5 m2/g to 515.3 m2/g. The gradient porous structure of the monolith is inherited from that of the preform, which depends on phase separation under TG in the resin mixtures. The pyrolysis mainly brings about the contraction of the pore size and wall thickness as well as the transformation of polymerized PF into glassy carbon.

  20. Monolithic Microwave Integrated Circuit (MMIC) Phased Array Demonstrated With ACTS

    Science.gov (United States)

    1996-01-01

    Monolithic Microwave Integrated Circuit (MMIC) arrays developed by the NASA Lewis Research Center and the Air Force Rome Laboratory were demonstrated in aeronautical terminals and in mobile or fixed Earth terminals linked with NASA's Advanced Communications Technology Satellite (ACTS). Four K/Ka-band experimental arrays were demonstrated between May 1994 and May 1995. Each array had GaAs MMIC devices at each radiating element for electronic beam steering and distributed power amplification. The 30-GHz transmit array used in uplinks to ACTS was developed by Lewis and Texas Instruments. The three 20-GHz receive arrays used in downlinks from ACTS were developed in cooperation with the Air Force Rome Laboratory, taking advantage of existing Air Force integrated-circuit, active-phased-array development contracts with the Boeing Company and Lockheed Martin Corporation. Four demonstrations, each related to an application of high interest to both commercial and Department of Defense organizations, were conducted. The location, type of link, and the data rate achieved for each of the applications is shown. In one demonstration-- an aeronautical terminal experiment called AERO-X--a duplex voice link between an aeronautical terminal on the Lewis Learjet and ACTS was achieved. Two others demonstrated duplex voice links (and in one case, interactive video links as well) between ACTS and an Army high-mobility, multipurpose wheeled vehicle (HMMWV, or "humvee"). In the fourth demonstration, the array was on a fixed mount and was electronically steered toward ACTS. Lewis served as project manager for all demonstrations and as overall system integrator. Lewis engineers developed the array system including a controller for open-loop tracking of ACTS during flight and HMMWV motion, as well as a laptop data display and recording system used in all demonstrations. The Jet Propulsion Laboratory supported the AERO-X program, providing elements of the ACTS Mobile Terminal. The successful

  1. Synthesis and characterization of novel stationary phases for small scale liquid chromatographic separations of proteins and nanoparticles

    Science.gov (United States)

    Hutanu, Daniela

    The emerging field of nanotechnology strictly requires the micro-scaling of the available separation technology and the design of novel devices for separations of molecules of interest. The separation of proteins and nanoparticles is challenging due to their relatively large size, non-specific adherence to surfaces and instability in many solvents. This dissertation presents the synthesis and characterization of novel stationary phases for use in separations of proteins or nanoparticles in both capillary and microchip formats. In order to separate blood proteins with high specificity, a DNA aptamer selected for alpha-thrombin was employed as an affinity component of the stationary phases. Silica surfaces and organic monoliths were modified with the aptamer via an azlactone linkage and have demonstrated highly efficient separations of thrombin from a mixture in the microscale. The high efficiency of the protein separation (HETP = 276 mum, RS = 1.7) is comparable with macroscale results using antibodies as the affinity factor. Novel hybrid inorganic-organic polysilsesquioxane stationary phases were synthesized by way of surfactant templated polymerization of bridged alcoxy-silyl ethane monomers, in presence of sodium hydroxide. The novel materials were successful in size exclusion separation of polystyrene standards with molecular diameters of 0.3-2.4 nm. A hybrid inorganic-organic polysilsesquioxane sorbent also proved useful for small scale separations of triphenyl phosphine protected gold nanoparticles, based on a sorptive mechanism instead of a size exclusion mechanism. Polysilsesquioxanes were easily synthesized in-situ inside fused silica capillary columns and PMMA microchip channels in order to facilitate integration with a micro-reactor. The novel stationary phases proved efficient for separation of proteins and nanoparticles in the micro-scale format and can further be utilized for online purification and separation of these difficult compounds.

  2. [Enantioseparation behavior of chiral stationary phases AD, AS and OD].

    Science.gov (United States)

    Li, Liqun; Fan, Jun; Zhang, Jing; Chen, Xiaodong; Wang, Tai; He, Jianfeng; Zhang, Weiguang

    2016-01-01

    Over the past decades, HPLC enantioseparation with chiral stationary phases (CSPs) has been widely applied in chiral analysis and preparation of new pharmaceuticals, pesticides, food, etc. Herein, enantioseparation of 20 chiral compounds have been carried out on three polysaccharide-based CSPs (EnantioPak AD, AS and OD) with normal phases by HPLC, separately. The influences of skeletal structure and the kinds of derivative groups on separation behaviors of these CSPs have been studied in detail. As results indicated, except for compound 13, the other compounds were baseline separated on EnantioPak AD, with most of resolution over 2. 0; in addition, better separation for acidic or basic compounds was achieved through adding acidic/basic additives into the mobile phase of hexane-alcohol. For four aromatic alcohols (compounds 13-16), their retention in the EnantioPak AD column showed a weakening tendency with increase of carbon number in side chain group, and the reverse trend of their resolution was observed. Furthermore, EnantioPak AD showed much better separation performance for eight compounds (13-20) than the others. In short, these results have provided some references for further investigation of separation behavior and applications of polysaccharide-based CSPs.

  3. Study of stationary phase metabolism via isotopomer analysis of amino acids from an isolated protein.

    Science.gov (United States)

    Shaikh, Afshan S; Tang, Yinjie J; Mukhopadhyay, Aindrila; Martín, Héctor García; Gin, Jennifer; Benke, Peter I; Keasling, Jay D

    2010-01-01

    Microbial production of many commercially important secondary metabolites occurs during stationary phase, and methods to measure metabolic flux during this growth phase would be valuable. Metabolic flux analysis is often based on isotopomer information from proteinogenic amino acids. As such, flux analysis primarily reflects the metabolism pertinent to the growth phase during which most proteins are synthesized. To investigate central metabolism and amino acids synthesis activity during stationary phase, addition of fully (13)C-labeled glucose followed by induction of green fluorescent protein (GFP) expression during stationary phase was used. Our results indicate that Escherichia coli was able to produce new proteins (i.e., GFP) in the stationary phase, and the amino acids in GFP were mostly from degraded proteins synthesized during the exponential growth phase. Among amino acid biosynthetic pathways, only those for serine, alanine, glutamate/glutamine, and aspartate/asparagine had significant activity during the stationary phase.

  4. Study of Stationary Phase Metabolism Via Isotopomer Analysis of Amino Acids from an Isolated Protein

    Energy Technology Data Exchange (ETDEWEB)

    Shaikh, AfshanS.; Tang, YinjieJ.; Mukhopadhyay, Aindrila; Martin, Hector Garcia; Gin, Jennifer; Benke, Peter; Keasling, Jay D.

    2009-09-14

    Microbial production of many commercially important secondary metabolites occurs during stationary phase, and methods to measure metabolic flux during this growth phase would be valuable. Metabolic flux analysis is often based on isotopomer information from proteinogenic amino acids. As such, flux analysis primarily reflects the metabolism pertinent to the growth phase during which most proteins are synthesized. To investigate central metabolism and amino acids synthesis activity during stationary phase, addition of fully 13C-labeled glucose followed by induction of green fluorescent protein (GFP) expression during stationary phase was used. Our results indicate that Escherichia coli was able to produce new proteins (i.e., GFP) in the stationary phase, and the amino acids in GFP were mostly from degraded proteins synthesized during the exponential growth phase. Among amino acid biosynthetic pathways, only those for serine, alanine, glutamate/glutamine, and aspartate/asparagine had significant activity during the stationary phase.

  5. Fabrication of Porous Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate Monoliths via Thermally Induced Phase Separation

    Directory of Open Access Journals (Sweden)

    Takashi Tsujimoto

    2016-02-01

    Full Text Available This study deals with the fabrication of biodegradable porous materials from bacterial polyester, poly(3-hydroxybutyrate-co-3-hydroxyhexanoate (P3HB3HHx, via thermally induced phase separation. P3HB3HHx monoliths with topological porous structure were prepared by dissolution of P3HB3HHx in dimethyl sulfoxide (DMSO at 85 °C and subsequent quenching. The microstructure of the resulting P3HB3HHx monoliths was changed by the P3HB3HHx concentration of the polymer solution. Differential scanning calorimetry and polarized optical microscope analysis revealed that the P3HB3HHx monoliths crystallized during phase separation and the subsequent aging. The mechanical properties, such as compression modulus and stress, of the monoliths depended on the 3-hydroxyhexanoate content of P3HB3HHx. Furthermore, the P3HB3HHx monolith absorbed linseed oil in preference to water in a plant oil–water mixture. In combination with the biodegradable character of P3HB3HHx, the present study is expected to contribute to the development of bio-based materials.

  6. Phased-Array Monolithic PEM for FT Spectrometry With Applications in Explosive Detection and CB Defense

    Science.gov (United States)

    2008-12-01

    1 PHASED-ARRAY MONOLITHIC PEM FOR FT SPECTROMETRY WITH APPLICATIONS IN EXPLOSIVE DETECTION AND CB DEFENSE Tudor N. Buican* Semiotic ...PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) Semiotic Engineering Associates LLC Albuquerque, NM 87108 8. PERFORMING ORGANIZATION REPORT NUMBER 9

  7. Reprofiled anthelmintics abate hypervirulent stationary-phase Clostridium difficile

    Science.gov (United States)

    Gooyit, Major; Janda, Kim D.

    2016-01-01

    Prolonged use of broad-spectrum antibiotics disrupts the indigenous gut microbiota, which consequently enables toxigenic Clostridium difficile species to proliferate and cause infection. The burden of C. difficile infections was exacerbated with the outbreak of hypervirulent strains that produce copious amounts of enterotoxins and spores. In recent past, membrane-active agents have generated a surge of interest due to their bactericidal property with a low propensity for resistance. In this study, we capitalized on the antimicrobial property and low oral bioavailability of salicylanilide anthelmintics (closantel, rafoxanide, niclosamide, oxyclozanide) to target the gut pathogen. By broth microdilution techniques, we determined the MIC values of the anthelmintics against 16 C. difficile isolates of defined PCR-ribotype. The anthelmintics broadly inhibited C. difficile growth in vitro via a membrane depolarization mechanism. Interestingly, the salicylanilides were bactericidal against logarithmic- and stationary-phase cultures of the BI/NAP1/027 strain 4118. The salicylanilides were poorly active against select gut commensals (Bacteroides, Bifidobacterium and Lactobacillus species), and were non-hemolytic and non-toxic to mammalian cell lines HepG2 and HEK 293T/17 within the range of their in vitro MICs and MBCs. The salicylanilide anthelmintics exhibit desirable properties for repositioning as anti-C. difficile agents. PMID:27633064

  8. Separation performance of guanidinium-based ionic liquids as stationary phases for gas chromatography.

    Science.gov (United States)

    Qiao, Lizhen; Lu, Kai; Qi, Meiling; Fu, Ruonong

    2013-02-08

    Room temperature ionic liquids (RTILs) as stationary phases for gas chromatography (GC) have made great achievements in both research and applications over the last decades. Until now, all of the RTIL stationary phases reported have involved imidazolium, ammonium, pyrrolidinium, and phosphonium-based RTILs, and however, no publications are available using guanidinium-based ionic liquids (GBILs) as GC stationary phases except two preliminary reports from our group. In the present work, three hexaalkyl GBILs stationary phases, namely N, N,N',N'-tetramethyl-N″, N″-dioctylguanidinum hexafluophosphate (DOTMG-PF(6)), N,N,N',N'-tetramethyl-N″, N″-dioctylguanidinium bis (trifluoromethylsulfonyl) imide (DOTMG-NTf(2)), and N,N,N',N'-tetraoctyl-N″, N″-dimethylguanidinium bis (trifluoromethylsulfonyl) imide (TODMG-NTf(2)), were synthesized and used as stationary phases for GC separation after they were statically coated onto the inner walls of fused-silica capillary columns. The evaluation of DOTMG-PF(6) and TODMG-NTf(2) as GC stationary phases is reported here for the first time, whereas additional results on the DOTMG-NTf(2) stationary phase are added here on the basis of our previous report. In this work, McReynolds constants and Abraham solvation system constants are used to evaluate the average polarity and the solvation properties of the GBILs stationary phases for GC separation, respectively. The results show that the GBILs stationary phases exhibit medium polarity with an average polarity of 293-390, and that the major molecular interactions of the GBILs with analytes are dipole/polarizable interactions, H-bond basicity and dispersion forces, etc. After this, the separation performance and thermal stability of the GBILs stationary phases were evaluated, showing that these stationary phases achieve excellent separation for analytes of great variety covering hydrocarbons, alcohols, esters, aldehydes, ketones, amines, amides and aromatics, and exhibit

  9. Polar stationary phases based on poly(oligo ethylene glycol)diacrylates for capillary gas chromatography

    Science.gov (United States)

    Shiryaeva, V. E.; Popova, T. P.; Korolev, A. A.; Kanat'eva, A. Yu.; Kurganov, A. A.

    2017-08-01

    New stationary phases for capillary columns in GC are synthesized and studied. The phases are prepared by depositing oligo(ethylene glycol)diacrylates on the column walls and subsequent polymerization (crosslinking) in the presence of peroxide initiators. It is shown that stationary phases based on monomers with molecular weights of 10 kDa or higher exhibit separation properties similar to those of conventional stationary phases based on polyethylene glycol (PEG); however, their thermal stability is higher because they have a higher degree of crosslinking and a more ordered structure of the crosslinked polymers than the respective parameters of phases based on native PEG.

  10. Use of ionic liquids as stationary phases in hyphenated gas chromatography techniques.

    Science.gov (United States)

    Ragonese, Carla; Sciarrone, Danilo; Tranchida, Peter Quinto; Dugo, Paola; Mondello, Luigi

    2012-09-14

    In the past decades a consistent number of ionic liquids have been specifically synthesized and evaluated as stationary phase in gas chromatography. Ionic liquid, also defined as "molten salts", are a class of organic non-molecular solvents liquid at room temperature (RTILs) that satisfy most of the requirements of a GC stationary phase, among which a high viscosity, the possibility to tune the selectivity (by changing the cation-anion combination) and a high thermal stability. The choice of the proper stationary phase plays a key role in the improvement/optimization of a GC method, and although the use of IL as stationary phases is still not well-established, the general interest in their applications has greatly increased, thanks to their particular properties. The present contribution provides an overview on recent evaluations and applications of IL stationary phases, focusing in particular on the use of these novel tools in hyphenated GC-based techniques.

  11. Genomewide Mutational Diversity in Escherichia coli Population Evolving in Prolonged Stationary Phase.

    Science.gov (United States)

    Chib, Savita; Ali, Farhan; Seshasayee, Aswin Sai Narain

    2017-01-01

    Prolonged stationary phase is an approximation of natural environments presenting a range of stresses. Survival in prolonged stationary phase requires alternative metabolic pathways for survival. This study describes the repertoire of mutations accumulating in starving Escherichia coli populations in lysogeny broth. A wide range of mutations accumulates over the course of 1 month in stationary phase. Single nucleotide polymorphisms (SNPs) constitute 64% of all mutations. A majority of these mutations are nonsynonymous and are located at conserved loci. There is an increase in genetic diversity in the evolving populations over time. Computer simulations of evolution in stationary phase suggest that the maximum frequency of mutations observed in our experimental populations cannot be explained by neutral drift. Moreover, there is frequent genetic parallelism across populations, suggesting that these mutations are under positive selection. Finally, functional analysis of mutations suggests that regulatory mutations are frequent targets of selection. IMPORTANCE Prolonged stationary phase in bacteria, contrary to its name, is highly dynamic, with extreme nutrient limitation as a predominant stress. Stationary-phase cultures adapt by rapidly selecting a mutation(s) that confers a growth advantage in stationary phase (GASP). The phenotypic diversity of starving E. coli populations has been studied in detail; however, only a few mutations that accumulate in prolonged stationary phase have been described. This study documented the spectrum of mutations appearing in Escherichia coli during 28 days of prolonged starvation. The genetic diversity of the population increases over time in stationary phase to an extent that cannot be explained by random, neutral drift. This suggests that prolonged stationary phase offers a great model system to study adaptive evolution by natural selection.

  12. Recent Research and Development of Monolithic Column in Ion Chromatography%整体柱离子色谱的研究进展

    Institute of Scientific and Technical Information of China (English)

    陈倩; 于泓

    2011-01-01

    Monolithic columns have been employed in low pressure, fast and high efficient separation, for their many excellent characteristics. When applied to ion chromatography field, monolithic columns can achieve more effective analysis than traditional columns. Therefore, the method of ion chromatography based on monolithic columns attracts increasing interests. In this paper, the sorts of ion chromatography, classification of monolithic stationary phases, preparation ways of monolithic columns, performances and features of monolithic columns were introduced. The application of conventional monolithic columns in ion chromatography and the utilization of capillary monolithic columns in capillary ion chromatography were summarized according to the different assortments and properties of monolithic columns and ion chromatography, which included silica-based monolithic stationary phase and organic polymer-based monolithic stationary phase. The development of ion chromatography based on monolithic columns in the future was reviewed and prospected.%该文介绍了离子色谱的分类,整体柱的分类、制备及特点,并以此为依据归纳总结了常规整体柱在离子色谱中的应用和毛细管整体柱在毛细管离子色谱中的应用,其中包括硅胶基质整体柱和聚合物基质整体柱,评述并展望了整体柱离子色谱的发展前景.

  13. Novas fases estacionárias à base de sílica para cromatografia líquida de alta eficiência New stationary phases based on silica for high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    César R. Silva

    2004-04-01

    Full Text Available The present work reviews recent advances in the preparation of new reversed phase packing materials such as sterically protected, bidentate, hybrid organic-inorganic and monolithic phases and phases containing embedded polar groups. The bonding chemistry involved in the preparation of these phases as well as their advantages over conventional C8 and C18 reversed phases are discussed. Understanding the reasons behind the development of these newer column packings helps analysts select the best stationary phase for a given application.

  14. A design concept for an MMIC (Monolithic Microwave Integrated Circuit) microstrip phased array

    Science.gov (United States)

    Lee, Richard Q.; Smetana, Jerry; Acosta, Roberto

    1987-02-01

    A conceptual design for a microstrip phased array with monolithic microwave integrated circuit (MMIC) amplitude and phase controls is described. The MMIC devices used are 20 GHz variable power amplifiers and variable phase shifters recently developed by NASA contractors for applications in future Ka proposed design, which concept is for a general NxN element array of rectangular lattice geometry. Subarray excitation is incorporated in the MMIC phased array design to reduce the complexity of the beam forming network and the number of MMIC components required.

  15. A design concept for an MMIC (Monolithic Microwave Integrated Circuit) microstrip phased array

    Science.gov (United States)

    Lee, Richard Q.; Smetana, Jerry; Acosta, Roberto

    1987-01-01

    A conceptual design for a microstrip phased array with monolithic microwave integrated circuit (MMIC) amplitude and phase controls is described. The MMIC devices used are 20 GHz variable power amplifiers and variable phase shifters recently developed by NASA contractors for applications in future Ka proposed design, which concept is for a general NxN element array of rectangular lattice geometry. Subarray excitation is incorporated in the MMIC phased array design to reduce the complexity of the beam forming network and the number of MMIC components required.

  16. Development of a monolithic polymer pipette for solid-phase extraction of liquiritigenin in rat plasma

    Institute of Scientific and Technical Information of China (English)

    Hong Wu Zhang; Kang Li; Zhi Xian Liang; Feng Yang Wang; Qi Wen Lu

    2012-01-01

    A monolithic polymer column with mixed-mode interaction was prepared by in situ polymerization in a 1000 μ,L pipette.Two kinds of monomers,butyl methacrylate (BMA) and 2-(dimethylamino)ethyl methacrylate (DMAM) were applied to constructing the mixed-mode interaction of monolithic polymer column.Its solid-phase extraction properties for liquiritigenin (LQG) were evaluated by high performance liquid chromatography (HPLC) with a gradient elution procedure.After the extraction procedure was optimized,the maximum binding capacity and extraction recovery following the optimal extraction procedure were investigated.Calibration curve was expressed as A =65.9C + 4.53 (r2 =0.998) with a linear range of 0.151-1.80 μg/mL.The experimental results indicate that the monolithic polymer pipette presents good extraction efficiency for LQG.It can be envisaged that the developed monolithic polymer pipette possesses the potential for its application to the enrichment of other flavonoids compounds being siniilar to the structure of LQG.

  17. Synthesis of novel chiral stationary phases for high-performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    Xu Lin Tan; Shi Cong Hou; Qing Hua Bian; Min Wang

    2007-01-01

    Three novel chiral selectors 4a-c were synthesized from (S)-amino acids and (R)- 1-phenyl-2-(4-methylphenyl)ethylamine. 4a-c were connected to 3-aminopropylsilanized silica gel to be used as the chiral stationary phase for HPLC. Five amino acid derivatives and two pyrethroid insecticides were fairly resolved on these three new chiral stationary phases under normal phase condition.

  18. Porous polymer monolithic col

    Directory of Open Access Journals (Sweden)

    Lydia Terborg

    2015-05-01

    Full Text Available A new approach has been developed for the preparation of mixed-mode stationary phases to separate proteins. The pore surface of monolithic poly(glycidyl methacrylate-co-ethylene dimethacrylate capillary columns was functionalized with thiols and coated with gold nanoparticles. The final mixed mode surface chemistry was formed by attaching, in a single step, alkanethiols, mercaptoalkanoic acids, and their mixtures on the free surface of attached gold nanoparticles. Use of these mixtures allowed fine tuning of the hydrophobic/hydrophilic balance. The amount of attached gold nanoparticles according to thermal gravimetric analysis was 44.8 wt.%. This value together with results of frontal elution enabled calculation of surface coverage with the alkanethiol and mercaptoalkanoic acid ligands. Interestingly, alkanethiols coverage in a range of 4.46–4.51 molecules/nm2 significantly exceeded that of mercaptoalkanoic acids with 2.39–2.45 molecules/nm2. The mixed mode character of these monolithic stationary phases was for the first time demonstrated in the separations of proteins that could be achieved in the same column using gradient elution conditions typical of reverse phase (using gradient of acetonitrile in water and ion exchange chromatographic modes (applying gradient of salt in water, respectively.

  19. The search for more pH stable stationary phases for high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Collins, Carol H.; Silva, Cesar R.; Faria, Anizio M.; Collins, Kenneth E.; Jardim, Isabel Cristina S.F. [Universidade Estadual de Campinas (UNICAMP), SP (Brazil). Inst. de Quimica

    2009-07-01

    High performance liquid chromatographic (HPLC) separations are largely carried out using reversed phase conditions with stationary phases based on silica. A serious problem with these stationary phases is the tendency of silica to dissolve in high pH solutions often needed to separate basic compounds. The literature reports many different ways that have been tried to resolve this problem. This paper reports the results obtained in our laboratory with stationary phases prepared using silica supports having a layer of a metal oxide (zirconia or titania) attached on their surfaces, followed by immobilization of a polysiloxane or by organofunctionalization with a trimethoxyalkylsilane. Stability tests, also developed in our laboratory, indicate that the metal oxide layer increases the HPLC column lifetimes by making the stationary phase less susceptible to dissolution. (author)

  20. Tuning of mobile and stationary phase polarity for the separation of polar compounds by SFC.

    Science.gov (United States)

    Ibañez, E; Señoráns, F J

    2000-07-05

    The separation of polar compounds by supercritical-fluid chromatography (SFC) is reviewed. New developments in mobile and stationary phase tuning are reviewed for packed and packed capillary SFC. In terms of mobile phase polarity adjustment, new pure and multiple component fluids are presented. The approach of tuning the polarity of the stationary phase as a way of increasing the range of polar compounds analyzed by SFC using pure CO(2) is discussed using either silica-based or new materials as stationary phase. Chiral, liquid crystal and polymer-based stationary phases coated on particles are widely covered in this review as an interesting approach to separate polar compounds avoiding the major drawbacks associated to the use of modifiers in SFC.

  1. Transcriptional characterization of Salmonella TA100 in log and stationary phase: influence of growth phase on mutagenicity of MX.

    Science.gov (United States)

    Ward, William O; Swartz, Carol D; Hanley, Nancy M; DeMarini, David M

    2010-10-13

    The Salmonella mutagenicity assay can be performed using cells that are in different growth phases. Thus, the plate-incorporation assay involves plating stationary-phase cells with the mutagen, after which the cells undergo a brief lag phase and, consequently, are exposed to the mutagen and undergo mutagenesis while in the logarithmic (log) phase. In contrast, a liquid-suspension assay involves exposure of either log- or stationary-phase cells to the mutagen for a specified period of time, sometimes followed by a wash, resulting in the cells growing in medium in the absence of the mutagen. To explore global gene expression in Salmonella, and to test for possible effects of growth phase and transcriptional status on mutagenesis, we performed microarray analysis on cells of Salmonella strain TA100 exposed to the drinking-water mutagen MX in either the log or stationary phase. The genes in functional pathways involving amino acid transport and metabolism and energy metabolism were expressed differentially in log-phase cells, whereas genes in functional pathways involving protein trafficking, cell envelope, and two-component systems using common signal transduction were expressed differentially in stationary-phase cells. More than 90% of the ribosomal-protein biosynthesis genes were up-regulated in stationary- versus log-phase cells. MX was equally mutagenic to cells in log- and stationary-phase growth when the results were expressed as mutant frequencies (revertants/survivors/μM), but it was twice as mutagenic in stationary-phase cells when the results were expressed as mutant yields (revertants/nmole or revertants/μM). There was a complex transcriptional response underlying these results, with mucA/B being greatly up-regulated in log-phase cells but umuC/D up-regulated in stationary-phase cells. The transcriptional state of TA100 cells at the time of mutagen treatment may influence the outcome of mutagen treatment.

  2. Ceramic stationary gas turbine development. Final report, Phase 1

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1994-09-01

    This report summarizes work performed by Solar Turbines Inc. and its subcontractors during the period September 25, 1992 through April 30, 1993. The objective of the work is to improve the performance of stationary gas turbines in cogeneration through implementation of selected ceramic components.

  3. Novel guanidinium-based ionic liquids as stationary phases for capillary gas chromatography

    Institute of Scientific and Technical Information of China (English)

    Li Zhen Qiao; Kai Lu; Mei Ling Qi; Ruo Nong Fu

    2010-01-01

    The present study describes guanidinium-based ionic liquids(GBILs)as stationary phases for capillary gas chromatography(CGC)and to the best of our knowledge,no related reports are available up to now.In this study,a hexaalkylguanidinium ionic liquid(DOTMG-NTf2)was synthesized and coated statically onto capillary columns.Selectivity of the stationary phase was evaluated by separating Grob test mixture,test mixture,alcohols mixture,and fatty acid methyl esters mixture,and thermal stability was investigated as well.The present study demonstrates that GBILs as CGC stationary phases exhibit satisfactory selectivity and thermal stability and have a great potential as new candidates for CGC stationary phases.

  4. Monolithic GaAs Dual-Gate FET Phase Shifter.

    Science.gov (United States)

    1982-02-01

    and P. Hats-op. "Dual-gae MESPET phase stutter with gainappioodwsel the43*phas alltif (Fi. 6. Net apliier is atl 12 Hz." Elcron. Lett.. vol. 16. O.14...Park Redondo Beach, CA 90278 Watkins -Johnson Co. 1 E. J. Crescenzi, Jr./ Professor L. Eastman 1 K. Niclas Phillips Hall 3333 Hillview Ave. Cornell

  5. Magnetism-Enhanced Monolith-Based In-Tube Solid Phase Microextraction.

    Science.gov (United States)

    Mei, Meng; Huang, Xiaojia; Luo, Qing; Yuan, Dongxin

    2016-02-02

    Monolith-based in-tube solid phase microextraction (MB/IT-SPME) has received wide attention because of miniaturization, automation, expected loading capacity, and environmental friendliness. However, the unsatisfactory extraction efficiency becomes the main disadvantage of MB/IT-SPME. To overcome this circumstance, magnetism-enhanced MB/IT-SPME (ME-MB/IT-SPME) was developed in the present work, taking advantage of magnetic microfluidic principles. First, modified Fe3O4 nanoparticles were mixed with polymerization solution and in situ polymerized in the capillary to obtain a magnetic monolith extraction phase. After that, the monolithic capillary column was placed inside a magnetic coil that allowed the exertion of a variable magnetic field. The effects of intensity of magnetic field, adsorption and desorption flow rate, volume of sample, and desorption solvent on the performance of ME-MB/IT-SPME were investigated in detail. The analysis of six steroid hormones in water samples by the combination of ME-MB/IT-SPME with high-performance liquid chromatography with diode array detection was selected as a paradigm for the practical evaluation of ME-MB/IT-SPME. The application of a controlled magnetic field resulted in an obvious increase of extraction efficiencies of the target analytes between 70% and 100%. The present work demonstrated that application of different magnetic forces in adsorption and desorption steps can effectively enhance extraction efficiency of MB/IT-SPME systems.

  6. Gold nanoparticle decorated graphene oxide/silica composite stationary phase for high-performance liquid chromatography.

    Science.gov (United States)

    Liang, Xiaojing; Wang, Xusheng; Ren, Haixia; Jiang, Shengxiang; Wang, Licheng; Liu, Shujuan

    2014-06-01

    In the initial phase of this study, graphene oxide (GO)/silica was fabricated by assembling GO onto the silica particles, and then gold nanoparticles (GNPs) were used to modify the GO/silica to prepare a novel stationary phase for high-performance liquid chromatography. The new stationary phase could be used in both reversed-phase chromatography and hydrophilic interaction liquid chromatography modes. Good separations of alkylbenzenes, isomerides, amino acids, nucleosides, and nucleobases were achieved in both modes. Compared with the GO/silica phase and GNPs/silica phase, it is found that except for hydrophilicity, large π-electron systems, hydrophobicity, and coordination functions, this new stationary phase also exhibited special separation performance due to the combination of 2D GO with zero-dimensional GNPs.

  7. PEG-linked geminal dicationic ionic liquids as selective, high-stability gas chromatographic stationary phases.

    Science.gov (United States)

    Huang, Ke; Han, Xinxin; Zhang, Xiaotong; Armstrong, Daniel W

    2007-12-01

    It is known that room-temperature ionic liquids (RTILs) have wide applicability in many scientific and technological fields. In this work, a series of three new dicationic room-temperature ionic liquids functionalized with poly(ethylene glycol) (PEG) linkages were synthesized and characterized via a linear solvation model. The application of these ILs as new GC stationary phases was studied. The efficient separation of several mixtures containing compounds of different polarities and 24 components of a flavor and fragrance mixture indicated comparable or higher resolving power for the new IL stationary phases compared to the commercial polysiloxane and poly(ethylene glycol)-based stationary phases. In addition, the selectivities of the IL stationary phases could be quite unique. The separation of a homologous alkane and alcohol mixture displayed the "dual nature" of these ionic liquids as GC stationary phases. The thermal stability study showed the column robustness up to 350 degrees C. The high separation power, unique selectivity, high efficiency and high thermal stability of the new dicationic ionic liquids indicate that they may be applicable as a new type of robust GC stationary phase.

  8. Study of surface-bonded dicationic ionic liquids as stationary phases for hydrophilic interaction chromatography.

    Science.gov (United States)

    Qiao, Lizhen; Li, Hua; Shan, Yuanhong; Wang, Shuangyuan; Shi, Xianzhe; Lu, Xin; Xu, Guowang

    2014-02-21

    In the present study, several geminal dicationic ionic liquids based on 1,4-bis(3-allylimidazolium)butane and 1,8-bis(3-allylimidazolium)octane in combination with different anions bromide and bis(trifluoromethanesulphonyl)imide were prepared and then bonded to the surface of 3-mercaptopropyl modified silica materials through the "thiol-ene" click chemistry as stationary phases for hydrophilic interaction chromatography (HILIC). Compared with their monocationic analogues, the dicationic ionic liquids stationary phases presented effective retention and good selectivity for typical hydrophilic compounds under HILIC mode with the column efficiency as high as 130,000 plates/m. Moreover, the influence of different alkyl chain spacer between dications and combined anions on the retention behavior and selectivity of the dicationic ionic liquids stationary phases under HILIC mode was displayed. The results indicated that the longer linkage chain would decrease the hydrophilicity and retention on the dicationic ionic liquid stationary phase, and while differently combined anions had no difference due to the exchangeability under the common HILIC mobile phase with buffer salt. Finally, the retention mechanism was investigated by evaluating the effect of chromatographic factors on retention, including the water content in the mobile phase, the mobile phase pH and buffer salt concentration. The results showed that the dicationic ionic liquids stationary phases presented a mixed-mode retention behavior with HILIC mechanism and anion exchange.

  9. Enantioseparation of Racemic Naproxen Esters on Cellulose Tris (4-methylbenzoate) Chiral Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Several kinds of racemic naproxen ester were successfully separated on CTMB chiral stationary phase with hexane-ethanol (98:2, vol./vol.) as the mobile phase. The influence of mobile phase composition and structure of racemic naproxen ester on chiral separation was studied and the chiral recognition mechanism of CTMB was discussed.

  10. Enantioseparation of Racemic Naproxen Esters on Cellulose Tris(4—methylbenzoate) Chiral Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    BaoHaiSHAO; XiuZhuXU; 等

    2002-01-01

    Several kinds of racemic naproxen ester were successfully separated on CTMB chiral stationary phase with hexane-ethanol(98:2,vol./vol.) as the mobile phase. The influence of mobile phase composition and structure of racemic naproxen ester on chiral separation was studied and the chiral recognition mechanism of CTMB was discussed.

  11. Diffraction coupled phase-locked arrays of quantum cascade lasers with monolithically integrated Talbot cavities

    CERN Document Server

    Wang, Lei; Jia, Zhi-Wei; Zhao, Yue; Liu, Chuan-Wei; Liu, Ying-Hui; Zhai, Shen-Qiang; Zhuo, Ning; Liu, Feng-Qi; Xu, Xian-Gang

    2016-01-01

    Diffraction coupled arrays of quantum cascade laser are presented. The phase-locked behavior is achieved through monolithic integration of a Talbot cavity at one side of the laser array. The principle is based on fractional Talbot effect. By controlling length of Talbot cavity to be a quarter of Talbot distance (Zt/4), in-phase mode operation is selected. Measured far-field radiation patterns reflect stable in-phase mode operation under different injection currents, from threshold current to full power current. Diffraction-limited performance is shown from the lateral far-field, where three peaks can be obtained and main peak and side peak interval is 10.5{\\deg}. The phase-locked arrays with in-phase mode operation may be a feasible solution to get higher output power and maintain well beam quality meanwhile.

  12. Role of ppGpp in rpoS stationary-phase regulation in Escherichia coli.

    Science.gov (United States)

    Hirsch, Matthew; Elliott, Thomas

    2002-09-01

    The bacterial sigma factor RpoS is strongly induced under a variety of stress conditions and during growth into stationary phase. Here, we used rpoS-lac fusions in Escherichia coli to investigate control acting at the level of RpoS synthesis, which is especially evident when cells approach stationary phase in rich medium. Previous work has shown that the small molecule ppGpp is required for normal levels of RpoS in stationary phase. Despite the attraction of a model in which the ppGpp level controls stationary-phase induction of RpoS, careful measurement of rpoS-lac expression in a mutant lacking ppGpp showed similar effects during both exponential growth and stationary phase; the main effect of ppGpp was on basal expression. In addition, a modest regulatory defect was associated with the mutant lacking ppGpp, delaying the time at which full expression was achieved by 2 to 3 h. Deletion analysis showed that the defect in basal expression was distributed over several sequence elements, while the regulatory defect mapped to the region upstream of the rpoS ribosome-binding site (RBS) that contains a cis-acting antisense element. A number of other genes that have been suggested as regulators of rpoS were tested, including dksA, dsrA, barA, ppkx, and hfq. With the exception of the dksA mutant, which had a modest defect in Luria-Bertani medium, none of these mutants was defective for rpoS stationary-phase induction. Even a short rpoS segment starting at 24 nucleotides upstream of the AUG initiation codon was sufficient to confer substantial stationary-phase regulation, which was mainly posttranscriptional. The effect of RBS-proximal sequence was independent of all known trans-acting factors, including ppGpp.

  13. Phased array of high-power, coherent, monolithic flared amplifier master oscillator power amplifiers

    Science.gov (United States)

    Osinski, J. S.; Mehuys, D.; Welch, D. F.; Waarts, R. G.; Major, J. S., Jr.; Dzurko, K. M.; Lang, R. J.

    1995-01-01

    A monolithically integrated array of InGaAs/AlGaAs flared amplifiers driven by a single DBR laser through a power splitter network and individually addressed phase modulators is described. Phase adjustment of ≳2π per element by free-carrier effects is verified by monitoring the interference pattern of all four emitters, and typically requires <15 mA of current to obtain a 2π phase shift. Phase matching is achieved among all four diffraction-limited emitters at a pulsed output power of ≳5 W, and, combined with the proper external lensing, could therefore result in an ultranarrow, single-lobed far-field pattern whose width is determined by the extended aperture of the array.

  14. Efficiency enhancements in micellar liquid chromatography through selection of stationary phase and alcohol modifier.

    Science.gov (United States)

    Thomas, David P; Foley, Joe P

    2007-05-18

    Micellar liquid chromatography (MLC) remains hindered by reduced chromatographic efficiency compared to reversed phase liquid chromatography (RPLC) using hydro-organic mobile phases. The reduced efficiency has been partially explained by the adsorption of surfactant monomers onto the stationary phase, resulting in a slow mass transfer of the analyte within the interfacial region of the mobile phase and stationary phase. Using an array of 12 columns, the effects of various bonded stationary phases and silica pore sizes, including large-pore short alkyl chain, non-porous, superficially porous and perfluorinated, were evaluated to determine their impact on efficiency in MLC. Additionally, each stationary phase was evaluated using 1-propanol and 1-butanol as separate micellar mobile phase alcohol additives, with several columns also evaluated using 1-pentanol. A simplified equation for calculation of A' and C' terms from reduced plate height (h) versus reduced velocity (nu) plots was used to compare the efficiency data obtained with the different columns and mobile phases. Analyte diffusion coefficients needed for the h versus nu plots were determined by the Taylor-Aris dispersion technique. The use of a short alkyl chain, wide-pore silica column, specifically, Nucleosil C4, 1000A, was shown to have the most improved efficiency when using a micellar mobile phase compared to a hydro-organic mobile phase for all columns evaluated. The use of 1-propanol was also shown to provide improved efficiency over 1-butanol or 1-pentanol in most cases. In a second series of experiments, column temperatures were varied from 40 to 70 degrees C to determine the effect of temperature on efficiency for a subset of the stationary phases. Efficiency improvements ranging from 9% for a Chromegabond C8 column to 58% for a Zorbax ODS column were observed over the temperature range. Based on these observed improvements, higher column temperatures may often yield significant gains in column

  15. Proteomic Analysis of Stationary Phase in the Marine Bacterium 'Candidatus Pelagibacter ubique'

    Energy Technology Data Exchange (ETDEWEB)

    Sowell, Sarah M.; Norbeck, Angela D.; Lipton, Mary S.; Nicora, Carrie D.; Callister, Stephen J.; Smith, Richard D.; Barofsky, Douglas F.; Giovannoni, Stephen J.

    2008-05-01

    Candidatus Pelagibacter ubique, an abundant marine alphaproteobacterium, subsists in nature at low ambient nutrient concentrations and may often be exposed to nutrient limitation, but its genome revealed no evidence of global regulatory adaptations to stationary phase. We used high-resolution capillary liquid chromatography (LC) coupled online to an LTQ mass spectrometer to build an Accurate Mass and Time (AMT) tag library, and employed the AMT tag approach to quantitatively examine proteome differences between exponentially growing and stationary phase Cand. P. ubique cells cultivated in a seawater medium. The AMT tag library represented 72% of the predicted protein coding genes. Stationary phase protein abundance increased for OsmC, which mitigates oxidative damage, and for molecular chaperones, enzymes involved in methionine and cysteine biosynthesis, proteins involved in rho-dependent transcription termination, and the signal transduction enzymes CheY-FisH and ChvG. Our findings indicate that Cand. P. ubique responds adaptively to stationary phase by increasing the abundance of a suite of proteins that contribute to homeostasis, but does not undergo major proteome remodeling. We speculate that this limited response may enable Cand. P. ubique to cope with ambient conditions in which nutrients are often insufficient for short periods, and the ability to resume growth overrides the capacity for long term survival afforded by more comprehensive global stationary phase responses.

  16. The Proteome and Lipidome of Thermococcus kodakarensis across the Stationary Phase.

    Science.gov (United States)

    Gagen, Emma J; Yoshinaga, Marcos Y; Garcia Prado, Franka; Hinrichs, Kai-Uwe; Thomm, Michael

    2016-01-01

    The majority of cells in nature probably exist in a stationary-phase-like state, due to nutrient limitation in most environments. Studies on bacteria and yeast reveal morphological and physiological changes throughout the stationary phase, which lead to an increased ability to survive prolonged nutrient limitation. However, there is little information on archaeal stationary phase responses. We investigated protein- and lipid-level changes in Thermococcus kodakarensis with extended time in the stationary phase. Adaptations to time in stationary phase included increased proportion of membrane lipids with a tetraether backbone, synthesis of proteins that ensure translational fidelity, specific regulation of ABC transporters (upregulation of some, downregulation of others), and upregulation of proteins involved in coenzyme production. Given that the biological mechanism of tetraether synthesis is unknown, we also considered whether any of the protein-level changes in T. kodakarensis might shed light on the production of tetraether lipids across the same period. A putative carbon-nitrogen hydrolase, a TldE (a protease in Escherichia coli) homologue, and a membrane bound hydrogenase complex subunit were candidates for possible involvement in tetraether-related reactions, while upregulation of adenosylcobalamin synthesis proteins might lend support to a possible radical mechanism as a trigger for tetraether synthesis.

  17. The Proteome and Lipidome of Thermococcus kodakarensis across the Stationary Phase

    Directory of Open Access Journals (Sweden)

    Emma J. Gagen

    2016-01-01

    Full Text Available The majority of cells in nature probably exist in a stationary-phase-like state, due to nutrient limitation in most environments. Studies on bacteria and yeast reveal morphological and physiological changes throughout the stationary phase, which lead to an increased ability to survive prolonged nutrient limitation. However, there is little information on archaeal stationary phase responses. We investigated protein- and lipid-level changes in Thermococcus kodakarensis with extended time in the stationary phase. Adaptations to time in stationary phase included increased proportion of membrane lipids with a tetraether backbone, synthesis of proteins that ensure translational fidelity, specific regulation of ABC transporters (upregulation of some, downregulation of others, and upregulation of proteins involved in coenzyme production. Given that the biological mechanism of tetraether synthesis is unknown, we also considered whether any of the protein-level changes in T. kodakarensis might shed light on the production of tetraether lipids across the same period. A putative carbon-nitrogen hydrolase, a TldE (a protease in Escherichia coli homologue, and a membrane bound hydrogenase complex subunit were candidates for possible involvement in tetraether-related reactions, while upregulation of adenosylcobalamin synthesis proteins might lend support to a possible radical mechanism as a trigger for tetraether synthesis.

  18. Regulation of Indole Signalling during the Transition of E. coli from Exponential to Stationary Phase.

    Science.gov (United States)

    Gaimster, Hannah; Summers, David

    2015-01-01

    During the transition from exponential to stationary phase E. coli produces a substantial quantity of the small, aromatic signalling molecule indole. In LB medium the supernatant indole concentration reaches a maximum of 0.5-1 mM. At this concentration indole has been implicated in many processes inducing acid resistance and the modulation of virulence. It has recently been shown that cell-associated indole transiently reaches a very high concentration (approx. 60 mM) during stationary phase entry, presumably because indole is being produced more rapidly than it can leave the cell. It is proposed that this indole pulse inhibits growth and cell division, causing the culture to enter stationary phase before nutrients are completely exhausted, with benefits for survival in long-term stationary phase. This study asks how E. coli cells rapidly upregulate indole production during stationary phase entry and why the indole pulse has a duration of only 10-15 min. We find that at the start of the pulse tryptophanase synthesis is triggered by glucose depletion and that this is correlates with the up-regulation of indole synthesis. The magnitude and duration of the resulting indole pulse are dependent upon the availability of exogenous tryptophan. Indole production stops when all the available tryptophan is depleted and the cell-associated indole equilibrates with the supernatant.

  19. Ionic liquids as novel stationary phases in gas liquid chromatography: inverse or normal isotope effect?

    Science.gov (United States)

    Schmarr, Hans-Georg; Slabizki, Petra; Müntnich, Sabrina; Metzger, Carmen; Gracia-Moreno, Elisa

    2012-12-28

    The separation of deuterated and non-deuterated compounds in gas liquid partitioning chromatography (GLC) on silicone type stationary phase usually results in the inverse isotope effect. With ionic liquids (ILs) as stationary phase, however, this may show a totally different nature. The inverse isotope effect, in which heavier (deuterated) isotopic compounds (isotopologues) elute earlier, is to be expected when van der Waals (London) dispersion forces play a dominant role in the solute-stationary phase interaction. Such (apolar) interactions seem to play only a minor role when ILs are the stationary phases, leading to only a marginal inverse isotope effect, e.g. for the separation of 2,4,6-trichloroanisole and its [(2)H(5)]-isotopologue on 1,12-di(tripropylphosphonium) dodecane bis(trifluoromethansulfonyl) amide (commercialized as SLB-IL59, Supelco). Indeed, with the most polar stationary phase available (commercialized as SLB-IL111; Supelco), this separation showed a normal isotope effect. Further examples are presented and the nature of the isotope effect observed is discussed.

  20. Modified Diatomaceous earth as a principal stationary phase component in TLC.

    Science.gov (United States)

    Ergül, Soner; Kadan, Imdat; Savaşci, Sahin; Ergül, Suzan

    2005-09-01

    Modified natural diatomaceous earth (DE) is a principal component of the stationary phase in normal thin-layer chromatography (TLC) applications and is mixed with commercial silica gel 60GF254 (Si-60GF254). Modification is carried out by flux calcination and refluxing with acid. Natural DE, modified DEs [flux calcinated (FC)DE and FCDE-I), and Si-60GF254 are characterized by scanning electron microscopy and Fourier-transform-IR spectroscopy. Particle size, specific surface area, pore distribution, pore volume, and surface hydroxyl group density parameters of materials are determined by various techniques. FCDE-I and Si-60GF254 are investigated for their usefulness in the stationary phase of TLC both individually and in composition. Commercially available red and blue ink samples are run on layers of Si-60GF254 and FCDE-I individually, and on various FCDE-I and Si-60GF254 mixtures. Butanol-ethanol-2M ammonia (3:1:1, v/v) and butanol-acetic acid-water (12:3:5, v/v) mixtures are used as mobile phases. The polarities of stationary phases decrease, and the retention factor (Rf) values of ink components increase when the FCDE-I content of the stationary phase increases. The properties of the stationary phase can be optimized by adding FCDE-I to Si-60GF254. This study may be useful in understanding both the systematic effects of stationary phase properties [e.g., specific surface area and surface hydroxyl group density, aOH(s)] and those of the mobile phase (e.g., polarity and acidity) on Rf values and the separability of components.

  1. Comparison of liquid and supercritical fluid chromatography mobile phases for enantioselective separations on polysaccharide stationary phases.

    Science.gov (United States)

    Khater, Syame; Lozac'h, Marie-Anne; Adam, Isabelle; Francotte, Eric; West, Caroline

    2016-10-07

    Analysis and production of enantiomerically pure compounds is a major topic of interest when active pharmaceutical ingredients are concerned. Enantioselective chromatography has become a favourite both at the analytical and preparative scales. High-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) are dominating the scene and are often seen as complementary techniques. Nowadays, for economic and ecologic reasons, SFC may be preferred over normal-phase HPLC (NPLC) as it allows significant reductions in solvent consumption. However, the transfer of NPLC methods to SFC is not always straightforward. In this study, we compare the retention of achiral molecules and separation of enantiomers under supercritical fluid (carbon dioxide with ethanol or isopropanol) and liquid normal-phase (heptane with ethanol or isopropanol) elution modes with polysaccharide stationary phases in order to explore the differences between the retention and enantioseparation properties between the two modes. Chemometric methods (namely quantitative structure-retention relationships and discriminant analysis) are employed to compare the results obtained on a large set of analytes (171 achiral probes and 97 racemates) and gain some understanding on the retention and separation mechanisms. The results indicate that, contrary to popular belief, carbon dioxide - solvent SFC mobile phases are often weaker eluents than liquid mobile phases. It appears that SFC and NPLC elution modes provide different retention mechanisms. While some enantioseparations are unaffected, facilitating the transfer between the two elution modes, other enantioseparations may be drastically different due to different types and strength of interactions contributing to enantioselectivity.

  2. Imidazolium embedded C8 based stationary phase for simultaneous reversed-phase/hydrophilic interaction mixed-mode chromatography.

    Science.gov (United States)

    Qiao, Xiaoqiang; Zhang, Lu; Zhang, Niu; Wang, Xin; Qin, Xinying; Yan, Hongyuan; Liu, Haiyan

    2015-06-26

    A new imidazolium embedded C8 based stationary phase (SIL-MPS-VOL) was facilely prepared by two steps and characterized by Fourier transform infrared spectrometry and thermogravimetric analysis. Due to the introduction of quaternary imidazolium group to the traditional C8 stationary phase, the developed SIL-MPS-VOL column demonstrated both reversed-phase liquid chromatography (RPLC) and hydrophilic interaction liquid chromatography (HILIC) retention mechanisms. A series of hydrophobic and hydrophilic test samples, including benzene homologues, anilines, positional isomers, nucleosides and nucleotides, were used to evaluate the developed SIL-MPS-VOL stationary phase. A rapid separation time, high separation efficiency and planar selectivity were achieved, compared with the commercially available C8 column. Moreover, the developed stationary phase was further used to detect and separate of melamine in powdered infant formula and high polar component of secondary metabolites of Trichoderma, and improved separation efficiency was achieved, indicating the potential merits of the developed SIL-MPS-VOL stationary phase for simultaneous separation of complex hydrophobic and hydrophilic samples with high selectivity.

  3. Fast separation of hen egg white protein with a phosphorylcholine type zwitterionic ion chromatography stationary phase

    Institute of Scientific and Technical Information of China (English)

    Qian Qu; Xiu Juan Yu; Xi Wu; Fei Shi; Li Li Wang

    2012-01-01

    In this work,a kind of preparation method of zwitterionic ion chromatography (ZIC) stationary phase modified with phosphorylcholine (PC) was obtained by hydrolyzing after bonding phosphorylcholine dichloride to diol-silica to better explore the characteristics of the PC groups as ZIC stationary phase ligand in simultaneous separation of acidic proteins and basic proteins.The results showed that two kinds of acidic proteins and three kinds of basic proteins can be separated completely,meanwhile,hen egg white was separated and purified and three kinds of egg white components ovalbumin,G2 ovoglobulin and ovotransfemin proteins were separated completely by one single step on PC-ZIC column,the purity of all proteins reached above 95%.PC-ZIC stationary phase was successfully improved with better separation capacity and selectivity than previously reported in this paper.

  4. Metal-Organic Framework Thin Films as Stationary Phases in Microfabricated Gas-Chromatography Columns.

    Energy Technology Data Exchange (ETDEWEB)

    Read, Douglas [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Sillerud, Colin Halliday [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2016-01-01

    The overarching goal of this project is to integrate Sandia's microfabricated gas-chromatography ( GC) columns with a stationary phase material that is capable of retaining high-volatility chemicals and permanent gases. The successful integration of such a material with GCs would dramatically expand the repertoire of detectable compounds for Sandia's various microanalysis systems. One such promising class of candidate materials is metal-organic frameworks (MOFs). In this report we detail our methods for controlled deposition of HKUST-1 MOF stationary phases within GC columns. We demonstrate: the chromatographic separation of natural gas; a method for determining MOF film thickness from chromatography alone; and the first-reported GC x GC separation of natural gas -- in general -- let alone for two disparate MOF stationary phases. In addition we determine the fundamental thermodynamic constant for mass sorption, the partition coefficient, for HKUST-1 and several light hydrocarbons and select toxic industrial chemicals.

  5. [Characteristics of electroosmotic flow in open-tubular capillary electrochromatography with magnetic nanoparticle coating as mixed-mode stationary phase].

    Science.gov (United States)

    Qin, Sasa; Zhou, Chaoran; Zhu, Yaxian; Ren, Zhiyu; Zhang, Lingyi; Fu, Honggang; Zhang, Weibing

    2011-09-01

    A novel open-tubular capillary electrochromatography (OT-CEC) column with magnetic nanoparticle coating as mixed-mode stationary phase was prepared. The mixed-mode stationary phases were obtained by mixing C18 and amino modified magnetic nanoparticles with different ratios. The mixed modified magnetic nanoparticles as stationary phase were introduced into the capillary by using external magnetic force. The magnetic nanoparticle coating can be easily regenerated by removing the external magnetic field, and applied to other separation modes. The characteristics of electroosmotic flow (EOF) were theoretically investigated through the effect of physicochemical properties of different stationary phases on EOF. The experiment was conducted under different ratios of mixed-mode stationary phases and coating lengths, and it was verified that the theoretical conclusions accorded with the experimental results. It was shown that the EOF can be easily adjusted by changing the ratio of stationary phases or the number of permanent magnets.

  6. Cantilever RF-MEMS for monolithic integration with phased array antennas on a PCB

    Science.gov (United States)

    Aguilar-Armenta, C. J.; Porter, S. J.

    2015-12-01

    This article presents the development and operation of a novel electrostatic metal-to-metal contact cantilever radio-frequency microelectromechanical system (RF-MEMS) switch for monolithic integration with microstrip phased array antennas (PAAs) on a printed circuit board. The switch is fabricated using simple photolithography techniques on a Rogers 4003c substrate, with a footprint of 200 µm × 100 µm, based on a 1 µm-thick copper cantilever. An alternative wet-etching technique for effectively releasing the cantilever is described. Electrostatic and electromagnetic measurements show that the RF-MEMS presents an actuation voltage of 90 V for metal-to-metal contact, an isolation of -8.7 dB, insertion loss of -2.5 dB and a return loss of -15 dB on a 50 Ω microstrip line at 12.5 GHz. For proof-of-concept, a beam-steering 2 × 2 microstrip PAA, based on two 1-bit phase shifters suitable for the monolithic integration of the RF-MEMS, has been designed and measured at 12.5 GHz. Measurements show that the beam-steering system presents effective radiation characteristics with scanning capabilities from broadside towards 29° in the H-plane.

  7. Activity of Sulfa Drugs and Their Combinations against Stationary Phase B. burgdorferi In Vitro

    Directory of Open Access Journals (Sweden)

    Jie Feng

    2017-03-01

    Full Text Available Lyme disease is a most common vector-borne disease in the US. Although the majority of Lyme patients can be cured with the standard two- to four-week antibiotic treatment, at least 10%–20% of patients continue to suffer from prolonged post-treatment Lyme disease syndrome (PTLDS. While the cause for this is unclear, one possibility is that persisting organisms are not killed by current Lyme antibiotics. In our previous studies, we screened an FDA drug library and an NCI compound library on B. burgdorferi and found some drug hits including sulfa drugs as having good activity against B. burgdorferi stationary phase cells. In this study, we evaluated the relative activity of three commonly used sulfa drugs, sulfamethoxazole (Smx, dapsone (Dps, sulfachlorpyridazine (Scp, and also trimethoprim (Tmp, and assessed their combinations with the commonly prescribed Lyme antibiotics for activities against B. burgdorferi stationary phase cells. Using the same molarity concentration, dapsone, sulfachlorpyridazine and trimethoprim showed very similar activity against stationary phase B. burgdorferi enriched in persisters; however, sulfamethoxazole was the least active drug among the three sulfa drugs tested. Interestingly, contrary to other bacterial systems, Tmp did not show synergy in drug combinations with the three sulfa drugs at their clinically relevant serum concentrations against B. burgdorferi. We found that sulfa drugs combined with other antibiotics were more active than their respective single drugs and that four-drug combinations were more active than three-drug combinations. Four-drug combinations dapsone + minocycline + cefuroxime + azithromycin and dapsone + minocycline + cefuroxime + rifampin showed the best activity against stationary phase B. burgdorferi in these sulfa drug combinations. However, these four-sulfa-drug–containing combinations still had considerably less activity against B. burgdorferi stationary phase cells than the

  8. Mixed Stationary Liquid Phases for Gas-Liquid Chromatography.

    Science.gov (United States)

    Koury, Albert M.; Parcher, Jon F.

    1979-01-01

    Describes a laboratory technique for use in an undergraduate instrumental analysis course that, using the interpretation of window diagrams, prepares a mixed liquid phase column for gas-liquid chromatography. A detailed procedure is provided. (BT)

  9. Quasi-stationary phase change heat transfer on a fin

    Science.gov (United States)

    Orzechowski, Tadeusz; Stokowiec, Katarzyna

    2016-03-01

    The paper presents heat transfer research basing on a long fin with a circular cross-section. Its basis is welded to the pipe where the hot liquid paraffin, having a temperature of 70°C at the inflow, is pumped. The analyzed element is a recurrent part of a refrigeration's condenser, which is immersed in a paraffin. The temperature of the inflowing liquid is higher than the temperature of the melting process for paraffin, which allows the paraffin to liquify. The temperature at the basis of the rib changes and it is assumed that the heat transfer is quasi-stationary. On this basis the estimation of the mean value of heat transfer coefficient was conducted. The unsteady thermal field of the investigated system was registered with an infrared camera V50 produced by a Polish company Vigo System. This device is equipped with a microbolometric detector with 384 × 288 elements and the single pixel size 25 × 25 μm. Their thermal resolution is lower than 70 mK at a temperature of 30 °C. The camera operates at 7,5 ÷ 14 μm long-wave infrared radiation range. For a typical lens 35 mm the special resolution is 0.7 mrad. The result of the calculations is mean heat transfer coefficient for the considered time series. It is equal to 50 W m -2 K-1 and 47 W m -2 K-1 on the left and right side of the fin, respectively. The distance between the experimental data and the curve approximating the temperature distribution was assessed with the standard deviation, Sd = 0.04 K.

  10. Quasi-stationary phase change heat transfer on a fin

    Directory of Open Access Journals (Sweden)

    Orzechowski Tadeusz

    2016-01-01

    Full Text Available The paper presents heat transfer research basing on a long fin with a circular cross-section. Its basis is welded to the pipe where the hot liquid paraffin, having a temperature of 70°C at the inflow, is pumped. The analyzed element is a recurrent part of a refrigeration’s condenser, which is immersed in a paraffin. The temperature of the inflowing liquid is higher than the temperature of the melting process for paraffin, which allows the paraffin to liquify. The temperature at the basis of the rib changes and it is assumed that the heat transfer is quasi-stationary. On this basis the estimation of the mean value of heat transfer coefficient was conducted. The unsteady thermal field of the investigated system was registered with an infrared camera V50 produced by a Polish company Vigo System. This device is equipped with a microbolometric detector with 384 × 288 elements and the single pixel size 25 × 25 μm. Their thermal resolution is lower than 70 mK at a temperature of 30 °C. The camera operates at 7,5 ÷ 14 μm long-wave infrared radiation range. For a typical lens 35 mm the special resolution is 0.7 mrad. The result of the calculations is mean heat transfer coefficient for the considered time series. It is equal to 50 W m −2 K−1 and 47 W m −2 K−1 on the left and right side of the fin, respectively. The distance between the experimental data and the curve approximating the temperature distribution was assessed with the standard deviation, Sd = 0.04 K.

  11. Profiling of Burkholderia cepacia secretome at mid-logarithmic and early-stationary phases of growth.

    Directory of Open Access Journals (Sweden)

    Vanitha Mariappan

    Full Text Available BACKGROUND: Burkholderia cepacia is a Gram-negative pathogen that causes serious respiratory infections in immunocompromised patients and individuals with cystic fibrosis. This bacterium is known to release extracellular proteins that may be involved in virulence. METHODOLOGY/PRINCIPAL FINDINGS: In the present study, B. cepacia grown to mid-logarithmic and early-stationary phases were investigated on their ability to invade and survive intracellularly in A549 lung epithelial cells in order to discern the fate of these bacteria in the pathogenesis of B. cepacia lung infections in in vitro condition. The early-stationary phase B. cepacia was demonstrated to be more invasive than mid-logarithmic phase. In addition, culture supernatants of B. cepacia obtained from these phases of growth were also demonstrated to cause different cytotoxic potency on the A549 human lung epithelial cells. Profiling of the supernatants using the gel-based proteomics approach identified 43 proteins that were commonly released in both the growth phases and 40 proteins newly-released at the early-stationary phase. The latter proteins may account for the higher cytotoxic activity of the early-stationary culture supernatant compared to that obtained at the mid-logarithmic phase. Among the newly-released proteins in the early-stationary phase supernatant were flagellar hook-associated domain protein (FliD, flagellar hook-associated protein (FlgK, TonB-dependent siderophore (Fiu, Elongation factor G (FusA, phosphoglycerate kinase (Pgk and sulfatase (AslA which are known for their virulence. CONCLUSION/SIGNIFICANCE: Differences in the ability of B. cepacia to invade and survive intracellularly inside the epithelial cells at different phases of growth may improve our understanding of the varied disease progressions associated with B. cepacia infections. In addition, the identified culture supernatant proteins may be used as targets for the development of new strategies to

  12. Plasmid DNA Supercoiling and Gyrase Activity in Escherichia coli Wild-Type and rpoS Stationary-Phase Cells

    Science.gov (United States)

    Reyes-Domínguez, Yazmid; Contreras-Ferrat, Gabriel; Ramírez-Santos, Jesús; Membrillo-Hernández, Jorge; Gómez-Eichelmann, M. Carmen

    2003-01-01

    Stationary-phase cells displayed a distribution of relaxed plasmids and had the ability to recover plasmid supercoiling as soon as nutrients became available. Preexisting gyrase molecules in these cells were responsible for this recovery. Stationary-phase rpoS cells showed a bimodal distribution of plasmids and failed to supercoil plasmids after the addition of nutrients, suggesting that rpoS plays a role in the regulation of plasmid topology during the stationary phase. PMID:12533486

  13. Preparation of novel chiral stationary phase based on click chemistry for ligand exchange chromatography

    Institute of Scientific and Technical Information of China (English)

    Chun Mei Fu; Hong Yu Shi; Guang Sheng Qian; Zhang Wan Li

    2009-01-01

    Click chemistry was applied to immobilize L-proline derivative onto azide-modified silica gel to give a novel chiral stationary phase (denoted as click-CSP) for ligand exchange chromatography. The developed protocol combines the benefits of operational simplicity, exceptionally mild conditions and high surface loadings. The enantioselectivity α of some DE-amino acids on the click-CSP were found to be in the range from 1.13 to 3.46. The chromatographic resolutions of some DL-amino acids and the stability study firmly illustrate the potential of click chemistry for preparation chiral stationary phase for ligand exchange chromatography.

  14. Separation of basic compounds by capillary electrochromatography on an X-Terra RP18 stationary phase.

    Science.gov (United States)

    Valette, J C; Bizet, A C; Demesmay, C; Rocca, J L; Verdon, E

    2004-09-17

    In this work we demonstrate that the X-Terra RP18 stationary phase, specially designed for the analysis of basic compounds in liquid chromatography, may be successfully used in capillary electrochromatography. Although this packing material does not afford a sufficient electroosmotic flow with classical hydro-organic mobile phases, the addition of a surfactant that adsorbs onto the stationary phase allows to generate a sustainable electroosmosis flow (EOF), the direction of which depends on the charge of the surfactant. The way of manipulating the electroosmotic flow is described (nature and concentration of the added surfactant, proportion of the organic modifier in the mobile phase, pH). It is then demonstrated that high efficiencies can be reached with this packing material (up to 220,000 plates/m with a mean diameter particles of 3.5 microm) when it is operated at high linear velocities. Then the separations of different classes of compounds such as amphenicol antibiotics, macrolide antibiotics or basic test solutes with mobile phases with pH up to 10.8 are described. The influence of the addition of sodium dodcylsulfate (SDS) to the mobile phase on the retention is described and the selectivity of the X-Terra RP18 stationary phase is compared to that of a more traditional phase, i.e. Hypersil C18 stationary phase with SDS added to the mobile phase. However, it is shown that a good repeatability of the retention factors can only be obtained when the ionization of the compounds is totally suppressed since electrolysis of the buffered hydro-organic mobile phase occurs in the buffer reservoirs leading to a variation of the mobile phase pH and consequently to a modification of the ionization degree of the solutes having their pKa close to the mobile phase pH.

  15. Preparation and Characterization of Temperature-responsive Porous Monoliths

    Institute of Scientific and Technical Information of China (English)

    ZHANG, Rongyue; QI, Li; XIN, Peiyong; YANG, Gengliang; CHEN, Yi

    2009-01-01

    A new temperature-responsive porous monolith has been prepared by surface-initiated activators generated by electron transfer atom transfer radical polymerization (AGET ATRP) grafting poly(N-isopropylacrylamide) (PNIPAAm) within the pores of the porous polymer monolith. The grafting copolymerization was carried out by a method based on a continuous flow-through technique without special deoxygenation procedure needed in the general ATRP. The addition of ascorbic acid could counteract the oxidation effect of oxygen diffusing into the reaction system. The resulting grafted monolith was characterized by a mercury intrusion method and the size of macropore was 3.65 μm, which was suitable for flow through the monolith for HPLC. The thermally responsive property of the grafted monolith was evaluated by HPLC using steroids with various hydrophobicities as probes. Through determination of retention factor of each steroid on the grafted monolith at different temperatures using water as mobile phase, it was found that the slope of the plot of retention factor of each steroid versus the temperature changed around the low critical solution temperature (LCST, 32 ℃) of PNIPAAm in water. It was relative to the grafted PNIPAAm temperature sensitivity that a hydrophobic and hydrophilic alternation would take place around its LCST.Based on this thermally responsive property, the grafted monolith was used as stationary phase for HPLC and to separate the steroids using water as mobile phase by changing the column temperature. As a mobile phase, water is much better than organic solvents concerning the environment.

  16. Ionic liquids as stationary phases in gas chromatography--an LSER investigation of six commercial phases and some applications.

    Science.gov (United States)

    Weber, Waldemar; Andersson, Jan T

    2014-09-01

    The separation properties of six novel stationary phases for gas chromatography, commercially available from Sigma-Aldrich (Supelco) and based on ionic liquids (ILs), were investigated. The linear solvation energy relationship model (LSER) was used to describe the molecular interactions between these stationary phases and 30 solutes. The solutes belong to different groups of compounds, like haloalkanes, alcohols, ketones, aromatics, aliphatics, and others. A good description of different interactions, as described by the LSER model, could be achieved. The calculated values of system constants for the ionic liquid phases were compared with constants of commonly used standard phases like a 5 % phenyl/95 % dimethyl siloxane and a polyethylene glycol phase. The solute descriptors are in good agreement with those found by previous authors who have used the LSER model for 44 different ionic liquids as stationary phase. The experiments were carried out at two temperatures to evaluate the influence on the phase parameters and separation characteristics. The interactions of different functional groups with the IL phases are discussed. These novel IL phases are a promising replacement of or an addition to common polar phases. Based on the evaluated phase properties, several possibilities for applications of these novel phases are shown.

  17. CspC and CspD are essential for Caulobacter crescentus stationary phase survival.

    Science.gov (United States)

    Balhesteros, Heloise; Mazzon, Ricardo R; da Silva, Carolina A P T; Lang, Elza A S; Marques, Marilis V

    2010-09-01

    The cold shock response in bacteria involves the expression of low-molecular weight cold shock proteins (CSPs) containing a nucleic acid-binding cold shock domain (CSD), which are known to destabilize secondary structures on mRNAs, facilitating translation at low temperatures. Caulobacter crescentus cspA and cspB are induced upon cold shock, while cspC and cspD are induced during stationary phase. In this work, we determined a new coding sequence for the cspC gene, revealing that it encodes a protein containing two CSDs. The phenotypes of C. crescentus csp mutants were analyzed, and we found that cspC is important for cells to maintain viability during extended periods in stationary phase. Also, cspC and cspCD strains presented altered morphology, with frequent non-viable filamentous cells, and cspCD also showed a pronounced cell death at late stationary phase. In contrast, the cspAB mutant presented increased viability in this phase, which is accompanied by an altered expression of both cspC and cspD, but the triple cspABD mutant loses this characteristic. Taken together, our results suggest that there is a hierarchy of importance among the csp genes regarding stationary phase viability, which is probably achieved by a fine tune balance of the levels of these proteins.

  18. Method development for impurity profiling in SFC: The selection of a dissimilar set of stationary phases.

    Science.gov (United States)

    Galea, Charlene; Mangelings, Debby; Heyden, Yvan Vander

    2015-01-01

    Supercritical fluid chromatography (SFC) is drawing considerable interest as separation technique in the pharmaceutical industry. The technique is already well established in chiral separations both analytically and on a preparative scale. The use of SFC as a technique for drug impurity profiling is examined here. To define starting conditions in method development for drug impurity profiling, a set of dissimilar stationary phases is screened in parallel. The possibility to select a set of dissimilar columns using the retention factors (k-values) for a set of 64 drugs measured on 27 columns in SFC was examined. Experiments were carried out at a back-pressure of 150 bar and 25 °C with a mobile phase consisting of CO2 and methanol with 0.1% isopropylamine (5-40% over 10 min) at a flow rate of 3 mL/min. These k-values were then used to calculate correlation coefficients on the one hand and to perform a principal component analysis on the other. The Kennard and Stone algorithm, besides dendrograms and correlation-coefficient colour maps were used to select a set of 6 dissimilar stationary phases. The stationary phase characterization results from this study were compared to those from previous studies found in the literature. Retention mechanisms for compounds possessing different properties were also evaluated. The dissimilarity of the selected subset of 6 stationary phases was validated using mixtures of compounds with similar properties and structures, as one can expect in a drug impurity profile.

  19. Coating properties of a novel water stationary phase in capillary supercritical fluid chromatography.

    Science.gov (United States)

    Murakami, Jillian N; Thurbide, Kevin B

    2015-05-01

    The coating properties of a novel water stationary phase used in capillary supercritical fluid chromatography were investigated. The findings confirm that increasing the length or internal diameter of the type 316 stainless-steel column used provides a linear increase in the volume of stationary phase present. Under normal operating conditions, results indicate that about 4.9 ± 0.5 μL/m of water phase is deposited uniformly inside of a typical 250 μm internal diameter 316 stainless-steel column, which translates to an area coverage of about 6.3 ± 0.5 nL/mm(2) regardless of dimension. Efforts to increase the stationary phase volume present showed that etching the stainless-steel capillary wall using hydrofluoric acid was very effective for this. For instance, after five etching cycles, this volume doubled inside of both the type 304 and the type 316 stainless-steel columns examined. This in turn doubled analyte retention, while maintaining good peak shape and column efficiency. Overall, 316 stainless-steel columns were more resistant to etching than 304 stainless-steel columns. Results indicate that this approach could be useful to employ as a means of controlling the volume of water stationary phase that can be established inside of the stainless-steel columns used with this supercritical fluid chromatography technique.

  20. Evaluation of new ionic liquids as high stability selective stationary phases in gas chromatography.

    Science.gov (United States)

    González Álvarez, Jaime; Blanco Gomis, Domingo; Arias Abrodo, Pilar; Díaz Llorente, Daniel; Busto, Eduardo; Ríos Lombardía, Nicolás; Gotor Fernández, Vicente; Gutiérrez Álvarez, María Dolores

    2011-05-01

    Two ionic liquids (ILs), namely (S,S)-1-butyl-3-(2'-hydroxy-cyclohexyl)-3H-imidazol-1-ium tetrafluoroborate and (S,S)-1-butyl-3-(2'-acetyl-cyclohexyl)-3H-imidazol-1-ium tetrafluoroborate have been employed as stationary phases in capillary gas chromatography. These new phases exhibit a column efficiency of 1,600 and 2,100 plates m(-1) for IL 1 and IL 2, respectively, a wide operating temperature range and good thermal stability (bleeding temperature of 250 °C for IL 1 and 160 °C for IL 2). Inverse gas chromatography (GC) analyses were used to study the solvation properties of these ILs through a linear solvation energy model. The application of these ILs as new GC stationary phases was studied. These stationary phases exhibited unique selectivity for many organic substances, such as alkanes, ketones, esters, and aromatic compounds. The efficient separation of several mixtures containing compounds of different polarities and the good separation of fatty acid methyl esters (FAMEs) and cis/trans isomers indicate that these ILs may be applicable as a new type of GC stationary phases.

  1. Chromatographic comparison of bupivacaine imprinted polymers prepared in crushed monolith, microsphere, silica-based composite and capillary monolith formats.

    Science.gov (United States)

    Oxelbark, Joakim; Legido-Quigley, Cristina; Aureliano, Carla S A; Titirici, Maria-Magdalena; Schillinger, Eric; Sellergren, Börje; Courtois, Julien; Irgum, Knut; Dambies, Laurent; Cormack, Peter A G; Sherrington, David C; De Lorenzi, Ersilia

    2007-08-10

    A comprehensive comparison of five chromatographic stationary phases based on molecularly imprinted polymers is presented. Efficiency, imprinting factors, water compatibility and batch-to-batch reproducibility are discussed for crushed monolith, microspheres, two silica-based composites and capillary monoliths, all imprinted with the local anaesthetic bupivacaine. Synthesis protocol and chromatographic test conditions have been kept fixed within certain limits, in order to provide further insight into the strengths and weaknesses of the different formats. Excluding microparticles, all formats give satisfactory performance, especially in aqueous mobile phases. An assessment of batch-to-batch reproducibility in different mobile phases adds further value to this comparison study.

  2. [Using barium fluoride fine particles as stationary phase for TLC/FTIR analysis].

    Science.gov (United States)

    Liu, Xi; Pan, Qing-hua; Ding, Jie; Zhu, Qing; He, An-qi; Yue, Shi-juan; Li, Xiao-pei; Hu, Li-ping; Xia, Jin-ming; Liu, Cui-ge; Wei, Yong-ju; Yu, Jiang; Yang, Zhan-lan; Zhu, Xi; Xu, Yi-zhuang; Wu, Jin-guang

    2011-07-01

    In situ TLC/FTIR technique has tremendous potential in the analysis of complex mixtures. However, the progress in this technique was quite slow. The reason is that conventional stationary phase such as silica gel etc. has strong absorption in FTIR spectrum and thus brings about severe interference in the detection of samples. To solve the problem, the authors propose to use barium fluoride fine particles as stationary phase of TLC plate. The reasons are as follows: Barium fluoride wafer has been extensively used as infrared window in FTIR experiments and it has no absorbance in an IR region between 4 000 and 800 cm'. As a matter of fact, the atomic mass of barium and fluoride is quite large, thus the normal vibration of BaF2 lattice is limited in far-IR region and low frequency part of mid-IR region. Therefore, the interference caused by IR absorption of stationary phase can be resolved if BaF2 is used as stationary phase of TLC plate. Moreover, BaF2 is quite stable and insolvable in water and most organic solvents and it will not be dissolved by mobile phase or react with samples in TLC separation. Additionally, decreasing the particle size of BaF2 is very important in TLC/FTIR analysis technique. The reason is two-fold: First, decreasing the particle size of stationary phase is helpful to improving the efficiency of separation by TLC plate; second, decreasing the size of BaFz particle can improve the quality of FTIR spectra by alleviating the problem of light scattering. By optimizing the synthetic conditions, fine particles of barium fluoride were obtained. SEM results indicate that the size of the BaF2 particles is around 500 nm. FTIR spectrum of the BaF2 particles shows that no absorption of impurity was observed. Moreover, the elevation of baseline caused by light scattering is insignificant. The authors have developed a new technique named "settlement volatilization method" to prepare TLC plate without polymeric adhesive that may bring about significant

  3. Isolation of quiescent and nonquiescent cells from yeast stationary-phase cultures

    NARCIS (Netherlands)

    Allen, Chris; Büttner, Sabrina; Aragon, Anthony D.; Thomas, Jason A.; Meirelles, Osorio; Jaetao, Jason E.; Benn, Don; Ruby, Stephanie W.; Veenhuis, Marten; Madeo, Frank; Werner-Washburne, Margaret

    2006-01-01

    Quiescence is the most common and, arguably, most poorly understood cell cycle state. This is in part because pure populations of quiescent cells are typically difficult to isolate. We report the isolation and characterization of quiescent and nonquiescent cells from stationary-phase (SP) yeast

  4. Planar gas chromatography column on aluminum plate with multi-walled carbon nanotubes as stationary phase

    Science.gov (United States)

    Platonov, I. A.; Platonov, V. I.; Pavelyev, V. S.

    2016-04-01

    The high selectivity of the adsorption layer for low-boiling alkanes is shown, the separation factor (α) couple iso-butane / butane is 1.9 at a column temperature of 50 °C.The paper presents sorption and selective properties of planar gas chromatography column on aluminum plate with multi-walled carbon nanotubes as the stationary phase.

  5. Surface modification of polytetrafluoroethylene column for two-stationary phase separations by counter-current chromatography.

    Science.gov (United States)

    Quan, Kai-jun; Huang, Xin-yi; Li, Xiao-ting; Wang, Gao-hong; Liu, Yan-juan; Duan, Wen-da; Di, Duo-long

    2015-11-27

    To improve the separation capability of CCC, a novel solid-liquid two-stationary phases CCC (ASP-CCC) column was prepared employing graphene oxide (GO) conjugated poly-dopamine (PD) coating (GO/PD) as auxiliary stationary phase (ASP). The results of Scanning electron microscopy (SEM), contact angle and X-ray photoelectron spectroscopy (XPS) indicated that nanostructured GO and PD were successfully grafted on the inner wall of the PTFE column. Three alkaloid compounds were selected as the target analytes to evaluate the performance of the novel column. Because of the intermolecular force (hydrogen bond, electrostatic interaction and π-π interaction) between the ASP and model compounds, three analytes were well separated with this novel ASP-CCC column. Additionally, the novel column exhibited higher stationary phase retention ratio, about 8%, than original column without changing the chromatographic condition. Furthermore, the eluotropic sequence of analytes on novel column was in accordance with that in the original column. This suggested that the novel column is a CCC column with auxiliary stationary phase (ASP) in its own right, and the present separation mode is the combination of partition chromatography and adsorption chromatography.

  6. Analysis of Data on Xanthan Fermentation in Stationary Phase Using Black Box and Metabolic Network Models

    Institute of Scientific and Technical Information of China (English)

    马红武; 赵学明; 唐寅杰

    1999-01-01

    The xanthan fermentation data in the stationary phase was analyzed using the black box and the metabolic network models. The data consistency ls checked through the elemental balance in the black box model. In the metabolic network model, the metabolic flux distribution in the cell is calculated using the metabolic flux analysis method, then the maintenance coefficients is calculated.

  7. Shotgun proteomic monitoring of Clostridium acetobutylicum during stationary phase of butanol fermentation using xylose and comparison with the exponential phase.

    Science.gov (United States)

    Sivagnanam, Kumaran; Raghavan, Vijaya G S; Shah, Manesh; Hettich, Robert L; Verberkmoes, Nathan C; Lefsrud, Mark G

    2012-06-01

    Economically viable production of solvents through acetone-butanol-ethanol (ABE) fermentation requires a detailed understanding of Clostridium acetobutylicum. This study focuses on the proteomic profiling of C. acetobutylicum ATCC 824 from the stationary phase of ABE fermentation using xylose and compares with the exponential growth by shotgun proteomics approach. Comparative proteomic analysis revealed 22.9% of the C. acetobutylicum genome and 18.6% was found to be common in both exponential and stationary phases. The proteomic profile of C. acetobutylicum changed during the ABE fermentation such that 17 proteins were significantly differentially expressed between the two phases. Specifically, the expression of five proteins namely, CAC2873, CAP0164, CAP0165, CAC3298, and CAC1742 involved in the solvent production pathway were found to be significantly lower in the stationary phase compared to the exponential growth. Similarly, the expression of fucose isomerase (CAC2610), xylulose kinase (CAC2612), and a putative uncharacterized protein (CAC2611) involved in the xylose utilization pathway were also significantly lower in the stationary phase. These findings provide an insight into the metabolic behavior of C. acetobutylicum between different phases of ABE fermentation using xylose.

  8. Shotgun proteomic monitoring of Clostridium acetobutylicum during stationary phase of butanol fermentation using xylose and comparison with the exponential phase

    Energy Technology Data Exchange (ETDEWEB)

    Sivagnanam, Kumaran [McGill University, Montreal, Quebec; Raghavan, Vijaya G. S. [McGill University, Montreal, Quebec; Shah, Manesh B [ORNL; Hettich, Robert {Bob} L [ORNL; Verberkmoes, Nathan C [ORNL; Lefsrud, Mark G [McGill University, Montreal, Quebec

    2012-01-01

    Economically viable production of solvents through acetone butanol ethanol (ABE) fermentation requires a detailed understanding of Clostridium acetobutylicum. This study focuses on the proteomic profiling of C. acetobutylicum ATCC 824 from the stationary phase of ABE fermentation using xylose and compares with the exponential growth by shotgun proteomics approach. Comparative proteomic analysis revealed 22.9% of the C. acetobutylicum genome and 18.6% was found to be common in both exponential and stationary phases. The proteomic profile of C. acetobutylicum changed during the ABE fermentation such that 17 proteins were significantly differentially expressed between the two phases. Specifically, the expression of five proteins namely, CAC2873, CAP0164, CAP0165, CAC3298, and CAC1742 involved in the solvent production pathway were found to be significantly lower in the stationary phase compared to the exponential growth. Similarly, the expression of fucose isomerase (CAC2610), xylulose kinase (CAC2612), and a putative uncharacterized protein (CAC2611) involved in the xylose utilization pathway were also significantly lower in the stationary phase. These findings provide an insight into the metabolic behavior of C. acetobutylicum between different phases of ABE fermentation using xylose.

  9. Carbon nanotube based stationary phases for microchip chromatography

    DEFF Research Database (Denmark)

    Mogensen, Klaus Bo; Kutter, Jörg Peter

    2012-01-01

    The objective of this article is to provide an overview and critical evaluation of the use of carbon nanotubes and related carbon-based nanomaterials for microchip chromatography. The unique properties of carbon nanotubes, such as a very high surface area and intriguing adsorptive behaviour, have...... already been demonstrated in more classical formats, for improved separation performance in gas and liquid chromatography, and for unique applications in solid phase extraction. Carbon nanotubes are now also entering the field of microfluidics, where there is a large potential to be able to provide...... integrated, tailor-made nanotube columns by means of catalytic growth of the nanotubes inside the fluidic channels. An evaluation of the different implementations of carbon nanotubes and related carbon-based nanomaterials for microfluidic chromatography devices is given in terms of separation performance...

  10. Enantioseparation of Timolol on a Novel β-Cyclodextrin Derivative Chiral Stationary Phase in HPLC.

    Science.gov (United States)

    Zhou, Jie; Sun, Fang; Du, Qiuzheng; Zhao, Suzhen; Pei, Wenjuan

    2016-01-01

    A novel chiral stationary phase was prepared by bonding a novel β-cyclodextrin derivative on silica gel, and it was used for the separation of timolol in high efficiency liquid phase. In the reverse mode, the factors such as the proportion of chiral additives, flow rate, column temperature, repeatability and stability were investigated. The optimum chromatographic conditions are as follows: column temperature was 25°C, flow rate was 0.6 mL min(-1) and mobile phase was methanol-25 mM KH2PO4 (80/20, v/v). The chiral column has good reproducibility (Rs = 4.49, 4.51 and 4.40, respectively) and a certain degree of stability (Rs = 4.49, 3.01 and 0.72, respectively). This chiral stationary phase presented good chiral recognition performance toward timolol with good resolution (Rs = 4.49).

  11. Synthesis and purification of iodoaziridines involving quantitative selection of the optimal stationary phase for chromatography.

    Science.gov (United States)

    Boultwood, Tom; Affron, Dominic P; Bull, James A

    2014-05-16

    The highly diastereoselective preparation of cis-N-Ts-iodoaziridines through reaction of diiodomethyllithium with N-Ts aldimines is described. Diiodomethyllithium is prepared by the deprotonation of diiodomethane with LiHMDS, in a THF/diethyl ether mixture, at -78 °C in the dark. These conditions are essential for the stability of the LiCHI2 reagent generated. The subsequent dropwise addition of N-Ts aldimines to the preformed diiodomethyllithium solution affords an amino-diiodide intermediate, which is not isolated. Rapid warming of the reaction mixture to 0 °C promotes cyclization to afford iodoaziridines with exclusive cis-diastereoselectivity. The addition and cyclization stages of the reaction are mediated in one reaction flask by careful temperature control. Due to the sensitivity of the iodoaziridines to purification, assessment of suitable methods of purification is required. A protocol to assess the stability of sensitive compounds to stationary phases for column chromatography is described. This method is suitable to apply to new iodoaziridines, or other potentially sensitive novel compounds. Consequently this method may find application in range of synthetic projects. The procedure involves firstly the assessment of the reaction yield, prior to purification, by (1)H NMR spectroscopy with comparison to an internal standard. Portions of impure product mixture are then exposed to slurries of various stationary phases appropriate for chromatography, in a solvent system suitable as the eluent in flash chromatography. After stirring for 30 min to mimic chromatography, followed by filtering, the samples are analyzed by (1)H NMR spectroscopy. Calculated yields for each stationary phase are then compared to that initially obtained from the crude reaction mixture. The results obtained provide a quantitative assessment of the stability of the compound to the different stationary phases; hence the optimal can be selected. The choice of basic alumina, modified to

  12. Development of monolith with a carbon-nanofiber-washcoat as a structured catalyst support in liquid phase

    NARCIS (Netherlands)

    Jarrah, Nabeel A.; Ommen, van J.G.; Lefferts, L.

    2003-01-01

    Washcoats with improved mass transfer properties are necessary to circumvent concentration gradients in case of fast reactions in liquid phase, e.g. nitrate hydrogenation. A highly porous, high surface area (180 m2/g) and thin washcoat of carbon fibers, was produced on a monolith support by methane

  13. Glucose, cellobiose, lactose and raffinose used as chiral stationary phases in HPLC

    Institute of Scientific and Technical Information of China (English)

    Jian Yu Wang; Feng Zhao; Mei Zhang; Ya Peng; Li Ming Yuan

    2008-01-01

    This paper presents the enantioseparation using glucose,cellobiose,lactose and raffinose as chiral selector bonded to silica gel via an arm in HPLC.Surprisingly,they also possess high enantioseparation selectivity,may be used in normal-phase and reversedphase mode.and there is a big chiral discriminating complementary.This work indicates that oligosacchafides could soon become very attractive as a new class of chiral stationary phase for HPLC.

  14. Enantiomeric Separation of Four Chiral Compounds Using Immobilized Cellulose 3, 5-Dimethylphenylcarbamate as Chiral Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    Yong Fei MING; Liang ZHAO; Hong Li ZHANG; Yan Ping SHI; Yong Min LI; Li Ren CHEN

    2006-01-01

    A new chiral stationary phase of 3, 5-dimethylphenylcarbamates of cellulose,chemically bonded to 3-aminopropylsilica gel at the 6-positions of the glucose units, was prepared.The solvent versatility of the CSP was investigated for the enantioselective separation of four pairs of enantiomers using THF and chloroform as non-standard mobile phase eluent in HPLC. The influence of temperature on the resolution was investigated.

  15. A Low Phase Noise Fully Monolithic 6 GHz Differential Coupled NMOS LC-VCO

    Science.gov (United States)

    Moalla, Dorra Mellouli; Cordeau, David; Mnif, Hassene; Paillot, Jean-Marie; Loulou, Mourad

    2016-01-01

    A fully monolithic 6 GHz low-phase noise Voltage-Controlled-Oscillator (VCO) is presented in this paper. It consists in two LC-NMOS differential VCOs coupled through a resistive network and is implemented on a 0.25 µm BiCMOS SiGe process. This proposed integrated VCO can be used also for phased-array applications to steer the beam over the entire spatial range. In this case, the radiation pattern of the phased antenna array is steered in a particular direction by establishing a constant phase progression in the oscillator chain which can be obtained by detuning the free-running frequencies of the two oscillators in the array. At 2.5 V power supply voltage and a power dissipation of 62.5 mW, the coupled VCO array features a measured worst case phase noise of -102.4 dBc/Hz and -125.64 dBc/Hz at 100 kHz and 1 MHz frequency offset respectively from a 6 GHz carrier. The tuning range is about 400 MHz, from 5.85 to 6.25 GHz, for a tuning voltage varying from 0 to 2.5 V.

  16. The use of the stationary phase method as a mathematical tool to determine the path of optical beams

    OpenAIRE

    Carvalho, Silvânia A.; Leo, Stefano de

    2015-01-01

    We use the stationary phase method to determine the path of optical beams which propagate through a dielectric block. In the presence of partial internal reflection, we recover the geometrical result obtained by using the Snell law. For total internal reflection, the stationary phase method overreaches the Snell law predicting the Goos-Haenchen shift.

  17. Effects of tea tree (Melaleuca alternifolia) oil on Staphylococcus aureus in biofilms and stationary growth phase.

    Science.gov (United States)

    Kwieciński, Jakub; Eick, Sigrun; Wójcik, Kinga

    2009-04-01

    Tea tree oil (TTO) is known for its antimicrobial activity. In this study, we determined whether TTO is effective against Staphylococcus aureus in biofilms and how TTO activity is affected by the S. aureus growth phase. All clinical strains tested were killed by TTO both as planktonic cells and as biofilms. The minimum biofilm eradication concentration was usually two times higher than the minimum bactericidal concentration, yet it was never higher than 1% v/v. The fastest killing of biofilm occurred during the first 15min of contact with TTO and was not influenced by increasing TTO concentration above 1% v/v. Planktonic stationary phase cells exhibited decreased susceptibility to TTO compared with exponential phase cells. The killing rate for stationary phase cells was also less affected by increasing TTO concentration than that for exponential phase cells. These data show that TTO efficiently kills S. aureus in the stationary growth phase and within biofilms and is therefore a promising tool for S. aureus eradication.

  18. Preparation and Evaluation of Immobilized SE-30 Coated Stationary Phases for CEC

    Institute of Scientific and Technical Information of China (English)

    NingBaoming; ZhengJie; ZhangGuodong; XuBingjiu

    2001-01-01

    A new type of stationary phase for capillary electrochromatography (CEC), immobilized SE-30 coated silica gel, was developed and the columns packed with this new phase were prepared and evaluated. It was found that this phase could be used to make frits for the micro-columns as well as to pack the bulk of the column. By sticking together the particles in the whole of the column bed, this new packing resulted in columns with stable performance, even under rigorous conditions: more than 380 consecutive separations were effected with these columns with the mobile phase pH of 11.7.

  19. Preparation of Monolithic Column and Its Application in p-CEC

    Institute of Scientific and Technical Information of China (English)

    Jun Jiao YANG; Kai ZHANG; Ru Yu GAO; Yu Kui ZHANG; Chao YAN

    2004-01-01

    A novel monolithic stationary phase having long alkyl chain ligands was introduced and evaluated in pressurized-capillary electrochromatography of small neutral and charged compounds.The monolithic column was prepared by the in situ copolymerization of ethylene dimethacrylate,1-hexadecene,allyl alcohol and 2-acrylamido-2-methyl-1-propanesulfonic acid in a quaternary porogenic solvent mixture consisting of 1,4-butanediol,cyclohexanol,dodecanol and water.

  20. Characterization of five chemistries and three particle sizes of stationary phases used in supercritical fluid chromatography.

    Science.gov (United States)

    Khater, S; West, C; Lesellier, E

    2013-12-06

    Sub-2-microns particles employed as supporting phases are known to favor column efficiency. Recently a set of columns based on sub-2-microns particles for use with supercritical fluid mobile phases have been introduced by Waters. Five different stationary phase chemistries are available: BEH silica, BEHEthyl-pyridine, X Select CSH Fluorophenyl, HSS C18 SB and BEH Shield RP18. This paper describes the characterization of 15 stationary phases, the five different chemistries, and three particle sizes, 1.7 (or 1.8), 3.5 and 5 microns, with the same carbon dioxide–methanol mobile phase and a set of more than a hundred compounds. The interactions established in the 15 different chromatographic systems used in supercritical fluid chromatography (SFC) are assessed with linear solvation energy relationships (LSERs).The results show the good complementarity of the five column chemistries, and their comparative location inside a classification map containing today around 70 different commercial phases. Among the five different chemistries, the HSS C18 SB phase displays a rather unusual behavior in regards of classical C18 phases, as it displays significant hydrogen–bonding interactions. Besides, it appears, as expected, that the BEH Ethyl–pyridine phase has weak interactions with basic compounds. The effect of particle size was studied because smaller particles induce increased inlet and internal pressure. For compressible fluids,this pressure change modifies the fluid density, i.e. the apparent void volume and the eluting strength.These changes could modify the retention and the selectivity of compounds in the case of method trans-fer, by using different particle sizes, from 5 down to 1.7 m. A hierarchical cluster analysis shows that stationary phase clusters were based on the phase chemistry rather than on the particle size, meaning that method transfer from 5 to 1.7 microns can be achieved in the subcritical domain i.e. by using a weakly compressible fluid.

  1. Preparation and evaluation of novel chiral stationary phases based on quinine derivatives comprising crown ether moieties.

    Science.gov (United States)

    Wang, Dongqiang; Zhao, Jianchao; Wu, Haixia; Wu, Haibo; Cai, Jianfeng; Ke, Yanxiong; Liang, Xinmiao

    2015-01-01

    The C9-position of quinine was modified by meta- or para-substituted benzo-18-crown-6, and immobilized on 3-mercaptopropyl-modified silica gel through the radical thiol-ene addition reaction. These two chiral stationary phases were evaluated by chiral acids, amino acids, and chiral primary amines. The crown ether moiety on the quinine anion exchanger provided a ligand-exchange site for primary amino groups, which played an important role in the retention and enantioselectivity for chiral compounds containing primary amine groups. These two stationary phases showed good selectivity for some amino acids. The complex interaction between crown ether and protonated primary amino group was investigated by the addition of inorganic salts such as LiCl, NH4Cl, NaCl, and KCl to the mobile phase. The resolution results showed that the simultaneous interactions between two function moieties (quinine and crown ether) and amino acids were important for the chiral separation.

  2. HPLC Enantioseparation of Phenylcarbamic Acid Derivatives by Using Macrocyclic Chiral Stationary Phases

    Directory of Open Access Journals (Sweden)

    Hroboňová Katarína

    2016-06-01

    Full Text Available The HPLC by using chiral stationary phases based on macrocyclic antibiotics, dimethylphenyl carbamate cyklofructan 7 and β-cyclodextrin in terms of polar-organic separation mode (mobile phase methanol/acetonitrile/acetic acid/triethylamine were used for enantioseparation of alkoxy derivatives of phenylcarbamic acid. The effect of the analyte structures on the efficiency of enantioseparation was investigated. The most suitable stationary phase was teicoplanin aglycone, where the separations of the enantiomers were obtained (the resolution value from 0.65 to 2.90, depending on the structure of the analyte. Significant effect on the resolution of the enantiomers has position of alkoxy substituent in the hydrophobic part of the molecule. The enantiorecognition was achieved for 3-alkoxysubstituted derivatives.

  3. Sol-gel synthesis of macroporous barium zirconate monoliths from ionic precursors via a phase separation route

    Science.gov (United States)

    Guo, Xingzhong; Wang, Zichen; Song, Jie; Yang, Hui

    2017-03-01

    Monolithic macroporous barium zirconate derived from ionic precursors has been successfully prepared via a phase separation route in the presence of poly(ethylene oxide) (PEO) and propyleneoxide (PO). Poly(ethylene oxide) (PEO) acts as a phase separation inducer, while propyleneoxide (PO) acts as a gelation accelerant in the sol-gel process. Appropriate choice of poly(ethylene oxide) (PEO) and propyleneoxide (PO) allows the production of continuous macroporous monolithic gel with a porosity of ca. 63% and a macropore size of 1.8 μm. Some BaCl2 recrystallizes in the dried gel, and subsequently tetragonal ZrO2 phase precipitates after heat-treated at 800 °C. The crystalline phase barium zirconate forms after heat treatment at 1100 °C in air, while the macroporous structure is preserved with a slight increase of porosity and a decrease of macropore size.

  4. Determination of volatile compounds in cider apple juices using a covalently bonded ionic liquid coating as the stationary phase in gas chromatography.

    Science.gov (United States)

    Pello-Palma, Jairo; González-Álvarez, Jaime; Gutiérrez-Álvarez, María Dolores; Dapena de la Fuente, Enrique; Mangas-Alonso, Juan José; Méndez-Sánchez, Daniel; Gotor-Fernández, Vicente; Arias-Abrodo, Pilar

    2017-04-01

    A chromatographic method for the separation of volatile compounds in Asturian cider apple juices has been developed. For this separation purpose, a monocationic imidazolium-based ionic liquid bearing a reactive terminal iodine atom was synthesized by a quaternization-anion exchange chemical sequence. Next, the gas chromatography (GC) stationary phase was prepared by covalently linking the imidazolium monolith to the reactive silanol groups of the inner capillary wall at 70 °C. This coated GC column exhibited good thermal stability (290 °C), as well as good efficiency (2000 plates/m) in the separation of volatile compounds from Asturian apple cider juices, and was characterized using the Abraham solvation parameter model. The intra-day and inter-day precision of the chromatographic method was evaluated, obtaining relative standard deviations from 3.7 to 12.9% and from 7.4 to 18.0%, respectively. Furthermore, recoveries from 82.5 to 122% were achieved. Graphical Abstract Covalent bonding of an ionic liquid to inner column wall led to a great improvement of the separation efficiencies of stationary phases in gas chromatography.

  5. Preparation of a biomimetic polyphosphorylcholine monolithic column for immobilized artificial membrane chromatography.

    Science.gov (United States)

    Zhao, XiangLong; Chen, WeiJia; Zhou, ZhengYin; Wang, QiQin; Liu, ZhengHua; Moaddel, Ruin; Jiang, ZhengJin

    2015-08-14

    The present work aims to prepare a novel phosphatidylcholine functionalized monolithic stationary phase by in situ co-polymerization of 12-methacryloyl dodecylphosphocholine (MDPC) and ethylene dimethacrylate (EDMA) for immobilized artificial membrane chromatography. Scanning electron microscopy, energy dispersive X-ray spectroscopy, FT-IR spectroscopy, pore size distribution analysis, ζ-potential analysis and micro-HPLC were used to evaluate the monolithic structure and physicochemical properties. Satisfactory morphology, high mechanical stability, good permeability and chromatographic performance were obtained on the optimized monolithic columns. A typical reverse-phase retention mechanism was observed over a wide range of organic solvent content (acetonitrilecolumn (IAM.PC.DD2) and poly(MDPC-co-EDMA) monolith. This novel poly(MDPC-co-EDMA) monolith exhibited good potential for studying the drug-membrane interaction.

  6. A monolithic K-band phase-locked loop for microwave radar application

    Science.gov (United States)

    Zhou, Guangyao; Ma, Shunli; Li, Ning; Ye, Fan; Ren, Junyan

    2017-02-01

    A monolithic K-band phase-locked loop (PLL) for microwave radar application is proposed and implemented in this paper. By eliminating the tail transistor and using optimized high-Q LC-tank, the proposed voltage-controlled oscillator (VCO) achieves a tuning range of 18.4 to 23.3 GHz and reduced phase noise. Two cascaded current-mode logic (CML) divide-by-two frequency prescalers are implemented to bridge the frequency gap, in which inductor peaking technique is used in the first stage to further boost allowable input frequency. Six-stage TSPC divider chain is used to provide programmable division ratio from 64 to 127, and a second-order passive loop filter with 825 kHz bandwidth is also integrated on-chip to minimize required external components. The proposed PLL needs only approximately 18.2 μs settling time, and achieves a wide tuning range from 18.4 to 23.3 GHz, with a typical output power of -0.84 dBm and phase noise of -91.92 dBc/Hz @ 1 MHz. The chip is implemented in TSMC 65 nm CMOS process, and occupies an area of 0.56 mm2 without pads under a 1.2 V single voltage supply. Project supported by the National High-Tech Research and Development Program of China (No. 2013AA014101).

  7. The extracellular proteome of Rhizobium etli CE3 in exponential and stationary growth phase

    Directory of Open Access Journals (Sweden)

    Mendoza-Hernández Guillermo

    2010-10-01

    Full Text Available Abstract Background The extracellular proteome or secretome of symbiotic bacteria like Rhizobium etli is presumed to be a key element of their infection strategy and survival. Rhizobia infect the roots of leguminous plants and establish a mutually beneficial symbiosis. To find out the possible role of secreted proteins we analyzed the extracellular proteome of R. etli CE3 in the exponential and stationary growth phases in minimal medium, supplemented with succinate-ammonium. Results The extracellular proteins were obtained by phenol extraction and identified by LC-ESI MS/MS. We identified 192 and 191 proteins for the exponential and stationary phases respectively. Using the software Signal P, we predicted signal peptides for 12.95% and 35.60% of the proteins identified in the exponential and stationary phases, respectively, which could therefore be secreted by the Sec pathway. For the exponential growth phase, we found in abundance proteins like the ribosomal proteins, toxins and proteins belonging to the group "defence mechanisms". For the stationary growth phase, we found that the most abundant proteins were those with unknown function, and in many of these we identified characteristic domains of proteases and peptidases. Conclusions Our study provided the first dataset of the secretome of R. etli and its modifications, which may lead to novel insights into the adaptive response of different stages of growth. In addition, we found a high number of proteins with unknown function; these proteins could be analyzed in future research to elucidate their role in the extracellular proteome of R. etli.

  8. Evaluation of thermally pretreated silica stationary phases under hydrophilic interaction chromatography conditions.

    Science.gov (United States)

    Mignot, Mélanie; Périat, Aurélie; Peulon-Agasse, Valérie; Cardinael, Pascal; Veuthey, Jean-Luc; Guillarme, Davy

    2016-05-01

    Three novel hydrophilic interaction chromatography columns packed with bare silica 2.6 μm superficially porous particles were evaluated. These stationary phases undergo a different pretreatment temperature (400, 525, and 900°C) that might influence their kinetic performance and thermodynamic properties. In the first instance, we demonstrated that the performance of these columns was inferior to the commercial ones in the low plate count range (10 000 plates), but was more favorable for N values beyond 40 000 plates. Thanks to its high permeability and reasonable flow resistance (φ = 695), together with a minimum reduced heights equivalent to a theoretical plate value of only 2.4, the stationary phase pretreated at 400°C was particularly attractive for N > 70 000 plates with a remarkably low impedance value (E = 2488). In a second step, the impact of pretreatment temperature was evaluated using two mixtures of polar substances, namely nucleobases and derivatives, as well as nicotine and derivatives. Retentions and selectivities achieved on the tested stationary phases were appropriate, but selectivity differences were minor when modifying pretreatment temperature from 400 to 525°C. When we increased the pretreatment temperature up to 900°C, the surface chemistry was more seriously modified. Finally, the columns presented a good stability even at high temperature (70°C), especially for the phases pretreated at 400 and 525°C.

  9. Recent development in liquid chromatography stationary phases for separation of Traditional Chinese Medicine components.

    Science.gov (United States)

    Jin, Hongli; Liu, Yanfang; Guo, Zhimou; Wang, Jixia; Zhang, Xiuli; Wang, Chaoran; Liang, Xinmiao

    2016-10-25

    Traditional Chinese Medicine (TCM) is an ancient medical practice which has been used to prevent and cure diseases for thousands of years. TCMs are frequently multi-component systems with mainly unidentified constituents. The study of the chemical compositions of TCMs remains a hotspot of research. Different strategies have been developed to manage the significant complexity of TCMs, in an attempt to determine their constituents. Reversed-phase liquid chromatography (RPLC) is still the method of choice for the separation of TCMs, but has many problems related to limited selectivity. Recently, enormous efforts have been concentrated on the development of efficient liquid chromatography (LC) methods for TCMs, based on selective stationary phases. This can improve the resolution and peak capacity considerably. In addition, high-efficiency stationary phases have been applied in the analysis of TCMs since the invention of ultra high-performance liquid chromatography (UHPLC). This review describes the advances in LC methods in TCM research from 2010 to date, and focuses on novel stationary phases. Their potential in the separation of TCMs using relevant applications is also demonstrated. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Staphylococcus aureus aconitase inactivation unexpectedly inhibits post-exponential-phase growth and enhances stationary-phase survival.

    Science.gov (United States)

    Somerville, Greg A; Chaussee, Michael S; Morgan, Carrie I; Fitzgerald, J Ross; Dorward, David W; Reitzer, Lawrence J; Musser, James M

    2002-11-01

    Staphylococcus aureus preferentially catabolizes glucose, generating pyruvate, which is subsequently oxidized to acetate under aerobic growth conditions. Catabolite repression of the tricarboxylic acid (TCA) cycle results in the accumulation of acetate. TCA cycle derepression coincides with exit from the exponential growth phase, the onset of acetate catabolism, and the maximal expression of secreted virulence factors. These data suggest that carbon and energy for post-exponential-phase growth and virulence factor production are derived from the catabolism of acetate mediated by the TCA cycle. To test this hypothesis, the aconitase gene was genetically inactivated in a human isolate of S. aureus, and the effects on physiology, morphology, virulence factor production, virulence for mice, and stationary-phase survival were examined. TCA cycle inactivation prevented the post-exponential growth phase catabolism of acetate, resulting in premature entry into the stationary phase. This phenotype was accompanied by a significant reduction in the production of several virulence factors and alteration in host-pathogen interaction. Unexpectedly, aconitase inactivation enhanced stationary-phase survival relative to the wild-type strain. Aconitase is an iron-sulfur cluster-containing enzyme that is highly susceptible to oxidative inactivation. We speculate that reversible loss of the iron-sulfur cluster in wild-type organisms is a survival strategy used to circumvent oxidative stress induced during host-pathogen interactions. Taken together, these data demonstrate the importance of the TCA cycle in the life cycle of this medically important pathogen.

  11. Separation of piracetam derivatives on polysaccharide-based chiral stationary phases.

    Science.gov (United States)

    Kažoka, H; Koliškina, O; Veinberg, G; Vorona, M

    2013-03-15

    High-performance liquid chromatography was used for the enantiomeric separation of two chiral piracetam derivatives. The suitability of six commercially available polysaccharide-based chiral stationary phases (CSPs) under normal phase mode for direct enantioseparation has been investigated. The influence of the CSPs as well the nature and content of an alcoholic modifier in the mobile phase on separation and elution order was studied. It was established that CSP Lux Amylose-2 shows high chiral recognition ability towards 4-phenylsubstituted piracetam derivatives. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Description and Evaluation of Chiral Interactive Sites on Bonded Cyclodextrin Stationary Phases for Liquid Chromatography

    Science.gov (United States)

    Beesley, Thomas E.

    Development of chiral separations has been essential to the drug discovery and development process. The solubility requirements for a number of methods and/or the mobile phase requirements for application of certain detection systems have opened up many opportunities for cyclodextrin-based CSPs for liquid chromatography. Even though a few chiral stationary phases cover a wide area of enantioselectivity, they do not meet the entire needs of the industry. Cyclodextrin phases offer some unique mechanisms and opportunities to resolve chiral separation problems especially in the aqueous reversed-phase and non-aqueous polar organic modes. This chapter addresses the need to understand the chiral stationary phase structure, the mechanisms at work, and the role mobile phase composition plays in driving those mechanisms to produce enantioselectivity. In addition, the development of certain derivatives has played an essential part in expanding that basic role for certain chiral separations. What these derivatives contribute in concert with the basic structure is a critical part of the understanding to the effective use of these phases. During this study it was determined that the role of steric hindrance has been vastly underestimated, both to the extent that it has occurred and to its effectiveness for obtaining enantioselectivity. References to the entire 20-year history of the cyclodextrin phase development and application literature up to this current date have been reviewed and incorporated.

  13. Correlations between the zeta potentials of silica hydride-based stationary phases, analyte retention behaviour and their ionic interaction descriptors.

    Science.gov (United States)

    Kulsing, Chadin; Yang, Yuanzhong; Munera, Caesar; Tse, Colby; Matyska, Maria T; Pesek, Joseph J; Boysen, Reinhard I; Hearn, Milton T W

    2014-03-19

    In this study, the zeta potentials of type-B silica, bare silica hydride, the so-called Diamond Hydride™ and phenyl substituted silica hydride stationary phases have been measured in aqueous-organic media and correction procedures developed to account for the more negative zeta potential values in media containing different acetonitrile contents. Retention studies of 16 basic, acidic and neutral compounds were also performed with these four stationary phases with mobile phases containing 0.1% (v/v) formic acid and various acetonitrile-water compositions ranging from 0-90% (v/v) acetonitrile. The retention properties of these analytes were correlated to the corrected stationary phase zeta potentials measured under these different mobile phase conditions with R(2) values ranging from 0.01 to 1.00, depending on the stationary phase and analyte type. Using linear solvation energy relationships, stationary phase descriptors for each stationary phase have been developed for the different mobile phase conditions. Very high correlations of the zeta potentials with the ionic interaction descriptors were obtained for the type-B silica and the Diamond Hydride™ phases and good correlation with bare silica hydride material whilst there was no correlation observed for the phenyl substituted silica hydride phase. The nature of the retention mechanisms which gives rise to these different observations is discussed. The described methods represent a useful new approach to characterize and assess the retention properties of silica-hydride based chromatographic stationary phases of varying bonded-phase coverage and chemistries, as would be broadly applicable to other types of stationary phase used in the separation sciences.

  14. Hydrophobic polymer monoliths as novel phase separators: Application in continuous liquid-liquid extraction systems

    NARCIS (Netherlands)

    Peroni, D.; Vanhoutte, D.; Vilaplana, F.; Schoenmakers, P.; de Koning, S.; Janssen, H.-G.

    2012-01-01

    Hydrophobic macroporous polymer monoliths are shown to be interesting materials for the construction of "selective solvent gates". With the appropriate surface chemistry and porous properties the monoliths can be made permeable only for apolar organic solvents and not for water. Different poly(butyl

  15. New hydrophilic interaction/reversed-phase mixed-mode stationary phase and its application for analysis of nonionic ethoxylated surfactants.

    Science.gov (United States)

    Liu, Xiaodong; Pohl, Christopher

    2008-05-16

    We have developed a new stationary phase that combines both hydrophilic interaction and reversed-phase characteristics. The new phase is based on high-purity, porous and spherical silica gel functionalized with a silyl ligand consisting of both hydrophilic and hydrophobic functionalities. This phase can be operated in both HILIC mode (high organic solvent) and RPLC mode (low organic solvent). An optimal balance of hydrophilic and hydrophobic moieties on the silica surface provides unique chromatographic properties that make it useful for determination of alkyl chain distribution and degree of ethoxylation (EO number) of nonionic ethoxylated surfactants.

  16. Homochiral metal-organic framework used as a stationary phase for high-performance liquid chromatography.

    Science.gov (United States)

    Kong, Jiao; Zhang, Mei; Duan, Ai-Hong; Zhang, Jun-Hui; Yang, Rui; Yuan, Li-Ming

    2015-02-01

    Metal-organic frameworks are promising porous materials. Chiral metal-organic frameworks have attracted considerable attention in controlling enantioselectivity. In this study, a homochiral metal-organic framework [Co(2) (D-cam)(2) (TMDPy)] (D-cam = D-camphorates, TMDPy = 4,4'-trimethylenedipyridine) with a non-interpenetrating primitive cubic net has been used as a chiral stationary phase in high-performance liquid chromatography. It has allowed the successful separation of six positional isomers and six chiral compounds. The good selectivity and baseline separation, or at least 60% valley separation, confirmed its excellent molecular recognition characteristics. The relative standard deviations for the retention time of run-to-run and column-to-column were less than 1.8 and 3.1%, respectively. These results demonstrate that [Co(2) (D-cam)(2) (TMDPy)] may represent a promising chiral stationary phase for use in high-performance liquid chromatography.

  17. Nutrient control for stationary phase cellulase production in Trichoderma reesei Rut C-30.

    Science.gov (United States)

    Callow, Nicholas V; Ray, Christopher S; Kelbly, Matthew A; Ju, Lu-Kwang

    2016-01-01

    This work describes the use of nutrient limitations with Trichoderma reesei Rut C-30 to obtain a prolonged stationary phase cellulase production. This period of non-growth may allow for dependable cellulase production, extended fermentation periods, and the possibility to use pellet morphology for easy product separation. Phosphorus limitation was successful in halting growth and had a corresponding specific cellulase production of 5±2 FPU/g-h. Combined with the addition of Triton X-100 for fungal pellet formation and low shear conditions, a stationary phase cellulase production period in excess of 300 h was achieved, with a constant enzyme production rate of 7±1 FPU/g-h. While nitrogen limitation was also effective as a growth limiter, it, however, also prevented cellulase production.

  18. Comparative Optical Separation of Racemic Ibuprofen by Using Chiral Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    Dalkeun; PARK; Joong; Kee; LEE; 等

    2002-01-01

    Ibprofen is widely used as a non-steroidal anti-inflammatory drug and poduced as racemic mixture.Its pharmacological activity resides only is S-(+)-enantiomer,and R-(-)-enantiomer is not only inactive but also has many side effects.Thus it is necessary to separate Renantiomer from racemic ibuprofen.We studied optical separation of racemic Ibuprofen with chiral high performance liquid chromatography(HPLC).,Out of three different chiral stationary phases,which were selected on the basis of structure and availability,two were found to be effective.There was optimum eluent composition for each stationary phase for good resolution in optical separation.Resolution decreased with increase of eluent flow rate,but effect of injection volume on resolution was insignificant at high eluent flow rate.

  19. Effect of Single-walled Carbon Nanotubes on Cellulose Phenylcarbamate Chiral Stationary Phases

    Institute of Scientific and Technical Information of China (English)

    CHANG Yin-xia; REN Chao-xing; RUAN Qiong; YUAN Li-ming

    2007-01-01

    Single-walled carbon nanotubes(SWNTs) have a high adsorption ability and nanoscale interactions. Cellulose trisphenylcarbamates possess high enantioseparation ability in high-performance liquid chromatography(HPLC). Single-walled carbon nanotubes mixed with cellulose trisphenylcarbamate are coated on the silica gel as chiral stationary phases and higher enantioseparation factors are obtained. After a single-walled carbon nanotube is linked to the 6-position of cellulose 2,3-bisphenylcarbamate, its enantioseparation resolution increases compared to that of the cellulose trisphenylcarbamate. It is the first time that SWNTs have been applied to enantioseparation. The results indicate that the single-walled carbon nanotubes are good promoters of chiral recognition. This method can be used to improve the enantioseparation efficiency of the polysaccharide chiral stationary phases.

  20. Experimental comparison of chiral metal-organic framework used as stationary phase in chromatography.

    Science.gov (United States)

    Xie, Sheng-Ming; Zhang, Mei; Fei, Zhi-Xin; Yuan, Li-Ming

    2014-10-10

    Chiral metal-organic frameworks (MOFs) are a new class of multifunctional material, which possess diverse structures and unusual properties such as high surface area, uniform and permanent cavities, as well as good chemical and thermal stability. Their chiral functionality makes them attractive as novel enantioselective adsorbents and stationary phases in separation science. In this paper, the experimental comparison of a chiral MOF [In₃O(obb)₃(HCO₂)(H₂O)] solvent used as a stationary phase was investigated in gas chromatography (GC), high-performance liquid chromatography (HPLC) and capillary electrochromatography (CEC). The potential relationship between the structure and components of chiral MOFs with their chiral recognition ability and selectivity are presented. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Polymeric imidazolium ionic liquids as valuable stationary phases in gas chromatography: chemical synthesis and full characterization.

    Science.gov (United States)

    González-Álvarez, Jaime; Blanco-Gomis, Domingo; Arias-Abrodo, Pilar; Díaz-Llorente, Daniel; Ríos-Lombardía, Nicolás; Busto, Eduardo; Gotor-Fernández, Vicente; Gutiérrez-Álvarez, María Dolores

    2012-04-06

    Seven new functionalized polymerizable ionic liquids were chemically prepared, and later applied for the preparation of polymeric stationary phases in gas chromatography. These coated GC columns, which exhibited good thermal stabilities (240-300°C) and very high efficiencies (3120-4200 plates/m), have been characterized using the Abraham solvation parameter model. The chromatographic behavior of these polymeric IL columns has been deeply studied observing excellent selectivities in the separation of many organic substances such as alkanes, ketones, alcohols, amines or esters in mixtures of polar and non polar solvents or fragrances. Remarkably, the challenging separation of xylene isomers has been possible using a bis(trifluoromethylsulfonyl)amide based imidazolium IL coated column as a gas chromatography stationary phase.

  2. Temporal transcriptomic analysis of Desulfovibrio vulgaris Hildenborough transition into stationary phase growth during electrondonor depletion

    Energy Technology Data Exchange (ETDEWEB)

    Clark, M.E.; He, Q.; He, Z.; Huang, K.H.; Alm, E.J.; Wan, X.-F.; Hazen, T.C.; Arkin, A.P.; Wall, J.D.; Zhou, J.-Z.; Fields, M.W.

    2006-08-01

    Desulfovibrio vulgaris was cultivated in a defined medium, and biomass was sampled for approximately 70 h to characterize the shifts in gene expression as cells transitioned from the exponential to the stationary phase during electron donor depletion. In addition to temporal transcriptomics, total protein, carbohydrate, lactate, acetate, and sulfate levels were measured. The microarray data were examined for statistically significant expression changes, hierarchical cluster analysis, and promoter element prediction and were validated by quantitative PCR. As the cells transitioned from the exponential phase to the stationary phase, a majority of the down-expressed genes were involved in translation and transcription, and this trend continued at the remaining times. There were general increases in relative expression for intracellular trafficking and secretion, ion transport, and coenzyme metabolism as the cells entered the stationary phase. As expected, the DNA replication machinery was down-expressed, and the expression of genes involved in DNA repair increased during the stationary phase. Genes involved in amino acid acquisition, carbohydrate metabolism, energy production, and cell envelope biogenesis did not exhibit uniform transcriptional responses. Interestingly, most phage-related genes were up-expressed at the onset of the stationary phase. This result suggested that nutrient depletion may affect community dynamics and DNA transfer mechanisms of sulfate-reducing bacteria via the phage cycle. The putative feoAB system (in addition to other presumptive iron metabolism genes) was significantly up-expressed, and this suggested the possible importance of Fe{sup 2+} acquisition under metal-reducing conditions. The expression of a large subset of carbohydrate-related genes was altered, and the total cellular carbohydrate levels declined during the growth phase transition. Interestingly, the D. vulgaris genome does not contain a putative rpoS gene, a common attribute

  3. Copolymerization preparation of cationic cyclodextrin chiral stationary phases for drug enantioseparation in chromatography

    OpenAIRE

    sprotocols

    2015-01-01

    Authors: Ren-Qi Wang, Teng-Teng Ong, Ke Huang, Weihua Tang & Siu-Choon Ng ### Abstract We described a facile and effective protocol wherein radical copolymerization is employed to covalently bond cationic β-cyclodextrin (β-CD) onto silica particles with extended linkage, resulting in a chiral stationary phase (IMPCSP) that can be used for the enantioseparation of racemic drugs in both high-performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC). Start...

  4. The stationary phase point method for transitional scattering: diffractive radio scintillation for pulsar

    CERN Document Server

    Zhang, C M

    2007-01-01

    The stationary phase point (SPP) method in one-dimensional case is introduced to treat the diffractive scintillation. From weak scattering, where the SPP number N=1, to strong scattering (N$\\gg$1), via transitional scattering regime (N$\\sim$2,3), we find that the modulation index of intensity experiences the monotonically increasing from 0 to 1 with the scattering strength, characterized by the ratio of Fresnel scale $\\rf$ to diffractive scale $\\rdiff$.

  5. Polystyrene-divinylbenzene-glycidyl methacrylate stationary phase grafted with poly (amidoamine) dendrimers for ion chromatography.

    Science.gov (United States)

    Guo, Dandan; Lou, Chaoyan; Zhang, Peimin; Zhang, Jiajie; Wang, Nani; Wu, Shuchao; Zhu, Yan

    2016-07-22

    In this work, a novel ion exchange stationary phase based on different generations of poly (amidoamine) dendrimers (PAMAM) was developed for the determination of inorganic anions and carbohydrates. Synthesis of the PAMAM was carried out with the polymerization reaction of ethylenediamine and methyl acrylate. The synthesized PAMAM was then grafted to the polystyrene-divinylbenzene-glycidyl methacrylate (PS-GMA) to form PAMAM-based beads. These beads were finally modified with 1,4-butanediol diglycidyl ether (BDDE) to generate the anion exchanger, which were characterized by scanning electron microscopy (SEM), brunauer-emmett-teller (BET), fourier transform infrared spectroscopy (FTIR), and elemental analysis. Elemental analysis, breakthrough curves and capacity factors showed that more epoxy groups and higher PAMAM generations in stationary phase could result in higher anion exchange capacity. The efficiency, durability and stability of the proposed anion exchanger were investigated by using six inorganic anions (fluoride, chloride, nitrite, bromide, nitrate and sulfate) and four carbohydrates (trehalose, glucose, maltotriose and galacturonic acid) as analytes, respectively. The reliability of the proposed ion chromatographic stationary phase was demonstrated by determining the content of galacturonic acid in polysaccharides from Poria cocos and Atractylodes macrocephala. The relative standard deviations of retention time, peak height, and peak area for galacturonic acid were 0.39%, 1.22%, and 2.02%, respectively. The spiked recoveries were in the range of 88.29%-100.51% for plant polysaccharides. Due to the good structural homogeneity, intense internal porosity, biological compatibility and high density of active groups in PAMAM, this grafted stationary phase showed good ion-exchange characteristics, especially in biological charged molecules.

  6. Stationary phase selection and comprehensive two-dimensional gas chromatographic analysis of trace biodiesel in petroleum-based fuel.

    Science.gov (United States)

    Seeley, John V; Bates, Carly T; McCurry, James D; Seeley, Stacy K

    2012-02-24

    The GC×GC solvation parameter model has been used to identify effective stationary phases for the separation of fatty acid methyl esters (FAMEs) from petroleum hydrocarbons. This simple mathematical model was used to screen the 1225 different combinations of 50 stationary phases. The most promising pairs combined a poly(methyltrifluoropropylsiloxane) stationary phase with a poly(dimethyldiphenylsiloxane) stationary phase. The theoretical results were experimentally tested by equipping a GC×GC instrument with a DB-210 primary stationary phase and an HP-50+ secondary stationary phase. This instrument was used to analyze trace levels of FAMEs in kerosene. The FAMEs were fully separated from the petroleum hydrocarbons on the secondary dimension of the 2-D chromatogram. The resulting GC×GC method was shown to be capable of accurately quantifying FAME levels as low as 2 ppm (w/w). These results demonstrate the utility of the solvation parameter model for identifying optimal stationary phases for high resolution GC×GC separations. Furthermore, this work presents an effective method for determining the level of biodiesel contamination in aviation fuel and other petroleum-based fuels. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. Hierarchical CaCO3 chromatography: a stationary phase based on biominerals.

    Science.gov (United States)

    Sato, Kosuke; Oaki, Yuya; Takahashi, Daisuke; Toshima, Kazunobu; Imai, Hiroaki

    2015-03-23

    In biomineralization, acidic macromolecules play important roles for the growth control of crystals through a specific interaction. Inspired by this interaction, we report on an application of the hierarchical structures in CaCO3 biominerals to a stationary phase of chromatography. The separation and purification of acidic small organic molecules are achieved by thin-layer chromatography and flash chromatography using the powder of biominerals as the stationary phase. The unit nanocrystals and their oriented assembly, the hierarchical structure, are suitable for the adsorption site of the target organic molecules and the flow path of the elution solvents, respectively. The separation mode is ascribed to the specific adsorption of the acidic molecules on the crystal face and the coordination of the functional groups to the calcium ions. The results imply that a new family of stationary phase of chromatography can be developed by the fine tuning of hierarchical structures in CaCO3 materials. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Enhanced dipicolinic acid production during the stationary phase in Bacillus subtilis by blocking acetoin synthesis.

    Science.gov (United States)

    Toya, Yoshihiro; Hirasawa, Takashi; Ishikawa, Shu; Chumsakul, Onuma; Morimoto, Takuya; Liu, Shenghao; Masuda, Kenta; Kageyama, Yasushi; Ozaki, Katsuya; Ogasawara, Naotake; Shimizu, Hiroshi

    2015-01-01

    Bacterial bio-production during the stationary phase is expected to lead to a high target yield because the cells do not consume the substrate for growth. Bacillus subtilis is widely used for bio-production, but little is known about the metabolism during the stationary phase. In this study, we focused on the dipicolinic acid (DPA) production by B. subtilis and investigated the metabolism. We found that DPA production competes with acetoin synthesis and that acetoin synthesis genes (alsSD) deletion increases DPA productivity by 1.4-fold. The mutant showed interesting features where the glucose uptake was inhibited, whereas the cell density increased by approximately 50%, resulting in similar volumetric glucose consumption to that of the parental strain. The metabolic profiles revealed accumulation of pyruvate, acetyl-CoA, and the TCA cycle intermediates in the alsSD mutant. Our results indicate that alsSD-deleted B. subtilis has potential as an effective host for stationary-phase production of compounds synthesized from these intermediates.

  9. Ionic liquids as stationary phases in gas chromatography: determination of chlorobenzenes in soils.

    Science.gov (United States)

    González Paredes, Rosa María; García Pinto, Carmelo; Pérez Pavón, José Luis; Moreno Cordero, Bernardo

    2014-06-01

    The present research focuses on the evaluation of different ionic liquid (IL) stationary phases in gas chromatography. The different IL columns were evaluated in terms of peak resolution (Rs) and peak symmetry for the separation of the chlorobenzenes. The determination of chlorobenzenes in soil samples by means of the optimal IL stationary phase (SLB-IL82) is proposed as an application. Soil pretreatment was based on a simplified quick, easy, cheap, effective, rugged, and safe extraction procedure and a large injection volume via a programed temperature vaporizer working in solvent vent mode. The retention time of the chlorobenzenes increased as the polarity of the IL column decreased. SLB-IL82 is the stationary phase that provides the best values as regards Rs and asymmetry factor. Soil sample blanks were spiked with the analytes before subjecting the sample to the extraction process. The existence of a matrix effect was checked and the analytical characteristics of the method were determined in a fortified garden soil sample. The method provided good linearity, good repeatability and reproducibility values, and the LODs were in the 0.1-4.7 μg/kg range. Two fortified soil samples were applied to validate the proposed methodology.

  10. A novel hybrid metal-organic framework-polymeric monolith for solid-phase microextraction.

    Science.gov (United States)

    Lin, Chen-Lan; Lirio, Stephen; Chen, Ya-Ting; Lin, Chia-Her; Huang, Hsi-Ya

    2014-03-17

    This study describes the fabrication of a novel hybrid metal-organic framework- organic polymer (MOF-polymer) for use as a stationary phase in fritless solid-phase microextraction (SPME) for validating analytical methods. The MOF-polymer was prepared by using ethylene dimethacrylate (EDMA), butyl methacrylate (BMA), and an imidazolium-based ionic liquid as porogenic solvent followed by microwave-assisted polymerization with the addition of 25 % MOF. This novel hybrid MOF-polymer was used to extract penicillin (penicillin G, penicillin V, oxacillin, cloxacillin, nafcillin, dicloxacillin) under different conditions. Quantitative analysis of the extracted penicillin samples using the MOF-organic polymer for SPME was conducted by using capillary electrochromatography (CEC) coupled with UV analysis. The penicillin recovery was 63-96.2 % with high reproducibility, sensitivity, and reusability. The extraction time with the proposed fabricated SPME was only 34 min.

  11. The Development and Characterization of Protein-Based Stationary Phases for Studying Drug-Protein and Protein-Protein Interactions

    OpenAIRE

    Sanghvi, Mitesh; Moaddel, Ruin; Wainer, Irving W.

    2011-01-01

    Protein-based liquid chromatography stationary phases are used in bioaffinity chromatography for studying drug-protein interactions, the determination of binding affinities, competitive and allosteric interactions, as well as for studying protein-protein interactions. This review addresses the development and characterization of protein-based stationary phase, and the application of these phases using frontal and zonal chromatography techniques. The approach will be illustrated using immobili...

  12. Synthesis and characterization of a multimode stationary phase: Congo red derivatized silica in nano-flow HPLC.

    Science.gov (United States)

    Zhang, Yi; Zhang, Yan; Wang, Guan; Chen, Wujuan; He, Pingang; Wang, Qingjiang

    2016-02-01

    A novel Congo red (CR) derivatized silica stationary phase was prepared and packed into a fused silica capillary tube for nano-flow HPLC. A variety of analytes including poly-aromatic hydrocarbons, parabens, acids, sulfonamides, bases, and nucleosides were successfully separated using the CR. In comparison with commercial ODS columns, this new stationary phase has a different separation mechanism (hydrophobically-assisted ion-exchange), which was evident in the separation of benzoic acid derivatives and sulfonamides. The successful application of CR-bonded silica stationary phase in the HILIC and PALC modes demonstrates the effectiveness of this potential chromatographic material in nano flow HPLC.

  13. Surface Confined Ionic Liquid-A New Stationary Phase for the Separation of Ephedrines in High-performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    Shu Juan LIU; Feng ZHOU; Xiao Hua XIAO; Liang ZHAO; Xia LIU; Sheng Xiang JIANG

    2004-01-01

    In this article, a new and effective stationary phase based on ionic liquid modified silica is first reported and used for the separation of ephedrines in high-performance liquid chromatography (HPLC). The separation results indicate the high efficiency and reproducibility of the stationary phase. The electrostatic interaction, ion-exchange interaction between the solutes and the stationary phase are considered to attribute the effective separation. Moreover, the free silanols on the surface of the silica are effectively masked by the immobilized ionic liquid, a result of which is to decrease the non-specific absorption.

  14. Synthesis, characterization, and application of a novel multifunctional stationary phase for hydrophilic interaction/reversed phase mixed-mode chromatography.

    Science.gov (United States)

    Aral, Hayriye; Çelik, K Serdar; Altındağ, Ramazan; Aral, Tarık

    2017-11-01

    A novel multifunctional stationary phase based on silica gel was synthesised starting from L- isoleucine and 4-phenylbutylamine and evaluated as a hydrophilic interaction/reversed-phase mixed-mode stationary phase for high-performance liquid chromatography (HPLC). The prepared stationary phase was characterized by elemental analysis, infrared spectroscopy (IR), scanning electron microscopy (SEM), Brunauer, Emmett and Teller (BET) and solid-state (13)C nuclear magnetic resonance (NMR). The mechanisms involved in the chromatographic separation are multi-interaction, including hydrophobic, π-π, hydrogen-bonding, dipole-dipole and ion-dipole interactions. Based on these interactions, successful separation could be achieved among several aromatic compounds having different polarities under both hydrophilic interaction liquid chromatography (HILIC) and reversed phase (RP) condition. Nucleotides/nucleosides were separated in the HILIC mode. The effects of different separation conditions, such as pH value, mobile-phase content, column temperature, buffer concentration and flow rate, on the separation of nucleotides/nucleosides in HILIC mode were investigated. The seven nucleotides/nucleosides were separated within 22min, while six of them were separated within 10min by isocratic elution. To determine the influence of the new multifunctional stationary phase under the RP condition, a number of moderately and weakly polar and nonpolar compounds, such as 10 substituted anilines and eight substituted phenols were separated successfully under the RP condition within 14 and 15min, respectively. Additionally, nine mixtures of polar/nonpolar test compounds were simultaneously separated within 19min, while seven of them were separated within 12min, under HILIC/RP mixed-mode conditions. Chromatographic parameters, such as the retention factor and peak asymmetry factor, were calculated for all of the analytes, while the theoretical plate number was calculated for analytes separated

  15. Enantioseparation of Six Antihistamines with Immobilized Cellulose Chiral Stationary Phase by HPLC

    Science.gov (United States)

    Zhou, Jie; Luo, Pei; Chen, Shanshan; Meng, Lingchang; Sun, Chong; Du, Qiuzheng; Sun, Fang

    2016-01-01

    A stereoselective high performance liquid chromatography method has been developed for the chiral separation of the enantiomers of six antihistamines, doxylamine, carbinoxamine, dioxopromethazine, oxomemazine, cetirizine and hydroxyzine. The effects of mobile phase additive, column temperature and flow rate on the retention time and resolution were studied. Enantiomeric separation of cetirizine, doxylamine and hydroxyzine were achieved on cellulose tris-(3,5-dichlorophenylcarbamate) immobilized on silica gel chiral stationary phase known as Chiralpak IC (RS = 3.74, RS = 1.85 and RS = 1.74, respectively). PMID:26657408

  16. Enantioseparation of Six Antihistamines with Immobilized Cellulose Chiral Stationary Phase by HPLC.

    Science.gov (United States)

    Zhou, Jie; Luo, Pei; Chen, Shanshan; Meng, Lingchang; Sun, Chong; Du, Qiuzheng; Sun, Fang

    2016-04-01

    A stereoselective high performance liquid chromatography method has been developed for the chiral separation of the enantiomers of six antihistamines, doxylamine, carbinoxamine, dioxopromethazine, oxomemazine, cetirizine and hydroxyzine. The effects of mobile phase additive, column temperature and flow rate on the retention time and resolution were studied. Enantiomeric separation of cetirizine, doxylamine and hydroxyzine were achieved on cellulose tris-(3,5-dichlorophenylcarbamate) immobilized on silica gel chiral stationary phase known as Chiralpak IC (RS = 3.74, RS = 1.85 and RS = 1.74, respectively).

  17. Development of N-ferrocenyl(benzoyl)amino-acid esters stationary phase for high performance liquid chromatography.

    Science.gov (United States)

    Zhou, Xiaohua; Li, Xiaole; Cao, Aijuan; Lijun, Qiao; Yu, Ajuan; Zhang, Shusheng; Wu, Yangjie

    2015-11-01

    A new stationary phase for high-performance liquid chromatography was prepared by covalently bonding N-ferrocenyl(benzoyl)amino-acid esters (L(1)) onto silica gel using 3-aminopropyltriethoxysilane as coupling reagent. The structure of new material was characterized by infrared spectroscopy, elemental analysis and thermogravimetric analysis. The chromatographic behavior of the phase was illustrated in reversed-phase (RP) and normal-phase (NP) modes using polycyclic aromatic hydrocarbons (PAHs), aromatics positional isomers, amines, 5-nitroimidazoles, organophosphorus pesticides and phenols as probes. Multiple mechanisms including hydrophobic, hydrogen bonding, π-π, dipole-dipole, charge-transfer and acid-base equilibrium interactions are involved. Thus, multi-interaction mechanisms and mixed-mode separation of the new phase can very likely guarantee its excellent chromatographic performance for the analysis of complex samples. The L(1) AminoSil column was successfully employed for the analysis of organophosphorus pesticides in vegetable.

  18. Theory for transitions between log and stationary phases: universal laws for lag time

    CERN Document Server

    Himeoka, Yusuke

    2016-01-01

    Quantitative characterization of bacterial growth has gathered substantial attention since Monod's pioneering study. Theoretical and experimental work has uncovered several laws for describing the log growth phase, in which the number of cells grows exponentially. However, microorganism growth also exhibits lag, stationary, and death phases under starvation conditions, in which cell growth is highly suppressed, while quantitative laws or theories for such phases are underdeveloped. In fact, models commonly adopted for the log phase that consist of autocatalytic chemical components, including ribosomes, can only show exponential growth or decay in a population, and phases that halt growth are not realized. Here, we propose a simple, coarse-grained cell model that includes inhibitor molecule species in addition to the autocatalytic active protein. The inhibitor forms a complex with active proteins to suppress the catalytic process. Depending on the nutrient condition, the model exhibits the typical transition a...

  19. Selective higher order fiber mode excitation using a monolithic setup of a phase plate at a fiber facet

    CERN Document Server

    Wilde, Johannes; Brüning, Robert; Duparré, Michael; Schröter, Siegmund

    2015-01-01

    Controlling the modal content coupled into an optical fiber can be desireable in many situations, e.g. for adjusting the sensitivity of the guided field distribution to external perturbations. For this purpose we used a monolithic setup of a phase plate at a fiber inpute facet to excite selectivly higher order modes, which theoretically can provide a mode purity of more than 99%. We investigated the capabilities of this approach by complete modal decomposition of the fiber output signals, considering the achievable mode purity with respect to several possible imperfections of the setup. The experiments are compared with detailed numerical simulations and show a high agreement. Additionally a comparison with a well known setup with free space phase plates was undertaken. This showed the monolithic setup to be energetically twice as efficient.

  20. Band-phase-randomized Surrogates to assess nonlinearity in non-stationary time series

    CERN Document Server

    Guarin, Diego; Orozco, Alvaro

    2011-01-01

    Testing for nonlinearity is one of the most important preprocessing steps in nonlinear time series analysis. Typically, this is done by means of the linear surrogate data methods. But it is a known fact that the validity of the results heavily depends on the stationarity of the time series. Since most physiological signals are non-stationary, it is easy to falsely detect nonlinearity using the linear surrogate data methods. In this document, we propose a methodology to extend the procedure for generating constrained surrogate time series in order to assess nonlinearity in non-stationary data. The method is based on the band-phase-randomized surrogates, which consists (contrary to the linear surrogate data methods) in randomizing only a portion of the Fourier phases in the high frequency band. Analysis of simulated time series showed that in comparison to the linear surrogate data method, our method is able to discriminate between linear stationarity, linear non-stationary and nonlinear time series. When apply...

  1. Preparation and Characterization of a New Quercetin-bonded Stationary Phase for High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    李来生; 方奕珊; 陈红; 张杨

    2012-01-01

    A quercetin-bonded silica gel stationary phase (QUSP) containing natural flavonoid ligand was first prepared via γ-glycidoxypropyltrimethoxysilane (KH-560) as a coupling reagent for high-performance liquid chromatography. Its chemical structure was characterized by Fourier infrared spectroscopy, elemental analysis, thermal thermogravimetry and 13C cross polarization/magic angle spinning nuclear magnetic resonance (CP/MAS NMR). The chromatographic property of QUSP was systematically evaluated by using neutral, basic and acidic aromatic com- pounds as probes. In order to clarify its retention mechanism, a comparative study of QUSP with conventional oc- tadecylsilyl-bonded stationary phase (ODS) was also carried out under the same conditions. The results showed that the new quercetin-bonded phase exhibited an excellent reversed-phase chromatographic property with relatively weak hydrophobicity. However, it has an advantage over ODS in the fast separation of polar aromatic compounds because the quercetin ligand could provide various sites besides hydrophobicity, such as hydrogen bonding, dipole-dipole, n-n staking and charge transfer interactions. QUSP was performed in the baseline separations of ion- ized polar basic or acidic compounds, including pyridines, anilines, pyrimidines, purines and phenols with symmet- ric peak shape in common mobile phases without buffer salt within relatively short time. The natural ligands from herbs are readily available and contain a variety of active sites, which facilitate the exploration of industrial chromatographic separation materials for green products.

  2. PREPARATION OF POROUS NANOCOMPOSITE SCAFFOLDS WITH HONEYCOMB MONOLITH STRUCTURE BY ONE PHASE SOLUTION FREEZE DRYING METHOD

    Institute of Scientific and Technical Information of China (English)

    Yang Xu; Duo Zhang; Zong-liang Wang; Zhan-tuan Gao; Pei-biao Zhang; Xue-si Chen

    2011-01-01

    Biodegradable porous nanocomposite scaffolds of poly(lactide-co-glycolide) (PLGA) and L-lactic acid (LAc) oligomer surface-grafted hydroxyapatite nanoparticles (op-HA) with a honeycomb monolith structure were fabricated with the single-phase solution freeze-drying method. The effects of different freezing temperatures on the properties of the scaffolds, such as microstructures, compressive strength, cell penetration and cell proliferation were studied. The highly porous and well interconnected scaffolds with a tunable pore structure were obtained. The effect of different freezing temperature (4℃, -20℃, -80℃ and -196℃) was investigated in relation to the scaffold morphology, the porosity varied from 91.2% to 83.0% and the average pore diameter varied from (167.2 ± 62.6) pm to (11.9 ± 4.2) μm while the σ10 increased significantly. The cell proliferation were decreased and associated with the above-mentioned properties. Uniform distribution of op-HA particles and homogeneous roughness of pore wall surfaces were found in the 4℃ frozen scaffold. The 4℃ frozen scaffold exhibited better cell penetration and increased cell proliferation because of its larger pore size, higher porosity and interconnection. The microstmctures described here provide a new approach for the design and fabrication of op-HA/PLGA based scaffold materials with potentially broad applicability for replacement of bone defects.

  3. Resolution of DL-Pantolactone with Ethyldiamine Bridged Dimer Permethyβ-Cyclodextrin as GC Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Ethyldiamine bridged dimer permethy-β-CD and other cyclodextrins were used as GC stationary phase to resolute DL-pantolactone.It is found that this CD dimer has a good selectivity for DL-pantolactone.

  4. Packed column supercritical fluid chromatography using stainless steel particles and water as a stationary phase.

    Science.gov (United States)

    Murakami, Jillian N; Thurbide, Kevin B

    2015-09-15

    Stainless steel (SS) particles were demonstrated as a novel useful support for a water stationary phase in packed column supercritical fluid chromatography using a CO2 mobile phase. Separations employed flame ionization detection, and the system was operated over a range of temperatures and pressures. Retention times reproduced well with RSD values of 2.6% or less. Compared to analogous separations employing a water stationary phase coated onto a SS capillary column, the packed column method provided separations that were about 10× faster, with nearly 8-fold larger analyte retention factors, while maintaining good peak shape and comparable column efficiency. Under normal operating conditions, the packed column contains about 131 ± 4 μL/m of water phase (around a 5% m/m coating), which is over 25× greater than the capillary column and also affords it a 20-fold larger sample capacity. Several applications of the packed column system are examined, and the results indicate that it is a useful alternative to the capillary column mode, particularly where analyte loads or sample matrix interference is a concern. Given its high sample capacity, this packed column method may also be useful to explore on a more preparative scale in the future.

  5. Sum of ranking differences to rank stationary phases used in packed column supercritical fluid chromatography.

    Science.gov (United States)

    West, Caroline; Khalikova, Maria A; Lesellier, Eric; Héberger, Károly

    2015-08-28

    The identification of a suitable stationary phase in supercritical fluid chromatography (SFC) is a major source of difficulty for those with little experience in this technique. Several protocols have been suggested for column classification in high-performance liquid chromatography (HPLC), gas chromatography (GC), and SFC. However, none of the proposed classification schemes received general acceptance. A fair way to compare columns was proposed with the sum of ranking differences (SRD). In this project, we used the retention data obtained for 86 test compounds with varied polarity and structure, analyzed on 71 different stationary phases encompassing the full range in polarity of commercial packed columns currently available to the SFC chromatographer, with a single set of mobile phase and operating conditions (carbon dioxide-methanol mobile phase, 25°C, 150bar outlet pressure, 3ml/min). First, a reference column was selected and the 70 remaining columns were ranked based on this reference column and the retention data obtained on the 86 analytes. As these analytes previously served for the calculation of linear solvation energy relationships (LSER) on the 71 columns, SRD ranks were compared to LSER methodology. Finally, an external comparison based on the analysis of 10 other analytes (UV filters) related the observed selectivity to SRD ranking. Comparison of elution orders of the UV filters to the SRD rankings is highly supportive of the adequacy of SRD methodology to select similar and dissimilar columns.

  6. Comparison of the mass transfer in totally porous and superficially porous stationary phases in liquid chromatography.

    Science.gov (United States)

    Kiss, Ibolya; Bacskay, Ivett; Kilár, Ferenc; Felinger, Attila

    2010-06-01

    The characterization of mass-transfer processes in a chromatographic column during a separation process is essential, since the influence of the mass-transfer kinetics on the shape of the chromatographic band profiles and on the efficiency of the separation is crucial. Several sources of mass transfer in a chromatographic bed have been identified and studied: the axial dispersion in the stream of mobile phase, the external mass-transfer resistance, intraparticle diffusion, and the kinetics of adsorption-desorption. We measured and compared the characteristics and performance of a new brand of shell particles and those of a conventional brand of totally porous silica particles. The shell stationary phase was made of 2.7-microm superficially porous particles (a 1.7-microm solid core is covered with a 0.5-microm-thick shell of porous silica). The other material consisted of totally porous particles of conventional 3.5-microm commercial silica. We measured the first and second central moments of the peaks of human insulin over a wide range of mobile phase velocities (from 0.02 to 1.3 mL/min) at 20 degrees C. The plate height equations were constructed and the axial dispersion, external mass transfer, as well as the intraparticle diffusion coefficients were calculated for the two stationary phases.

  7. Dynamic proteome changes of Shigella flexneri 2a during transition from exponential growth to stationary phase.

    Science.gov (United States)

    Zhu, Li; Liu, Xian-Kai; Zhao, Ge; Zhi, Yi-Dan; Bu, Xin; Ying, Tian-Yi; Feng, Er-Ling; Wang, Jie; Zhang, Xue-Min; Huang, Pei-Tang; Wang, Heng-Liang

    2007-05-01

    Shigella flexneri is an infectious pathogen that causes dysentery to human, which remains a serious threat to public health, particularly in developing countries. In this study, the global protein expression patterns of S. flexneri during transition from exponential growth to stationary phase in vitro were analyzed by using 2-D PAGE combined with MALDI-TOF MS. In a time-course experiment with five time points, the relative abundance of 49 protein spots varied significantly. Interestingly, a putative outer membrane protein YciD (OmpW) was almost not detected in the exponential growth phase but became one of the most abundant proteins in the whole stationary-phase proteome. Some proteins regulated by the global regulator FNR were also significantly induced (such as AnsB, AspA, FrdAB, and KatG) or repressed (such as AceEF, OmpX, SodA, and SucAB) during the growth phase transition. These proteins may be the key effectors of the bacterial cell cycle or play important roles in the cellular maintenance and stress responses. Our expression profile data provide valuable information for the study of bacterial physiology and form the basis for future proteomic analyses of this pathogen.

  8. Dynamic Proteome Changes of Shigella flexneri 2a During Transition from Exponential Growth to Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Shigella flexneri is an infectious pathogen that causes dysentery to human, which remains a serious threat to public health, particularly in developing countries. In this study, the global protein expression patterns of S. flexneri during transition from exponential growth to stationary phase in vitro were analyzed by using 2-D PAGE combined with MALDI-TOF MS. In a time-course experiment with five time points, the relative abundance of 49 protein spots varied significantly. In terestingly, a putative outer membrane protein YciD (OmpW) was almost not detected in the exponential growth phase but became one of the most abundant proteins in the whole stationary-phase proteome. Some proteins regulated by the global regulator FNR were also significantly induced (such as AnsB, AspA, FrdAB,and KatG) or repressed (such as AceEF, OmpX, SodA, and SucAB) during the growth phase transition. These proteins may be the key effectors of the bacterial cell cycle or play important roles in the cellular maintenance and stress responses.Our expression profile data provide valuable information for the study of bacterial physiology and form the basis for future proteomic analyses of this pathogen.

  9. Molecularly imprinted macroporous monolithic materials for protein recognition

    Institute of Scientific and Technical Information of China (English)

    Qi Liang Deng; Yan Li Li; Li Hua Zhang; Yu Kui Zhang

    2011-01-01

    Synthetic materials that can specifically recognize proteins will find wide application in many fields. In this report, bovine serum albumin was chosen as the template protein. Acrylamide and N, N'-methylenebisacrylamide were employed as the functional and cross-linker monomers, respectively. Molecularly imprinted macroporous monolithic materials that can preferentially bind the template protein in an aqueous environment were prepared by combination of molecular imprinting technique and freezing/thawing preparation method. The resulted imprinted macroporous monolithic columns were evaluated by utilizing as stationary phase in high performance liquid chromatography and solid-phase extraction materials. The experimental results indicated that the imprinted macroporous monolithic column exhibited good recognition for template protein, as compared with the control protein (hemoglobin), whereas the non-imprinted polymer (prepared under the same conditions except without addition template protein) had no selective properties.

  10. Monolithic liquid-chromatography columns for protein analysisprotein digest separation and integrated systems

    NARCIS (Netherlands)

    van de Meent, M.H.M.

    2010-01-01

    The objectives of the research described in this thesis are to evaluate the applicability of both silica-based and polymeric monolithic columns for protein analysis. The first part describes investigations into the effects of column length and stationary-phase chemistry on the separation of protein

  11. Direct tandem mass spectrometric analysis of amino acids in plasma using fluorous derivatization and monolithic solid-phase purification.

    Science.gov (United States)

    Tamashima, Erina; Hayama, Tadashi; Yoshida, Hideyuki; Imakyure, Osamu; Yamaguchi, Masatoshi; Nohta, Hitoshi

    2015-11-10

    In this study, we developed a novel direct tandem mass spectrometric method for rapid and accurate analysis of amino acids utilizing a fluorous derivatization and purification technique. Amino acids were perfluoroalkylated with 2H,2H,3H,3H-perfluoroundecan-1-al in the presence of 2-picoline borane via reductive amination. The derivatives were purified by perfluoroalkyl-modified silica-based monolithic solid-phase extraction (monolithic F-SPE), and directly analyzed by tandem mass spectrometry using electrospray ionization without liquid chromatographic separation. The perfluoroalkyl derivatives could be sufficiently distinguished from non-fluorous compounds, i.e. the biological matrix, due to their fluorous interaction. Thus, rapid and accurate determination of amino acids was accomplished. The method was validated with human plasma samples and applied to the analysis of amino acids in the plasma of mice with maple syrup urine disease or phenylketonuria.

  12. Proteomic Analysis of Stationary Phase in the Marine Bacterium "Candidatus Pelagibacter ubique"

    Energy Technology Data Exchange (ETDEWEB)

    Sowell, S. M.; Norbeck, A. D.; Lipton, M. S.; Nicora, C. D.; Callister, S. J.; Smith, R. D.; Barofsky, D. F.; Giovannoni, S. J.

    2008-05-09

    The α-proteobacterium ‘Candidatus Pelagibacter ubique’ str. HTCC1062, and most other members of the SAR11 clade, lack genes for assimilatory sulfate reduction, making them dependent on organosulfur compounds that occur naturally in seawater. To investigate how these cells adapt to sulfur limitation, batch cultures were grown in defined media containing either limiting or non-limiting amounts of dimethylsulfoniopropionate (DMSP) as the sole sulfur source. Protein and mRNA expression were measured during exponential growth, immediately prior to stationary phase, and in late stationary phase. Two distinct responses were observed: one as DMSP became exhausted, and another as cells acclimated to a sulfur-limited environment. The first response was characterized by increased transcription and translation of all Ca. P. ubique genes downstream of previously confirmed S-adenosyl methionine (SAM) riboswitches: bhmT, mmuM, and metY. Proteins encoded by these genes were up to 33 times more abundant as DMSP became limiting. Their predicted function is to shunt all available sulfur to methionine. The secondary response, observed during sulfur-depleted stationary phase, was a 6-10 fold increase in transcription of the heme c shuttle ccmC and two small genes of unknown function (SAR11_1163 and SAR11_1164). This bacterium's strategy for coping with sulfur stress appears to be intracellular redistribution to support methionine biosynthesis, rather than increasing organosulfur import. Many of the genes and SAM riboswitches involved in this response are located in a hypervariable genome region (HVR). One of these HVR genes, ordL, is located downstream of a conserved motif that evidence suggests is a novel riboswitch.

  13. Validation of stationary phases in (111)In-pentetreotide planar chromatography.

    Science.gov (United States)

    Moreno-Ortega, E; Mena-Bares, L M; Maza-Muret, F R; Hidalgo-Ramos, F J; Vallejo-Casas, J A

    2013-01-01

    Since Pall-German stopped manufacturing ITLC-SG, it has become necessary to validate alternative stationary phases. To validate different stationary phases versus ITLC-SG Pall-Gelman in the determination of the radiochemical purity (RCP) of (111)In-pentetreotide ((111)In-Octreoscan) by planar chromatography. We conducted a case-control study, which included 66 (111)In-pentetreotide preparations. We determined the RCP by planar chromatography, using a freshly prepared solution of 0,1M sodium citrate (pH 5) and the following stationary phases: ITLC-SG (Pall-Gelman) (reference method), iTLC-SG (Varian), HPTLC silica gel 60 (Merck), Whatman 1, Whatman 3MM and Whatman 17. For each of the methods, we calculated: PRQ, relative front values (RF) of the radiopharmaceutical and free (111)In, chromatographic development time, resolution between peaks. We compared the results obtained with the reference method. The statistical analysis was performed using the SPSS program. The p value was calculated for the study of statistical significance. The highest resolution is obtained with HPTLC silica gel 60 (Merck). However, the chromatographic development time is too long (mean=33.62minutes). Greater resolution is obtained with iTLC-SG (Varian) than with the reference method, with lower chromatographic development time (mean=3.61minutes). Very low resolutions are obtained with Whatman paper, essentially with Whatman 1 and 3MM. Therefore, we do not recommend their use. Although iTLC-SG (Varian) and HPTLC silica gel 60 (Merck) are suitable alternatives to ITLC-SG (Pall-Gelman) in determining the RCP of (111)In-pentetreotide, iTLC-SG (Varian) is the method of choice due to its lower chromatographic development time. Copyright © 2012 Elsevier España, S.L. and SEMNIM. All rights reserved.

  14. Stationary phase expression of the arginine biosynthetic operon argCBH in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Sun Yuan

    2006-02-01

    Full Text Available Abstract Background Arginine biosynthesis in Escherichia coli is elevated in response to nutrient limitation, stress or arginine restriction. Though control of the pathway in response to arginine limitation is largely modulated by the ArgR repressor, other factors may be involved in increased stationary phase and stress expression. Results In this study, we report that expression of the argCBH operon is induced in stationary phase cultures and is reduced in strains possessing a mutation in rpoS, which encodes an alternative sigma factor. Using strains carrying defined argR, and rpoS mutations, we evaluated the relative contributions of these two regulators to the expression of argH using operon-lacZ fusions. While ArgR was the main factor responsible for modulating expression of argCBH, RpoS was also required for full expression of this biosynthetic operon at low arginine concentrations (below 60 μM L-arginine, a level at which growth of an arginine auxotroph was limited by arginine. When the argCBH operon was fully de-repressed (arginine limited, levels of expression were only one third of those observed in ΔargR mutants, indicating that the argCBH operon is partially repressed by ArgR even in the absence of arginine. In addition, argCBH expression was 30-fold higher in ΔargR mutants relative to levels found in wild type, fully-repressed strains, and this expression was independent of RpoS. Conclusion The results of this study indicate that both derepression and positive control by RpoS are required for full control of arginine biosynthesis in stationary phase cultures of E. coli.

  15. Persistent Spins in the Linear Diffusion Approximation of Phase Ordering and Zeros of Stationary Gaussian Processes

    Science.gov (United States)

    Derrida, Bernard; Hakim, Vincent; Zeitak, Reuven

    1996-09-01

    The fraction r\\(t\\) of spins which have never flipped up to time t is studied within a linear diffusion approximation to phase ordering. Numerical simulations show that r\\(t\\) decays with time like a power law with a nontrivial exponent θ which depends on the space dimension. The dynamics is a special case of a stationary Gaussian process of known correlation function. The exponent θ is given by the asymptotic decay of the probability distribution of intervals between consecutive zero crossings. An approximation based on the assumption that successive zero crossings are independent random variables gives values of θ in close agreement with the results of simulations.

  16. Trigonal tricationic ionic liquids: a generation of gas chromatographic stationary phases.

    Science.gov (United States)

    Payagala, Tharanga; Zhang, Ying; Wanigasekara, Eranda; Huang, Ke; Breitbach, Zachary S; Sharma, Pritesh S; Sidisky, Leonard M; Armstrong, Daniel W

    2009-01-01

    Trigonal tricationic ionic liquids (ILs) are a new class of ILs that appear to be unique when used as gas chromatographic stationary phases. They consist of four core structures; (1) A = mesitylene core, (2) B = benzene core, (3) C = triethylamine core, and (4) D = tri(2-hexanamido)ethylamine core; to which three identical imidazolium or phosphonium cationic moieties were attached. These were coated on fused silica capillaries, and their gas chromatographic properties were evaluated. They were characterized using a linear solvation parameter model and a number of test mixtures. On the basis of the literature, it is known that both monocationic and dicationic ILs possess almost identical polarities, solvation characteristics, and chromatographic selectivities. However, some of the trigonal tricationic ILs were quite different. The different solvation parameters and higher apparent polarities appear to generate from the more rigid trigonal geometry of these ILs, as well as their ability to retain the positive charges in relatively close proximity to one another in some cases. Their unique selectivities, retention behaviors, and separation efficiencies were demonstrated using the Grob mixture, a flavor and fragrance test mixture, alcohols/alkanes test, and FAME isomer separations. Two ILs C1 (methylimidazolium substitution) and C4 (2-hydroxyethylimidazolium substitution) had higher apparent polarities than any know IL (mono, di, and tricationic ILs) or commercial stationary phases. The tri(2-hexanamido)ethylamine core IL series proved to be very interesting in that it not only showed the highest separation efficiency for all test mixtures, but it also is the first IL stationary phase (containing NTf(2)(-) anions) that eliminates peak tailing for alcohols and other H-bonding analytes. The thermal stabilities were investigated using three methods: thermogravimetric analysis (TGA) method, temperature programmed gas chromatographic method (TPGC), and isothermal gas

  17. Continuous wavelet transform for non-stationary vibration detection with phase-OTDR.

    Science.gov (United States)

    Qin, Zengguang; Chen, Liang; Bao, Xiaoyi

    2012-08-27

    We propose the continuous wavelet transform for non-stationary vibration measurement by distributed vibration sensor based on phase optical time-domain reflectometry (OTDR). The continuous wavelet transform approach can give simultaneously the frequency and time information of the vibration event. Frequency evolution is obtained by the wavelet ridge detection method from the scalogram of the continuous wavelet transform. In addition, a novel signal processing algorithm based on the global wavelet spectrum is used to determine the location of vibration. Distributed vibration measurements of 500 Hz and 500 Hz to 1 kHz sweep events over 20 cm fiber length are demonstrated using a single mode fiber.

  18. Methylation of yeast ribosomal protein S2 is elevated during stationary phase growth conditions.

    Science.gov (United States)

    Ladror, Daniel T; Frey, Brian L; Scalf, Mark; Levenstein, Mark E; Artymiuk, Jacklyn M; Smith, Lloyd M

    2014-03-14

    Ribosomes, as the center of protein translation in the cell, require careful regulation via multiple pathways. While regulation of ribosomal synthesis and function has been widely studied on the transcriptional and translational "levels," the biological roles of ribosomal post-translational modifications (PTMs) are largely not understood. Here, we explore this matter by using quantitative mass spectrometry to compare the prevalence of ribosomal methylation and acetylation for yeast in the log phase and the stationary phase of growth. We find that of the 27 modified peptides identified, two peptides experience statistically significant changes in abundance: a 1.9-fold decrease in methylation for k(Me)VSGFKDEVLETV of ribosomal protein S1B (RPS1B), and a 10-fold increase in dimethylation for r(DiMe)GGFGGR of ribosomal protein S2 (RPS2). While the biological role of RPS1B methylation has largely been unexplored, RPS2 methylation is a modification known to have a role in processing and export of ribosomal RNA. This suggests that yeast in the stationary phase increase methylation of RPS2 in order to regulate ribosomal synthesis. These results demonstrate the utility of mass spectrometry for quantifying dynamic changes in ribosomal PTMs.

  19. Characterization of hexacationic imidazolium ionic liquids as effective and highly stable gas chromatography stationary phases.

    Science.gov (United States)

    González-Álvarez, Jaime; Blanco-Gomis, Domingo; Arias-Abrodo, Pilar; Díaz-Llorente, Daniel; Ríos-Lombardía, Nicolás; Busto, Eduardo; Gotor-Fernández, Vicente; Gutiérrez-Álvarez, María D

    2012-01-01

    Polycationic ionic liquids (ILs) are an attractive class of ILs with great potential applicability as gas chromatography stationary phases. A family of hexacationic imidazolium ILs derived from the cycloalkanol family was chemically first prepared in a straightforward manner and then applied for analytical separation purposes. Four tuneable engineering vectors, namely cation ring size structure, anion nature, spatial disposition of cycloalkanol substituents and O-substitution, were considered as experimental parameters for the design of the desired ionic liquids. A total number of five new phases based on a common benzene core respectively exhibited column efficiencies around to 2500 plates/m, broad operating temperature ranges and also, even more importantly, good thermal stabilities (bleeding temperature between 260 and 365°C), finding variations in the selectivity and analytes elution orders depending on the IL structures. Their solvation characteristics were evaluated using the Abraham solvation parameter model, establishing clear correlations between their cation structure and retention capability with respect to certain analytes. The study of relationships between the ILs structure and solvation parameters gives us an idea of the IL stationary phase to be used for specific separations.

  20. Impact of mobile phase composition on the performance of porous polymeric monoliths in the elution of small molecules.

    Science.gov (United States)

    Causon, Tim J; Hilder, Emily F; Nischang, Ivo

    2012-11-09

    The influence of mobile phase solvent composition and consequently retention factor on the chromatographic performance for a set of small molecules was studied using a commercially available poly(styrene-co-divinyl benzene) analytical scale porous polymeric monolithic column as an example. Chromatographic elution performance was studied across retention factors from close to 0 up to 100 realized for a set of structurally similar small molecules in a binary reversed-phase solvent environment of acetonitrile and water. By altering the mobile phase composition from volume fractions of acetonitrile of just 10% (v/v) to only acetonitrile it was systematically shown that gel porosity of the monolithic column plays a dominant role in modulating mass transport and the associated chromatographic efficiency in a consistent manner. Up to a sixfold difference in plate height was recorded for the most strongly retained hydrophobic solute (ethylbenzene) at a constant, low flow velocity simply by varying the amount of acetonitrile in the mobile phase. Plate height curves recorded for the set of solutes that comprise benzene, toluene, ethylbenzene as well as phenol and benzyl alcohol further demonstrate the importance of functional group content of the solute and the modulated porous gel structure on mass transport. These results highlight some important practical considerations for characterizing the chromatographic properties of any polymeric monolithic column. First, it is imperative that any chromatographic performance characterization using plate height data explicitly considers the influence of mobile phase composition, retention factor, molecular size and functional groups of the probe solute. Second, as the physicochemical conditions of the material are directly reflected in the gel porosity, a range of different mobile phase compositions, retention factors and probe-specific effects must be investigated to yield a fair appraisal of the chromatographic performance.

  1. Transcriptional Characterization of Salmonella TAl00 in Growth and Stationary Phase: Mutagenesis of MX in Both Types of Cells

    Science.gov (United States)

    The Salmonella (Ames) mutagenicity assay can be performed using cells that are in different growth phases. Thus, the plate-incorporation assay involves plating stationary-phase cells with the mutagen, after which the cells undergo a brief lag phase and, consequently, are exposed ...

  2. Recent advances in the preparation and application of monolithic capillary columns in separation science

    Energy Technology Data Exchange (ETDEWEB)

    Hong, Tingting; Yang, Xi; Xu, Yujing [Department of Analytical Chemistry, China Pharmaceutical University, Nanjing, 210009 (China); Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, Nanjing, 210009 (China); Ji, Yibing, E-mail: jiyibing@msn.com [Department of Analytical Chemistry, China Pharmaceutical University, Nanjing, 210009 (China); Key Laboratory of Drug Quality Control and Pharmacovigilance, Ministry of Education, Nanjing, 210009 (China)

    2016-08-10

    Novel column technologies involving various materials and efficient reactions have been investigated for the fabrication of monolithic capillary columns in the field of analytical chemistry. In addition to the development of these miniaturized systems, a variety of microscale separation applications have achieved noteworthy results, providing a stepping stone for new types of chromatographic columns with improved efficiency and selectivity. Three novel strategies for the preparation of capillary monoliths, including ionic liquid-based approaches, nanoparticle-based approaches and “click chemistry”, are highlighted in this review. Furthermore, we present the employment of state-of-the-art capillary monolithic stationary phases for enantioseparation, solid-phase microextraction, mixed-mode separation and immobilized enzyme reactors. The review concludes with recommendations for future studies and improvements in this field of research. - Highlights: • Preparation of novel monolithic capillary columns have shown powerful potential in analytical chemistry field. • Various materials including ionic liquids and nanoparticles involved into capillary monolithic micro-devices are concluded. • Click chemistry strategy applied for preparing monolithic capillary columns is reviewed. • Recent strategies utilized in constructing different capillary monoliths for enantiomeric separation are summarized. • Advancement of capillary monoliths for complex samples analysis is comprehensively described.

  3. Poly(methacrylic acid-ethylene glycol dimethacrylate) monolith in-tube solid phase microextraction coupled to high performance liquid chromatography and analysis of amphetamines in urine samples.

    Science.gov (United States)

    Fan, Yi; Feng, Yu-Qi; Zhang, Jian-Tao; Da, Shi-Lu; Zhang, Min

    2005-05-13

    In-tube solid-phase microextraction (SPME) based on a poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column was investigated for the extraction of amphetamine, methamphetamine and their methylenedioxy derivatives. The monolithic capillary column showed high extraction efficiency towards target analytes, which could be attributed to its larger loading amount of extraction phase than conventional open-tubular extraction capillaries and the convective mass transfer procedure provided by its monolithic structure. The extraction mechanism was studied, and the results indicated that the extraction process of the target analytes was involved with hydrophobic interaction and ion-exchange interaction. The polymer monolith in-tube SPME-HPLC system with UV detection was successfully applied to the determination of amphetamine, methamphetamine and their methylenedioxy derivatives in urine samples, yielding the detection limits of 1.4 - 4.0 ng/mL. Excellent method reproducibility (RSD urine samples.

  4. Chromatographic selectivity of poly(alkyl methacrylate-co-divinylbenzene) monolithic columns for polar aromatic compounds by pressure-driven capillary liquid chromatography.

    Science.gov (United States)

    Lin, Shu-Ling; Wang, Chih-Chieh; Fuh, Ming-Ren

    2016-10-05

    In this study, divinylbenzene (DVB) was used as the cross-linker to prepare alkyl methacrylate (AlMA) monoliths for incorporating π-π interactions between the aromatic analytes and AlMA-DVB monolithic stationary phases in capillary LC analysis. Various AlMA/DVB ratios were investigated to prepare a series of 30% AlMA-DVB monolithic stationary phases in fused-silica capillaries (250-μm i.d.). The physical properties (such as porosity, permeability, and column efficiency) of the synthesized AlMA-DVB monolithic columns were investigated for characterization. Isocratic elution of phenol derivatives was first employed to evaluate the suitability of the prepared AlMA-DVB columns for small molecule separation. The run-to-run (0.16-1.20%, RSD; n = 3) and column-to-column (0.26-2.95%, RSD; n = 3) repeatabilities on retention times were also examined using the selected AlMA-DVB monolithic columns. The π-π interactions between the aromatic ring and the DVB-based stationary phase offered better recognition on polar analytes with aromatic moieties, which resulted in better separation resolution of aromatic analytes on the AlMA-DVB monolithic columns. In order to demonstrate the capability of potential environmental and/or food safety applications, eight phenylurea herbicides with single benzene ring and seven sulfonamide antibiotics with polyaromatic moieties were analyzed using the selected AlMA-DVB monolithic columns.

  5. Molecularly imprinted solid-phase extraction monolithic capillary column for selective extraction and sensitive determination of safranine T in wolfberry.

    Science.gov (United States)

    Su, Zihao; Zhai, Haiyun; Chen, Zuanguang; Zhou, Qing; Li, Jiangmei; Liu, Zhenping

    2014-02-01

    A method was developed to sensitively determine safranine T in wolfberry by molecularly imprinted solid-phase extraction (MISPE) coupled with high-performance liquid chromatography and laser-induced fluorescence detection (HPLC-LIF). The MISPE capillary monolithic column was prepared by water-bath in situ polymerization, using safranine T, methacrylic acid (MAA), and ethylene dimethacrylate (EDMA) as template, functional monomer, and cross-linker, respectively. The properties of the homemade MISPE capillary monolithic column, including capacity and specificity, were investigated under optimized conditions and the morphologies of inner polymers were characterized by scanning electron microscopy (SEM). The mean recoveries of safranine T in wolfberry ranged from 91.2 % to 92.9 % and the intraday and interday relative standard deviation (RSD) values all ranged from 3.4 % to 4.2 %. Good linearity was obtained over 0.001-1.0 μg mL(-1) (r = 0.9999) with a detection limit (S/N = 3) of 0.4 ng g(-1). Under the selected conditions, enrichment factors of over 90-fold were obtained and the extraction on the monolithic column effectively cleaned up the wolfberry matrix. The results demonstrated that the proposed MISPE-HPLC-LIF method could be applied to sensitively determine safranine T in wolfberry.

  6. Utilization of a diol-stationary phase column in ion chromatographic separation of inorganic anions.

    Science.gov (United States)

    Arai, Kaori; Mori, Masanobu; Kozaki, Daisuke; Nakatani, Nobutake; Itabashi, Hideyuki; Tanaka, Kazuhiko

    2012-12-28

    We describe the ion chromatographic separation of inorganic anions using a diol-stationary phase column (-CH(OH)CH(2)OH; diol-column) without charged functional groups. Anions were separated using acidic eluent as in typical anion-exchange chromatography. The retention volumes of anions on the diol-column increased with increasing H(+) concentration in the eluent. The anion-exchange capacities of diol-columns in the acidic eluent (pH 2.8) were larger than that of zwitterionic stationary phase column but smaller than that of an anion-exchange column. The separation of anions using the diol-column was strongly affected by the interaction of H(+) ions with the diol-functional groups and by the types of the eluents. In particular, the selection of the eluent was very important for controlling the retention time and resolution. Good separation was obtained using a diol-column (HILIC-10) with 5 mM phthalic acid as eluent. The limits of detection at a signal-to-noise ratio of 3 ranged from 1.2 to 2.7 μM with relative standard deviations (RSD, n=5) of 0.04-0.07% for the retention time and 0.4-2.0% for the peak areas. This method was successfully applied to the determination of H(2)PO(4)(-), Cl(-), and NO(3)(-) in a liquid fertilizer sample.

  7. Isolation and molecular characterization of a stationary phase promoter useful for gene expression in Gordonia.

    Science.gov (United States)

    Singh, Pooja; Chachan, Sahil; Singhi, Divya; Srivastava, Preeti

    2016-10-10

    Gordonia are gram-positive bacteria belonging to Actinomycetes family with a wide variety of industrial and environmental applications. The genetic toolbox, however, is limited for manipulation of these organisms. In the present study, a new promoter has been isolated from Gordonia sp. IITR 100 and characterized in detail. The promoter was found to be functional in Escherichia coli. The minimal promoter was identified in a 166bp fragment by deletion mapping. The putative -35 and -10 hexamer showed four and five nucleotide matches respectively with the E. coli consensus sequence. Three direct repeats and an imperfect inverted repeat upstream to -35 were found. The isolated promoter was found to be six times stronger than the Pkan promoter observed by cloning lacZ downstream to each of them in a plasmid in E. coli. The β-galactosidase activity was maximum at stationary phase and found to be ~800MU for Gordonia sp. IITR 100 and E. coli. This is the first report of a stationary phase promoter isolated and characterized from Gordonia.

  8. Battle of the Bacteria: Characterizing the Evolutionary Advantage of Stationary Phase Growth

    Directory of Open Access Journals (Sweden)

    Karin E. Kram

    2016-05-01

    Full Text Available Providing students with authentic research opportunities has been shown to enhance learning and increase retention in STEM majors. Accordingly, we have developed a novel microbiology lab module, which focuses on the molecular mechanisms of evolution in E. coli, by examining the growth advantage in stationary phase (GASP phenotype. The GASP phenotype is demonstrated by growing cells into long-term stationary phase (LTSP and then competing them against un-aged cells in a fresh culture. This module includes learning goals related to strengthening practical laboratory skills and improving student understanding of evolution. In addition, the students generate novel data regarding the effects of different environmental stresses on GASP and the relationship between evolution, genotypic change, mutation frequency, and cell stress. Pairs of students are provided with the experimental background, select a specific aspect of the growth medium to modify, and generate a hypothesis regarding how this alteration will impact the GASP phenotype. From this module, we have demonstrated that students are able to achieve the established learning goals and have produced data that has furthered our understanding of the GASP phenotype.

  9. Preparation of Medium Cation Exchange Stationary Phase of Polymeric Matrix and Their Chromatographic Properties

    Institute of Scientific and Technical Information of China (English)

    CHEN,Gang; GONG,Bo-Lin; BAI,Quan; GENG,Xin-Du

    2007-01-01

    Based on the monodisperse poly(glycidyl methacrylate-co-ethylenedimethacrylate) beads (PGMA/EDMA) with macropore as a medium, a new hydrophilic medium cation exchange (MCX) stationary phase for HPLC was synthesized by a new chemically modified method. The stationary phase was evaluated with the property of ion exchange, separability, reproducibility, hydrophilicity, effect of salt concentration, salt types, column loading and pH on the separation and retention of proteins in detail. It was found that it follows ion exchange chromatographic (IEC)retention mechanism. The measured bioactivity recovery for lysozyme was (96±5)%. The dynamic protein loading capacity of the synthesized MCX packings was 21.8 mg/g. Five proteins were almost completely separated within 6.0 min at a flow rate of 4 mL/min using the synthesized MCX resin. The MCX resin was also used for the rapid separation and purification of lysozyme from egg white with only one step. The purity and specific bioactivity of the purified lysozyme was found more than 95% and 70345 U/mg, respectively.

  10. Stationary point analysis of the one-dimensional lattice Landau gauge fixing functional, aka random phase XY Hamiltonian

    Science.gov (United States)

    Mehta, Dhagash; Kastner, Michael

    2011-06-01

    We study the stationary points of what is known as the lattice Landau gauge fixing functional in one-dimensional compact U(1) lattice gauge theory, or as the Hamiltonian of the one-dimensional random phase XY model in statistical physics. An analytic solution of all stationary points is derived for lattices with an odd number of lattice sites and periodic boundary conditions. In the context of lattice gauge theory, these stationary points and their indices are used to compute the gauge fixing partition function, making reference in particular to the Neuberger problem. Interpreted as stationary points of the one-dimensional XY Hamiltonian, the solutions and their Hessian determinants allow us to evaluate a criterion which makes predictions on the existence of phase transitions and the corresponding critical energies in the thermodynamic limit.

  11. Macroporous Monolithic Polymers: Preparation and Applications

    Directory of Open Access Journals (Sweden)

    Cecilia Inés Alvarez Igarzabal

    2009-12-01

    Full Text Available In the last years, macroporous monolithic materials have been introduced as a new and useful generation of polymers used in different fields. These polymers may be prepared in a simple way from a homogenous mixture into a mold and contain large interconnected pores or channels allowing for high flow rates at moderate pressures. Due to their porous characteristics, they could be used in different processes, such as stationary phases for different types of chromatography, high-throughput bioreactors and in microfluidic chip applications. This review reports the contributions of several groups working in the preparation of different macroporous monoliths and their modification by immobilization of specific ligands on the products for specific purposes.

  12. Preparation of new hybrid organic/inorganic polymeric chiral stationary phases for ligand-exchange chromatography

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Three new hybrid organic/inorganic polymeric ligand-exchange chiral stationary phases were developed by radical chain transfer reaction and surface grafting on silica gel, and successfully used for the enantioseparations of DL-amino acids and DL-hydroxyl acids. The resolutions were achieved by using water containing 2.0 × 10-4 mol/L of CuAc2 as a mobile phase, column temperature of 40 ℃, flow rate of 1.0 mL/min and detection at UV 254 mn. The elution order of D-isomer before L-isomer was observed for all DL-amino acids resolved except DL-Pro.

  13. Principle of stationary phase for propagating wave packets in the unidimensional scattering problem

    Energy Technology Data Exchange (ETDEWEB)

    Bernardini, A.E. [Universidade Federal de Sao Carlos, Departamento de Fisica, PO Box 676, Sao Carlos, SP (Brazil)

    2008-08-15

    We point out some incompatibilities which appear when one applies the stationary phase method for deriving phase times to obtain the spatial localization of wave packets scattered by a unidimensional potential barrier. We concentrate on the above barrier diffusion problem where the wave packet collision implies the possibility of multiple reflected and transmitted wave packets, which, depending on the boundary conditions, can overlap or stand in relative separation in space. We demonstrate that the indiscriminate use of the method for such a particular configuration leads to paradoxical results for which the correct interpretation, confirmed by analytical/numerical calculations, imposes the necessity of the appearance of multiple peaks as a consequence of multiple reflections by the barrier steps. (orig.)

  14. Adsorption of the Enantiomers of Tryptophan on Stationary Phase Bonded with Bovine Serum Albumin

    Institute of Scientific and Technical Information of China (English)

    ZHANG Fengbao; CHENG Ming; LI Shuang; LIU Liang; ZHANG Guoliang

    2006-01-01

    Frontal analysis is frequently applied to measuring single or multi-component adsorption isotherms. In this work, the competitive adsorption isotherm data of two enantiomers of tryptophan were obtained by competitive frontal analysis. The stationary phase in the column was silica-immobilized bovine serum albumin(BSA)by the derivative method, and the mobile phase was a phosphate buffer. These isotherm data were fitted by the competitive Bilangmuir model. This model can account for the behavior of both tryptophan enantiomers and these profiles were found to fit the experimental band profiles(square error is 0.999 6). The parameters obtained were used in numericai calculations to predict the band profiles of the racemic mixtures of tryptophan. The equilibriumdispersive model provides satisfactory prediction, with minor differences between the calculated and the experimental profiles.

  15. Chiral HPLC analysis of milnacipran and its FMOC-derivative on cellulose-based stationary phases.

    Science.gov (United States)

    Patti, Angela; Pedotti, Sonia; Sanfilippo, Claudia

    2008-02-01

    The HPLC enantioseparation of the last generation antidepressive drug milnacipran (+/-)-1 was investigated on different cellulose-based chiral stationary phases (CSPs). On carbamate-type columns, Chiralcel OD and OD-H (+/-)-1 could be separated with alpha value about 1.20 but the resolution was quite low because of the tailing of the peaks. Direct determination of (+/-)-1 with high selectivity and resolution was obtained on Chiralcel OJ in normal phase mode elution. Precolumn derivatization of milnacipran with Fmoc-Cl gave compound (+/-)-2 which was enantioseparated on all the investigated CSPs and allowed enhanced UV or fluorimetric detection. The Chiralpak IB, that could be considered the immobilized version of Chiralcel OD-H, was found completely ineffective in the chiral recognition of (+/-)-1 and moderately efficient in the separation of (+/-)-2.

  16. The phase delay and its complex time: From stationary states up to wave packets

    Energy Technology Data Exchange (ETDEWEB)

    Grossel, Ph., E-mail: philippe.grossel@univ-reims.fr

    2013-03-15

    Complex time is often invoked about tunneling effect where the classical phase delay is completed with a crucial filter effect. Usually the complex times are obtained by considering the flux-flux correlation function, but this can be obtained by a very simple approach using the search of the maximum of the generalized complex phase function, including the amplitude of the wave function. Various aspects of the phase delay are presented in the case of wave packets impinging on simple or resonant quantum barriers. Formal links with the classical mechanics give birth to quasi-trajectories of the quantum particle, totally compatible with the quantum mechanics. - Highlights: Black-Right-Pointing-Pointer The stationary phase method is extended in including the variations of the spectra. Black-Right-Pointing-Pointer The complex phase delay leads to a complex trajectory inside and out-side the barrier. Black-Right-Pointing-Pointer Examples of quasi-trajectories are given in case of different quantum barriers. Black-Right-Pointing-Pointer Phase delays are specified for resonant tunneling or above-barrier wave-packets. Black-Right-Pointing-Pointer The coherence between the quasi-trajectories and quantum mechanics is shown.

  17. Hybrid organic-inorganic silica monolith with hydrophobic/strong cation-exchange functional groups as a sorbent for micro-solid phase extraction.

    Science.gov (United States)

    Zheng, Ming-Ming; Ruan, Ge-Deng; Feng, Yu-Qi

    2009-11-06

    A hybrid organic-inorganic silica monolith with hydrophobic and strong cation-exchange functional groups was prepared and used as a sorbent for micro-solid phase extraction (micro-SPE). The hybrid silica monolith functionalized with octyl and thiol groups was conveniently synthesized by hydrolysis and polycondensation of a mixture of tetraethoxysilane (TEOS), n-octyltriethoxysilane (C8-TEOS) and 3-mercaptopropyltrimethoxysilane (MPTMS) via a two-step catalytic sol-gel process. Due to the favorable chemical reactivity of mercapto pendant moieties, the obtained hybrid monolith was oxidized using hydrogen peroxide (30%, w/w) to yield sulfonic acid groups, which provided strong cation-exchange sites. The obtained hybrid monolith was characterized by diffused infrared spectroscopy, elemental analysis, scanning electron microscopy and mercury intrusion porosimetry. The results show that the resulting monolith contains much higher carbon (31.6%) and sulfur (4.8%) contents than traditionally bonded silica materials. The extraction performance of the hybrid monolith was evaluated using sulfonamides as testing analytes by micro-SPE on-line coupled to HPLC. The results show that the hybrid monolith with hydrophobic and strong cation-exchange functional groups exhibits high extraction efficiency towards the testing analytes. The column-to-column RSD values were 1.3-9.8% for the extraction of SAs investigated. The extraction performance of the hybrid silica monolith remained practically unchanged after treated with acid (pH 1.0) and basic solutions (pH 10.5). Finally, the application of the hybrid monolith was demonstrated by micro-SPE of sulfonamide residues from milk followed by HPLC-UV analysis. The limits of detection (S/N=3) for eight SAs were found to be 1.0-3.0ng/mL in milk. The recoveries of eight SAs spiked in milk sample ranged from 80.2% to 115.6%, with relative standard deviations less than 11.8%.

  18. Poly(ethylene oxide)-bonded stationary phase for separation of inorganic anions in capillary ion chromatography.

    Science.gov (United States)

    Linda, Roza; Lim, Lee Wah; Takeuchi, Toyohide

    2013-06-14

    A tosylated-poly(ethylene oxide) (PEO) reagent was reacted with primary amino groups of an aminopropylsilica packing material (TSKgel NH2-60) in acetonitrile to form PEO-bonded stationary phase. The reaction was a single and simple step reaction. The prepared stationary phase was able to separate inorganic anions. The retention behavior of six common inorganic anions on the prepared stationary phase was examined under various eluent conditions in order to clarify its separation/retention mechanism. The elution order of the tested anions was iodate, bromate, bromide, nitrate, iodide, and thiocyanate, which was similar as observed in common ion chromatography. The retention of inorganic anions could be manipulated by ion exchange interaction which is expected that the eluent cation is coordinated among the PEO chains and it works as the anion-exchange site. Cations and anions of the eluent therefore affected the retention of sample anions. We demonstrated that the retention of the analyte anions decreased with increasing eluent concentration. The repeatability of retention time for the six anions was satisfactory on this column with relative standard deviation values from 1.1 to 4.3% when 10mM sodium chloride was used as the eluent. Compared with the unmodified TSKgel NH2-60, the prepared stationary phase retained inorganic anions more strongly and the selectivity was also improved. The present stationary phase was applied for the determination of inorganic anions contained in various water samples.

  19. Damage detection using transient trajectories in phase-space with extended random decrement technique under non-stationary excitations

    Science.gov (United States)

    Liu, Gang; Mao, Zhu; Todd, Michael

    2016-11-01

    This paper proposes a damage detection method based on the geometrical variation of transient trajectories in phase-space, and the proposed methodology is compatible with non-stationary excitations (e.g., earthquake-induced ground motion). The work presented assumes zero-mean non-stationary excitation, and extends the random decrement technique to convert non-stationary response signals of the structure into free-vibration data. Transient trajectories of the structure are reconstructed via the embedding theorem from the converted free-vibration data, and trajectories are mapped successively into phase-space to enhance statistical analysis. Based upon the characterized system dynamics in terms of phase-space, the time prediction error is adopted as the damage index. To identify the presence and severity of damage in a statistically rigorous way, receiver operating characteristic curves and the Bhattacharyya distance are employed. The results from both numerical simulations and experiments validate the proposed framework, when the test structures are subject to non-stationary excitations. The extension achieved in this paper enables the phase-space damage detection approach to be compatible with non-stationary scenarios, such as traffic, wind, and earthquake loadings. Moreover, the results indicate that this phase-state-based method is able to identify damage-induced nonlinearity in response, which is an intrinsic characteristic associated with most structural damage types.

  20. Integrated Time and Phase Synchronization Strategy for a Multichannel Spaceborne-Stationary Bistatic SAR System

    Directory of Open Access Journals (Sweden)

    Feng Hong

    2016-07-01

    Full Text Available The spatial separation of the transmitter and receiver in Bistatic Synthetic Aperture Radar (BiSAR makes it a promising and useful supplement to a classical Monostatic SAR system (MonoSAR. This paper proposes a novel integrated time and phase synchronization strategy for a multichannel spaceborne-stationary BiSAR system. Firstly, the time synchronization strategy is proposed, which includes Pulse Repetition Frequency (PRF generation under noisy conditions, multichannel calibration and the alignment of the recorded data with the orbital data. Furthermore, the phase synchronization strategy, which fully considers the deteriorative factors in the BiSAR configuration, is well studied. The contribution of the phase synchronization strategy includes two aspects: it not only compensates the phase error, but also improves the Signal to Noise Ratio (SNR of the obtained signals. Specifically, all direct signals on different PRF time can be reconstructed with the shift and phase compensation operation using a reference signal. Besides, since the parameters of the reference signal can be estimated only once using the selected practical direct signal and a priori information, the processing complexity is well reduced. Final imaging results with and without compensation for real data are presented to validate the proposed synchronization strategy.

  1. Separation of transition and heavy metals using stationary phase gradients and thin layer chromatography.

    Science.gov (United States)

    Stegall, Stacy L; Ashraf, Kayesh M; Moye, Julie R; Higgins, Daniel A; Collinson, Maryanne M

    2016-05-13

    Stationary phase gradients for chelation thin layer chromatography (TLC) have been investigated as a tool to separate a mixture of metal ions. The gradient stationary phases were prepared using controlled rate infusion (CRI) from precursors containing mono-, bi-, and tri-dentate ligands, specifically 3-aminopropyltriethoxysilane, N-[3-(trimethoxysilyl)propyl] ethylenediamine, and N-[3-(trimethoxysilyl)propyl] diethylenetriamine. The presence and the extent of gradient formation were confirmed using N1s X-ray photoelectron spectroscopy (XPS). XPS results showed that the degree of modification was dependent on the aminosilane precursor, its concentration, and the rate of infusion. The separation of four transition and heavy metals (Co(2+), Pb(2+), Cu(2+), and Fe(3+)) on gradient and uniformly modified plates was compared using a mobile phase containing a stronger chelating agent, ethylenediaminetetraacetic acid (EDTA). The retention of the metal ions was manipulated by varying the surface concentration of the chelating ligands. The order of retention on unmodified plates and on plates modified with a monodentate ligand was Fe(3+)>Cu(2+)∼Pb(2+)∼Co(2+), while the order of retention on plates modified with bi- and tri-dentate ligands was Fe(3+)>Cu(2+)>Pb(2+)∼Co(2+). Fe(3+) and Cu(2+) were much more sensitive to the concentration of chelating ligand on the surface (displaying lower Rf values with increasing ligand concentration) than Pb(2+) and Co(2+). Complete separation was achieved using a high concentration of the tridentate ligand coupled with a longer time for modification, yielding a retention order of Fe(3+)>Cu(2+)>Co(2+)>Pb(2+). Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Novel RpoS-Dependent Mechanisms Strengthen the Envelope Permeability Barrier during Stationary Phase.

    Science.gov (United States)

    Mitchell, Angela M; Wang, Wei; Silhavy, Thomas J

    2017-01-15

    Gram-negative bacteria have effective methods of excluding toxic compounds, including a largely impermeable outer membrane (OM) and a range of efflux pumps. Furthermore, when cells become nutrient limited, RpoS enacts a global expression change providing cross-protection against many stresses. Here, we utilized sensitivity to an anionic detergent (sodium dodecyl sulfate [SDS]) to probe changes occurring to the cell's permeability barrier during nutrient limitation. Escherichia coli is resistant to SDS whether cells are actively growing, carbon limited, or nitrogen limited. In actively growing cells, this resistance depends on the AcrAB-TolC efflux pump; however, this pump is not necessary for protection under either carbon-limiting or nitrogen-limiting conditions, suggesting an alternative mechanism(s) of SDS resistance. In carbon-limited cells, RpoS-dependent pathways lessen the permeability of the OM, preventing the necessity for efflux. In nitrogen-limited but not carbon-limited cells, the loss of rpoS can be completely compensated for by the AcrAB-TolC efflux pump. We suggest that this difference simply reflects the fact that nitrogen-limited cells have access to a metabolizable energy (carbon) source that can efficiently power the efflux pump. Using a transposon mutant pool sequencing (Tn-Seq) approach, we identified three genes, sanA, dacA, and yhdP, that are necessary for RpoS-dependent SDS resistance in carbon-limited stationary phase. Using genetic analysis, we determined that these genes are involved in two different envelope-strengthening pathways. These genes have not previously been implicated in stationary-phase stress responses. A third novel RpoS-dependent pathway appears to strengthen the cell's permeability barrier in nitrogen-limited cells. Thus, though cells remain phenotypically SDS resistant, SDS resistance mechanisms differ significantly between growth states. Gram-negative bacteria are intrinsically resistant to detergents and many

  3. Design of monoliths through their mechanical properties.

    Science.gov (United States)

    Podgornik, Aleš; Savnik, Aleš; Jančar, Janez; Krajnc, Nika Lendero

    2014-03-14

    Chromatographic monoliths have several interesting properties making them attractive supports for analytics but also for purification, especially of large biomolecules and bioassemblies. Although many of monolith features were thoroughly investigated, there is no data available to predict how monolith mechanical properties affect its chromatographic performance. In this work, we investigated the effect of porosity, pore size and chemical modification on methacrylate monolith compression modulus. While a linear correlation between pore size and compression modulus was found, the effect of porosity was highly exponential. Through these correlations it was concluded that chemical modification affects monolith porosity without changing the monolith skeleton integrity. Mathematical model to describe the change of monolith permeability as a function of monolith compression modulus was derived and successfully validated for monoliths of different geometries and pore sizes. It enables the prediction of pressure drop increase due to monolith compressibility for any monolith structural characteristics, such as geometry, porosity, pore size or mobile phase properties like viscosity or flow rate, based solely on the data of compression modulus and structural data of non-compressed monolith. Furthermore, it enables simple determination of monolith pore size at which monolith compressibility is the smallest and the most robust performance is expected. Data of monolith compression modulus in combination with developed mathematical model can therefore be used for the prediction of monolith permeability during its implementation but also to accelerate the design of novel chromatographic monoliths with desired hydrodynamic properties for particular application.

  4. Preparation of a monolith functionalized with zinc oxide nanoparticles and its application in the enrichment of fluoroquinolone antibiotics.

    Science.gov (United States)

    Liu, Dan; Ma, Jiutong; Jin, Yan; Li, Xiqian; Zhou, Xiao; Jia, Qiong; Zhou, Weihong

    2015-01-01

    This study describes the enrichment ability of ZnO-modified methacrylic acid-co-ethylene dimethacrylate polymer monoliths as stationary phases for the simultaneous determination of antibiotics (ofloxacin, ciprofloxacin, enoxacin, and pefloxacin) combined with high-performance liquid chromatography. The prepared monolith was characterized by scanning electron microscopy, X-ray photoelectron spectroscopy, Fourier-transformed infrared spectroscopy, and thermogravimetric analysis. The polymer monolith microextraction method has been applied to the enrichment of fluoroquinolone antibiotics and satisfactory results were obtained in the analysis of water samples. Compared with the conventional methacrylic acid based monolith, the developed monolith exhibited a higher enrichment capacity because of the introduction of zinc oxide into the preparation process. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Green synthesis of a typical chiral stationary phase of cellulose-tris(3, 5-dimethylphenylcarbamate)

    Science.gov (United States)

    2013-01-01

    Background At present, the study on the homogeneous-phase derivatization of cellulose in ionic liquid is mainly focused on its acetylation. To the best of our knowledge, there has been no such report on the preparation of cellulose-tris(3,5-dimethylphenylcarbamate) (CDMPC) with ionic liquid 1-allyl-3-methyl-imidazolium chloride (AmimCl) so far. Results With ionic liquid 1-allyl-3-methylimidazolium chloride (AmimCl) as a reaction solvent, cellulose-tris(3,5-dimethylphenylcarbamate) (CDMPC) was synthesized by the reaction of 3,5-dimethylphenyl isocyanate and soluble microcrystalline cellulose in a homogeneous phase. The synthesized CDMPC was then coated onto the surfaces of aminopropyl silica gel to prepare a chiral stationary phase (CSP). The prepared CSP was successfully used in chiral separation of seven racemic pesticides by high performance liquid chromatography (HPLC). Good chiral separation was obtained using n-hexane and different modifiers as the mobile phases under the optimal percentage and column temperature, with the resolution of metalaxyl, diniconazole, flutriafol, paclobutrazol, hexaconazole, myclobutanil and hexythiazox of 1.73, 1.56, 1.26, 1.00, 1.18, 1.14 and 1.51, respectively. The experimental results suggested it was a good choice using a green solvent of AmimCl for cellulose functionalization. Conclusion CDMPC was successfully synthesized as the chiral selector by reacting 3, 5-dimethylphenyl isocyanate with dissolved microcrystalline cellulose in a green ionic liquid of AmimCl. PMID:23890199

  6. Growth potential of exponential- and stationary-phase Salmonella Typhimurium during sausage fermentation.

    Science.gov (United States)

    Birk, T; Henriksen, S; Müller, K; Hansen, T B; Aabo, S

    2016-11-01

    Raw meat for sausage production can be contaminated with Salmonella. For technical reasons, meat is often frozen prior to mincing but it is unknown how growth of Salmonella in meat prior to freezing affects its growth potential during sausage fermentation. We investigated survival of exponential- and stationary-phase Salmonella Typhimurium (DT12 and DTU292) during freezing at -18°C and their subsequent growth potential during 72h sausage fermentation at 25°C. After 0, 7 and >35d of frozen storage, sausage batters were prepared with NaCl (3%) and NaNO2 (0, 100ppm) and fermented with and without starter culture. With no starter culture, both strains grew in both growth phases. In general, a functional starter culture abolished S. Typhimurium growth independent of growth phase and we concluded that ensuring correct fermentation is important for sausage safety. However, despite efficient fermentation, sporadic growth of exponential-phase cells of S. Typhimurium was observed drawing attention to the handling and storage of sausage meat.

  7. Preparation, characterization, and analytical applications of a novel polymer stationary phase with embedded or grafted carbon fibers.

    Science.gov (United States)

    Zhong, Yingying; Zhou, Wenfang; Zhang, Peimin; Zhu, Yan

    2010-09-15

    A polymer-based chromatographic stationary phase with embedded or grafted multi-walled carbon nanotubes (MWCNTs) has been developed. Three different synthetic methods were utilized to combine the nano-fibers with the substrate of polystyrene-divinylbenzene (PS-DVB). After optimizing the synthetic conditions, this novel polystyrene-divinylbenzene-carbon nanotube (PS-DVB-CNT) stationary phase was characterized by scanning electron microscopy, Raman spectroscopy, thermogravimetric analysis, chemical adsorption and desorption measurement, and mechanical stability test. Compared to PS-DVB particles, PS-DVB-CNT particles have certain improvement in physical and chromatographic performances because the addition of MWCNTs has altered the structures of the particles. The novel stationary phase owns satisfactory resolution, wide pH endurance, and long lifetime, which can be used as an extent to normal HPLC.

  8. Improved Procedure for Preparation of Covalently Bonded Cellulose Tris-phenylcarbamate Chiral Stationary Phases

    Institute of Scientific and Technical Information of China (English)

    秦峰; 陈小明; 刘月启; 邹汉法; 王俊德

    2005-01-01

    The classical method for preparation of covalently boned cellulose derivative chiral stationary phases (CSP) with diisocyanate as spacer was improved. Diisocyanate was firstly allowed to react with 3-aminopropyltriethoxysilane, and the resulting product was then applied as the spacer reagent to immobilize cellulose derivatives onto silica gel. Influences of the amount and the length of the spacer on the optical resolution ability of the CSP were investigated. Comparing improved procedure to classical diisocyanate method, the cross-linking between the glucose units of the cellulose derivatives was avoided to the most extent. With the improved procedure, regio-nonselective ways could be adooted to prepare covalently bonded CSP, which showed an advantage for the rapid preparation.

  9. A novel amide stationary phase for hydrophilic interaction liquid chromatography and ion chromatography.

    Science.gov (United States)

    Shen, Guobin; Zhang, Feifang; Yang, Bingcheng; Chu, Changhu; Liang, Xinmiao

    2013-10-15

    A novel amide stationary phase (ASP) for hydrophilic interaction liquid chromatography (HILIC) has been prepared via the Click chemistry method. It was based on the strategy that the amino group of Asparagine was easily transferred to the corresponding azido group and then clicked onto terminal alkyne-silica gel in the presence of Cu(I)-based catalyst. For the tested polar compounds including nucleosides and nucleic acid bases, ASP-based column has demonstrated good performance in terms of separation efficiency and column stability, and the retention mechanism was found to match well the typical HILIC retention. In addition, the ASP described here showed much better selectivity in separation of inorganic anions under ion chromatography mode relative to other kinds of commercial ASP.

  10. Fundamental and practical studies on high-performance liquid affinity chromatography of biopolymers with novel stationary phases

    Energy Technology Data Exchange (ETDEWEB)

    Bacolod, M.D.

    1992-01-01

    Rigid microparticulate stationary phases having surface-bound metal chelating functions were developed and evaluated in high performance metal chelate affinity chromatography of proteins. Silica- and polystyrene-divinylbenzene-based metal chelate sorbents were produced in wide pore and in non-porous type of column packings. A major effort has been placed on development of non-porous highly crosslinked polystyrene-divinylbenzene (PSDVB). These PSDVB microparticles were produced by a two-step swelling polymerization, and exhibited excellent mechanical strength over a wide range of flow-rates and composition used in high performance liquid chromatography (HPLC). Simple and reproducible hydrophilic coatings were developed for the surface modification of hydrophobic PSDVB supports. A tetradentate metal chelating ligand, ethylenediamine-N, N[prime]-diacetic acid (EDDA), was covalently bound to the surface of the various supports. Sorbents having iminodiacetic acid (IDA) metal chelating functions were also evaluated. The hydrophilic character and surface coverage of various stationary phases were assessed chromatographically. Studies concerning the effects of eluent pH as well as the nature and concentration of salts on retention and selectivity with different metal chelate stationary phases having various immobilized metal ions were carried out. Elution schemes were developed for rapid separation of proteins in metal chelate affinity chromatography. EDDA stationary phases in metal forms can be viewed as complementary to IDA stationary phases since they afforded different selectivity and retentivity toward proteins. Hydrophilic PSDVB could be functionalized with IDA or EDDA metal chelating ligands or lectins. The non-porous metal chelate stationary phases afforded rapid separation of proteins by the development of multiple gradient systems, which permitted higher column peak capacity, enabling the separation of a greater number of proteins in a single chromatographic run.

  11. Separations of corticosteroids using electrochemically modulated liquid chromatography: Selectivity enhancements at a porous graphitic carbon stationary phase

    Energy Technology Data Exchange (ETDEWEB)

    Ting, E.Y.; Porter, M.D. [Ames Lab., IA (United States)]|[Iowa State Univ., Ames, IA (United States)

    1997-02-15

    Electrochemically modulated liquid chromatography has been applied to the separation of a mixture of structurally similar corticosteroids (i.e., prednisone, prednisolone, cortisone, and hydrocortisone) using a porous graphitic carbon stationary phase. Changes in the voltage applied to the column markedly affected the efficiency as well as the elution order of the separation, with the mixture fully resolved at large negative values of applied potential. Mechanistic aspects in terms of the influence of changes in the applied voltage on the extent of the interactions between these analytes and the stationary phase are briefly discussed. 19 refs., 2 figs.

  12. Novel chiral stationary phases based on peptoid combining a quinine/quinidine moiety through a C9-position carbamate group.

    Science.gov (United States)

    Wu, Haibo; Wang, Dongqiang; Song, Guangjun; Ke, Yanxiong; Liang, Xinmiao

    2014-04-01

    By connecting a quinine or quinidine moiety to the peptoid chain through the C9-position carbamate group, we synthesized two new chiral selectors. After immobilizing them onto 3-mercaptopropyl-modified silica gel, two novel chiral stationary phases were prepared. With neutral, acid, and basic chiral compounds as analytes, we evaluated these two stationary phases and compared their chromatographic performance with chiral columns based on quinine tert-butyl carbamate and the previous peptoid. From the resolution of neutral and basic analytes under normal-phase mode, it was found that the new stationary phases exhibited much better enantioselectivity than the quinine tert-butyl carbamate column; the peptoid moiety played an important role in enantiorecognition, which controlled the elution orders of enantiomers; the assisting role of the cinchona alkaloid moieties was observed in some separations. Under acid polar organic phase mode, it was proved that cinchona alkaloid moieties introduced excellent enantiorecognitions for chiral acid compounds; in some separations, the peptoid moiety affected enantioseparations as well. Overall, chiral moieties with specific enantioselectivity were demonstrated to improve the performance of peptoid chiral stationary phase efficiently.

  13. Nanocellulose Derivative/Silica Hybrid Core-Shell Chiral Stationary Phase: Preparation and Enantioseparation Performance

    Directory of Open Access Journals (Sweden)

    Xiaoli Zhang

    2016-05-01

    Full Text Available Core-shell silica microspheres with a nanocellulose derivative in the hybrid shell were successfully prepared as a chiral stationary phase by a layer-by-layer self-assembly method. The hybrid shell assembled on the silica core was formed using a surfactant as template by the copolymerization reaction of tetraethyl orthosilicate and the nanocellulose derivative bearing triethoxysilyl and 3,5-dimethylphenyl groups. The resulting nanocellulose hybrid core-shell chiral packing materials (CPMs were characterized and packed into columns, and their enantioseparation performance was evaluated by high performance liquid chromatography. The results showed that CPMs exhibited uniform surface morphology and core-shell structures. Various types of chiral compounds were efficiently separated under normal and reversed phase mode. Moreover, chloroform and tetrahydrofuran as mobile phase additives could obviously improve the resolution during the chiral separation processes. CPMs still have good chiral separation property when eluted with solvent systems with a high content of tetrahydrofuran and chloroform, which proved the high solvent resistance of this new material.

  14. Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase

    Science.gov (United States)

    Borisova, Marina; Gaupp, Rosmarie; Duckworth, Amanda; Schneider, Alexander; Dalügge, Désirée; Mühleck, Maraike; Deubel, Denise; Unsleber, Sandra; Yu, Wenqi; Muth, Günther; Bischoff, Markus; Götz, Friedrich

    2016-01-01

    ABSTRACT Peptidoglycan recycling is a metabolic process by which Gram-negative bacteria reutilize up to half of their cell wall within one generation during vegetative growth. Whether peptidoglycan recycling also occurs in Gram-positive bacteria has so far remained unclear. We show here that three Gram-positive model organisms, Staphylococcus aureus, Bacillus subtilis, and Streptomyces coelicolor, all recycle the sugar N-acetylmuramic acid (MurNAc) of their peptidoglycan during growth in rich medium. They possess MurNAc-6-phosphate (MurNAc-6P) etherase (MurQ in E. coli) enzymes, which are responsible for the intracellular conversion of MurNAc-6P to N-acetylglucosamine-6-phosphate and d-lactate. By applying mass spectrometry, we observed accumulation of MurNAc-6P in MurNAc-6P etherase deletion mutants but not in either the isogenic parental strains or complemented strains, suggesting that MurQ orthologs are required for the recycling of cell wall-derived MurNAc in these bacteria. Quantification of MurNAc-6P in ΔmurQ cells of S. aureus and B. subtilis revealed small amounts during exponential growth phase (0.19 nmol and 0.03 nmol, respectively, per ml of cells at an optical density at 600 nm [OD600] of 1) but large amounts during transition (0.56 nmol and 0.52 nmol) and stationary (0.53 nmol and 1.36 nmol) phases. The addition of MurNAc to ΔmurQ cultures greatly increased the levels of intracellular MurNAc-6P in all growth phases. The ΔmurQ mutants of S. aureus and B. subtilis showed no growth deficiency in rich medium compared to the growth of the respective parental strains, but intriguingly, they had a severe survival disadvantage in late stationary phase. Thus, although peptidoglycan recycling is apparently not essential for the growth of Gram-positive bacteria, it provides a benefit for long-term survival. PMID:27729505

  15. A new imidazolium-embedded C{sub 18} stationary phase with enhanced performance in reversed-phase liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Qiu Hongdeng [Department of Applied Chemistry and Biochemistry, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Lanzhou Institute of Chemical Physics, Chinese Academy of Science, Lanzhou 730000 (China); Mallik, Abul K. [Department of Applied Chemistry and Biochemistry, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Takafuji, Makoto [Department of Applied Chemistry and Biochemistry, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Kumamoto Institute for Photo-Electro Organics (Phoenics), Kumamoto 862-0901 (Japan); Liu Xia; Jiang Shengxiang [Lanzhou Institute of Chemical Physics, Chinese Academy of Science, Lanzhou 730000 (China); Ihara, Hirotaka, E-mail: ihara@kumamoto-u.ac.jp [Department of Applied Chemistry and Biochemistry, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Kumamoto Institute for Photo-Electro Organics (Phoenics), Kumamoto 862-0901 (Japan)

    2012-08-13

    Highlights: Black-Right-Pointing-Pointer Imidazolium-embedded C{sub 18} stationary phase was prepared and characterized. Black-Right-Pointing-Pointer Enhanced chromatographic selectivity was observed in SiImC{sub 18} column. Black-Right-Pointing-Pointer Seven nucleosides and bases were separated using only water as eluent within 8 min. Black-Right-Pointing-Pointer Multiple-interactions induced by embedded polar imidazolium was investigated. - Abstract: In this paper, a new imidazolium-embedded C{sub 18} stationary phase (SiImC{sub 18}) for reversed-phase high-performance liquid chromatography is described. 1-Allyl-3-octadecylimidazolium bromide ionic liquid compound having a long alkyl chain and reactive groups was newly prepared and grafted onto 3-mercaptopropyltrimethoxysilane-modified silica via a surface-initiated radical-chain transfer addition reaction. The SiImC{sub 18} obtained was characterized by elemental analysis, infrared spectroscopy, thermogravimetric analysis, diffuse reflectance infrared Fourier transform, and solid-state {sup 13}C and {sup 29}Si cross-polarization/magic angle spinning nuclear magnetic resonance spectroscopy. The selectivity toward polycyclic aromatic hydrocarbons relative to that toward alkylbenzenes exhibited by SiImC{sub 18} was higher than the corresponding selectivity exhibited by a conventional octadecyl silica (ODS) column, which could be explained by electrostatic {pi}-{pi} interaction cationic imidazolium and electron-rich aromatic rings. On the other hand, SiImC{sub 18} also showed high selectivity for polar compounds, which was based on the multiple interaction and retention mechanisms of this phase with different analytes. 1,6-Dinitropyrene and 1,8-dinitropyrene, which form a positional isomer pair of dipolar compounds, were separated successfully with the SiImC{sub 18} phase. Seven nucleosides and bases (i.e. cytidine, uracil, uridine, thymine, guanosine, xanthosine, and adenosine) were separated using only water as

  16. Chiral separation of novel diazenes on a polysaccharide-based stationary phase in the reversed-phase mode.

    Science.gov (United States)

    Vojtylová, Terézia; Hamplová, Věra; Galewski, Zbigniew; Korbecka, Izabela; Sýkora, David

    2017-01-31

    Chiral high-performance liquid chromatography separation of two recently synthesized liquid crystalline materials C1 and C2 was studied in the reversed-phase mode. Both materials have an azo-moiety and one chiral centre in their molecular structures. They were available in racemic and pure S forms. For the enantiomeric separations, a Chiralpak AY-RH stationary phase based on amylose tris(5-chloro-2-methylphenylcarbamate) coated on 5 μm silica was used. The compounds were analyzed in both of their possible forms, the more thermodynamically stable E form and the labile Z form. The conditions and time scale of the UV-induced E to Z transition were briefly evaluated. Under the optimized conditions, we were able to baseline separate S and R enantiomers of both of the studied materials not only in their E forms but also in their Z forms. In comparison to the separation in the normal-phase mode, which we have reported recently, the resolution in the reversed-phase mode is significantly better. Interestingly, peak reversal was noticed for the S and R enantiomers when the separation was carried out with E versus Z forms of both compounds. This article is protected by copyright. All rights reserved.

  17. Simultaneous separation of hydrophobic and hydrophilic peptides with a silica hydride stationary phase using aqueous normal phase conditions.

    Science.gov (United States)

    Boysen, Reinhard I; Yang, Yuanzhong; Chowdhury, Jamil; Matyska, Maria T; Pesek, Joseph J; Hearn, Milton T W

    2011-11-04

    The application of a silica hydride modified stationary phase with low organic loading has been investigated as a new type of chromatographic material suitable for the separation and analysis of peptides with electrospray ionization mass spectrometric detection. Retention maps were established to delineate the chromatographic characteristics of a series of peptides with physical properties ranging from strongly hydrophobic to very hydrophilic and encompassing a broad range of pI values (pI 5.5-9.4). The effects of low concentrations of two additives (formic acid and acetic acid) in the mobile phase were also investigated with respect to their contribution to separation selectivity and retention under comparable conditions. Significantly, strong retention of both the hydrophobic and the hydrophilic peptides was observed when high-organic low-aqueous mobile phases were employed, thus providing a new avenue to achieve high resolution peptide separations. For example, simultaneous separation of hydrophobic and hydrophilic peptides was achieved under aqueous normal phase (ANP) chromatographic conditions with linear gradient elution procedures in a single run, whilst further gradient optimization enabled improved peak efficiencies of the more strongly retained hydrophobic and hydrophilic peptides.

  18. Poly(L-lactic acid)-modified silica stationary phase for reversed-phase and hydrophilic interaction liquid chromatography.

    Science.gov (United States)

    Ohyama, Kaname; Takasago, Shizuka; Kishikawa, Naoya; Kuroda, Naotaka

    2015-03-01

    Poly(L-lactic acid) is a linear aliphatic thermoplastic polyester that can be produced from renewable resources. A poly(L-lactic acid)-modified silica stationary phase was newly prepared by amide bond reaction between amino groups on aminopropyl silica and carboxylic acid groups at the end of the poly(L-lactic acid) chain. The poly(L-lactic acid)-silica column was characterized in reversed-phase liquid chromatography and hydrophilic interaction liquid chromatography with the use of different mobile phase compositions. The poly(L-lactic acid)-silica column was found to work in both modes, and the retention of test compounds depending on acetonitrile content exhibited "U-shaped" curves, which was an indicator of reversed-phase liquid chromatography/hydrophilic interaction liquid chromatography mixed-mode retention behavior. In addition, carbonyl groups included into the poly(L-lactic acid) backbone work as an electron-accepting group toward a polycyclic aromatic hydrocarbon and provide π-π interactions.

  19. Preparation of a novel carboxyl stationary phase by "thiol-ene" click chemistry for hydrophilic interaction chromatography.

    Science.gov (United States)

    Peng, Xi-Tian; Liu, Tao; Ji, Shu-Xian; Feng, Yu-Qi

    2013-08-01

    A novel carboxyl-bonded silica stationary phase was prepared by "thiol-ene" click chemistry. The resultant Thiol-Click-COOH phase was evaluated under hydrophilic interaction liquid chromatography (HILIC) mobile phase conditions. A comparison of the chromatographic performance of Thiol-Click-COOH and pure silica columns was performed according to the retention behaviors of analytes and the charged state of the stationary phases. The results indicated that the newly developed Thiol-Click-COOH column has a higher surface charge and stronger hydrophilicity than the pure silica column. Furthermore, the chromatographic behaviors of five nucleosides on the Thiol-Click-COOH phase were investigated in detail. Finally, a good separation of 13 nucleosides and bases, and four water-soluble vitamins was achieved.

  20. High-power coherent phased array of monolithic flared amplifier-master oscillator power amplifiers (MFA-MOPAs)

    Science.gov (United States)

    Osinski, Jules S.; Mehuys, David G.; Welch, David F.; Dzurko, Kenneth M.; Lang, Robert J.

    1995-04-01

    A monolithically integrated array of InGaAs/AlGaAs flared amplifiers driven by a single DBR laser through a power splitter network and individually addressed phase modulators is described. Phase adjustment of > 2(pi) per element by free-carrier effects is verified by monitoring the interference pattern of 4 emitters, and typically requires 5 W, and, combined with the proper external lensing, could therefore result in an ultra-narrow, single-lobed far-field pattern whose width is determined by the extended aperture of the array. This architecture is capable of providing single-mode, diffraction-limited performance from each emitter and is scalable to unprecedented power levels. Over 20 W of pulsed, spectrally coherent emission is generated at 955 nm from a 4-element array, and 39 W is obtained from an 8-element array.

  1. Nanocellulose 3, 5-Dimethylphenylcarbamate Derivative Coated Chiral Stationary Phase: Preparation and Enantioseparation Performance.

    Science.gov (United States)

    Zhang, Xiaoli; Wang, Litao; Dong, Shuqing; Zhang, Xia; Wu, Qi; Zhao, Liang; Shi, Yanping

    2016-05-01

    Nanocrystalline cellulose (NCC) with high surface area and high ordered crystalline structure was prepared from microcrystalline cellulose (MCC) under the hydrolysis of sodium hypochlorite. NCC was further reacted with 3,5-dimethylphenyl isocyanate to obtain the nanocellulose derivative, and then coated successfully on the surface of silica gel to a prepared NCC-coated chiral stationary phase (CSP) as a new kind of chiral separation material. Similarly, MCC derivative-coated CSP was also prepared as contrast. The chiral separation performance of NCC-based CSP was evaluated and compared with MCC-based CSP by high-performance liquid chromatography. Moreover, the effects of the alcohol modifiers, mobile phase additives, and flow rates on chiral separations were investigated in detail. The results showed that 10 chiral compounds were separated on NCC-based CSP with better peak shape and higher column efficiency than MCC-based CSP, which confirmed that NCC-based CSP was a promising packing material for the resolution of chiral compounds.Chirality 28:376-381, 2016. © 2016 Wiley Periodicals, Inc.

  2. Comparison of the performance of chiral stationary phase for separation of fluoxetine enantiomers

    Institute of Scientific and Technical Information of China (English)

    ZHOU Jie; YANG Yi-wen; WEI Feng; WU Ping-dong

    2007-01-01

    Separation of fluoxetine enantiomers on five chiral stationary phases (chiralcel OD-H, chiralcel OJ-H, chiralpak AD-H, cyclobond I 2000 DM and kromasil CHI-TBB) was investigated. The optimal mobile phase compositions of fluoxetine separation on each column were hexane/isopropanol/diethyl amine (98/2/0.2, v/v/v), hexane/isopropanol/diethyl amine (99/1/0.1,v/v/v), hexane/isopropanol/diethyl amine (98/2/0.2, v/v/v), methanol/0.2% triethylamine acetic acid (TEAA) (25/75, v/v; pH 3.8)and hexane/isopropanol/diethyl amine (98/2/0.2, v/v/v), respectively. Experimental results demonstrated that baseline separation (RS>1.5) of fluoxetine enantiomers was obtained on chiralcel OD-H, chiralpak AD-H, and cyclobond I 2000 DM while the best separation was obtained on the last one. The eluate orders of fluoxetine enantiomers on the columns were determined. The first eluate by chiralcel OJ-H and kromasil CHI-TBB is the S-enantiomer, while by chiralpak AD-H and cyclobond I 2000 DM is the R-enantiomer.

  3. Characterization and classification of stationary phases in HPLC and SFC – a review.

    Science.gov (United States)

    Galea, Charlene; Mangelings, Debby; Vander Heyden, Yvan

    2015-07-30

    Packed column supercritical fluid chromatography (pSFC) is an attractive technique in drug discovery related analysis because it offers several advantages over the more commonly used high-performance liquid chromatography (HPLC) technique. The environmental-friendly CO2 mobile phase, the high-throughput capacity, the increased efficiency and the lower operational costs give SFC additional benefits over HPLC in analysis related to drug development. The latter technique is well established and has been used for decades in the pharmaceutical industry. On the other hand, SFC is still in its infancy, even though the technique has been known for decades and researchers are still discovering the possibilities and limitations of this technique. Column characterization aims to obtain a quantitative understanding of the properties of a column that influence the selectivity of a separation. Columns have been extensively characterized in HPLC using several approaches. However, limited column characterization has been done in SFC. Determining column properties allows the rapid selection of appropriate columns for method development for a particular application. Hence, they also show which columns will provide similar or dissimilar separations. The aim of this review is to compare and contrast approaches used to characterize stationary phases in HPLC and SFC, and to highlight points that still may need to be researched further.

  4. Stationary-phase production of the antibiotic actinorhodin in Streptomyces coelicolor A3(2) is transcriptionally regulated

    NARCIS (Netherlands)

    Gramajo, Hugo C.; Takano, Eriko; Bibb, Mervyn J.

    1993-01-01

    Production of actinorhodin, a polyketide antibiotic made by Streptomyces coelicolor A3(2), normally occurs only in stationary-phase cultures. S1 nuclease protection experiments showed that transcription of actII-ORF4, the activator gene required for expression of the biosynthetic structural genes, i

  5. Lipidic ionic liquid stationary phases for the separation of aliphatic hydrocarbons by comprehensive two-dimensional gas chromatography.

    Science.gov (United States)

    Nan, He; Zhang, Cheng; O'Brien, Richard A; Benchea, Adela; Davis, James H; Anderson, Jared L

    2017-01-20

    Lipidic ionic liquids (ILs) possessing long alkyl chains as well as low melting points have the potential to provide unique selectivity as well as wide operating ranges when used as stationary phases in gas chromatography. In this study, a total of eleven lipidic ILs containing various structural features (i.e., double bonds, linear thioether chains, and cyclopropanyl groups) were examined as stationary phases in comprehensive two dimensional gas chromatography (GC×GC) for the separation of nonpolar analytes in kerosene. N-alkyl-N'-methyl-imidazolium-based ILs containing different alkyl side chains were used as model structures to investigate the effects of alkyl moieties with different structural features on the selectivities and operating temperature ranges of the IL-based stationary phases. Compared to a homologous series of ILs containing saturated side chains, lipidic ILs exhibit improved selectivity toward the aliphatic hydrocarbons in kerosene. The palmitoleyl IL provided the highest selectivity compared to all other lipidic ILs as well as the commercial SUPELCOWAX 10 column. The linoleyl IL containing two double bonds within the alkyl side chain showed the lowest chromatographic selectivity. The lipidic IL possessing a cyclopropanyl group within the alkyl moiety exhibited the highest thermal stability. The Abraham solvation parameter model was used to evaluate the solvation properties of the lipidic ILs. This study provides the first comprehensive examination into the relation between lipidic IL structure and the resulting solvation characteristics. Furthermore, these results establish a basis for applying lipidic ILs as stationary phases for solute specific separations in GC×GC.

  6. Synthesis and characterization of mesoporous silica modified with chiral auxiliaries for their potential application as chiral stationary phase.

    Science.gov (United States)

    Mayani, Vishal J; Abdi, S H R; Kureshy, R I; Khan, N H; Agrawal, Santosh; Jasra, R V

    2008-05-16

    Novel chiral stationary phase (CSP) based on chiral aminoalcohol immobilized on ordered mesoporous silica SBA-15 1a and standard silica 1b and their copper complexes 1a' and 1b', respectively, was synthesized as potential material for chiral ligand exchange chromatography (CLEC). Microanalysis, inductively coupled plasma spectroscopy (ICP), thermo-gravimetric analysis (TGA), cross polarized magic angle spinning (CP-MAS) (13)C NMR, Powder X-ray diffraction (PXRD), FTIR, N(2) adsorption isotherm, scanning electron microscopy (SEM), transmitted electron microscope (TEM) and solid reflectance UV-vis spectroscopy were used to characterize these materials. All the chiral stationary phases thus synthesized were used for the separation of different racemic compounds such as mandelic acid, 2,2'-dihydroxy-1,1'-binaphthalene BINOL) and diethyl tartrate by simple medium-pressure column chromatography. Successful enantio-separation of racemic mandelic acid was achieved with all the stationary phases but 1a and 1b gave slightly better resolution than their copper complexes 1a' and 1b'. Remarkably these materials are stable under the given experimental conditions and can be used repeatedly for several cycles of enantioresolution. It was observed that the porosity and surface area of the stationary phase play an important role in the chiral separation.

  7. Recombinant Staphylococcal protein A with cysteine residue for preparation of affinity chromatography stationary phase and immunosensor applications

    Directory of Open Access Journals (Sweden)

    Gorbatiuk O. B.

    2015-04-01

    Full Text Available Aim. Engineering of recombinant Staphylococcal protein A with cysteine residue (SPA-Cys for preparation of affinity chromatography stationary phase and formation of bioselective element of immunosensor. Methods. DNA sequences encoding IgG-binding region of SPA, His-tag and cysteine were genetically fused and expressed in E. coli. SPA-Cys was immobilized on maleimide-functionalized silica beads for affinity chromatography stationary phase preparation and on a gold sensor surface as a bioselective element of immunosensor. Results. SPA-Cys was expressed at a high-level in a soluble form. The target protein was purified and showed a high IgG-binding activity. The capacity of the obtained SPA-Cys-based affinity chromatography stationary phase was 10–12 mg of IgG /ml. The purity of eluted IgG was more than 95 % in one-step purification procedure. The developed SPA-Cys-based bioselective element of immunosensor selectively interacted with human IgG and did not interact with the control proteins. Conclusions. The recombinant Staphylococcal protein A with cysteine residue was successfully used for the preparation of affinity chromatography stationary phase and formation of the bioselective element of immunosensor.

  8. Effect of benzalkonium chloride on viability and energy metabolism in exponential- and stationary-growth-phase cells of Listeria monocytogenes

    NARCIS (Netherlands)

    Luppens, S.B.I.; Abee, T.; Oosterom, J.

    2001-01-01

    The difference in killing exponential- and stationary-phase cells of Listeria monocytogenes by benzalkonium chloride (BAC) was investigated by plate counting and linked to relevant bioenergetic parameters. At a low concentration of BAC (8 mg liter-1), a similar reduction in viable cell numbers was o

  9. Pre-stack time migration based on stationary-phase stacking in the dip-angle domain

    Science.gov (United States)

    Xu, Jincheng; Zhang, Hao; Zhang, Jianfeng; Li, Zhengwei; Liu, Wei

    2017-03-01

    The Kirchhoff-type migration approach often suffers from migration noise, aliasing artifacts due to operator error, or weak noise from a truncated aperture or pre-stacked data. These noises can be attenuated by using stationary-phase migration, which only stacks the reflection energy within the Fresnel zone rather than along the whole migration aperture, and therefore obtains a higher signal-to-noise ratio (SNR) for the migration results. This paper proposes a new implementation for the pre-stack time migration (PSTM) approach, which is based on stationary-phase stacking in the dip-angle domain. This implementation generates a pair of migrated dip-angle gathers in the image domain using PSTM. We can obtain the dip-angle field corresponding to the contribution of the Fresnel zones from the migrated dip-angle gathers, which allows us to remove noise outside the Fresnel zones and significantly improves the SNR of the image gathers. The proposed stationary-phase PSTM could effectively handle the problem of low SNR in migrated images, especially in the presence of steeply dipping structures. We test the method by applying stationary-phase PSTM to an overthrust model example and a three-dimensional field data set, and both examples demonstrate that the resulting images are of good quality with the method.

  10. Characterisation of reversed-phase liquid chromatography stationary phases for the analysis of basic pharmaceuticals : eluent properties and comparison of empirical test methods

    NARCIS (Netherlands)

    Vervoort, RJM; Ruyter, E; Debets, AJJ; Claessens, HA; Cramers, CA; de Jong, GJ

    2001-01-01

    The reversed-phase liquid chromatographic analysis of basic pharmaceuticals can be problematic. Both the properties of the eluent and the stationary phase can influence the chromatographic performance. Therefore selection of suitable experimental conditions for the analysis of basic compounds can be

  11. HPLC analysis of synthetic polymers on short monolithic columns.

    Science.gov (United States)

    Maksimova, Elena; Vlakh, Evgenia; Sinitsyna, Ekaterina; Tennikova, Tatiana

    2013-12-01

    Ultrashort monolithic columns (disks) were thoroughly studied as efficient stationary phases for precipitation-dissolution chromatography of synthetic polymers. Gradient elution mode was applied in all chromatographic runs. The mixtures of different flexible chain homopolymers, such as polystyrenes, poly(methyl methacrylates), and poly(tert-butylmethacrylates) were separated according to their molecular weights on both commercial poly(styrene-co-divinylbenzene) disks (12 id × 3 mm and 5 × 5 mm) and lab-made monolithic columns (4.6 id × 50 mm) filled with supports of different hydrophobicity. The experimental conditions were optimized to reach fast and highly efficient separation. It was observed that, similar to the separation of monoliths of other classes of (macro)molecules (proteins, DNA, oligonucleotides), the length of column did not affect the peak resolution. A comparison of the retention properties of the poly(styrene-co-divinylbenzene) disk-shaped monoliths with those based on poly(lauryl methacrylate-co-ethylene dimethacrylate), poly(butyl methacrylate-co-ethylene dimethacrylate), and poly(glycidyl methacrylate-co-ethylene dimethacrylate) supports demonstrated the obvious effect of surface chemistry on the resolution factor. Additionally, the results of the discussed chromatographic mode on the fast determination of the molecular weights of homopolymers used in this study were compared to those established by SEC on columns packed with sorbent beads of a similar nature to the monoliths. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Characterization of the properties of stationary phases for liquid chromatography in aqueous mobile phases using aromatic sulphonic acids as the test compounds.

    Science.gov (United States)

    Jandera, P; Bocian, S; Molíková, M; Buszewski, B

    2009-01-09

    We investigated the effects of the concentration of naphthalene sulphonic acids (NSAs) as anionic test compounds in the injected sample and of the salt additives to the mobile phase on ion-exclusion. The retention behaviour of NSAs sensitively reflects even minor changes in the ionic and hydrophobic interactions and can be useful for predicting the effects of the stationary phases in reversed-phase chromatography of polar and ionic compounds, both small ones and biopolymers, e.g., oligonucleotides. We studied chromatographic properties of several stationary phases intended for separations in aqueous mobile phases: a C18 column end-capped with polar hydrophilic groups, a densely bonded C8 column doubly end-capped with short alkyl groups, a short alkyl stationary phase designed to keep full pore accessibility in highly-aqueous mobile phases and a Bidentate column with "bridged" C18 groups attached to the silica hydride support. The chemistry and pore structure of various types of column packing materials and of the salt additives to the mobile phase affect the proportion of the pore volume non-accessible to anions due to ion-exclusion and consequently the peak asymmetry and hydrophobic selectivity in reversed-phase chromatography of organic acids. We also addressed the problems connected with the determination of column hold-up volume in aqueous mobile phases. The accessibility of the stationary phase for anionic compounds in contact with the sample zone is affected by ion-exclusion due to repulsive interactions with the negatively charged surface in the pores of the stationary phase. The accessible part of the stationary phase increases and consequently the migration velocity along the column decreases with increasing concentration of the sample in the zone moving along the column. Because of a limited access to the stationary phase, its capacity can be easily overloaded. The combination of the column overload and ion-exclusion effects may result in fronting or

  13. Mechanical properties examined by nanoindentation for selected phases relevant to the development of monolithic uranium-molybdenum metallic fuels

    Science.gov (United States)

    Newell, Ryan; Park, Youngjoo; Mehta, Abhishek; Keiser, Dennis; Sohn, Yongho

    2017-04-01

    Nanomechanical properties, specifically the reduced modulus and hardness of several intermetallic and solid solution phases are reported to assist the development of the U-10 wt% Mo (U-10Mo) monolithic fuel system for research and test reactors. Findings from this study and reported values of mechanical properties provide data critical for understanding and predicting the structural behavior of the fuel system during fabrication and irradiation. The phases examined are products of interdiffusion and reaction between (1) the AA6061 cladding and the Zr diffusion barrier, namely (Al,Si)3Zr and Al3Zr, (2) the U-10Mo fuel and the Zr diffusion barrier, namely UZr2, Mo2Zr, and α-U, and (3) the U (or U-10Mo) and Mo, namely a mixture gradient of α- and γ-phases. The UC inclusions observed within the fuel alloy were also examined. Only phases present in thick or continuous microstructure on cross-sectioned fuel plates and diffusion couples were investigated for reduced modulus and hardness. Concentration-dependence of room-temperature reduced modulus in U solid solution with 0-10 wt% Mo was semi-quantitatively modeled based on mixture of α- and γ-phases and solid solutioning within the γ-phase.

  14. Deconvoluting the effects of buffer salt concentration in hydrophilic interaction chromatography on a zwitterionic stationary phase.

    Science.gov (United States)

    West, Caroline; Auroux, Emeline

    2016-08-26

    Quantitative structure-retention relationships (QSRRs) furnish a detailed and reliable description of the role and extent of different molecular interactions that can be established between the analytes and the chromatographic system. Among QSRRs, the solvation parameter model using Abraham descriptors has gained acceptance as a general tool to explore the factors affecting retention in chromatographic systems. We have previously shown how a modified version of the solvation parameter model, with two extra terms to take account of interactions occurring with ionic and ionizable species (with positive and/or negative charges), could be applied to the characterization of hydrophilic interaction chromatographic (HILIC) systems. In the present study, we will show how this methodology can be used to evaluate the effects of increasing buffer salt concentration on retention and separation in a HILIC system. A commercial stationary phase possessing a sulfobetaine zwitterionic bonded ligand (Nucleodur HILIC) was used with a mobile phase composed of 80% acetonitrile and 20% pwwH4 ammonium acetate buffer, with aqueous buffer concentrations varying from 10 to 100mM, resulting in overall concentrations ranging from 2 to 20mM in the mobile phase. Retention factors were measured for a selection of 76 probe analytes. The chosen compounds are small molecules presenting a wide diversity of molecular structures and are relevant to biomedical and pharmaceutical applications. The QSRR models obtained allow for a rationalization of the interactions contributing to retention and separation in the HILIC system considered and shed some light on the effect of varying buffer salt concentration, namely the progressive transition from ion-exchange and electrostatic-repulsion mechanisms to hydrophilic partitioning.

  15. A new imidazolium-embedded C18 stationary phase with enhanced performance in reversed-phase liquid chromatography.

    Science.gov (United States)

    Qiu, Hongdeng; Mallik, Abul K; Takafuji, Makoto; Liu, Xia; Jiang, Shengxiang; Ihara, Hirotaka

    2012-08-13

    In this paper, a new imidazolium-embedded C(18) stationary phase (SiImC(18)) for reversed-phase high-performance liquid chromatography is described. 1-Allyl-3-octadecylimidazolium bromide ionic liquid compound having a long alkyl chain and reactive groups was newly prepared and grafted onto 3-mercaptopropyltrimethoxysilane-modified silica via a surface-initiated radical-chain transfer addition reaction. The SiImC(18) obtained was characterized by elemental analysis, infrared spectroscopy, thermogravimetric analysis, diffuse reflectance infrared Fourier transform, and solid-state (13)C and (29)Si cross-polarization/magic angle spinning nuclear magnetic resonance spectroscopy. The selectivity toward polycyclic aromatic hydrocarbons relative to that toward alkylbenzenes exhibited by SiImC(18) was higher than the corresponding selectivity exhibited by a conventional octadecyl silica (ODS) column, which could be explained by electrostatic π-π interaction cationic imidazolium and electron-rich aromatic rings. On the other hand, SiImC(18) also showed high selectivity for polar compounds, which was based on the multiple interaction and retention mechanisms of this phase with different analytes. 1,6-Dinitropyrene and 1,8-dinitropyrene, which form a positional isomer pair of dipolar compounds, were separated successfully with the SiImC(18) phase. Seven nucleosides and bases (i.e. cytidine, uracil, uridine, thymine, guanosine, xanthosine, and adenosine) were separated using only water as the mobile phase within 8min, which is difficult to achieve when using conventional hydrophobic columns such as ODS. The combination of electrostatic and hydrophobic interactions is important for the effective separation of such basic compounds without the use of any organic additive as the eluent in the octadecylimidazolium column.

  16. Separation of Immunoglobulin in Conjunction with High Performance Liquid Chromatography Using Poly(vinyl ester resin) Monolith

    Institute of Scientific and Technical Information of China (English)

    YANG,Guanqun; YANG,Gengliang; LIU,Haiyan; BAI,Ligai; FENG,Xiaojuan; YANG,Xinru

    2009-01-01

    Using vinyl ester resin both as the monomer and the crosslinker,2,2-azobis(2-methylpropionitrile) (AIBN) as initiator,poly(vinyl ester resin) monolithic column was prepared to separate the immunoglobulin G (lgG) and yolk of eggs (lgY) from human plasma and egg yolk by HPLC respectively.The influence of concentration and pH of mobile phase on the chromatographic performance was investigated.Scanning electron microcopy showed that the poly(vinyl ester resin) monolithic stationary phase had a through-pore structure.Dynamic capacity of IgG on the polymeric monolithic column was investigated.The monolithic column had good flow-through properties and high resolution.The maximum adsorptive amount of IgG was 144 μg·g-1.

  17. Stationary-point approach to the phase transition of the classical XY chain with power-law interactions.

    Science.gov (United States)

    Kastner, Michael

    2011-03-01

    The stationary points of the potential energy function V of the classical XY chain with power-law pair interactions (i.e., interactions decaying like r{-α} with the distance) are analyzed. For a class of "spinwave-type" stationary points, the asymptotic behavior of the Hessian determinant of V is computed analytically in the limit of large system size. The computation is based on the Toeplitz property of the Hessian and makes use of a Szegö-type theorem. The results serve to illustrate a recently discovered relation between phase transitions and the properties of stationary points of classical many-body potentials. In agreement with this relation, the exact phase transition potential energy of the model can be read off from the behavior of the Hessian determinant for exponents α between zero and one. For α between one and two, the phase transition is not manifest in the behavior of the determinant, and it might be necessary to consider larger classes of stationary points.

  18. Monolithic column incorporated with lanthanide metal-organic framework for capillary electrochromatography.

    Science.gov (United States)

    Zhang, Li-Shun; Du, Pei-Yao; Gu, Wen; Zhao, Qing-Li; Huang, Yan-Ping; Liu, Zhao-Sheng

    2016-08-26

    A new lanthanide metal-organic frameworks NKU-1 have successfully incorporated into poly (BMA-co-EDMA) monolith and evaluated by capillary electrochromatography (CEC). Lanthanide metal-organic frameworks [Eu2(ABTC)1.5(H2O)3(DMA)] (NKU-1) were synthesized by self-assembly of Eu(III) ions and 3,3',5,5'-azo benzene tetracarboxylic acid ligands have been fabricated into poly(BMA-co-EDMA) monoliths. 1-Butyl-3-methylimidazolium tetrafluoroborate and N,N-dimethylformamide were developed as binary porogen obtaining homogeneous dispersibility for NKU-1 and high permeability for monolithic column. The successful incorporation of NKU-1 into poly(BMA-co-EDMA) was confirmed and characterized by FT-IR spectra, scanning electron microscopy, X-ray diffraction, energy dispersive spectrometer area scanning, and transmission electron microscopy. Separation ability of the NKU-1-poly (BMA-co-EDMA) monoliths was demonstrated by separating four groups of analytes in CEC, including alkylbenzenes, polycyclic aromatic hydrocarbon, aniline series and naphthyl substitutes. Compared with bare monolithic (column efficiency of 100,000plates/m), the NKU-1-poly (BMA-co-EDMA) monoliths have displayed greater column efficiency (maximum 210,000plates/m) and higher permeability, as well as less peak tailing. The results showed that the NKU-1-poly (BMA-co-EDMA) monoliths are promising stationary phases for CEC separations.

  19. Purification of Astaxanthin from Laminariajaponica by Ionic Liquid-based Monolithic Cartridge

    Institute of Scientific and Technical Information of China (English)

    YOON Chang-hwan; ZHU Tao; ROW Kyung-ho

    2012-01-01

    An effective and accurate method was developed for the extraction of astaxanthin from Laminariajaponica with subsequent separation by ionic liquid-based monolithic cartridge.The optimized extraction conditions including extraction solvent(ethanol),extraction time(90 min) and ultrasonic power(75 W) were obtained by systematical investigation.Chromatographic analysis was performed on a C18 column with ultraviolet(UV) detection at 476nm,and a solution consisting of methanol/acetonitrile/H20/dichloromethane(83:6:6:5,volume ratio) was used as the mobile phase at a flow rate of 0.7 mL/min.After ionic liquid-based monolithic solid phase extraction,17.82 μg/g astaxanthin was obtained from Laminariajaponica.This ionic liquid-based monolithic cartridge exhibits high affinity and selectivity for astaxanthin,and it can be potentially used as the stationary phase of high performance liquid chromatography( HPLC).

  20. Postpolymerization modification of a hydroxy monolith precursor. Part I. Epoxy alkane and octadecyl isocyanate modified poly (hydroxyethyl methacrylate-co-pentaerythritol triacrylate) monolithic capillary columns for reversed-phase capillary electrochromatography.

    Science.gov (United States)

    Khadka, Shantipriya; El Rassi, Ziad

    2016-12-01

    A novel precursor monolithic capillary column referred to as "hydroxy monolith" or OHM was prepared by the in situ copolymerization of hydroxyethylmethacrylate (HEMA) with pentaerythritol triacrylate (PETA) yielding the neutral poly(HEMA-co-PETA) monolith. The neutral precursor OHM capillary thus obtained was subjected to postpolymerization modifications of the hydroxyl functional groups present on its surface with 1,2-epoxyalkanes catalyzed by boron trifluoride (BF3 ) ultimately providing Epoxy OHM C-m capillary column at varying alkyl chain lengths where m = 8, 12, 14, and 16 for RP-CEC. Also, the same precursor OHM was grafted with octadecyl isocyanate yielding Isocyanato OHM C-18 column to provide an insight into the effect of the nature of the linkage to the surface hydroxyl groups of the OHM precursor. While the epoxide reaction leaves on the surface of the OHM precursor hydroxy-ether linkages, the isocyanato reaction leaves carbamate linkages on the same surface of the OHM precursor. This study revealed that changing the alkyl chain length resulted in changing the column phase ratio (ϕ) and also the solute distribution constant (K). While increasing the surface alkyl chain length increased steeply the solute hydrophobic selectivity, i.e. methylene group selectivity, the nature of the ligand linkage produced different retention for the same solutes and affected the selectivity of slightly polar solutes. The various monoliths proved very useful for RP-CEC of different small solutes at varying polarity over a wide range of mobile phase composition. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Stationary phase modulation in liquid chromatography through the serial coupling of columns: A review.

    Science.gov (United States)

    Alvarez-Segura, T; Torres-Lapasió, J R; Ortiz-Bolsico, C; García-Alvarez-Coque, M C

    2016-06-01

    Liquid chromatography with single columns often does not succeed in the analysis of complex samples, in terms of resolution and analysis time. A relatively simple solution to enhance chromatographic resolution is the modulation of the stationary phase through the serial coupling of columns. This can be implemented with any type of column using compatible elution conditions and conventional instruments. This review describes the key features of column coupling and published procedures, where two or more columns were coupled in series to solve separation problems. In all reports, the authors could not resolve their samples with single columns, whereas significant enhancement in chromatographic performance was obtained when the columns were combined. Particularly interesting is the reduction in the analysis time in the isocratic mode, which alleviates the "general elution problem" of liquid chromatography, and may represent a stimulus for the proposal of new procedures, especially in combination with mass spectrometric, electrochemical and refractometric detection. Developments proposed to make the serial coupling of columns useful in routine and research laboratories are outlined, including optimisation strategies that facilitate the selection of the appropriate column combination and elution conditions (solvent content, flow rate or temperature) in both isocratic and gradient modes. The availability of zero dead volume couplers, able to connect standard columns, and the commercialisation of short columns with multiple lengths, have expanded the possibilities of success.

  2. Site-Specific Ser/Thr/Tyr Phosphoproteome of Sinorhizobium meliloti at Stationary Phase.

    Science.gov (United States)

    Liu, Tao; Tian, Chang Fu; Chen, Wen Xin

    2015-01-01

    Sinorhizobium meliloti, a facultative microsymbiont of alfalfa, should fine-tune its cellular processes to live saprophytically in soils characterized with limited nutrients and diverse stresses. In this study, TiO2 enrichment and LC-MS/MS were used to uncover the site-specific Ser/Thr/Tyr phosphoproteome of S. meliloti in minimum medium at stationary phase. There are a total of 96 unique phosphorylated sites, with a Ser/Thr/Tyr distribution of 63:28:5, in 77 proteins. Phosphoproteins identified in S. meliloti showed a wide distribution pattern regarding to functional categories, such as replication, transcription, translation, posttranslational modification, transport and metabolism of amino acids, carbohydrate, inorganic ion, succinoglycan etc. Ser/Thr/Tyr phosphosites identified within the conserved motif in proteins of key cellular function indicate a crucial role of phosphorylation in modulating cellular physiology. Moreover, phosphorylation in proteins involved in processes related to rhizobial adaptation was also discussed, such as those identified in SMa0114 and PhaP2 (polyhydroxybutyrate synthesis), ActR (pH stress and microaerobic adaption), SupA (potassium stress), chaperonin GroEL2 (viability and potentially symbiosis), and ExoP (succinoglycan synthesis and secretion). These Ser/Thr/Tyr phosphosites identified herein would be helpful for our further investigation and understanding of the role of phosphorylation in rhizobial physiology.

  3. Site-Specific Ser/Thr/Tyr Phosphoproteome of Sinorhizobium meliloti at Stationary Phase.

    Directory of Open Access Journals (Sweden)

    Tao Liu

    Full Text Available Sinorhizobium meliloti, a facultative microsymbiont of alfalfa, should fine-tune its cellular processes to live saprophytically in soils characterized with limited nutrients and diverse stresses. In this study, TiO2 enrichment and LC-MS/MS were used to uncover the site-specific Ser/Thr/Tyr phosphoproteome of S. meliloti in minimum medium at stationary phase. There are a total of 96 unique phosphorylated sites, with a Ser/Thr/Tyr distribution of 63:28:5, in 77 proteins. Phosphoproteins identified in S. meliloti showed a wide distribution pattern regarding to functional categories, such as replication, transcription, translation, posttranslational modification, transport and metabolism of amino acids, carbohydrate, inorganic ion, succinoglycan etc. Ser/Thr/Tyr phosphosites identified within the conserved motif in proteins of key cellular function indicate a crucial role of phosphorylation in modulating cellular physiology. Moreover, phosphorylation in proteins involved in processes related to rhizobial adaptation was also discussed, such as those identified in SMa0114 and PhaP2 (polyhydroxybutyrate synthesis, ActR (pH stress and microaerobic adaption, SupA (potassium stress, chaperonin GroEL2 (viability and potentially symbiosis, and ExoP (succinoglycan synthesis and secretion. These Ser/Thr/Tyr phosphosites identified herein would be helpful for our further investigation and understanding of the role of phosphorylation in rhizobial physiology.

  4. Solid-phase extraction based on ground methacrylate monolith modified with gold nanoparticles for isolation of proteins.

    Science.gov (United States)

    Vergara-Barberán, María; Lerma-García, María Jesús; Simó-Alfonso, Ernesto Francisco; Herrero-Martínez, José Manuel

    2016-04-21

    In this study, a novel polymeric material functionalized with gold nanoparticles (AuNPs) was prepared as solid-phase extraction (SPE) sorbent for isolation of proteins. The sorbent was synthesized from a powdered poly(glycidyl-co-ethylene dimethacrylate) monolith, and modified with ammonia, followed by immobilization of AuNPs on the pore surface of the material. To evaluate the performance of this SPE support, proteins were selected as test solutes, being the extraction conditions and other parameters (loading capacity and regenerative ability of sorbent) established. The results indicated that this sorbent could be employed to selectively capture proteins according to their pI, on the basis of the strong affinity of these biomacromolecules towards to AuNPs surface. The applicability of this sorbent was demonstrated by isolating protein species of interest (bovine serum albumin, cytochrome c and lectins in European mistletoe leaves), followed by SDS-PAGE analysis.

  5. Numerical study of the phase separation in binary lipid membrane containing protein inclusions under stationary shear flow.

    Science.gov (United States)

    Chen, Xiao-Bo; Shi, Hui-Ji; Niu, Li-Sha

    2009-03-26

    The phase separation of lipids is believed to be responsible for the formation of lipid rafts in biological cell membrane. In the present work, a continuum model and a particle model are constructed to study the phase separation in binary lipid membrane containing inclusions under stationary shear flow. In each model, employing the cell dynamical system (CDS) approach, the kinetic equations of the confusion-advection process are numerically solved. Snapshot figures of the phase morphology are performed to intuitively display such phase evolving process. Considering the effects from both the inclusions and the shear flow, the time growth law of the characteristic domain size is discussed.

  6. Graphene aerogel based monolith for effective solid-phase extraction of trace environmental pollutants from water samples.

    Science.gov (United States)

    Han, Qiang; Liang, Qionglin; Zhang, Xiaoqiong; Yang, Liu; Ding, Mingyu

    2016-05-20

    Graphene aerogel (GA), a typical kind of three-dimensional (3D) macroscopic assembly, not only provides inherently excellent properties of graphene sheets (GS), but also exhibits interesting characteristics of the 3D macroporous architecture including large and tunable pore volumes, high specific surface areas and fast mass transport kinetics. Thus, it is rational to expect GA to be an efficient adsorbent for solid-phase extraction (SPE). In this paper, a novel GA monolith based solid-phase extraction method was investigated in the application of environment analysis. The GA monolith based SPE cartridge was fabricated directly in the empty cartridge through template-free "sol-cryo" method. Due to the efficient mass transfer, more adsorption sites as well as effective retention for the analytes, the adsorption property of GA for bisphenol A revealed better performance than that of GS. What's more, GA also outperformed in loading and eluting for target analysis. On the basis of the above advantages, the obtained cartridge was applied for the separation of environmental pollutants from water samples. Taking endocrine disrupting chemicals and polychlorinated biphenyls as the polar and weak polar model analytes, optimizing several parameters influencing the recoveries, limits of detection in the range of 0.01-0.11ngmL(-1) and 0.19-1.53ngL(-1) for the two series of compounds were provided by the established methods. The satisfied sensitivity was accessed and recoveries ranging from 76.3 to 112.5% were obtained for all the analytes when the proposed methods were applied in real water samples analysis. The results revealed the potential of GA as an effective sorbent in sample preparation processes.

  7. [Separation mechanism of chiral stationary phase based on quinine and crown ether for the direct stereoselective separation of amino acids].

    Science.gov (United States)

    Wu, Haixia; Wang, Dongqiang; Zhao, Jianchao; Ke, Yanxiong; Liang, Xinmiao

    2016-01-01

    A novel chiral stationary phase combining quinine and crown ether (QN-CR CSP) was developed to separate amino acid enantiomers. This CSP showed good enantioselectivity for some amino acids. Since the synergistic effect of ion exchange and complexation in chiral recognition of amino acids, a new adsorption isotherm was built. Using the method of frontal analysis by characteristic point (FACP), the adsorption isotherms of tryptophan (Trp) under different mobile phase conditions were determined and fitted the proposed adsorption isotherm model well. With the increase of the competition between metal cationic and amino to crown ether, the equilibrium constant of complexing adsorption was found increased. The chiral separation ability was decreased. The adsorption isotherm improved the understanding of the retention behavior of amino acids on QN-CR CSP, which was also benefit to optimize the structure of the stationary phase.

  8. A micro gas chromatography with separation capability enhanced by polydimethylsiloxane stationary phase functionalized by carbon nanotubes and graphene.

    Science.gov (United States)

    Li, Yubo; Zhang, Runzhou; Wang, Tao; Wang, Youhao; Wang, Yonghuan; Li, Lingfeng; Zhao, Weijun; Wang, Xiaozhi; Luo, Jikui

    2016-07-01

    Polydimethylsiloxane (PDMS) stationary phases functionalized with multi-walled carbon nanotubes (MWCNTs) and graphene, respectively, for the columns in micro gas chromatography are presented in this paper. To exploit the merits of MWCNTs and graphene in terms of their high specific surface area, low surface energy and chemical inertness, experimental conditions for separation (heating rate and final temperature of temperature programming, flow rate of carrier gas and the volume of samples injection) are investigated, and separations of both polar and nonpolar compound mixtures under these conditions are performed. Compared with PDMS-only coated stationary phases, the functionalization of the phases with carbon nano-materials improves the performance of columns in separation, repeatability, stability and revolution significantly.

  9. Study on the Chromatographic Behavior of Bases on Dodecylamine-N,N-dimethylenephosphonic Acid-Modified Zirconia Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    Yao Lifeng; Feng Yuqi; Da Shilu

    2006-01-01

    A new zirconia-based stationary phase(DPZ)was prepared by modifying zirconia with dodecylamine-N,N-dimethylenephosphonic acid(DDPA).DDPA was adsorbed on zirconia with only one phosphonic group,with the other being free.Besides the hydrophobic interaction provided by nonpolar dodecyl,DPZ also has dipolar interaction,ion-exchange or electrostatic repellent interaction provided by the free phosphonic group and amino group at different conditions.Separation of bases on this stationary phase was achieved with satisfaction owing to the various retention mechanisms.The influence of methanol content,pH value,ion types,and ionic strength of mobile phase are studied in detail.

  10. Crosslinked structurally-tuned polymeric ionic liquids as stationary phases for the analysis of hydrocarbons in kerosene and diesel fuels by comprehensive two-dimensional gas chromatography.

    Science.gov (United States)

    Zhang, Cheng; Park, Rodney A; Anderson, Jared L

    2016-04-01

    Structurally-tuned ionic liquids (ILs) have been previously applied as the second dimension column in comprehensive two-dimensional gas chromatography (GC×GC) and have demonstrated high selectivity in the separation of individual aliphatic hydrocarbons from other aliphatic hydrocarbons. However, the maximum operating temperatures of these stationary phases limit the separation of analytes with high boiling points. In order to address this issue, a series of polymeric ionic liquid (PIL)-based stationary phases were prepared in this study using imidazolium-based IL monomers via in-column free radical polymerization. The IL monomers were functionalized with long alkyl chain substituents to provide the needed selectivity for the separation of aliphatic hydrocarbons. Columns were prepared with different film thicknesses to identify the best performing stationary phase for the separation of kerosene. The bis[(trifluoromethyl)sulfonyl]imide ([NTf2](-))-based PIL stationary phase with larger film thickness (0.28μm) exhibited higher selectivity for aliphatic hydrocarbons and showed a maximum allowable operating temperature of 300°C. PIL-based stationary phases containing varied amount of IL-based crosslinker were prepared to study the effect of the crosslinker on the selectivity and thermal stability of the resulting stationary phase. The optimal resolution of aliphatic hydrocarbons was achieved when 50% (w/w) of crosslinker was incorporated into the PIL-based stationary phase. The resulting stationary phase exhibited good selectivity for different groups of aliphatic hydrocarbons even after being conditioned at 325°C. Finally, the crosslinked PIL-based stationary phase was compared with SUPELCOWAX 10 and DB-17 columns for the separation of aliphatic hydrocarbons in diesel fuel. Better resolution of aliphatic hydrocarbons was obtained when employing the crosslinked PIL-based stationary phase as the second dimension column.

  11. Antioxidant defence system during exponential and stationary growth phases of Phycomyces blakesleeanus: response to oxidative stress by hydrogen peroxide.

    Science.gov (United States)

    de Castro, Cristina; del Valle, Pilar; Rúa, Javier; García-Armesto, María Rosario; Gutiérrez-Larraínzar, Marta; Busto, Félix; de Arriaga, Dolores

    2013-04-01

    An analysis of the components of the antioxidant defence system in exponential and stationary growth phases of filamentous fungus Phycomyces blakesleeanus and the response to the oxidative stress hydrogen peroxide were performed. There is a strong positive correlation between mycelial antioxidant capacity and the contents of gallic acid, d-erythroascorbate (d-EAA) or d-erythroascorbate monoglucoside (d-EAAG). These secondary metabolites are specifically synthesized by this fungus and reach maximal values in the stationary growth phase, suggesting that they can play some role in the antioxidant defence system of this fungus. There is a differential expression of the two more notable antioxidant activities, catalase (CAT) and superoxide dismutase (SOD), depending of the growth stage of P. blakesleeanus, CAT being expressed in the exponential and SOD in the stationary phase. Phycomyces blakesleeanus showed a high resistance to the oxidative stress caused by H2O2 (50 and 200 mM) which was higher in exponential phase. This higher resistance can be explained by the presence of CAT, glutathione peroxidase (GPx), and the probable contribution of glutathione-S-transferase (GST) and high levels of reduced form of glutathione (GSH). The transition to stationary phase was accompanied with a higher physiological oxidative damage illustrated by the higher protein carbonylation. In this growth stage the resistance of the fungus to the oxidative stress caused by H2O2 could be explained by the presence of SOD, GPx, and the probable contribution of GST as well as of secondary metabolites, mainly d-EAA and d-EAAG. These results highlight a specific response to oxidative stress by H2O2 depending on the growth phase of P. blakesleeanus.

  12. [Preparation and evaluation of 2,6-di-O-pentyl-beta-cyclodextrin bonded silica stationary phase for high performance liquid chromatography].

    Science.gov (United States)

    Liu, Liwen; Luo, Aiqin; Dai, Rongji; Ge, Xiaoxia; Yang, Shaoning

    2004-11-01

    In order to improve the chiral separation capability of the conventional beta-cyclodextrin bonded-silica gel stationary phase, 2,6-di-O-pentyl-beta-cyclodextrin bonded stationary phase (PCDS) was prepared via a long spacer. The resulted bonded-silica stationary phase was characterized by three methods, namely Fourier transform infrared, Molisch color reaction, X-ray optical electrical energy spectrogram. The chromatographic performances of PCDS were investigated by using liquid chromatography with toluene, dimethyl phthalate, and phenanthrene as solutes, and their retention mechanism was investigated and discussed. The results show that the introduction of pentyl to beta-cyclodextrin leads to enhancement of the retention of the solutes. The chiral separation capability of the new bonded-silica stationary phase was evaluated by using liquid chromatography with some chiral drugs. Some of the enantiomers such as chlorphenamine maleate and bupropion hydrochloride were separated by heptakis (2,6-di-O-pentyl)-beta-cyclodextrin bonded silica stationary.

  13. Silica gel microspheres decorated with covalent triazine-based frameworks as an improved stationary phase for high performance liquid chromatography.

    Science.gov (United States)

    Zhao, Wenjie; Hu, Kai; Hu, Chenchen; Wang, Xiaoyu; Yu, Ajuan; Zhang, Shusheng

    2017-03-03

    A new stationary phase for high performance liquid chromatography (HPLC) applications based on silica gel microspheres decorated with covalent triazine-based frameworks (CTF-SiO2) composite has been reported here. In this new hybrid material, sheet-like covalent triazine-based frameworks (CTF) were grown onto the supporting silica spheres, in order to achieve improved chromatographic separation and selectivity. The new material was characterized by infrared spectroscopy, elemental analysis and thermogravimetric analysis. The chromatographic performance and retention mechanism of the new stationary phase were investigated in reversed-phase mode and compared against standard C18 and cyano-modified silica (CN-SiO2) columns. A variety of different probe molecules were analyzed, including mono-substituted benzenes, polycyclic aromatic hydrocarbons, phenols, anilines and bases. The synergism of triazine and aromatic moieties provided several different retention mechanisms, thus improving the selectivity in the CTF-SiO2 composite. The good column packing properties of the uniform silica microspheres combined with the separation ability of the CTF frameworks make the new CTF-SiO2 material a potentially useful stationary phase for the analysis of complex samples.

  14. Immunoassay of paralytic shellfish toxins by moving magnetic particles in a stationary liquid-phase lab-on-a-chip.

    Science.gov (United States)

    Kim, Myoung-Ho; Choi, Suk-Jung

    2015-04-15

    In this study, we devised a stationary liquid-phase lab-on-a-chip (SLP LOC), which was operated by moving solid-phase magnetic particles in the stationary liquid phase. The SLP LOC consisted of a sample chamber to which a sample and reactants were added, a detection chamber containing enzyme substrate solution, and a narrow channel connecting the two chambers and filled with buffer. As a model system, competitive immunoassays of saxitoxin (STX), a paralytic shellfish toxin, were conducted in the SLP LOC using protein G-coupled magnetic particles (G-MPs) as the solid phase. Anti-STX antibodies, STX-horseradish peroxidase conjugate, G-MPs, and a STX sample were added to the sample chamber and reacted by shaking. While liquids were in the stationary state, G-MPs were transported from the sample chamber to the detection chamber by moving a magnet below the LOC. After incubation to allow the enzymatic reaction to occur, the absorbance of the detection chamber solution was found to be reciprocally related to the STX concentration of the sample. Thus, the SLP LOC may represent a novel, simple format for point-of-care testing applications of enzyme-linked immunosorbent assays by eliminating complicated liquid handling steps.

  15. Nuclear magnetic resonance and high-performance liquid chromatographic evaluation of polymer-based stationary phases immobilized on silica.

    Science.gov (United States)

    Meyer, Christoph; Skogsberg, Urban; Welsch, Norbert; Albert, Klaus

    2005-06-01

    Three poly(ethylene-co-acrylic) acid copolymers (-CH(2)CH(2)-)(x)[CH(2)CH(CO(2)H)-](y) with different chain lengths and mass fractions of acrylic acid were covalently immobilized as stationary phases on silica via two variants of spacer molecules (3-aminopropyltriethoxysilane and 3-glycidoxypropyltrimethoxysilane). Different mobilities of the alkyl chains in the stationary phases were observed using (13)C solid-state NMR spectroscopy. The stationary phases with more rigid trans-ordered alkyl chains had better selectivity for geometric beta-carotene and xanthophyll isomers (provitamin A derivatives). Also, all the separations of the analytes were affected by polar interactions with the chromatographic sorbent. This was further proved by separating more polar cis/trans retinoic acid isomers (vitamin A derivatives). (13)C high-resolution/magic-angle spinning (HR/MAS) NMR measurements of the chromatographic sorbents suspended in the mobile phase confirmed a dependence of molecular shape recognition ability on alkyl chain conformation.

  16. Regenerated silica gel as stationary phase on vacuum column chromatography to purify temulawak’s extracts

    Energy Technology Data Exchange (ETDEWEB)

    Cahyono, Bambang; Maduwu, Ratna Dewi; Widayat,; Suzery, Meiny [Organic Chemistry Laboratory, Departement of Chemistry, Diponegoro University Jln Prof. Soedharto SH, Tembalang, Semarang 50275, Indonesia Tel / Fax: (024) 7460058 (Indonesia)

    2015-12-29

    Commercial silica gel only used once by many researchers and affected high cost for purification process, also less support the green chemistry program. This research focused in regeneration silica gel that used purification of temulawak’s extracts (Curcuma xanthorrhiza Roxb) by vacuum column chromatography. Sample extracts (contains 10.1195±0.5971% of curcuminoids) was purified by vacuum column chromatography (pressure: 45 kPa, column: 100mm on length and 16mm on diameter). Ethanol 96% and acetone were compared as eluent. The amount of solvent and yield of curcuminoids used as indicator purification. The silica gel was regenerated with heating in 600°C for 8 hours The silica gels were analyzed by IR spectroscopy and X-ray diffraction. Furthermore, regenerated silica gel was used as the stationary phase in vacuum column chromatography under the same conditions with the previous purification. All the purification experiments were performed in three repetitions. Based on regression equation, y=0.132x+0.0011 (r{sup 2}=0.9997) the yield of curcuminoids on purified products using ethanol as the eluent was improved 4.26% (to 14.3724±0.5749%) and by acetone was improved 3,03% (to 13.1450 ±0.6318%). The IR spectrum of both silica gel showed the same vibration profile and also there were three crystallinity peaks missing on its X-ray diffraction. Regenerated silica gel has the same performance with new silica gel in purification of temulawak’s extract: by ethanol has increased 4.08% (14.1947±0.7415%) and 2.93% (13.0447±0.4822) by acetone. In addition, all purification products showed similar TLC profiles. Purification using regenerated silica gel as the adsorbent on vacuum column chromatography has exactly same potential with the new silica gel.

  17. Chiral stationary phases based on chitosan bis(methylphenylcarbamate)-(isobutyrylamide) for high-performance liquid chromatography.

    Science.gov (United States)

    Tang, Sheng; Bin, Qin; Chen, Wei; Bai, Zheng-Wu; Huang, Shao-Hua

    2016-04-01

    A series of chitosan bis(methylphenylcarbamate)-(isobutyrylamide) derivatives were synthesized by carbamylating chitosan isobutyrylamide with different methylphenyl isocyanates. Then the prepared chitosan derivatives were coated onto 3-aminopropyl silica particles, resulting in a series of new chiral stationary phases (CSPs) for high-performance liquid chromatography. It was observed that the chiral recognition abilities of these coated-type CSPs depended very much on the substituents on the phenyl moieties of the chitosan derivatives, the eluent composition, as well as the structure of racemates. As a typical example, the eluent tolerance of the prepared CSP with the best enantioseparation ability was investigated in detail, and the results revealed that the CSP exhibited extraordinary solvent tolerance and could still work without significant loss in enantioseparation capability after being flushed with chloroform (100%), ethyl acetate (100%) and even THF/n-hexane (70/30, v/v), while the traditional coated-type CSPs based on the cellulose and amylose derivatives, such as cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) and amylose tris(3,5-dimethylphenylcarbamate) (ADMPC), might be dissolved or highly swollen in these eluents. Therefore, the application of the resultant CSPs could address the problem of the dissolution and high swelling of traditional coated-type CSPs in some unusual eluents, broadening the possibility of eluent choice. In addition, a comparison of the prepared CSPs with the well known CDMPC- and ADMPC- based CSPs concerning the chiral recognition ability was also made. Separation performances achieved on the as-prepared CSPs in different eluents were found to be even superior to CDMPC- and ADMPC-based CSPs for the tested chiral compounds. In summary, we could safely draw the conclusion that the CSPs derived from chitosan isobutyrylamide derivatives were capable of excellent chiral recognition ability, and meanwhile possessed satisfactory

  18. Enantiomeric separation of iridium (III) complexes using HPLC chiral stationary phases based on amylose derivatives

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hee Eun; Seo, Na Hyeon; Hyun, Myung Ho [Dept. of Chemistry and Chemistry Institute for Functional Materials, Pusan National University, Busan (Korea, Republic of)

    2016-12-15

    Cyclometalated iridium (III) complexes formed with three identical cyclometalating (C-N) ligands (homoleptic) or formed with two cyclometalating (C-N) ligands and one ancillary (LX) ligand (heteroleptic) have been known as highly phosphorescent materials and, thus, they have been utilized as efficient phosphorescent dopants in organic light emitting diodes (OLEDs) 1–3 or as effective phosphorescent chemosensors. 4–7 Cylometalated iridium (III) complexes are chiral compounds consisting of lambda (Λ, left-handed) and delta (Δ, right-handed) isomers. Racemic cyclometa- lated iridium (III) complexes emit light with no net polarization, but optically active cyclometalated iridium (III) complexes emit circularly polarized light. 8,9 Circularly polarized light can be used in various fields including highly efficient three dimensional electronic devices, photo nic devices for optical data storage, biological assays, and others. 8,9 In order to obtain optically active cylometalated iridium (III) complexes and to determine the enantiomeric composition of optically active cylometalated iridium (III) complexes, liquid chromatogr aphic enantiomer separation method on chiral stationary phases (CSPs) has been used. For example, Okamoto and coworkers first reported the high performance liquid chromatographic (HPLC) direct enantiomeric separation of two homoleptic cylometalated iridium (III) complexes on immobilized amylose tris(3,5- dimethylphenylcarbamate) (Chiralpak IA), coated cellulose tris(3,5-dimethylphenylcarbamate) (Chiralc el OD), and coated cellulose tris(4-methylbenzoate) (Chiralce l OJ). 10 Supercritical fluid chromatography (SFC) was also used by Bernhard and coworkers for the enantiomeric separation of cylometalated iridium (III) complexes on coated amylose tris(3,5-dimethylphenylcarbamate) (Chiralpak AD-H). 8 However, the general use of the HPLC method for the direct enantiomeric separation of homoleptic.

  19. Dynamic Behavior of Clobazam on High-Performance Liquid Chromatography Chiral Stationary Phases.

    Science.gov (United States)

    Sabia, Rocchina; De Martino, Michela; Cavazzini, Alberto; Villani, Claudio

    2016-01-01

    Clobazam, a 1,5-benzodiazepin-2,4-dione, is a chiral molecule because its ground state conformation features a nonplanar seven-membered ring lacking reflection symmetry elements. The two conformational enantiomers of clobazam interconvert at room temperature by a simple ring-flipping process. Variable temperature HPLC on the Pirkle type (R)-N-(3,5-dinitronenzoyl)phenylglycine and (R,R)-Whelk-O1 chiral stationary phases (CSPs) allowed us to separate for the first time the conformational enantiomers of clobazam and to observe peak coalescence-decoalescence phenomena due to concomitant separation and interconversion processes occurring on the same time scale. Clobazam showed temperature dependent dynamic high-performance liquid chromatography (HPLC) profiles with interconversion plateaus on the two CSPs indicative of on-column enantiomer interconversion. (enantiomerization) in the column temperature range between Tcol = 10°C and Tcol = 30°C, whereas on-column interconversion was absent at temperature close to or lower than Tcol = 5°C. Computer simulation of exchange-deformed HPLC profiles using a program based on the stochastic model yielded the apparent rate constants for the on-column enantiomerization and the corresponding free energy activation barriers. At Tcol = 20°C the averaged enantiomerization barriers, ΔG(‡), for clobazam were found in the range 21.08-21.53 kcal mol(-1) on the two CSPs. The experimental dynamic chromatograms and the corresponding interconversion barriers reported in this article are consistent with the literature data measured by DNMR at higher temperatures and in different solvents.

  20. Regenerated silica gel as stationary phase on vacuum column chromatography to purify temulawak's extracts

    Science.gov (United States)

    Cahyono, Bambang; Maduwu, Ratna Dewi; Widayat, Suzery, Meiny

    2015-12-01

    Commercial silica gel only used once by many researchers and affected high cost for purification process, also less support the green chemistry program. This research focused in regeneration silica gel that used purification of temulawak's extracts (Curcuma xanthorrhiza Roxb) by vacuum column chromatography. Sample extracts (contains 10.1195±0.5971% of curcuminoids) was purified by vacuum column chromatography (pressure: 45 kPa, column: 100mm on length and 16mm on diameter). Ethanol 96% and acetone were compared as eluent. The amount of solvent and yield of curcuminoids used as indicator purification. The silica gel was regenerated with heating in 600°C for 8 hours The silica gels were analyzed by IR spectroscopy and X-ray diffraction. Furthermore, regenerated silica gel was used as the stationary phase in vacuum column chromatography under the same conditions with the previous purification. All the purification experiments were performed in three repetitions. Based on regression equation, y=0.132x+0.0011 (r2=0.9997) the yield of curcuminoids on purified products using ethanol as the eluent was improved 4.26% (to 14.3724±0.5749%) and by acetone was improved 3,03% (to 13.1450 ±0.6318%). The IR spectrum of both silica gel showed the same vibration profile and also there were three crystallinity peaks missing on its X-ray diffraction. Regenerated silica gel has the same performance with new silica gel in purification of temulawak's extract: by ethanol has increased 4.08% (14.1947±0.7415%) and 2.93% (13.0447±0.4822) by acetone. In addition, all purification products showed similar TLC profiles. Purification using regenerated silica gel as the adsorbent on vacuum column chromatography has exactly same potential with the new silica gel.

  1. A mechanistic model of ion-exchange chromatography on polymer fiber stationary phases.

    Science.gov (United States)

    Winderl, Johannes; Hahn, Tobias; Hubbuch, Jürgen

    2016-12-02

    Fibers are prominent among novel stationary phase supports for preparative chromatography. Several recent studies have highlighted the potential of fiber-based adsorbents for high productivity downstream processing in both batch and continuous mode, but so far the development of these materials and of processes employing these materials has solely been based on experimental data. In this study we assessed whether mechanistic modeling can be performed on fiber-based adsorbents. With a column randomly filled with short cut hydrogel grafted anion exchange fibers, we tested whether tracer, linear gradient elution, and breakthrough data could be reproduced by mechanistic models. Successful modeling was achieved for all of the considered experiments, for both non-retained and retained molecules. For the fibers used in this study the best results were obtained with a transport-dispersive model in combination with a steric mass action isotherm. This approach accurately accounted for the convection and dispersion of non-retained tracers, and the breakthrough and elution behaviors of three different proteins with sizes ranging from 6 to 160kDa were accurately modeled, with simulation results closely resembling the experimental data. The estimated model parameters were plausible both from their physical meaning, and from an analysis of the underlying model assumptions. Parameters were determined within good confidence levels; the average confidence estimate was below 7% for confidence levels of 95%. This shows that fiber-based adsorbents can be modeled mechanistically, which will be valuable for the future design and evaluation of these novel materials and for the development of processes employing such materials.

  2. Numerical simulation of the phase separation in binary lipid membrane under the effect of stationary shear flow.

    Science.gov (United States)

    Chen, Xiao-Bo; Niu, Li-Sha; Shi, Hui-Ji

    2008-06-01

    A numerical simulation of the phase separation in binary lipid membrane under the effect of stationary shear flow is performed. We numerically solved the modified two-dimensional time-dependent Ginzburg-Landau (TDGL) equations with an external velocity term, employing the CDS (i.e., Cell Dynamical System) technique. In the present simulation, stationary shear flows with different shear rates are taken into account. The evolution process of the phase separation is illustrated macroscopically via the snapshot figures and simulated scattering patterns at several typical moments. For each case, the growth exponents of the characteristic domain sizes in both directions parallel and perpendicular to the flow are studied, and the domain area as well. Also, the behavior of the excess viscosity has been investigated, which is a peculiar rheological indicator of such a membrane system with domain structures.

  3. Preparation of a pipette tip-based molecularly imprinted solid-phase microextraction monolith by epitope approach and its application for determination of enkephalins in human cerebrospinal fluid.

    Science.gov (United States)

    Li, Hua; Li, Dan

    2015-11-10

    In this study, a novel molecularly imprinted polymer (MIP) monolith for highly selective extraction of enkephalins was synthesized and prepared in a micropipette tip using epitope imprinting technique. The synthesized MIPs were characterized by scanning electron microscope (SEM) and infrared spectroscopy. A molecularly imprinted solid-phase microextraction (MISPME) method was developed for extraction of enkephalins in aqueous solutions. The parameters affecting MISPME were optimized. The results indicated that this MIP monolith exhibited specific recognition capability, high enrichment efficiency and excellent reusability for enkephalins. MALDI-TOF MS analysis demonstrated that this MIP monolith can act as a useful tool for highly selective purification and enrichment of enkephalin, a kind of low abundance protein, from high-abundance proteins in human cerebrospinal fluids (CSF). Employed this MIP monolith as solid-phase microextraction column, quantitative assay of enkephalins in human CSF was developed by HPLC-ultraviolet (UV) detection in this work. The detection limits were 0.05-0.08nM. This MISPME/HPLC-UV method was used to quantify Met-enkephalin and Leu-enkephalin levels in the CSF of patients with cancer pain.

  4. Evaluation of a medium-polarity ionic liquid stationary phase in the analysis of flavor and fragrance compounds.

    Science.gov (United States)

    Ragonese, Carla; Sciarrone, Danilo; Tranchida, Peter Quinto; Dugo, Paola; Dugo, Giovanni; Mondello, Luigi

    2011-10-15

    The present research is focused on the evaluation, in terms of efficiency and polarity, of a recently introduced gas chromatography (GC) column, coated with a 1,12-di(tripropylphosphonium) dodecane bis(trifluoromethansulfonyl) amide ionic-liquid stationary phase (SLB-IL59) and its application to the analysis of a complex essential oil. The ionic liquid column demonstrated very good efficiency, in terms of plate number, and a polarity close to that of the 100% poly(ethyleneglycol) stationary phase. In this preliminary evaluation, the SLB-IL59 30 m column was subjected to bleeding measurements, by means of conventional gas chromatography/mass spectrometry (GC/MS) and, in addition, of comprehensive 2D GC. The SLB-IL59 column (30 m × 0.20 μm d(f), 0.25 mm i.d.) was then evaluated in the analysis of typical essential oil constituents, in the form of pure standard compounds. Resolution toward several analytes was measured and the results were compared to those obtained with both apolar [silphenylene polymer, equivalent to poly(5% diphenyl/95% dimethylsiloxane)] and medium-polarity [100% poly(ethyleneglycol)] stationary phases, namely, the most common columns employed in the analysis of essential oils; peak symmetry, for different essential oil constituents, was also measured and expressed through tailing factors (at 10% of peak height). The final part of the investigation was devoted to the GC/MS analysis of lemon essential oil, with GC-flame ionization detection (FID) used for quantification. Linear retention indices of all the identified compounds were determined, and the data obtained were compared to those attained on the apolar and "wax" columns. The results obtained in the present investigation reveal the great potential of this novel stationary phase, as a medium-polarity alternative, in the analysis of essential oils.

  5. The extracellular proteome of Rhizobium etli CE3 in exponential and stationary growth phase

    OpenAIRE

    Mendoza-Hernández Guillermo; Encarnación Sergio; Meneses Niurka

    2010-01-01

    Abstract Background The extracellular proteome or secretome of symbiotic bacteria like Rhizobium etli is presumed to be a key element of their infection strategy and survival. Rhizobia infect the roots of leguminous plants and establish a mutually beneficial symbiosis. To find out the possible role of secreted proteins we analyzed the extracellular proteome of R. etli CE3 in the exponential and stationary growth phases in minimal medium, supplemented with succinate-ammonium. Results The extra...

  6. Schizosaccharomyces pombe Homologs of Human DJ-1 Are Stationary Phase-Associated Proteins That Are Involved in Autophagy and Oxidative Stress Resistance.

    Directory of Open Access Journals (Sweden)

    Yang Su

    Full Text Available The Parkinson's disease protein DJ-1 is involved in various cellular functions including detoxification of dicarbonyl compounds, autophagy and oxidative stress response. DJ-1 homologs are widely found in both prokaryotes and eukaryotes, constituting a superfamily of proteins that appear to be involved in stress response. Schizosaccharomyces pombe contains six DJ-1 homologs, designated Hsp3101-Hsp3105 and Sdj1 (previously named SpDJ-1. Here we show that deletion of any one of these six genes somehow affects autophagy during prolonged stationary phase. Furthermore, deletions of each of these DJ-1 homologs result in reduced stationary phase survival. Deletion of sdj1 also increases the sensitivity of stationary-phase cells to oxidative stress induced by hydrogen peroxide (H2O2 whereas overexpression of sdj1 has the opposite effect. Consistent with their role in stationary phase, expression of hsp3101, hsp3102, hsp3105 and sdj1, and to a lesser extent hsp3103 and hsp3104, is increased in stationary phase. The induction of hsp3101, hsp3102, hsp3105 and sdj1 involves the Sty1-regulated transcription factor Atf1 but not the transcription factor Pap1. Our results firmly establish that S. pombe homologs of DJ-1 are stationary-phase associated proteins and are likely involved in autophagy and antioxidant defense in stationary phase of S. pombe cells.

  7. Development of a novel amide-silica stationary phase for the reversed-phase HPLC separation of different classes of phytohormones.

    Science.gov (United States)

    Aral, Hayriye; Aral, Tarık; Ziyadanoğulları, Berrin; Ziyadanoğulları, Recep

    2013-11-15

    A novel amide-bonded silica stationary phase was prepared starting from N-Boc-phenylalanine, cyclohexylamine and spherical silica gel (4 µm, 60 Å). The amide ligand was synthesised with high yield. The resulting amide bonded stationary phase was characterised by SEM, IR and elemental analysis. The resulting selector bearing a polar amide group is used for the reversed-phase chromatography separation of different classes of thirteen phytohormones (plant hormones). The chromatographic behaviours of these analytes on the amide-silica stationary phase were compared with those of RP-C18 column under same conditions. The effects of different separation conditions, such as mobile phase, pH value, flow rate and temperature, on the separation and retention behaviours of the 13 phytohormones in this system were studied. The optimum separation was achieved using reversed-phase HPLC gradient elution with an aqueous mobile phase containing pH=6.85 potassium phosphate buffer (20 mM) and acetonitrile with a 22 °C column temperature. Under these experimental conditions, the 12 phytohormones could be separated and detected at 230 or 270 nm within 26 min.

  8. 6S RNA regulation of relA alters ppGpp levels in early stationary phase.

    Science.gov (United States)

    Cavanagh, Amy T; Chandrangsu, Pete; Wassarman, Karen M

    2010-12-01

    6S RNA is a small, non-coding RNA that interacts directly with σ(70)-RNA polymerase and regulates transcription at many σ(70)-dependent promoters. Here, we demonstrate that 6S RNA regulates transcription of relA, which encodes a ppGpp synthase. The 6S RNA-dependent regulation of relA expression results in increased ppGpp levels during early stationary phase in cells lacking 6S RNA. These changes in ppGpp levels, although modest, are sufficient to result in altered regulation of transcription from σ(70)-dependent promoters sensitive to ppGpp, including those promoting expression of genes involved in amino acid biosynthesis and rRNA. These data place 6S RNA as another player in maintaining appropriate gene expression as cells transition into stationary phase. Independent of this ppGpp-mediated 6S RNA-dependent regulation, we also demonstrate that in later stationary phase, 6S RNA continues to downregulate transcription in general, and specifically at a subset of the amino acid promoters, but through a mechanism that is independent of ppGpp and which we hypothesize is through direct regulation. In addition, 6S RNA-dependent regulation of σ(S) activity is not mediated through observed changes in ppGpp levels. We suggest a role for 6S RNA in modulating transcription of several global regulators directly, including relA, to downregulate expression of key pathways in response to changing environmental conditions.

  9. Preparation of a hierarchically porous AlPO4 monolith via an epoxide-mediated sol–gel process accompanied by phase separation

    Directory of Open Access Journals (Sweden)

    Wenyan Li, Yang Zhu, Xingzhong Guo, Kazuki Nakanishi, Kazuyoshi Kanamori and Hui Yang

    2013-01-01

    Full Text Available Monolithic aluminum phosphate (AlPO4 with a macro–mesoporous structure has been successfully prepared via the sol–gel process accompanied by phase separation in the presence of poly(ethylene oxide (PEO. Gelation of the system has been mediated by propylene oxide (PO, while PEO induces a phase separation. The dried gel is amorphous, whereas the crystalline tridymite phase precipitates upon heating above 1000 °C. Heat treatment does not spoil the macroporous morphology of the AlPO4 monoliths. Nitrogen adsorption–desorption measurements revealed that the skeletons of the dried gels possess a mesostructure with a median pore size of about 30 nm and a surface area as high as 120 m2 g−1. Hydrothermal treatment before heat treatment can increase the surface area to 282 m2 g−1.

  10. Preparation of a hierarchically porous AlPO4 monolith via an epoxide-mediated sol-gel process accompanied by phase separation

    Science.gov (United States)

    Li, Wenyan; Zhu, Yang; Guo, Xingzhong; Nakanishi, Kazuki; Kanamori, Kazuyoshi; Yang, Hui

    2013-08-01

    Monolithic aluminum phosphate (AlPO4) with a macro-mesoporous structure has been successfully prepared via the sol-gel process accompanied by phase separation in the presence of poly(ethylene oxide) (PEO). Gelation of the system has been mediated by propylene oxide (PO), while PEO induces a phase separation. The dried gel is amorphous, whereas the crystalline tridymite phase precipitates upon heating above 1000 °C. Heat treatment does not spoil the macroporous morphology of the AlPO4 monoliths. Nitrogen adsorption-desorption measurements revealed that the skeletons of the dried gels possess a mesostructure with a median pore size of about 30 nm and a surface area as high as 120 m2 g-1. Hydrothermal treatment before heat treatment can increase the surface area to 282 m2 g-1.

  11. Thermodynamics of the sorption of organic compounds on polyethylene glycol 400-permethylated β-cyclodextrin stationary phase and its enantioselectivity in gas chromatography

    Science.gov (United States)

    Kuraeva, Yu. G.; Onuchak, L. A.; Evdokimova, M. A.

    2016-08-01

    The thermodynamic characteristics of sorption of 24 organic compounds of various classes from the gas phase on the binary stationary phase based on polyethylene glycol 400 and permethylated β-cyclodextrin were determined. The influence of geometrical structure and optical activity of organic compounds on the possibility of forming sorbate-macrocycle complexes was examined. It was found that the studied stationary phase shows the enantioselectivity towards low-polar terpenes under the conditions of gas chromatography.

  12. Monolithic spectrometer

    Science.gov (United States)

    Rajic, Slobodan; Egert, Charles M.; Kahl, William K.; Snyder, Jr., William B.; Evans, III, Boyd M.; Marlar, Troy A.; Cunningham, Joseph P.

    1998-01-01

    A monolithic spectrometer is disclosed for use in spectroscopy. The spectrometer is a single body of translucent material with positioned surfaces for the transmission, reflection and spectral analysis of light rays.

  13. Preparation, characterization and application of a reversed phase liquid chromatography/hydrophilic interaction chromatography mixed-mode C18-DTT stationary phase.

    Science.gov (United States)

    Wang, Qing; Long, Yao; Yao, Lin; Xu, Li; Shi, Zhi-Guo; Xu, Lanying

    2016-01-01

    A mixed-mode chromatographic stationary phase, C18-DTT (dithiothreitol) silica (SiO2) was prepared through "thiol-ene" click chemistry. The obtained material was characterized by fourier transform infrared spectroscope, nitrogen adsorption analysis and contact angle analysis. Chromatographic performance of the C18-DTT was systemically evaluated by studying the effect of acetonitrile content, pH, buffer concentration of the mobile phase and column temperature. It was demonstrated that the novel stationary phase possessed reversed phase liquid chromatography (RPLC)/hydrophilic interaction liquid chromatography (HILIC) mixed-mode property. The stop-flow test revealed that C18-DTT exhibited excellent compatibility with 100% aqueous mobile phase. Additionally, the stability and column-to-column reproducibility of the C18-DTT material were satisfactory, with relative standard deviations of retention factor of the tested analytes (verapamil, fenbufen, guanine, tetrandrine and nicotinic acid) in the range of 1.82-3.72% and 0.85-1.93%, respectively. Finally, the application of C18-DTT column was demonstrated in the separation of non-steroidal anti-inflammatory drugs, aromatic carboxylic acids, alkaloids, nucleo-analytes and polycyclic aromatic hydrocarbons. It had great resolving power in the analysis of various compounds in HILIC and RPLC chromatographic conditions and was a promising RPLC/HILIC mixed-mode stationary phase.

  14. Multi-mode application of graphene quantum dots bonded silica stationary phase for high performance liquid chromatography.

    Science.gov (United States)

    Wu, Qi; Sun, Yaming; Zhang, Xiaoli; Zhang, Xia; Dong, Shuqing; Qiu, Hongdeng; Wang, Litao; Zhao, Liang

    2017-04-07

    Graphene quantum dots (GQDs), which possess hydrophobic, hydrophilic, π-π stacking and hydrogen bonding properties, have great prospect in HPLC. In this study, a novel GQDs bonded silica stationary phase was prepared and applied in multiple separation modes including normal phase, reversed phase and hydrophilic chromatography mode. Alkaloids, nucleosides and nucleobases were chosen as test compounds to evaluate the separation performance of this column in hydrophilic chromatographic mode. The tested polar compounds achieved baseline separation and the resolutions reached 2.32, 4.62, 7.79, 1.68 for thymidine, uridine, adenosine, cytidine and guanosine. This new column showed satisfactory chromatographic performance for anilines, phenols and polycyclic aromatic hydrocarbons in normal and reversed phase mode. Five anilines were completely separated within 10min under the condition of mobile phase containing only 10% methanol. The effect of water content, buffer concentration and pH on chromatographic separation was further investigated, founding that this new stationary phase showed a complex retention mechanism of partitioning, adsorption and electrostatic interaction in hydrophilic chromatography mode, and the multiple retention interactions such as π-π stacking and π-π electron-donor-acceptor interaction played an important role during the separation process. This GQDs bonded column, which allows us to adjust appropriate chromatography mode according to the properties of analytes, has possibility in actual application after further research.

  15. 信息动态%Preparation of Polysaccharide Derivatives-based Composite Chiral Stationary Phases and Their Chiral Recognition

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    Two coated-type composite chiral stationary phases (CSPs) were prepared based on cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) and amylose tris (3,5-dimethylphenylcarbamate)(ADMPC) by coating the corresponding derivatives onto 3-aminopropyl silica gel separately and then mixing or by coating the mixed derivatives onto silica gel. The CSPs containing only CDMPC or ADMPC were also prepared for comparison. The mixing method does not significantly influence the enantioselectivities. The composite CSPs generally show chiral recognition abilities intermediate between those of the two individual phases, while some racemates were poorer and at the same time one was better resolved on the composite CSPs.

  16. Preparation and Evaluation of a Novel Cellulose Tris(N-3,5-dimethylphenylcarbamate) Chiral Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    GE,Jin; ZHAO,Liang; SHI,Yan-Ping

    2008-01-01

    A novel cellulose tris(N-3,5-dimethylphenylcarbamate) (CDMPC) chiral stationary phase (CSP) was prepared by coating CDMPC on TiO2/SiO2, which was prepared by coating titania nanoparticles on silica through a self-assemble technique. At first, 2-hydroxyl-phenyl acetonitrile and α-phenylethanol were separated on this new CSP to evaluate the chiral separation ability. Then, two pesticides, matalaxyl and diclofop-methyl were separated.The influence of the mobile phase composition on the enantioselectivity was discussed, and the repeatability and stability of the CSP were studied too.

  17. Preparation and application of a novel molecularly imprinted solid-phase microextraction monolith for selective enrichment of cholecystokinin neuropeptides in human cerebrospinal fluid.

    Science.gov (United States)

    Ji, Xiang; Li, Dan; Li, Hua

    2015-08-01

    A novel molecularly imprinted polymer (MIP) monolith for highly selective extraction of cholecystokinin (CCK) neuropeptides was prepared in a micropipette tip. The MIPs were synthesized by epitope imprinting technique and the polymerization conditions were investigated and optimized. The synthesized MIPs were characterized by infrared spectroscopy, elemental analyzer and scanning electron microscope. A molecularly imprinted solid-phase microextraction (MI-μ-SPE) method was developed for the extraction of CCK neuropeptides in aqueous solutions. The parameters affecting MI-μ-SPE were optimized. The results indicated that this MIP monolith exhibited specific recognition capability and high enrichment efficiency for CCK neuropeptides. In addition, it showed excellent reusability. This MIP monolith was used for desalting and enrichment of CCK4, CCK5 and CCK8 from human cerebrospinal fluid prior to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis, and the results show that this MIP monolith can be a useful tool for effective purification and highly selective enrichment of multiple homologous CCK neuropeptides in cerebrospinal fluid simultaneously. By employing MI-μ-SPE combined with HPLC-ESI-MS/MS analysis, endogenous CCK4 in human cerebrospinal fluid was quantified.

  18. Porous silicon as a stationary phase for shear-driven chromatography

    NARCIS (Netherlands)

    Clicq, D.; Tjerkstra, R.W.; Gardeniers, J.G.E.; Berg, van den Albert; Baron, G.V.; Desmet, G.

    2004-01-01

    We report on the possibility to strongly increase the mass loadability and retention capacity of shear-driven chromatography (SDC) channels by growing a thin porous silicon layer on the stationary wall part. The thickness of the produced porous silicon layers was found to increase linearly with the

  19. High-internal-phase-emulsion polymeric monolith coupled with liquid chromatography-electrospray tandem mass spectrometry for enrichment and sensitive detection of trace cytokinins in plant samples.

    Science.gov (United States)

    Du, Fuyou; Sun, Lin; Zhen, Xian; Nie, Honggang; Zheng, Yanjie; Ruan, Guihua; Li, Jianping

    2015-08-01

    High-internal-phase-emulsion polymers (polyHIPEs) show great promise as solid-phase-extraction (SPE) materials because of the tremendous porosity and highly interconnected framework afforded by the high-internal-phase-emulsion (HIPE) technique. In this work, polyHIPE monolithic columns as novel SPE materials were prepared and applied to trace enrichment of cytokinins (CKs) from complex plant samples. The polyHIPE monoliths were synthesized via the in-situ polymerization of the continuous phase of a HIPE containing styrene (STY) and divinylbenzene (DVB) in a stainless column, and revealed highly efficient and selective enrichment ability for aromatic compounds. Under the optimized experimental conditions, a method using a monolithic polyHIPE column combined with liquid chromatography-electrospray tandem mass spectrometry (LC-MS-MS) was developed for the simultaneous extraction and sensitive determination of trans-zeatin (tZ), meta-topolin (mT), kinetin (K), and kinetin riboside (KR). The proposed method had good linearity, with correlation coefficients (R (2)) from 0.9957 to 0.9984, and low detection limits (LODs, S/N = 3) in the range 2.4-47 pg mL(-1) for the four CKs. The method was successfully applied to the determination of CKs in real plant samples, and obtained good recoveries ranging from 68.8 % to 103.0 % and relative standard deviations (RSDs) lower than 16 %.

  20. Monolithic microwave integrated circuits

    Science.gov (United States)

    Pucel, R. A.

    Monolithic microwave integrated circuits (MMICs), a new microwave technology which is expected to exert a profound influence on microwave circuit designs for future military systems as well as for the commercial and consumer markets, is discussed. The book contains an historical discussion followed by a comprehensive review presenting the current status in the field. The general topics of the volume are: design considerations, materials and processing considerations, monolithic circuit applications, and CAD, measurement, and packaging techniques. All phases of MMIC technology are covered, from design to testing.

  1. Lipophilicity measurement of drugs by reversed phase HPLC over Wide pH range using an alkaline-resistant silica-based stationary phase, XBridge Shield RP(18).

    Science.gov (United States)

    Liu, Xiangli; Hefesha, Hossam; Tanaka, Hideji; Scriba, Gerhard; Fahr, Alfred

    2008-10-01

    We propose a reversed phase HPLC (RP-HPLC) with an alkaline-resistant silica-based stationary phase, XBridge Shield RP(18), for the determination of the lipophilicity of drugs with diverse chemical nature ranging from acidic to basic. A set of 40 model compounds with well-defined solvatochromic parameters was selected to allow a broad distribution of structural properties. The chromatographic results showed that the lipophilicity index log k(w) obtained with XBridge Shield RP(18) was well correlated with experimental log P(oct) values (r(2)=0.96). Linear solvation free-energy relationship (LSER) analyses revealed that the retention mechanism of the stationary phase and 1-octanol/water partitioning were controlled by almost the same balance of intermolecular forces (hydrophobicity as expressed by the van der Waals volume V(w), H-bond acceptor basicity beta, and dipolarity/polarizability pi*). The results showed that XBridge Shield RP(18) phase overcomes the shortcomings of the silica-based stationary phases, the application of which to lipophilicity measurements had been limited to neutral and acidic compounds.

  2. Growth advantage in stationary-phase (GASP) phenotype in long-term survival strains of Geobacter sulfurreducens.

    Science.gov (United States)

    Helmus, Ruth A; Liermann, Laura J; Brantley, Susan L; Tien, Ming

    2012-01-01

    Geobacter sulfurreducens exists in the subsurface and has been identified in sites contaminated with radioactive metals, consistent with its ability to reduce metals under anaerobic conditions. The natural state of organisms in the environment is one that lacks access to high concentrations of nutrients, namely electron donors and terminal electron acceptors (TEAs). Most studies have investigated G. sulfurreducens under high-nutrient conditions or have enriched for it in environmental systems via acetate amendments. We replicated the starvation state through long-term batch culture of G. sulfurreducens, where both electron donor and TEA were scarce. The growth curve revealed lag, log, stationary, death, and survival phases using acetate as electron donor and either fumarate or iron(III) citrate as TEA. In survival phase, G. sulfurreducens persisted at a constant cell count for as long as 23 months without replenishment of growth medium. Geobacter sulfurreducens demonstrated an ability to acquire a growth advantage in stationary-phase phenotype (GASP), with strains derived from subpopulations from death- or survival phase being able to out-compete mid-log-phase populations when co-cultured. The molecular basis for GASP was not because of any detectable mutation in the rpoS gene (GSU1525) nor because of a mutation in a putative homolog to Escherichia coli lrp, GSU3370.

  3. High performance liquid chromatographic separation of eight drugs collected in Chinese Pharmacopoeia 2010 on amylose ramification chiral stationary phase

    Directory of Open Access Journals (Sweden)

    Yan Wang

    2012-10-01

    Full Text Available The enantiomers separation of eight pharmaceutical racemates collected in Chinese Pharmacopoeia 2010 (Ch.P2010, including nitrendipine, felodipine, omeprazole, praziquantel, sulpiride, clenbuterol hydrochloride, verapamil hydrochloride and chlorphenamine maleate, was performed on chiral stationary phase of amylose ramification by high performance liquid chromatography (HPLC on Chiralpak AD-H column and Chiralpak AS-H column with the mobile phase consisted of isopropanol and n-hexane. The detection wavelength and the flow rate were set at 254 nm and 0.7 mL/min, respectively. The effects of proportion of organic additives, alcohol displacer and temperature on the separation were investigated. The results indicated that eight chiral drugs were separated on chiral stationary phase of amylase ramification in normal phase chromatographic system. The chromatographic retention and resolution of enantiomers were adjusted by factors, including the changes of the concentration of alcohol displacer in mobile phase, organic alkaline modifier and column temperature. It was shown that the resolution was improved with reducing concentration of alcohol displacer. When the concentration of organic alkaline modifier was 0.2%, the resolution and the peak shape were fairly good. Most racemates mentioned above had the best resolution at column temperature of 25 °C. The best temperature should be kept unchanged in the process of separation so as to obtain stable separation results.

  4. Enantioselective and diastereoselective separation of synthetic pyrethroid insecticides on a novel chiral stationary phase by high-performance liquid chromatography.

    Science.gov (United States)

    Tan, Xulin; Hou, Shicong; Wang, Min

    2007-07-01

    A novel chiral packing material for high-performance liquid chromatography (HPLC) was prepared by connecting (R)-1-phenyl-2-(4-methylphenyl) ethylamine (PTE) amide derivative of (S)-isoleucine to aminopropyl silica gel through 2-amino-3,5-dinitro-1-carboxamido-benzene unit. This chiral stationary phase was applied to the enantioselective and diastereoselective separation of five pyrethroid insecticides by HPLC under normal phase condition. To achieve satisfactory baseline separation an optimization of the variables of mobile phase composition was required. The two enantiomers of fenpropathrin and four stereoisomers of fenvalerate were baseline separated using hexane-1,2-dichloroethane-2-propanol as mobile phase. The results show that the enantioselectivity of CSP is better than Pirkle type 1-A column for these compounds. Only partial separations for the cypermethrin and cyfluthrin stereoisomers were observed. Seven peaks and eight peaks were observed for cypermethrin and cyfluthrin, respectively. The elution orders were assigned by using different stereoisomer-enriched products.

  5. Prediction of the zeta potentials and ionic descriptors of a silica hydride stationary phase with mobile phases of different pH and ionic strength.

    Science.gov (United States)

    Kulsing, Chadin; Yang, Yuanzhong; Matyska, Maria T; Pesek, Joseph J; Boysen, Reinhard I; Hearn, Milton T W

    2015-02-15

    In this study, the zeta potentials of a silica hydride stationary phase (Diamond Hydride™) in the presence of different water-acetonitrile mixtures (from 0-80% (v/v) acetonitrile) of different ionic strengths (from 0-40mM) and pH values (from pH 3.0-7.0) have been investigated. Debye-Hückel theory was applied to explain the effect of changes in the pH and ionic strength of these aqueous media on the negative zeta potential of this stationary phase. The experimental zeta potentials of the Diamond Hydride™ particles as a function of acetonitrile content up to 50% (v/v) correlated (R(2)=0.998) with the predicted zeta potential values based on this established theory, when the values of the dissociation constant of all related species, as well as viscosity, dielectric constant and refractive index of the aqueous medium were taken into consideration. Further, the retention behavior of basic, acidic and neutral analytes was investigated under mobile phase conditions of higher pH and lower ionic strength. Under these conditions, the Diamond Hydride™ stationary phase surface became more negative, as assessed from the increasingly more negative zeta potentials, resulting in the ion exchange characteristics becoming more dominant and the basic analytes showing increasing retention. Ionic descriptors were derived from these chromatographic experiments based on the assumption that linear solvation energy relationships prevail. The results were compared with predicted ionic descriptors based on the different calculated zeta potential values resulting in an overall correlation of R(2)=0.888. These studies provide fundamental insights into the impact on the separation performance of changes in the zeta potential of the Diamond Hydride™ surface with the results relevant to other silica hydride and, potentially, to other types of stationary phase materials.

  6. Preparation and chromatographic evaluation of a newly designed steviol glycoside modified-silica stationary phase in hydrophilic interaction liquid chromatography and reversed phase liquid chromatography.

    Science.gov (United States)

    Liang, Tu; Fu, Qing; Shen, Aijin; Wang, Hui; Jin, Yu; Xin, Huaxia; Ke, Yanxiong; Guo, Zhimou; Liang, Xinmiao

    2015-04-03

    A diterpene glycoside compound, rebaudioside A (commonly abbreviated as RA), was immobilized onto porous silica surface through "thiol-ene" click chemistry strategy. The successful immobilization of the RA on the silica support was confirmed by FT-IR and elemental analysis. Chromatographic characteristics of the new stationary phase, named Click TE-RA, were evaluated by a set of diverse analytes such as carbohydrates, nucleosides, and organic acids in hydrophilic interaction liquid chromatography (HILIC) mode. The effects of water content, buffer pH and concentration were investigated and a typical HILIC retention feature of Click TE-RA was observed at high organic modifier content. The Click TE-RA stationary phase was further studied by a series of glycoside compounds. Tunable retention mechanisms from hydrophilic to hydrophobic interactions were observed. Separation of very polar compounds including oligosaccharides, nucleic acid bases and nucleosides using Click TE-RA in HILIC mode was successfully accomplished. In addition, separation of saponins both in HILIC and reversed-phase liquid chromatography (RPLC) modes was performed, demonstrating the presence of orthogonality between two different modes on Click TE-RA column. The multiple interactions induced by polar sugar group and hydrophobic aglycone group allowed this Click TE-RA to serve as a multi-mode stationary phase in two-dimensional liquid chromatography.

  7. Preparation and evaluation of surface-bonded tricationic ionic liquid silica as stationary phases for high-performance liquid chromatography.

    Science.gov (United States)

    Qiao, Lizhen; Shi, Xianzhe; Lu, Xin; Xu, Guowang

    2015-05-29

    Two tricationic ionic liquids were prepared and then bonded onto the surface of supporting silica materials through "thiol-ene" click chemistry as new stationary phases for high-performance liquid chromatography. The obtained columns of tricationic ionic liquids were evaluated respectively in the reversed-phase liquid chromatography (RPLC) mode and hydrophilic interaction liquid chromatography (HILIC) mode, and possess ideal column efficiency of 80,000 plates/m in the RPLC mode with naphthalene as the test solute. The tricationic ionic liquid stationary phases exhibit good hydrophobic and shape selectivity to hydrophobic compounds, and RPLC retention behavior with multiple interactions. In the HILIC mode, the retention and selectivity were evaluated through the efficient separation of nucleosides and bases as well as flavonoids, and the typical HILIC retention behavior was demonstrated by investigating retention changes of hydrophilic solutes with water volume fraction in mobile phase. The results show that the tricationic ionic liquid columns possess great prospect for applications in analysis of hydrophobic and hydrophilic samples.

  8. A humic acid stationary phase for the high performance liquid chromatography separation of buckminsterfullerenes: Theoretical and practical aspects

    Energy Technology Data Exchange (ETDEWEB)

    Casadei, Nicolas [Equipe des Sciences Separatives et Biopharmaceutiques (2SB)-EA 3924, Laboratoire de Chimie Analytique, Faculte de Medecine Pharmacie, Place Saint Jacques, 25030 Besancon Cedex (France); Thomassin, Mireille [Equipe des Sciences Separatives et Biopharmaceutiques (2SB)-EA 3924, Laboratoire de Chimie Analytique, Faculte de Medecine Pharmacie, Place Saint Jacques, 25030 Besancon Cedex (France); Guillaume, Yves-Claude [Equipe des Sciences Separatives et Biopharmaceutiques (2SB)-EA 3924, Laboratoire de Chimie Analytique, Faculte de Medecine Pharmacie, Place Saint Jacques, 25030 Besancon Cedex (France)]. E-mail: yves.guillaume@univ-fcomte.fr; Andre, Claire [Equipe des Sciences Separatives et Biopharmaceutiques (2SB)-EA 3924, Laboratoire de Chimie Analytique, Faculte de Medecine Pharmacie, Place Saint Jacques, 25030 Besancon Cedex (France)

    2007-04-11

    The influence of the mobile phase composition and column temperature on the chromatographic separation of five buckminsterfullerenes (C60, C70, C76, C78, C84) on a stationary phase based on silica gel with chemically bonded humic acid (Bonded humic acid column (BHAC)) was studied. The retention behavior of the fullerenes was measured under isocratic conditions with different mobile phase compositions, ranging from 0.05-0.70 (v/v) of toluene in cyclohexane. The column temperature was analysed in the range 35-75 deg. C. The retention factors of the five fullerenes do not depend linearly on the toluene fraction but follow a quadratic relationship. The best chromatographic conditions for baseline separation of the five fullerenes were selected. The retention of the fullerenes on the HA stationary phase was strongly affected by temperature. Positive values of thermodynamic parameters (changes of enthalpy and entropy) were due to the abnormal solubility behaviour of fullerenes in toluene in the temperature range 35-75 deg. C. The information obtained in this work makes this BHAC very simple to prepare and low cost, useful for fullerene research applications.

  9. On-chip optical phase locking of single growth monolithically integrated Slotted Fabry Perot lasers.

    Science.gov (United States)

    Morrissey, P E; Cotter, W; Goulding, D; Kelleher, B; Osborne, S; Yang, H; O'Callaghan, J; Roycroft, B; Corbett, B; Peters, F H

    2013-07-15

    This work investigates the optical phase locking performance of Slotted Fabry Perot (SFP) lasers and develops an integrated variable phase locked system on chip for the first time to our knowledge using these lasers. Stable phase locking is demonstrated between two SFP lasers coupled on chip via a variable gain waveguide section. The two lasers are biased differently, one just above the threshold current of the device with the other at three times this value. The coupling between the lasers can be controlled using the variable gain section which can act as a variable optical attenuator or amplifier depending on bias. Using this, the width of the stable phase locking region on chip is shown to be variable.

  10. Evaluation of innovative stationary phase ligand chemistries and analytical conditions for the analysis of basic drugs by supercritical fluid chromatography.

    Science.gov (United States)

    Desfontaine, Vincent; Veuthey, Jean-Luc; Guillarme, Davy

    2016-03-18

    Similar to reversed phase liquid chromatography, basic compounds can be highly challenging to analyze by supercritical fluid chromatography (SFC), as they tend to exhibit poor peak shape, especially those with high pKa values. In this study, three new stationary phase ligand chemistries available in sub -2 μm particle sizes, namely 2-picolylamine (2-PIC), 1-aminoanthracene (1-AA) and diethylamine (DEA), were tested in SFC conditions for the analysis of basic drugs. Due to the basic properties of these ligands, it is expected that the repulsive forces may improve peak shape of basic substances, similarly to the widely used 2-ethypyridine (2-EP) phase. However, among the 38 tested basic drugs, less of 10% displayed Gaussian peaks (asymmetry between 0.8 and 1.4) using pure CO2/methanol on these phases. The addition of 10mM ammonium formate as mobile phase additive, drastically improved peak shapes and increased this proportion to 67% on 2-PIC. Introducing the additive in the injection solvent rather than in the organic modifier, gave acceptable results for 2-PIC only, with 31% of Gaussian peaks with an average asymmetry of 1.89 for the 38 selected basic drugs. These columns were also compared to hybrid silica (BEH), DIOL and 2-EP stationary phases, commonly employed in SFC. These phases commonly exhibit alternative retention and selectivity. In the end, the two most interesting ligands used as complementary columns were 2-PIC and BEH, as they provided suitable peak shapes for the basic drugs and almost orthogonal selectivities.

  11. Synthesis of novel glucose-based polymers and their applications as chiral stationary phases for high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    Tomoyuki IKAI; Takayuki YAMADA

    2016-01-01

    Two novel polymers containing glucose units as the main-chain that only differ in terms of their regioregularity were synthesized to evaluate their chiral recognition abilities as chiral stationary phases( CSPs) for high performance liquid chromatography( HPLC). The regioregular polymer( poly-5)shows clear resolution ability for the racemate of cobalt(Ⅲ)acetylacetonate( Co( acac)3 ),whereas the corresponding regioirregular polymer(poly-3)does not show any chiral recognition for Co(acac)3. The regioregular polymer main-chain seems to play an important role not only in providing an efficient interaction with the racemate but also in expressing the chiral recognition ability as a CSP for HPLC.

  12. 6S RNA regulation of relA alters ppGpp levels in early stationary phase

    OpenAIRE

    Cavanagh, Amy T.; Chandrangsu, Pete; Wassarman, Karen M.

    2010-01-01

    6S RNA is a small, non-coding RNA that interacts directly with ? 70-RNA polymerase and regulates transcription at many ? 70-dependent promoters. Here, we demonstrate that 6S RNA regulates transcription of relA, which encodes a ppGpp synthase. The 6S RNA-dependent regulation of relA expression results in increased ppGpp levels during early stationary phase in cells lacking 6S RNA. These changes in ppGpp levels, although modest, are sufficient to result in altered regulation of transcription fr...

  13. Synthesis and chromatographic evaluation of new polymeric chiral stationary phases based on three (1S,2S)-(-)-1,2-diphenylethylenediamine derivatives in HPLC and SFC.

    Science.gov (United States)

    Payagala, Tharanga; Wanigasekara, Eranda; Armstrong, Daniel W

    2011-03-01

    Three new polymeric chiral stationary phases were synthesized based on (1S,2S)-1,2-bis(2,4,6-trimethylphenyl)ethylenediamine, (1S,2S)-1,2-bis(2-chlorophenyl)ethylenediamine, and (1S,2S)-1,2-di-1-naphthylethylenediamine via a simple free-radical-initiated polymerization in solution. These monomers are structurally related to (1S,2S)-1,2-diphenylethylenediamine which is the chiral monomer used for the commercial P-CAP-DP polymeric chiral stationary phase (CSP). The performance of these three new chiral stationary phases were evaluated in normal phase high-performance liquid chromatography (HPLC) and supercritical fluid chromatography and the results were compared with those of the P-CAP-DP column. All three new phases showed enantioselectivity for a large number of racemates with a variety of functional groups, including amines, amides, alcohols, amino acids, esters, imines, thiols, and sulfoxides. In normal phase, 68 compounds were separated with 28 baseline separations (Rs ≥ 1.5) and in SFC, 65 compounds were separated with 24 baseline separations. In total 72 out of 100 racemates were separated by these CSPs with 37 baseline separations. Complimentary separation capabilities were observed for many analytes. The new polymeric CSPs showed similar or better enantioselectivities compared with the commercial column in both HPLC and SFC. However, faster separations were achieved on the new stationary phases. Also, it was shown that these polymeric stationary phases have good sample loading capacities while maintaining enantioselectivity.

  14. A Monolithic Radiation-Pressure Driven, Low Phase Noise Silicon Nitride Opto-Mechanical Oscillator

    CERN Document Server

    Tallur, Siddharth; Bhave, Sunil A

    2011-01-01

    Cavity opto-mechanics enabled radiation pressure (RP) driven oscillators shown in the past offer an all optical Radio Frequency (RF) source without the need for external electrical feedback. However these oscillators require external tapered fiber or prism coupling and non-standard fabrication processes. In this work, we present a CMOS compatible fabrication process to design high optical quality factor opto-mechanical resonators in silicon nitride. The ring resonators designed in this process demonstrate low phase noise RP driven oscillations. Using integrated grating couplers and waveguide to couple light to the micro-resonator eliminates 1/f^3 and other higher order phase noise slopes at close-to-carrier frequencies present in previous demonstrations. We present an RP driven OMO operating at 41.97MHz with a signal power of -11dBm and phase noise of -85dBc/Hz at 1kHz offset with only 1/f^2 noise down to 10Hz offset from carrier.

  15. Chromatographic retention behaviour, modelling and separation optimisation of the quaternary ammonium salt isometamidium chloride and related compounds on a range of reversed-phase liquid chromatographic stationary phases.

    Science.gov (United States)

    Schad, Gesa J; Euerby, Melvin R; Skellern, Graham G; Tettey, Justice N A

    2012-07-01

    This paper describes the reversed-phase liquid chromatographic behaviour of the trypanocidal quaternary ammonium salt isometamidium chloride and its related compounds on a range of liquid chromatographic phases possessing alkyl and phenyl ligands on the same inert silica. In a parallel study with various extended polar selectivity phases which possessed different hydrophobic/silanophilic (hydrogen bonding) activity ratios, the chromatographic retention/selectivities of the quaternary ammonium salts was shown to be due to a co-operative mechanism between hydrophobic and silanophilic interactions. The highly aromatic and planar isometamidium compounds were found to be substantially retained on stationary phases containing aromatic functionality via strong π-π interactions. The chemometric approach of principal component analysis was used to characterise the chromatographic behaviour of the isometamidium compounds on the differing phases and to help identify the dominant retention mechanism(s). Two-dimensional (temperature/gradient) retention modelling was employed to develop and optimise a rapid liquid chromatography method for the separation of the six quaternary ammonium salts within 2.5 min which would be suitable for bioanalysis using liquid chromatography-mass spectrometry. This is the first reported systematic study of the relationship between stationary phase chemistries and retention/selectivity for a group of quaternary ammonium salts.

  16. Separation performance of cucurbit[7]uril in ionic liquid-based sol-gel coating as stationary phase for capillary gas chromatography.

    Science.gov (United States)

    Wang, Xiaogang; Qi, Meiling; Fu, Ruonong

    2014-12-05

    Here we report the separation performance of a new stationary phase of cucurbit[7]uril (CB7) incorporated into an ionic liquid-based sol-gel coating (CB7-SG) for capillary gas chromatography (GC). The CB7-SG stationary phase showed an average polarity of 455, suggesting its polar nature. Abraham system constants revealed that its major interactions with analytes include H-bond basicity (a), dipole-dipole (s) and dispersive (l) interactions. The CB7-SG stationary phase achieved baseline separation for a wide range of analytes with symmetrical peak shapes and showed advantages over the conventional polar stationary phase that failed to resolve some critical analytes. Also, it exhibited different retention behaviors from the conventional stationary phase in terms of retention times and elution order. Most interestingly, in contrast to the conventional polar phase, the CB7-SG stationary phase exhibited longer retentions for analytes of lower polarity but relatively comparable retentions for polar analytes such as alcohols and phenols. The high resolving ability and unique retention behaviors of the CB7-SG stationary phase may stem from the comprehensive interactions of the aforementioned interactions and shape selectivity. Moreover, the CB7-SG column showed good peak shapes for analytes prone to peak tailing, good thermal stability up to 280°C and separation repeatability with RSD values in the range of 0.01-0.11% for intra-day, 0.04-0.41% for inter-day and 2.5-6.0% for column-to-column, respectively. As demonstrated, the proposed coating method can simultaneously address the solubility problem with CBs for the intended purpose and achieve outstanding GC separation performance.

  17. The VrrA sRNA controls a stationary phase survival factor Vrp of Vibrio cholerae.

    Science.gov (United States)

    Sabharwal, Dharmesh; Song, Tianyan; Papenfort, Kai; Wai, Sun Nyunt

    2015-01-01

    Small non-coding RNAs (sRNAs) are emerging regulatory elements in bacteria. The Vibrio cholerae sRNA VrrA has previously been shown to down-regulate outer membrane proteins (OmpA and OmpT) and biofilm matrix protein (RbmC) by base-pairing with the 5' region of the corresponding mRNAs. In this study, we present an additional target of VrrA in V. cholerae, the mRNA coding for the ribosome binding protein Vrp. Vrp is homologous to ribosome-associated inhibitor A (RaiA) of Escherichia coli which facilitates stationary phase survival through ribosome hibernation. We show that VrrA down-regulates Vrp protein synthesis by base-pairing to the 5' region of vrp mRNA and that the regulation requires the RNA chaperone protein, Hfq. We further demonstrate that Vrp is highly expressed during stationary phase growth and associates with the ribosome of V. cholerae. The effect of the Vrp protein in starvation survival is synergistic with that of the VC2530 protein, a homolog of the E. coli hibernation promoting factor HPF, suggesting a combined role for these proteins in ribosome hibernation in V. cholerae. Vrp and VC2530 are important for V. cholerae starvation survival under nutrient deficient conditions. While VC2530 is down-regulated in cells lacking vrrA, mutation of vrp results in VC2530 activation. This is the first report indicating a regulatory role for an sRNA, modulating stationary factors involved in bacterial ribosome hibernation.

  18. Wide-band residual phase-noise measurements on 40-GHz monolithic mode-locked lasers

    DEFF Research Database (Denmark)

    Larsson, David; Hvam, Jørn Märcher

    2005-01-01

    We have performed wide-band residual phase-noise measurements on semiconductor 40-GHz mode-locked lasers by employing electrical waveguide components for the radio-frequency circuit. The intrinsic timing jitters of lasers with one, two, and three quantum wells (QW) are compared and our design...... prediction, concerning noise versus number of QWs, for the first time corroborated by experiments. A minimum jitter of 44 fs is found, by extrapolating to the Nyquist frequency, for the one-QW device having nearly transform-limited pulses of 1.2 ps. This jitter is nearly three times lower than for a three...

  19. Determination of urinary 8-hydroxy-2′-deoxyguanosine by capillary electrophoresis with molecularly imprinted monolith in-tube solid phase microextraction

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Urinary 8-hydroxy-2'-deoxyguanosine(8-OHdG) is an excellent marker of oxidative DNA damage.In this study,employing guanosine as dummy template a novel molecularly imprinted(MIP) monolithic capillary column had been synthesized,and that was used as medium of in-tube solid phase microextraction(SPME).Coupled with capillary electrophoresis-electrochemical detection(CE-ECD),the system of extraction and detection of 8-OHdG in urinary sample had been developed.Because of its greater phase ratio combined with c...

  20. Green chromatography separation of analytes of greatly differing properties using a polyethylene glycol stationary phase and a low-toxic water-based mobile phase.

    Science.gov (United States)

    Šatínský, Dalibor; Brabcová, Ivana; Maroušková, Alena; Chocholouš, Petr; Solich, Petr

    2013-07-01

    A simple, rapid, and environmentally friendly HPLC method was developed and validated for the separation of four compounds (4-aminophenol, caffeine, paracetamol, and propyphenazone) with different chemical properties. A "green" mobile phase, employing water as the major eluent, was proposed and applied to the separation of analytes with different polarity on polyethylene glycol (PEG) stationary phase. The chromatography separation of all compounds and internal standard benzoic acid was performed using isocratic elution with a low-toxicity mobile phase consisting of 0.04% (v/v) triethylamine and water. HPLC separation was carried out using a PEG reversed-phase stationary phase Supelco Discovery HS PEG column (15 × 4 mm; particle size 3 μm) at a temperature of 30 °C and flow rate at 1.0 mL min(-1). The UV detector was set at 210 nm. In this study, a PEG stationary phase was shown to be suitable for the efficient isocratic separation of compounds that differ widely in hydrophobicity and acid-base properties, particularly 4-aminophenol (log P, 0.30), caffeine (log P, -0.25), and propyphenazone (log P, 2.27). A polar PEG stationary phase provided specific selectivity which allowed traditional chromatographic problems related to the separation of analytes with different polarities to be solved. The retention properties of the group of structurally similar substances (aromatic amines, phenolic compounds, and xanthine derivatives) were tested with different mobile phases. The proposed green chromatography method was successfully applied to the analysis of active substances and one degradation impurity (4-aminophenol) in commercial preparation. Under the optimum chromatographic conditions, standard calibration was carried out with good linearity correlation coefficients for all compounds in the range (0.99914-0.99997, n = 6) between the peak areas and concentration of compounds. Recovery of the sample preparation was in the range 100 ± 5% for all compounds

  1. Retention behavior of amino acids and peptides on protoporphyrin-silica stationary phases with varying metal ion centers.

    Science.gov (United States)

    Xiao, J; Meyerhoff, M E

    1996-09-01

    Various metalloprotoporphyrins (MProP) covalently linked to silica supports are examined as novel immobilized metal ion affinity chromatography (IMAC) stationary phases for separations of amino acids/peptides. Under reversed-phase HPLC conditions, the MProP-silicas exhibit high affinity toward L-histidine via metal-nitrogen axial ligation interactions, with an increasing degree of histidine retention highly dependent on the specific metal ion (M) in the center of the protoporphyrin (ProP) structure: Fe(III) > Ni(II) > Cu(II) > Zn (II) approximately Cd(II). Aromatic amino acids (i.e., L-trytophan and L-phenylalanine) are also retained on MProP columns through pi-pi interactions with the immobilized porphyrins, with the greatest affinity for L-trytophan observed on CuProP-silica columns. Peptides rich in L-histidine and L-tryptophan residues are selectively retained on most of the MProP-silica phases examined; however, the addition of an organic modifier and/or lowering the pH of the mobile phase can be used independently to attenuate the pi-pi and metal ion-nitrogen ligation interactions, respectively. Reproducible separations of His-Phe and trytophan releasing hormone are achieved on a FeProP-silica column even after extensive washing with 50 mM EDTA, demonstrating a fundamental advantage of the new MProP-silica over existing IMAC stationary phases, in which the metal ion is anchored weakly to the support via immobilized iminodiacetate and related ligands.

  2. Preparation and application of methylcalix[4]resorcinarene-bonded silica particles as chiral stationary phase in high-performance liquid chromatography.

    Science.gov (United States)

    Tan, Huey Min; Soh, Shu Fang; Zhao, Jia; Yong, E L; Gong, Yinhan

    2011-01-01

    Two new types of methylcalix[4]resorcinarene-bonded stationary phases, (3-(C-methylcalix[4]resorcinarene)-2-hydroxypropoxy)-propylsilyl-appended silica particles (MCR-HPS) and bromoacetate-substituted MCR-HPS particles (BAMCR-HPS), have been synthesized and used as chiral stationary phases for high-performance liquid chromatography (HPLC) for the first time. The synthetic stationary phases are characterized by means of elemental analysis and Fourier-transform infrared spectroscopy. The chromatographic behavior of MCR-HPS and BAMCR-HPS was studied with several disubstituted benzenes and some chiral drug compounds under both normal phase and reversed-phase conditions. The results show that MCR-HPS has excellent selectivity for the separation of aromatic positional isomers and BAMCR-HPS exhibits excellent performance for separation of enantiomers of chiral compounds.

  3. An immobilized carboxyl containing metal-organic framework-5 stationary phase for open-tubular capillary electrochromatography.

    Science.gov (United States)

    Bao, Tao; Tang, Pingxiu; Mao, Zhenkun; Chen, Zilin

    2016-07-01

    A novel capillary column with metal-organic framework-5 (MOF-5) as the stationary phase was prepared for open-tubular capillary electrochromatography (OT-CEC). To grow MOF-5, the fused-silica capillary was functionalized firstly using 3-aminopropyltriethoxysilane and glutaraldehyde as covalent linkers; and then MOF-5 would be immobilized on the inner wall of COOH-terminated capillary by epitaxial growth, to produce a MOF-5-modified capillary. The successful growth of MOF-5 has been characterized and confirmed by scanning electron microscopy, X-Ray diffraction and Fourier transform infrared spectra. The influence of pH value and methanol on electroosmotic flow (EOF) of the MOF-5-modified capillary column was investigated. The EOF showed a pH-dependent from anode to cathode. The immobilization of MOF-5 improved the interactions between analytes and layer on inner wall of the capillary. Excellent separations of substituted benzenes and acidic and basic analytes were obtained on the fabricated capillary columns. The MOF-5-modified capillary columns exhibited good repeatability, with relative standard deviations for intra-day, inter-day runs and column-to-column less than 1.87%, 3.53%, and 8.49%, respectively. Our successful application of MOF-5 paved the way for introducing series of dicarboxylate-based isoreticular MOFs to OT-CEC as novel stationary phase.

  4. High-resolution profiling of stationary-phase survival reveals yeast longevity factors and their genetic interactions.

    Directory of Open Access Journals (Sweden)

    Erika Garay

    2014-02-01

    Full Text Available Lifespan is influenced by a large number of conserved proteins and gene-regulatory pathways. Here, we introduce a strategy for systematically finding such longevity factors in Saccharomyces cerevisiae and scoring the genetic interactions (epistasis among these factors. Specifically, we developed an automated competition-based assay for chronological lifespan, defined as stationary-phase survival of yeast populations, and used it to phenotype over 5,600 single- or double-gene knockouts at unprecedented quantitative resolution. We found that 14% of the viable yeast mutant strains were affected in their stationary-phase survival; the extent of true-positive chronological lifespan factors was estimated by accounting for the effects of culture aeration and adaptive regrowth. We show that lifespan extension by dietary restriction depends on the Swr1 histone-exchange complex and that a functional link between autophagy and the lipid-homeostasis factor Arv1 has an impact on cellular lifespan. Importantly, we describe the first genetic interaction network based on aging phenotypes, which successfully recapitulated the core-autophagy machinery and confirmed a role of the human tumor suppressor PTEN homologue in yeast lifespan and phosphatidylinositol phosphate metabolism. Our quantitative analysis of longevity factors and their genetic interactions provides insights into the gene-network interactions of aging cells.

  5. High-resolution profiling of stationary-phase survival reveals yeast longevity factors and their genetic interactions.

    Science.gov (United States)

    Garay, Erika; Campos, Sergio E; González de la Cruz, Jorge; Gaspar, Ana P; Jinich, Adrian; Deluna, Alexander

    2014-02-01

    Lifespan is influenced by a large number of conserved proteins and gene-regulatory pathways. Here, we introduce a strategy for systematically finding such longevity factors in Saccharomyces cerevisiae and scoring the genetic interactions (epistasis) among these factors. Specifically, we developed an automated competition-based assay for chronological lifespan, defined as stationary-phase survival of yeast populations, and used it to phenotype over 5,600 single- or double-gene knockouts at unprecedented quantitative resolution. We found that 14% of the viable yeast mutant strains were affected in their stationary-phase survival; the extent of true-positive chronological lifespan factors was estimated by accounting for the effects of culture aeration and adaptive regrowth. We show that lifespan extension by dietary restriction depends on the Swr1 histone-exchange complex and that a functional link between autophagy and the lipid-homeostasis factor Arv1 has an impact on cellular lifespan. Importantly, we describe the first genetic interaction network based on aging phenotypes, which successfully recapitulated the core-autophagy machinery and confirmed a role of the human tumor suppressor PTEN homologue in yeast lifespan and phosphatidylinositol phosphate metabolism. Our quantitative analysis of longevity factors and their genetic interactions provides insights into the gene-network interactions of aging cells.

  6. Identification of new compounds with high activity against stationary phase Borrelia burgdorferi from the NCI compound collection.

    Science.gov (United States)

    Feng, Jie; Shi, Wanliang; Zhang, Shuo; Zhang, Ying

    2015-06-03

    Lyme disease is the leading tick-borne disease in the USA. Whereas the majority of Lyme disease patients with early disease can be cured with standard treatment, some patients suffer from chronic fatigue and joint and muscular pain despite treatment, a syndrome called posttreatment Lyme disease syndrome. Although the cause is unclear, ineffective killing of Borrelia burgdorferi persisters by current Lyme disease antibiotics is one possible explanation. We took advantage of our recently developed high-throughput viability assay and screened the National Cancer Institute compound library collection consisting of 2526 compounds against stationary phase B. burgdorferi. We identified the top 30 new active hits, including the top six anthracycline antibiotics daunomycin 3-oxime, dimethyldaunomycin, daunomycin, NSC299187, NSC363998 and nogalamycin, along with other compounds, including prodigiosin, mitomycin, nanaomycin and dactinomycin, as having excellent activity against B. burgdorferi stationary phase culture. The anthracycline or anthraquinone compounds, which are known to have both anti-cancer and antibacterial activities, also had high activity against growing B. burgdorferi with low minimum inhibitory concentration. Future studies on the structure-activity relationship and mechanisms of action of anthracyclines/anthraquinones are warranted. In addition, drug combination studies with the anthracycline class of compounds and the current Lyme antibiotics to eradicate B. burgdorferi persisters in vitro and in animal models are needed to determine if they improve the treatment of Lyme disease.

  7. Ultra high performance liquid chromatography versus high performance liquid chromatography: stationary phase selectivity for generic carotenoid screening.

    Science.gov (United States)

    Bijttebier, Sebastiaan; D'Hondt, Els; Noten, Bart; Hermans, Nina; Apers, Sandra; Voorspoels, Stefan

    2014-03-07

    Aim of study was to find the most suitable LC column for generic carotenoid screening. To represent the diversity of carotenoids in nature and to optimize chromatographic separation, a set of carotenoid standards was carefully chosen to account for the various classes of carotenoids. The HPLC C30 column has since long been the 'golden standard' in the chromatographic separation of carotenoids. Since approximately one decade, new UHPLC technology has led to much shorter analysis times, smaller peak widths and higher chromatographic resolution. However, there are currently no UHPLC columns on the market containing the specific stationary phase chemistry of the HPLC C30 column. Therefore during this study, we investigated the separation of carotenoids on a set of UHPLC columns and compared it to their separation on the HPLC C30 column. Comparison of carotenoids separations on the different stationary phases with objective column comparison parameters clearly indicated that the HPLC C30 column is an overall better performer in the separation of carotenoids. This is due to the lack of UHPLC column chemistries that are adapted for carotenoid analysis. However, analysis time on the HPLC C30 column takes about four times longer compared to UHPLC analysis. Therefore, with the range of columns that are commercially available nowadays, a choice has to be made between very high selectivity (HPLC C30 column) and analysis times that are adapted to modern laboratory requirements (UHPLC technology). Therefore, carotenoid separations would be even more performing if an appropriate UHPLC C30 column would be available.

  8. Polystyrene-divinylbenzene stationary phases agglomerated with quaternized multi-walled carbon nanotubes for anion exchange chromatography.

    Science.gov (United States)

    Huang, Zhongping; Wu, Hongwei; Wang, Fengli; Yan, Wenwu; Guo, Weiqiang; Zhu, Yan

    2013-06-14

    This work explores the potential of multi-walled carbon nanotubes as an agglomerated material for ion chromatography stationary phases for the separation of inorganic anions. Polyelectrolytes with quaternary ammonium groups were introduced onto the carbon nanotube surface, based on condensation polymerization of 1,4-butanediol diglycidyl ether (BDDE) and methylamine (MA). Quaternized multi-walled carbon nanotubes (Q-MWCNTs) were electrostatically adsorbed onto the surface of sulfonated polystyrene-divinylbenzene (PS-DVB) beads to generate the anion exchanger, which were confirmed by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). A 100mm×4.0mm i.d. column was packed with Q-MWCNTs agglomerated PS-DVB particles, with a capacity of 56μequiv./column. Separation of inorganic anions, such as F(-), Cl(-), NO2(-), Br(-), NO3(-), SO4(2-) and PO4(3-) were performed. The stationary phase was rigid, chemically stable and showed good ion-exchange characteristics.

  9. Purification of Monoclonal Antibodies Using a Fiber Based Cation-Exchange Stationary Phase: Parameter Determination and Modeling

    Directory of Open Access Journals (Sweden)

    Jan Schwellenbach

    2016-10-01

    Full Text Available Monoclonal antibodies (mAb currently dominate the market for protein therapeutics. Because chromatography unit operations are critical for the purification of therapeutic proteins, the process integration of novel chromatographic stationary phases, driven by the demand for more economic process schemes, is a field of ongoing research. Within this study it was demonstrated that the description and prediction of mAb purification on a novel fiber based cation-exchange stationary phase can be achieved using a physico-chemical model. All relevant mass-transport phenomena during a bind and elute chromatographic cycle, namely convection, axial dispersion, boundary layer mass-transfer, and the salt dependent binding behavior in the fiber bed were described. This work highlights the combination of model adaption, simulation, and experimental parameter determination through separate measurements, correlations, or geometric considerations, independent from the chromatographic cycle. The salt dependent binding behavior of a purified mAb was determined by the measurement of adsorption isotherms using batch adsorption experiments. Utilizing a combination of size exclusion and protein A chromatography as analytic techniques, this approach can be extended to a cell culture broth, describing the salt dependent binding behavior of multiple components. Model testing and validation was performed with experimental bind and elute cycles using purified mAb as well as a clarified cell culture broth. A comparison between model calculations and experimental data showed a good agreement. The influence of the model parameters is discussed in detail.

  10. Quinine-Based Zwitterionic Chiral Stationary Phase as a Complementary Tool for Peptide Analysis: Mobile Phase Effects on Enantio- and Stereoselectivity of Underivatized Oligopeptides.

    Science.gov (United States)

    Ianni, Federica; Sardella, Roccaldo; Carotti, Andrea; Natalini, Benedetto; Lindner, Wolfgang; Lämmerhofer, Michael

    2016-01-01

    Peptide stereoisomer analysis is of importance for quality control of therapeutic peptides, the analysis of stereochemical integrity of bioactive peptides in food, and the elucidation of the stereochemistry of peptides from a natural chiral pool which often contains one or more D-amino acid residues. In this work, a series of model peptide stereoisomers (enantiomers and diastereomers) were analyzed on a zwitterionic ion-exchanger chiral stationary phase (Chiralpak ZWIX(+) 5 µm), in order to investigate the retention and separation performance for such compounds on this chiral stationary phase and elucidate its utility for this purpose. The goal of the study focused on 1) investigations of the effects of the sample matrix used to dissolve the peptide samples; 2) optimization of the mobile phase (enabling deriving information on factors of relevance for retention and separation); and 3) derivation of structure-selectivity relationships. It turned out that small di- and tripeptides can be well resolved under optimized conditions, typically with resolutions larger than 1.5. The optimized mobile phase often consisted of methanol-tetrahydrofuran-water (49:49:2; v/v/v) with 25 mM formic acid and 12.5 mM diethylamine. This work proposes some guidance on which mobile phases can be most efficiently used for peptide stereoisomer separations on Chiralpak ZWIX. Chirality 28:5-16, 2016. © 2015 Wiley Periodicals, Inc.

  11. Stationary States in Saturated Two-Photon Processes and Generation of Phase-Averaged Mixtures of Even and Odd Quantum States

    CERN Document Server

    Dodonov, V V

    1998-01-01

    We consider a relaxation of a single mode of the quantized field in a presence of one- and two-photon absorption and emission processes. Exact stationary solutions of the master equation for the diagonal elements of the density matrix in the Fock basis are found in the case of completely saturated two-photon emission. If two-photon processes dominate over single-photon ones, the stationary state is a mixture of phase averaged even and odd coherent states.

  12. Online micro-solid-phase extraction based on boronate affinity monolithic column coupled with high-performance liquid chromatography for the determination of monoamine neurotransmitters in human urine.

    Science.gov (United States)

    Yang, Xiaoting; Hu, Yufei; Li, Gongke

    2014-05-16

    Quantification of monoamine neurotransmitters is very important in diagnosing and monitoring of patients with neurological disorders. We developed an online analytical method to selectively determine urinary monoamine neurotransmitters, which coupled the boronate affinity monolithic column micro-solid-phase extraction with high-performance liquid chromatography (HPLC). The boronate affinity monolithic column was prepared by in situ polymerization of vinylphenylboronic acid (VPBA) and N,N'-methylenebisacrylamide (MBAA) in a stainless capillary column. The prepared monolithic column showed good permeability, high extraction selectivity and capacity. The column-to-column reproducibility was satisfactory and the enrichment factors were 17-243 for four monoamine neurotransmitters. Parameters that influence the online extraction efficiency, including pH of sample solution, flow rate of extraction and desorption, extraction volume and desorption volume were investigated. Under the optimized conditions, the developed method exhibited low limit of detection (0.06-0.80μg/L), good linearity (with R(2) between 0.9979 and 0.9993). The recoveries in urine samples were 81.0-105.5% for four monoamine neurotransmitters with intra- and inter-day RSDs of 2.1-8.2% and 3.7-10.6%, respectively. The online analytical method was sensitive, accurate, selective, reliable and applicable to analysis of trace monoamine neurotransmitters in human urine sample.

  13. Monolithic molecularly imprinted solid-phase extraction for the selective determination of trace cytokinins in plant samples with liquid chromatography-electrospray tandem mass spectrometry.

    Science.gov (United States)

    Du, Fuyou; Ruan, Guihua; Liang, Shenghua; Xie, Fuwei; Liu, Huwei

    2012-08-01

    Cytokinins (CTKs) are a class of growth-regulating hormones involved in various physiological and developmental processes. More novel analytical methods for the accurate identification and quantitative determination of trace CTKs in plants have been desired to better elucidate the roles of CTKs. In this work, a novel method based on monolithic molecularly imprinted solid-phase extraction followed by liquid chromatography-electrospray tandem mass spectrometry (mMI-SPE-LC-MS/MS) was developed for accurate determination of four CTKs in plant samples. The molecularly imprinted polymer monolith was prepared by using kinetin as the template in syringes and exhibited specific recognition ability for the four CTKs in comparison with that of non-imprinted polymer monolith. Several factors affecting the extraction performance of mMI-SPE, including the pH of loading sample solution, the nature and volume of elution solvent, the flow rate of sample loading, and sample volume, were investigated, respectively. Under the optimized conditions, the proposed mMI-SPE-LC-MS/MS method was successfully applied in the selective extraction and determination of four CTKs in plant tissues, and it offers detection limits (S/N = 3) of 104, 113, 130, and 89 pg/mL and mean recoveries of 85.9%, 79.3%, 73.5%, and 70.1% for kinetin, kinetin glucoside, trans-zeatin, and meta-topolin (mT), respectively, with the corresponding RSDs less than 15%.

  14. Fast analysis of glycosides based on HKUST-1-coated monolith solid-phase microextraction and direct analysis in real-time mass spectrometry.

    Science.gov (United States)

    Li, Xianjiang; Wang, Xin; Ma, Wen; Ai, Wanpeng; Bai, Yu; Ding, Li; Liu, Huwei

    2017-04-01

    Glycosides are a kind of highly important natural aromatic precursors in tobacco leaves. In this study, a novel HKUST-1-coated monolith dip-it sampler was designed for the fast and sensitive analysis of trace glycosides using direct analysis in real-time mass spectrometry. This device was prepared in two steps: in situ polymerization of monolith in a glass capillary of dip-it and layer-by-layer growth of HKUST-1 on the surface of monolith. Sufficient extraction was realized by immersing the tip to solution and in situ desorption was carried out by plasma direct analysis in real time. Compared with traditional solid-phase microextraction protocols, sample desorption was not needed anymore, and only extraction conditions were needed to be optimized in this method, including the gas temperature of direct analysis in real time, extraction time, and CH3 COONH4 additive concentration. This method enabled the simultaneous detection of six kinds of glycosides with the limits of detection of 0.02-0.05 μg/mL and the linear ranges covering two orders of magnitude with the limits of quantitation of 0.05-0.1 μg/mL. Moreover, the developed method was applied for the glycosides analysis of three tobacco samples, which only took about 2 s for every sample.

  15. A Model for Solving the Maxwell Quasi Stationary Equations in a 3-Phase Electric Reduction Furnace

    Directory of Open Access Journals (Sweden)

    S. Ekrann

    1982-10-01

    Full Text Available A computer code has been developed for the approximate computation of electric and magnetic fields within an electric reduction furnace. The paper describes the numerical methods used to solve Maxwell's quasi-stationary equations, which are the governing equations for this problem. The equations are discretized by a staggered grid finite difference technique. The resulting algebraic equations are solved by iterating between computations of electric and magnetic quantities. This 'outer' iteration converges only when the skin depth is larger or of about the same magnitude as the linear dimensions of the computational domain. In solving for electric quantities with magnetic quantities being regarded as known, and vice versa, the central computational task is the solution of a Poisson equation for a scalar potential. These equations are solved by line successive overrelaxation combined with a rebalancing technique.

  16. Molecularly imprinted coated graphene oxide solid-phase extraction monolithic capillary column for selective extraction and sensitive determination of phloxine B in coffee bean

    Energy Technology Data Exchange (ETDEWEB)

    Zhai, Haiyun, E-mail: zhaihaiyun@126.com [College of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006 (China); Su, Zihao [College of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006 (China); Chen, Zuanguang, E-mail: chenzg@mail.sysu.edu.cn [School of Pharmaceutical Science, Sun Yat-sen University, Guangzhou 510006 (China); Liu, Zhenping; Yuan, Kaisong; Huang, Lu [College of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006 (China)

    2015-03-20

    Highlights: • A new GO-MISPE monolithic capillary column was prepared. • The column showed ability of impurities removal and excellent selectivity. • Phloxine B existed in real sample was enriched more than 90 times. • The GO-MISPE column presented good recovery and high stability. • The method was prospered to analyze phloxine B and LOD achieved 0.3 ng g{sup −1}. - Abstract: A method was developed to sensitively determine phloxine B in coffee bean by molecularly imprinted polymers (MIPs) coated graphene oxide (GO) solid-phase extraction (GO-MISPE) coupled with high-performance liquid chromatography and laser-induced fluorescence detection (HPLC–LIF). The GO-MISPE capillary monolithic column was prepared by water-bath in situ polymerization, using GO as supporting material, phloxine B, methacrylic acid (MAA), and ethylene dimethacrylate (EDMA) as template, functional monomer, and cross-linker, respectively. The properties of the homemade GO-MISPE capillary monolithic column, including capacity and specificity, were investigated under optimized conditions. The GO-MIPs were characterized by scanning electron microscopy (SEM) and Fourier transform-infrared spectroscopy (FT-IR). The mean recoveries of phloxine B in coffee bean ranged from 89.5% to 91.4% and the intra-day and inter-day relative standard deviation (RSD) values all ranged from 3.6% to 4.7%. Good linearity was obtained over 0.001–2.0 μg mL{sup −1} (r = 0.9995) with the detection limit (S/N = 3) of 0.075 ng mL{sup −1}. Under the selected conditions, enrichment factors of over 90-fold were obtained and extraction on the monolithic column effectively cleaned up the coffee bean matrix. The results demonstrated that the proposed GO-MISPE HPLC–LIF method can be applied to sensitively determine phloxine B in coffee bean.

  17. Recent development of oligosaccharides chiral stationary phases and their applications in chromatography%寡糖手性固定相研究进展

    Institute of Scientific and Technical Information of China (English)

    赵峰; 袁黎明; 熊汝琴

    2015-01-01

    In this paper ,The development of the Oligosaccharide chiral stationary phase for the HPLC chiral stationa‐ry phase is reviewed ,the study found that these sugars as the hplc chiral stationary phase is effect better ,more promis‐ing ,and the prospect of the chiral stationary phase is reviewed .%介绍了葡萄糖、纤维二塘、棉子糖、乳糖等用来作为高效液相色谱手性固定相的研究进展,研究发现这几种糖类作为高效液相色谱的手性固定相拆分效果比较好,比较有前途。同时展望了高效液相色谱手性固定相的发展前景。

  18. Enantiomer Separation of α-Dimethyl Dicarboxylate Biphenyl and Related Biphenyl Compounds by Normal Phase HPLC on Polysaccharide Based Chiral Stationary Phases

    Institute of Scientific and Technical Information of China (English)

    LIU,Yue-Qi(刘月启); HAN,Xiao-Qian(韩小茜); Qi,Bang-Feng(齐邦峰); LIU,Chun-Hui(柳春辉); LI,Yong-Min(李永民); CHEN,Li-Ren(陈立仁)

    2002-01-01

    Cellulose tris(4-methylphenylcarbamate), amylose tris(3,5-dimethylpphenylcarbamate) and amylose tris (phenylcarbamate)were prepared by the method reported by Okamoto and were coated onto an aminopropylated mesoporous spherical silica gel. These final products were used as chiral stationary phases of high performance liquid chromatography for the eighteen structurally related biphenyl conmpounds. The resolution was made using normal-phase methodology with a mobile phase consisting of n-hexane-alcohol (ethanol, 1-propanol, 2-propanol or 1-butanol). The effects of various aliphatic alcohols in the mobile phase were studied. The structural features of the solutes that influence their k'were discussed. A dominant effect of trifluoroaetic acid on chiral separation of aacidicdic solutes was noted.

  19. Enantioseparation of Novel Chiral Tetrahedral Clusters on an Amylose Tris-(3,5-dimethylphenylcarbamate) Chiral Stationary Phase by Normal Phase HPLC

    Institute of Scientific and Technical Information of China (English)

    LI,Wen-Zhi(李文智); WANG,Xia(王霞); ZHANG,Wei-Qiang(张伟强); CHEN Li-Ren(陈立仁); LI,Yong-Min(李永民); MA,Chun-Lin(马春林); YIN,Yuan-Qi(殷元骐)

    2004-01-01

    Amylose tris(3,5-dimethylphenylcarbamate) (ADMPC) coated on a kind of small particle silica gel was prepared. On this ADMPC chiral stationary phase (CSP), the direct enantiomeric separation of six novel chiral transition metal tetrahedral clusters has firstly been achieved using n-hexane as the mobile phase containing various alcohols as modifiers. The effect of mobile phase modifiers and the structural variation of the solutes on their retention factors (k′) and resolutions (Rs) were investigated. The result suggests that not only the structure and concentration of alcohol in mobile phase, but also the structural differences in racemates can have a pronounced effect on enantiomeric separation. ADMPC-CSP is a suitable CSP for the optical resolution of chiral tetrahedral cluster by HPLC.

  20. Evaluation and comparison of n-alkyl chain and polar ligand bonded stationary phases for protein separation in reversed-phase liquid chromatography.

    Science.gov (United States)

    Ding, Ling; Guo, Zhimou; Xiao, Yuansheng; Xue, Xingya; Zhang, Xiuli; Liang, Xinmiao

    2014-09-01

    Protein retention is very sensitive to the change of solvent composition in reversed-phase liquid chromatography for so called "on-off" mechanism, leading to difficulty in mobile phase optimization. In this study, a novel 3-chloropropyl trichlorosilane ligand bonded column was prepared for protein separation. The differences in retention characteristics between the 3-chloropropyl trichlorosilane ligand bonded column and n-alkyl chain modified (C2, C4, C8) stationary phases were elucidated by the retention equation l nk=a+cC(B). Retention parameters (a and c) of nine standard proteins with different molecular weights were calculated by using homemade software. Results showed that retention times of nine proteins were similar on four columns, but the 3-chloropropyl trichlorosilane ligand bonded column obtained the lowest retention parameter values of larger proteins. It meant that their retention behavior affected by acetonitrile concentration would be different due to lower |c| values. More specifically, protein elution windows were broader, and retentions were less sensitive to the change of acetonitrile concentration on the 3-chloropropyl trichlorosilane ligand bonded column than that on other columns. Meanwhile, the 3-chloropropyl trichlorosilane ligand bonded column displayed distinctive selectivity for some proteins. Our results indicated that stationary phase with polar ligand provided potential solutions to the "on-off" problem and optimization in protein separation.

  1. Differential expression of secretion machinery during bacterial growth: SecY and SecF decrease while SecA increases during transition from exponential phase to stationary phase.

    Science.gov (United States)

    Yang, Chun-Kai; Lu, Chung-Dar; Tai, Phang C

    2013-12-01

    Transcription of many house-keeping genes, including secY and some other sec genes, decreases in the transition from the exponential phase to the stationary phase (feast to famine) in Bacillus subtilis. Unexpectedly and in contradiction to earlier reports, enhanced transcription was observed for another group of sec genes, including secA which codes for an essential ATPase for protein secretion. Consistent with the transcription data, the SecA protein of B. subtilis increases significantly in the stationary phase. Immunoblot analyses of Sec proteins during the transition in Escherichia coli also revealed the pronounced decreases of SecY and SecF and the increase of SecA, resulting in drastic increases of SecA/SecY and SecA/SecF ratios from exponential to stationary phases. The differential expression of Sec proteins in the stationary phase suggests the possibility of specific physiological functions.

  2. Comparison of analytical protein separation characteristics for three amine-based capillary-channeled polymer (C-CP) stationary phases.

    Science.gov (United States)

    Jiang, Liuwei; Marcus, R Kenneth

    2016-02-01

    Capillary-channeled polymer (C-CP) fiber stationary phases are finding utility in the realms of protein analytics as well as downstream processing. We have recently described the modification of poly(ethylene terephthalate) (PET) C-CP fibers to affect amine-rich phases for the weak anion-exchange (WAX) separation of proteins. Polyethylenimine (PEI) is covalently coupled to the PET surface, with subsequent cross-linking imparted by treatment with 1,4-butanediol diglycidyl ether (BUDGE). These modifications yield vastly improved dynamic binding capacities over the unmodified fibers. We have also previously employed native (unmodified) nylon 6 C-CP fibers as weak anion/cation-exchange (mixed-mode) and hydrophobic interaction chromatography (HIC) phases for protein separations. Polyamide, nylon 6, consists of amide groups along the polymer backbone, with primary amines and carboxylic acid end groups. The analytical separation characteristics of these three amine-based C-CP fiber phases are compared here. Each of the C-CP fiber columns in this study was shown to be able to separate a bovine serum albumin/hemoglobin/lysozyme mixture at high mobile phase linear velocity (∼70 mm s(-1)) but with different elution characteristics. These differences reflect the types of protein-surface interactions that are occurring, based on the active group composition of the fiber surfaces. This study provides important fundamental understanding for the development of surface-modified C-CP fiber columns for protein separation.

  3. Development of a V-shape bis(tetraoxacalix[2]arene[2]triazine) stationary phase for high performance liquid chromatography.

    Science.gov (United States)

    Hu, Kai; Feng, Suxiang; Wu, Mingxia; Wang, Shuang; Zhao, Wenjie; Jiang, Qiong; Yu, Ajuan; Zhang, Shusheng

    2014-12-01

    A new stationary phase for high-performance liquid chromatography was prepared by covalently bonding a V-shape cage heteroatom-bridged calixarene onto silica gel using 3-aminopropyltriethoxysilane as coupling reagent. The structure of the new material was characterized by infrared spectroscopy, elemental analysis and thermogravimetric analysis. Linear solvation energy relationship method was successfully employed to evaluate the new phase with a set of 34 solutes. The retention characteristic of the new phase shows significant similarities with ODS, as well as distinctive features. Multiple mechanisms including hydrophobic, hydrogen bonding, π-π and n-π interaction are involved. The chromatographic behavior of the phase was illustrated by using alkylbenzenes, aromatics positional isomers and flavonoids as probes. Moreover, inorganic anions were individually separated in anion-exchange mode by using the same column. Thus, multi-interaction mechanisms and mixed-mode separation of the new phase can very likely guarantee its excellent chromatographic performance for the analysis of complex samples. The column has been successfully employed for the analysis of clenbuterol in animal urine, and it is demonstrated to be suitable and a competitive alternative analytical method for the determination of clenbuterol.

  4. The Optical Resolution of Chiral Tetrahedrone-type Clusters Contai- ning SCoFeM (M=Mo or W) Using High Performance Liquid Chromatography Chiral Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Amylose tris (phenylcarbamate) chiral stationary phase (ATPC-CSP) was prepared and used for optical resolution of clusters 1 and 2. n-Hexane/2-propanol ( 99/1; v/v) were found to be the most suitable mobile phase on ATPC-CSP.

  5. Monolithic silica-based capillary reversed-phase liquid chromatography/electrospray mass spectrometry for plant metabolomics

    NARCIS (Netherlands)

    Tolstikov, V.V.; Lommen, A.; Nakanishi, K.; Tanaka, N.; Fiehn, O.

    2003-01-01

    Application of C18 monolithic silica capillary columns in HPLC coupled to ion trap mass spectrometry detection was studied for probing the metabolome of the model plant Arabidopsis thaliana. It could be shown that the use of a long capillary column is an easy and effective approach to reduce ionizat

  6. Preparation, characterization and application of N-methylene phosphonic acid chitosan grafted magnesia–zirconia stationary phase

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Qing; Chen, Jie; Huang, Kun; Zhang, Xin; Xu, Li [Tongji School of Pharmacy, Huazhong University of Science and Technology, Wuhan 430030 (China); Shi, Zhi-guo, E-mail: shizg@whu.edu.cn [Department of Chemistry, Wuhan University, Wuhan 430072 (China)

    2015-01-07

    Highlights: • N-methylene phosphonic acid chitosan grafted MgO–ZrO{sub 2} was prepared. • It exhibited superior HILIC chromatographic performance to the bare MgO–ZrO{sub 2}. • Monosaccharides, phospholipids and peptides were successfully separated. • It was a promising HILIC stationary phase. - Abstract: A hydrophilic stationary phase (SP) was prepared through grafting N-methylene phosphonic acid chitosan on magnesia–zirconia particles (P-CTS-MgO–ZrO{sub 2}) via Lewis acid–base interaction. The resulting material was characterized by thermogravimetric analysis, Fourier transform infrared spectroscopy, scanning electron microscope and nitrogen adsorption analysis. The chromatographic performance of P-CTS-MgO–ZrO{sub 2} was systemically evaluated by studying effect of acetonitrile content, pH and buffer concentration in the mobile phase. The results demonstrated that the novel SP provided hydrophilic, electrostatic-repulsion and ion-exchange interactions. Compared to the bare MgO–ZrO{sub 2}, P-CTS-MgO–ZrO{sub 2} exhibited superior peak shape, reasonable resolution and reduced analysis time in separation of basic analytes. Besides, remarkable resolving power of acids, i.e. six non-steroidal anti-inflammatory drugs which failed to be eluted from the bare MgO–ZrO{sub 2}, was obtained with the theoretical plate number (N/m) of 4653–31313, asymmetry factor <1.21 and the resolution of 1.6–3.4. Finally, P-CTS-MgO–ZrO{sub 2} SP was applied to separate monosaccharides, phospholipids and peptides. P-CTS-MgO–ZrO{sub 2} was a promising hydrophilic SP for wide applications.

  7. Direct enantioseparation of underivatized aliphatic 3-hydroxyalkanoic acids with a quinine-based zwitterionic chiral stationary phase.

    Science.gov (United States)

    Ianni, Federica; Pataj, Zoltán; Gross, Harald; Sardella, Roccaldo; Natalini, Benedetto; Lindner, Wolfgang; Lämmerhofer, Michael

    2014-10-10

    While aliphatic 2-hydroxyalkanoic acids have been more or less successfully enantioseparated with various chiral stationary phases by HPLC and GC, analogous applications on underivatized aliphatic 3-hydroxyalkanoic acids are completely absent in the scientific literature. With the aim of closing this gap, the enantioseparation of 3-hydroxybutyric acid, 3-hydroxydecanoic acid and 3-hydroxymyristic acid has been performed with two ion-exchange type chiral stationary phases (CSPs): one containing the anion-exchange type tert-butyl carbamoyl quinine chiral selector motif (Chiralpak QN-AX), and the other carrying the new zwitterionic variant based on trans-(S,S)-2-aminocyclohexanesulfonic acid-derivatized quinine carbamate (Chiralpak ZWIX(+)) as the chiral selector and enantiodiscriminating element, respectively. The zwitterionic enantiorecognition material provided better results in terms of enantioselectivity and resolution compared to the anion-exchanger CSP at reduced retention times due to the intramolecular counterion effect imposed by the sulfonic acid moiety and its competition with the 3-hydroxyalkanoic acid analyte for ionic interaction at the quininium-anion exchanger site. It is thus recommended as the CSP of first choice for enantioseparations of the class of aliphatic 3-hydroxyalkanoic acids. With use of polar organic eluent composed of ACN/MeOH/AcOH - 95/5/0.05 (v/v/v), a good compromise in terms of analysis time and enantioresolution quality was accomplished. The major experimental variables have been investigated for optimization of the resolution and allowed to derive information on the enantiorecognition mechanism. Corresponding Chiralpak ZWIX(-), based on pseudo-enantiomeric selector derived from quinidine and trans-(R,R)-2-aminocyclohexanesulfonic acid with opposite configurations provided reversed enantiomer elution orders. It has further to be stressed that these separations can be obtained with mass spectrometry compatible mobile phases.

  8. Comparison of Membrane Chromatography and Monolith Chromatography for Lactoferrin and Bovine Serum Albumin Separation

    Directory of Open Access Journals (Sweden)

    Chalore Teepakorn

    2016-09-01

    Full Text Available These last few decades, membranes and monoliths have been increasingly used as stationary phases for chromatography. Their fast mass transfer is mainly based on convection, which leads to reduced diffusion, which is usually observed in resins. Nevertheless, poor flow distribution, which causes inefficient binding, remains a major challenge for the development of both membrane and monolith devices. Moreover, the comparison of membranes and monoliths for biomolecule separation has been very poorly investigated. In this paper, the separation of two proteins, bovine serum albumin (BSA and lactoferrin (LF, with similar sizes, but different isoelectric points, was investigated at a pH of 6.0 with a BSA-LF concentration ratio of 2/1 (2.00 mg·mL−1 BSA and 1.00 mg·mL−1 LF solution using strong cation exchange membranes and monoliths packed in the same housing, as well as commercialized devices. The feeding flow rate was operated at 12.0 bed volume (BV/min for all devices. Afterward, bound LF was eluted using a phosphate-buffered saline solution with 2.00 M NaCl. Using membranes in a CIM housing from BIA Separations (Slovenia with porous frits before and after the membrane bed, higher binding capacities, sharper breakthrough curves, as well as sharper and more symmetric elution peaks were obtained. The monolith and commercialized membrane devices showed lower LF binding capacity and broadened and non-symmetric elution peaks.

  9. Characterization of polymer monoliths containing embedded nanoparticles by scanning transmission X-ray microscopy (STXM).

    Science.gov (United States)

    Arrua, R Dario; Hitchcock, Adam P; Hon, Wei Boon; West, Marcia; Hilder, Emily F

    2014-03-18

    The structural and chemical homogeneity of monolithic columns is a key parameter for high efficiency stationary phases in liquid chromatography. Improved characterization techniques are needed to better understand the polymer morphology and its optimization. Here the analysis of polymer monoliths by scanning transmission X-ray microscopy (STXM) is presented for the first time. Poly(butyl methacrylate-co-ethyleneglycoldimethacrylate) [poly(BuMA-co-EDMA)] monoliths containing encapsulated divinylbenzene (DVB) nanoparticles were characterized by STXM, which gives a comprehensive, quantitative chemical analysis of the monolith at a spatial resolution of 30 nm. The results are compared with other methods commonly used for the characterization of polymer monoliths [scanning electron microscopy (SEM), transmission electron microscopy (TEM), mercury porosimetry, and nitrogen adsorption]. The technique permitted chemical identification and mapping of the nanoparticles within the polymeric scaffold. Residual surfactant, which was used during the manufacture of the nanoparticles, was also detected. We show that STXM can give more in-depth chemical information for these types of materials and therefore lead to a better understanding of the link between polymer morphology and chromatographic performance.

  10. Heat and pulsed electric field resistance of pigmented and non-pigmented enterotoxigenic strains of Staphylococcus aureus in exponential and stationary phase of growth.

    Science.gov (United States)

    Cebrián, G; Sagarzazu, N; Pagán, R; Condón, S; Mañas, P

    2007-09-30

    The survival of four enterotoxigenic strains of Staphylococcus aureus (with different pigment content) to heat and to pulsed electric fields (PEF) treatments, and the increase in resistance to both processing stresses associated with entrance into stationary phase was examined. Survival curves to heat (58 degrees C) and to PEF (26 kV/cm) of cells in the stationary and in the exponential phase of growth were obtained. Whereas a wide variation in resistance to heat treatments was detected amongst the four strains, with decimal reduction time values at 58 degrees C (D(58 degrees C)) ranging from 0.93 to 0.20 min, the resistance to PEF was very similar. The occurrence of a higher tolerance to heat in stationary phase was coincident with a higher content in carotenoid pigmentation in S. aureus colonies. However, cells of the most heat resistant (pigmented) and the most heat sensitive (non-pigmented) strains in the mid-exponential phase of growth showed similar resistance to heat and to PEF. Therefore the increase in thermotolerance upon entrance into stationary phase of growth was more marked for the pigmented strains. Recovery in anaerobic conditions particularly enhanced survival to heat treatments in a non-pigmented strain. Strain CECT 4630, which possess a deficient sigma B activity, showed low heat resistance, low pigmentation, and reduced increase in thermotolerance in stationary phase. These results indicate that the magnitude of the development of a higher heat resistance in S. aureus in stationary phase is positively related to the carotenoid content of the strain. The development of tolerance to pulsed electric field was less relevant and not linked to the carotenoid content.

  11. Enantioseparation and impurity determination of the enantiomers of novel phenylethanolamine derivatives by high performance liquid chromatography on amylose stationary phase

    Energy Technology Data Exchange (ETDEWEB)

    Yang Jing [Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang 110016 (China); Guan Jin [Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang 110016 (China); School of Applied Chemistry, Shenyang Institute of Chemical Technology, Shenyang 110142 (China); Pan Li [School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016 (China); Jiang Kun [Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang 110016 (China); Cheng Maosheng [School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016 (China); Li Famei [Department of Analytical Chemistry, Shenyang Pharmaceutical University, Shenyang 110016 (China)], E-mail: lifamei@syphu.edu.cn

    2008-03-10

    Simple and efficient analytical HPLC methods using Chiralpak AS-H as chiral stationary phase were developed for direct enantioseparation of 11 novel phenylethanolamine derivatives. The chromatographic experiments were performed in normal phase mode with n-hexane-ethanol-triethylamine (TEA) as mobile phase. Excellent baseline enantioseparation was obtained for most of compounds. The effects of the concentration of organic modifiers and column temperature were studied for the enantiomeric separation. The mechanism of chiral recognition was discussed based on the relationship between the thermodynamic parameters and structures of compounds. It was found that the enantioseparations were all enthalpy driven, and the tert-butyl groups of compounds had significant influence on the chiral recognition. Trantinterol enantiomers were resolved (R{sub s} = 2.73) within 14 min using n-hexane-ethanol-TEA (98:2:0.1, v/v/v) as mobile phase with a flow rate of 0.8 mL min{sup -1} at 30 deg. C. The optimized method was validated for linearity, precision, accuracy and stability in solution and proved to be robust. The limits of detection (LOD) and quantification (LOQ) for (+)-trantinterol were 0.15 and 0.46 {mu}g mL{sup -1}. The method was applied for enantiomeric impurity determination of (-)-trantinterol bulk samples.

  12. Two-dimensional high-performance thin-layer chromatography of tryptic bovine albumin digest using normal- and reverse-phase systems with silanized silica stationary phase.

    Science.gov (United States)

    Gwarda, Radosław Łukasz; Dzido, Tadeusz Henryk

    2013-10-18

    Among many advantages of planar techniques, two-dimensional (2D) separation seems to be the most important for analysis of complex samples. Here we present quick, simple and efficient two-dimensional high-performance thin-layer chromatography (2D HPTLC) of bovine albumin digest using commercial HPTLC RP-18W plates (silica based stationary phase with chemically bonded octadecyl ligands of coverage density 0.5μmol/m(2) from Merck, Darmstadt). We show, that at low or high concentration of water in the mobile phase comprised methanol and some additives the chromatographic systems with the plates mentioned demonstrate normal- or reversed-phase liquid chromatography properties, respectively, for separation of peptides obtained. These two systems show quite different separation selectivity and their combination into 2D HPTLC process provides excellent separation of peptides of the bovine albumin digest. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Growth potential of exponential- and stationary-phase Salmonella Typhimurium during sausage fermentation

    DEFF Research Database (Denmark)

    Birk, Tina; Henriksen, Sidsel; Müller, K.

    2016-01-01

    Raw meat for sausage production can be contaminated with Salmonella. For technical reasons, meat is often frozen prior to mincing but it is unknown how growth of Salmonella in meat prior to freezing affects its growth potential during sausage fermentation. We investigated survival of exponential...... starter culture. With no starter culture, both strains grew in both growth phases. In general, a functional starter culture abolished S. Typhimurium growth independent of growth phase and we concluded that ensuring correct fermentation is important for sausage safety. However, despite efficient...... fermentation, sporadic growth of exponential-phase cells of S. Typhimurium was observed drawing attention to the handling and storage of sausage meat....

  14. Monolithic metal-organic framework MIL-53(Al)-polymethacrylate composite column for the reversed-phase capillary liquid chromatography separation of small aromatics.

    Science.gov (United States)

    Yusuf, Kareem; Badjah-Hadj-Ahmed, Ahmed Yacine; Aqel, Ahmad; ALOthman, Zeid Abdullah

    2016-03-01

    A monolithic capillary column containing a composite of metal-organic framework MIL-53(Al) incorporated into hexyl methacrylate-co-ethylene dimethacrylate was prepared to enhance the separation of mixtures of small aromatic compounds by using capillary liquid chromatography. The addition of 10 mg/mL MIL-53(Al) microparticles increased the micropore content in the monolithic matrix and increased the Brunauer-Emmett-Teller surface area from 26.92 to 85.12 m(2) /g. The presence of 1,4-benzenedicarboxylate moieties within the structure of MIL-53(Al) as an organic linker greatly influenced the separation of aromatic mixtures through π-π interactions. High-resolution separation was obtained for a series of alkylbenzenes (with resolution factors in the range 0.96-1.75) in less than 8 min, with 14 710 plates/m efficiency for propylbenzene, using a binary polar mobile phase of water/acetonitrile in isocratic mode. A reversed-phase separation mechanism was indicated by the increased retention factor and resolution as the water percentage in the mobile phase increased. A stability study on the composite column showed excellent mechanical stability under various conditions. The higher resolution and faster separation observed at increased temperature indicated an exothermic separation, whereas the negative values for the free energy change of transfer indicated a spontaneous process.

  15. Polydopamine-supported immobilization of covalent-organic framework-5 in capillary as stationary phase for electrochromatographic separation.

    Science.gov (United States)

    Bao, Tao; Tang, Pingxiu; Kong, Deying; Mao, Zhenkun; Chen, Zilin

    2016-05-06

    Covalent-organic frameworks (COFs) are attractive materials for their fascinating properties, such as rigid structures, exceptional thermal stabilities, low densities, and permanent porosity with specific surface areas, which indicate potential for application in chromatography similar to related metal-organic frameworks (MOFs). However, the utilization of COFs in analytical chemistry is far behind as compared to that of the MOFs due to the challenging work of their immobilization. Here, we have successfully demonstrated the growth of the boron COF-5 on the inner wall of the fused silica capillary by a developed polydopamine-supported method. Combined with the layer-by-layer strategy, multilayer COF-5-coated capillary was obtained. The formation of COF-5 on polydopamine-coated substrate has been confirmed by scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction. A novel stationary phase of COF-5 was developed on the basis of successful growth of COF-5 on polydopamine for open-tubular capillary electrochromatography (OT-CEC). Baseline separation of neutral, acidic and basic analytes was achieved on multilayer COF-5-coated capillary column. The fabricated capillary columns showed high column efficiency (154,060 plates/m for methylbenzene), excellent stability and repeatability. The precision (relative standard deviation (RSD), n=3) of retention time, peak height, and peak area for tested neutral compounds were in the range of 1.2-1.3%, 1.8-4.2%, and 0.9-2.4%, respectively. To the best of our knowledge, it was the first demonstration that COF-5 was developed as a novel stationary phase. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. A versatile polydopamine platform for facile preparation of protein stationary phase for chip-based open tubular capillary electrochromatography enantioseparation.

    Science.gov (United States)

    Liu, Chun-Ming; Liang, Ru-Ping; Wang, Xiao-Ni; Wang, Jing-Wu; Qiu, Jian-Ding

    2013-06-14

    A novel, simple, and economical method for the preparation of chiral stationary phases for chip-based enantioselective open tubular capillary electrochromatography (OT-CEC) using polydopamine (PDA) coating as an adhesive layer was reported for the first time. After the poly(dimethylsiloxane) (PDMS) microfluidic chip was filled with dopamine (DA) solution, PDA film was gradually formed and deposited on the inner wall of microchannel as permanent coating via the oxidation of DA by the oxygen dissolved in the solution. Due to possessing plentiful catechol and amine functional groups, PDA coating can serve as a versatile multifunctional platform for further secondary reactions, leading to tailoring of the coatings for protein bioconjugation by the thiols and amines via Michael addition or Schiff base reactions. Bovine serum albumin (BSA), acting as a target protein, was then stably and homogeneously immobilized in the PDA-coated PDMS microchannel to fabricate a novel protein stationary phase. Compared with the native PDMS microchannels, the modified surfaces exhibited much better wettability, more stable and enhanced electroosmotic mobility, and less nonspecific adsorption. The water contact angle and electroosmotic flow of PDA/BSA-coated PDMS substrate were measured to be 44° and 2.83×10(-4)cm(2)V(-1)s(-1), compared to those of 112° and 2.10×10(-4)cm(2)V(-1)s(-1) from the untreated one, respectively. Under a mild condition, d- and l-tryptophan were efficiently separated with a resolution of 1.68 within 130s utilizing a separation length of 37mm coupled with in-column amperometric detection on the PDA/BSA-coated PDMS microchips. This present versatile platform, facile conjugation of biomolecules onto microchip surfaces via mussel adhesive protein inspired coatings, may offer new processing strategies to prepare a biomimetic surface design on microfluidic chips, which is promising in high-throughput and complex biological analysis. Copyright © 2013 Elsevier B

  17. Histidine-modified organic-silica hybrid monolithic column for mixed-mode per aqueous and ion-exchange capillary electrochromatography.

    Science.gov (United States)

    Tang, Sheng; Liu, Shujuan; Liang, Xiaojing; Tang, Xiaofen; Wu, Xingcai; Guo, Yong; Liu, Xia; Jiang, Shengxiang

    2015-06-01

    A novel organic-silica hybrid monolith was prepared through the binding of histidine onto the surface of monolithic matrix for mixed-mode per aqueous and ion-exchange capillary electrochromatography. The imidazolium and amino groups on the surface of the monolithic stationary phase were used to generate an anodic electro-osmotic flow as well as to provide electrostatic interaction sites for the charged compounds at low pH. Typical per aqueous chromatographic behavior was observed in water-rich mobile phases. Various polar and hydrophilic analytes were selected to evaluate the characteristics and chromatographic performance of the obtained monolith. Under per aqueous conditions, the mixed-mode mechanism of hydrophobic and ion-exchange interactions was observed and the resultant monolithic column proved to be very versatile for the efficient separations of these polar and hydrophilic compounds (including amides, nucleosides and nucleotide bases, benzoic acid derivatives, and amino acids) in highly aqueous mobile phases. The successful applications suggested that the histidine-modified organic-silica hybrid monolithic column could offer a wide range of retention behaviors and flexible selectivities toward polar and hydrophilic compounds.

  18. Improvement in Liquid Chromatographic Performance of Organic Polymer Monolithic Capillary Columns with Controlled Free-Radical Polymerization.

    Science.gov (United States)

    Gama, Mariana R; Aggarwal, Pankaj; Liu, Kun; Lee, Milton L; Bottoli, Carla B G

    2016-12-19

    Capillary columns containing butyl or lauryl methacrylate monoliths were prepared using two different free-radical polymerization methods: conventional free-radical polymerization and controlled/living free-radical polymerization, both initiated thermally, and these methods were compared for the first time. Both monolith morphology and chromatographic efficiency were compared for the synthesized stationary phases using scanning electronic microscopy (SEM) and capillary liquid chromatography, respectively. Columns prepared using controlled method gave better chromatographic performance for both monomers tested. The lauryl-based monolith showed 7-fold improvement in chromatographic efficiency with a plate count of 42,000 plates/m (corrected for dead volume) for a non-retained compound. Columns fabricated using controlled polymerization appeared more homogenous radially with fused small globular morphologies, evaluated by SEM, and lower column permeability. The columns were compared with respect to resolving power of a series of alkylbenzenes under isocratic and gradient elution conditions.

  19. UV-polymerized butyl methacrylate monoliths with embedded carboxylic single-walled carbon nanotubes for CEC applications.

    Science.gov (United States)

    Navarro-Pascual-Ahuir, María; Lucena, Rafael; Cárdenas, Soledad; Ramis-Ramos, Guillermo; Valcárcel, Miguel; Herrero-Martínez, José Manuel

    2014-10-01

    The preparation of polymeric monoliths with embedded carboxy-modified single-walled carbon nanotubes (c-SWNTs) and their use for capillary electrochromatography (CEC) is described. Carbon nanotube composites were obtained by preparing a polymerization mixture in the presence of increasing c-SWNT concentrations, followed by UV initiation. The novel stationary phases were studied by optical microscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Raman spectroscopy. Using short UV-polymerization times, the optimized porogenic solvent (a binary mixture of 1,4-butanediol and 2-propanol) gave rise to polymeric beds with homogenously dispersed embedded c-SWNTs. The CEC features of these monoliths were evaluated using polycyclic aromatic hydrocarbons (PAHs), non-steroidal anti-inflammatory drugs (NSAIDs) and chiral compounds. The monolith prepared in the presence of c-SWNTs showed enhanced resolution of the text mixtures, including a remarkable capability to separate enantiomers.

  20. Kinetic performance appraisal of poly(styrene-co-divinylbenzene) monolithic high-performance liquid chromatography columns for biomolecule analysis.

    Science.gov (United States)

    Causon, Tim J; Shellie, Robert A; Hilder, Emily F

    2010-06-11

    A broad appraisal of the kinetic performance of organic polymeric monolithic columns is reported using commercially available poly(styrene-co-divinylbenzene) monolithic columns (Dionex ProSwift RP-1S). Analysis of a protein digest sample at elevated temperatures (> or =80 degrees C) indicated no apparent analyte degradation using an inert polymeric stationary phase. Comparison between low molecular weight solute and peptide separations highlighted the markedly different mass transport processes observed on macroporous monolithic beds and an improved C term at elevated temperature in both instances. The current usefulness of this column format for biomolecule analysis was further studied via employment of a kinetic performance characterisation for the first time to provide direction for column development servicing this application.

  1. High-performance liquid chromatographic separation of rolipram, bupivacaine and omeprazole using a tartardiamide-based stationary phase influence of flow rate and temperature on the enantioseparation.

    Science.gov (United States)

    da Silva Junior, Ivanildo José; Sartor, João Paulo; Rosa, Paulo César Pires; de Veredas, Vinícius; Barreto Júnior, Amaro Gomes; Santana, Cesar Costapinto

    2007-08-24

    Chromatographic separation of the chiral drugs rolipram, bupivacaine and omeprazole on a tartardiamide-based stationary phase commercially named Kromasil CHI-TBB is shown in this work. The effect of temperature on the chromatographic separation of the chiral drugs using the Kromasil CHI-TBB stationary phase was determined quantitatively so as to contribute toward the design for the racemic mixtures of the named compound by using chiral columns. A decrease in the retention and selectivity factors was observed, when the column temperature increased. Van't Hoff plots provided the thermodynamic data. The variation of the thermodynamic parameters enthalpy and entropy are clearly negative meaning that the separation is enthalpy controlled.

  2. Silica-based 2-(N,N-dimethylamino)-1,3-propanediol hydrophilic interaction liquid chromatography stationary phase for separating cephalosporins and carbapenems.

    Science.gov (United States)

    Yin, Wei; Cheng, Lingping; Chai, Huihui; Guo, Ruiqiang; Liu, Renhua; Chu, Changhu; Palasota, John A; Cai, Xiaohui

    2015-08-01

    A silica-based stationary phase bearing both hydrophilic hydroxyl and amino groups was developed by covalently bonding a small molecular N,N-dimethylamino 1,3-propanediol moiety onto silica beads via copper(I)-catalyzed Huisgen azide-alkyne 1,3-dipolar cycloaddition (CuAAC). This new stationary phase showed good HILIC characteristics and high column efficiency (the theoretical plate number is up to 37000 plates m(-1) in the case of inosine) in the separation of polar compounds, such as nucleosides and bases, organic acids, cephalosporins, and carbapenems.

  3. Preparation of a silica stationary phase co-functionalized with Wulff-type phenylboronate and C12 for mixed-mode liquid chromatography.

    Science.gov (United States)

    Li, Hengye; Zhang, Xuemeng; Zhang, Lin; Wang, Xiaojin; Kong, Fenying; Fan, Dahe; Li, Lei; Wang, Wei

    2017-04-15

    A silica stationary phase was designed and synthesized through the co-functionalization of silica with Wulff-type phenylboronate and C12 for mixed-mode liquid chromatography applications. The as-synthesized stationary phase was characterized by elemental analysis and Fourier Transform-InfraRed Spectroscopy (FT-IR). Retention mechanisms, including boronate affinity (BA), reversed-phase (RP) and anion-exchange (AE), were involved. Retention mechanism switching was easily realized by adjustment of the mobile phase constitution. Cis-diol compounds could be selectively captured under neutral conditions in BA mode and off-line separated in RP mode. Neutral, basic, acidic and amphiprotic compounds were chromatographed on the column in RP chromatography, while inorganic anions were chromatographed in AE chromatography to characterize the mixed-mode nature of the prepared stationary phase. In addition, the RP performance was compared with an octadecyl silica column in terms of column efficiency (N/m), asymmetry factor (Af), retention factor (k) and resolution (Rs). The prepared stationary phase offered multiple interactions with analytes in addition to hydrophobic interactions under RP elution conditions. Based on the mixed-mode properties, off-line 2D-LC, for selective capture and separation of urinary nucleosides, was successfully realized on a single column, demonstrating its powerful application potential for complex samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Strong cation exchange monoliths for HPLC by Reactive Gelation.

    Science.gov (United States)

    Brand, Bastian; Krättli, Martin; Storti, Giuseppe; Morbidelli, Massimo

    2011-08-01

    Polymeric monolithic stationary phases for HPLC can be produced by Reactive Gelation. Unlike the conventional method of using porogens, such novel process consists of a number of separate steps, thus enabling a better control of the quality of the final material. A suspension of polymer nanoparticles in water is produced and subsequently swollen with hydrophobic monomers. The particles are then destabilised (usually by salt addition) to make them aggregate into a large percolating structure, the so-called monolith. Finally, the added monomer can then be polymerised to harden the structure. In this work, a polystyrene latex is used as the base material and functionalised by introduction of epoxide groups on the surface and subsequent reaction to sulphonic acid groups, yielding a SO3(-) density of 0.7 mmol/g dry material. Morphological investigations show 54% porosity made of 300 nm large pores. Van Deemter measurements of a large protein show no practical influence of diffusion limitations on the plate number. Finally, a preliminary separation of a test protein mixture is shown, demonstrating the potential of using ion-exchange chromatography on Reactive Gelation monoliths.

  5. Two-Dimensional Micro-TLC Phenolic Fingerprints of Selected Mentha sp. on Cyano-Bonded Polar Stationary Phase.

    Science.gov (United States)

    Hawrył, Mirosław A; Niemiec, Małgorzata A; Słomka, Kamil; Waksmundzka-Hajnos, Monika; Szymczak, Grażyna

    2016-01-01

    Micro-thin-layer chromatography in two-dimensional (2D-mTLC) mode in normal- (NP) and reversed-phase (RP) systems by use of cyanopropyl-bonded stationary phases was applied to make fingerprints of 11 species of Mentha genus and two finished pharmaceutical products. Non-aqueous eluents were used in the NP systems. Mixtures of acetonitrile with water and methanol with water were used in the RP chromatographic systems. Optimization of one-dimensional systems was performed by determining RM vs. composition of mobile phase dependencies for standards occurring in various Mentha sp. On the basis of these dependencies, the most selective chromatographic systems for each run were chosen. Then most selective eluents were applied to optimize two-dimensional systems by creating RF in NP systems vs. RF in RP systems correlations. The best two-dimensional systems were chosen on the basis of R(2) values for RF vs. RF correlations (the lowest values of R(2) coefficients). The 2D-mTLC optimized systems were applied to separate phenolic compounds and make fingerprints of the examined plant materials.

  6. Chromatographic analysis of some drugs employed in erectile dysfunction therapy: qualitative and quantitative studies using calixarene stationary phase.

    Science.gov (United States)

    Hashem, Hisham; Ibrahim, Adel Ehab; Elhenawee, Magda

    2014-10-01

    In this study, the effect of change in chromatographic process variables on the retention behavior of four drugs employed in erectile dysfunction therapy on a calixarene stationary phase is described. Three of these drugs are known to treat erectile dysfunction, namely, sildenafil citrate, tadalafil, and apomorphine hydrochloride, and one drug that is used as opioid analgesic, tramadol hydrochloride, which is quiet widely misused to treat premature ejaculation. The results indicate the importance of considering the structure and pKa values of drugs to be separated along with mobile phase composition. A new optimized, rapid, and accurate liquid chromatography method is also established for simultaneous determination of sildenafil citrate, tadalafil, and apomorphine hydrochloride in pharmaceutical preparations and bulk powders. The chromatographic separation of the three pharmaceuticals was achieved on a calixarene column in less than 10 min using a binary mobile phase of 35% acetonitrile and 65% 50 mM sodium perchlorate pH2.5 at 1 mL/min flow rate. The method was validated for system efficiency, linearity, accuracy, precision, limits of detection and quantitation, specificity, stability, and robustness. Statistical analysis proved that the method enabled reproducible and selective quantification of all three analytes in bulk drugs and in pharmaceutical preparations.

  7. Characterization and classification of pseudo-stationary phases in micellar electrokinetic chromatography using chemometric methods.

    Science.gov (United States)

    Fu, Cexiong; Khaledi, Morteza G

    2014-03-04

    Two types of chemometric methods, principal component analysis (PCA) and cluster analysis, are employed to characterize and classify a total of 70 pseudostationary phases (54 distinct systems and 16 decoy systems) in micellar electrokinetic chromatography (MEKC). PCA excels at removing redundant information for micellar phase characterization and retaining principal determinants for phase classification. While PCA is useful in the characterization of micelle selectivities, it is ineffective in defining the grouping of micellar phases. Hierarchical clustering yields a complete dendrogram of cluster structures but provides only limited cluster characterizations. The combination of these two chemometric methods leads to a comprehensive interpretation of the micellar phase classification. Moreover, the k-means analysis can further discern subtle differences among those closely located micellar phases. All three chemometric methods result in similar classifications with respect to the similarities and differences of the 70 micelle systems investigated. These systems are categorized into 3 major clusters: fluoro-surfactants represent cluster I, identified as strong hydrogen bond donors and dipolar but weak hydrogen bond acceptors. Cluster II includes sulfonated acrylamide/acrylate copolymers and surfactants with trimethylammonium head groups, characterized by strong hydrophobicity (v) and weak hydrogen bond acidity (b). The last cluster consists of two subclusters: clusters III and IV. Cluster III includes siloxane-based polymeric micelles, exhibiting weak hydrophobicity and medium hydrogen bond acidity and basicity (a), and the cluster IV micellar systems are characterized by their strong hydrophobicity and medium hydrogen bond acidity and basicity but rather weak dipolarity. Cluster III differs from cluster IV by its slightly weaker hydrophobicity and hydrogen bond donating capability. The classification by chemometric methods is in good agreement with the

  8. Chiral anion exchangers applied to capillary electrochromatography enantioseparation of oppositely charged chiral analytes: investigation of stationary and mobile phase parameters.

    Science.gov (United States)

    Lämmerhofer, M; Tobler, E; Lindner, W

    2000-07-28

    Weak anion-exchange (WAX) type chiral stationary phases (CSPs) based on tert.-butyl carbamoyl quinine as chiral selector (SO) and different types of silica particles (porous and non-porous) as chromatographic support are evaluated in packed capillary electrochromatography (CEC). Their ability to resolve the enantiomers of negatively charged chiral analytes, e.g., N-derivatized amino acids, in the anion-exchange mode and their electrochromatographic characteristics are described in dependence of several mobile phase parameters (pH, buffer type and concentration, organic modifier type and concentration) and other experimental variables (electric field strength, capillary temperature). The inherent "zwitterionic" surface character of such silica-based WAX type CSPs (positively charged SO and negatively charged residual silanols) allows the reversal of the electroosmotic flow (EOF) towards the anode at pH values below the isoelectric point (pI) of the modified surface, whereas a cathodic EOF results at pH values above the pI. Since for negatively charged analytes also an electrophoretic transport increment has to be considered, which can be either in or against the EOF direction, several distinct modes of elution have been observed under different stationary phase and mobile phase conditions: (i) co-electrophoretic elution of the negatively charged solutes with the anodic EOF in the negative polarity mode, (ii) counter-electrophoretic elution with the cathodic EOF in the positive polarity mode, and (iii) electrophoretically dominated elution in the negative polarity mode with a cathodic EOF directed to the injection end of the capillary. Useful enantioseparations of chiral acids have been obtained with all three modes. Enantioselectivity values as high as under pressure-driven conditions and theoretical plate numbers up to 120000 per meter could be achieved under electrically driven conditions. A repeatability study yielded RSD values below 2% for retention times and

  9. COVALENTLY BONDING CHIRAL POLYURETHANE ON AMINATED SILICA GEL: A NEW STRATEGY TO PREPARE CHIRAL STATIONARY PHASE FOR HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Institute of Scientific and Technical Information of China (English)

    Shao-hua Huang; Zheng-wu Bai; Chuan-qi Yin; Shi-rong Li

    2006-01-01

    Two polyurethanes of different molecular weights were prepared by the copolymerization of phenyl diisocyanate and diisopropyl tartrate. The polyurethanes having terminal isocyanate groups were reacted with 3-aminopropyl silica gel to afford two chiral stationary phases. The (M-)n of the two polyurethanes were 4057 g/mol and 6442 g/mol. The polyurethanes and the corresponding chiral stationary phases were characterized by FT-IR, 1H NMR and elemental analysis. The loading capacities of the polyurethanes on silica gel were 0.68 mmol units/g and 0.61 mmol units/g, respectively. The separation performance and the influence of additives, triethylamine and trichloroacetic acid, on the separation of chiral compounds were investigated by HPLC. The chiral stationary phase prepared from polyurethane with (M-)n of 4057 g/mol demonstrated better enantioseparation capability than that with (M-)n of 6442 g/mol. Additionally, it was found that the addition of triethylamine and trichloroacetic acid in the mobile phases significantly improved the enantioseparation for these two chiral stationary phases.

  10. Miniaturized monolithic columns for the electrochromatographic separation and SERS detection of molecules of exobiological interest

    Science.gov (United States)

    Carbonnier, Benjamin; Guerrouache, Mohamed

    Development of miniaturized separation and detection media represents one of the major challenges in the field of modern analytical chemistry dedicated to space exploration. To date, gas chromatography-mass spectrometry has been selected as the method of choice for exobiology flight experiments for seeking for organic molecules and especially potential chemical indicators of life. [1] Liquid phase separation methods have also been developed with for instance, the so-called Mars Organic Analyzer (MOA) capillary electrophoresis (CE) microchip.[2] Although CE offers the advantages of easy miniaturization and high separation efficiency it suffers from a lack of selectivity towards a broad range of analytes with varied chemical nature. In this respect, we propose the use of capillary columns filled with monolithic stationary phases for the electrochromatographic separation of organic molecules of exobiology interest. Polymer monoliths have attracted a great deal of interest in analytical science over the last years as (electro)chromatographic stationary phases [3], immunosensors [4]. Beyond the intrinsic properties of monolithic polymers, i.e. fast mass transport between the monolithic support and the surrounding fluid and high permeability, other major advantages are their easy in situ preparation and tuning of surface functionality. Indeed, monoliths can be simply prepared through free radical copolymerization of a homogeneous mixture made of monomers, cross-linkers, porogenic solvents and initiator. UV-initiation process has been exploited to the synthesis of a discrete section of monolith as a flow-through active element within the confines of micro channels [5,6] while two-step strategies have been reported for the design of varied adsorbent starting with a generic monolith [7,8]. Although a nearly limitless range of monolithic supports can be prepared by this traditional method, the resulting monoliths exhibit unique function. In this contribution, we describe an

  11. Comprehensive profiling of ribonucleosides modification by affinity zirconium oxide-silica composite monolithic column online solid-phase microextraction - Mass spectrometry analysis.

    Science.gov (United States)

    Jiang, Han-Peng; Chu, Jie-Mei; Lan, Meng-Dan; Liu, Ping; Yang, Na; Zheng, Fang; Yuan, Bi-Feng; Feng, Yu-Qi

    2016-09-02

    More than 140 modified ribonucleosides have been identified in RNA. Determination of endogenous modified ribonucleosides in biological fluids may serve as non-invasive disease diagnostic strategy. However, detection of the modified ribonucleosides in biological fluids is challenging, especially for the low abundant modified ribonucleosides due to the serious matrix interferences of biological fluids. Here, we developed a facile preparation strategy and successfully synthesized zirconium oxide-silica (ZrO2/SiO2) composite capillary monolithic column that exhibited excellent performance for the selective enrichment of cis-diol-containing compounds. Compared with the boronate-based affinity monolith, the ZrO2/SiO2 monolith showed ∼2 orders of magnitude higher extraction capacity and can be used under physiological pH (pH 6.5-7.5). Using the prepared ZrO2/SiO2 composite monolith as the trapping column and reversed-phase C18 column as the analytical column, we further established an online solid-phase microextraction (SPME) in combination with liquid chromatography-mass spectrometry (online SPME-LC-MS/MS) analysis for the comprehensive profiling of ribonucleosides modification in human urine. Our results showed that 68 cis-diol-containing ribosylated compounds were identified in human urine, which is, to the best of our knowledge, the highest numbers of cis-diol-containing compounds were determined in a single analysis. It is worth noting that four modified ribonucleosides were discovered in the human urine for the first time. In addition, the quantification results from the pooled urine samples showed that compared to healthy controls, the contents of sixteen ribose conjugates in the urine of gastric cancer, eleven in esophagus cancer and seven in lymphoma increased more than two folds. Among these ribose conjugates, four ribose conjugates increased more than two folds in both gastric cancer and esophagus cancer; three ribose conjugates increased more than two

  12. Aluminum based metal-organic framework-polymer monolith in solid-phase microextraction of penicillins in river water and milk samples.

    Science.gov (United States)

    Lirio, Stephen; Liu, Wan-Ling; Lin, Chen-Lan; Lin, Chia-Her; Huang, Hsi-Ya

    2016-01-08

    In this study, aluminum based metal-organic framework (Al-MOF)-organic polymer monoliths were prepared via microwave-assisted polymerization of ethylene dimethacrylate (EDMA), butyl methacrylate (BMA) with different weight percentages of Al-MOF (MIL-53; 37.5-62.5%) and subsequently utilized as sorbent in solid-phase microextraction (SPME) of penicillins (penicillin G, penicillin V, oxacillin, cloxacillin, dicloxacillin, nafcillin). The Al-MOF-polymer was characterized using Fourier transform infrared (FTIR) spectroscopy, powder X-ray diffraction (XRD), scanning electron microscopy (SEM) and SEM-energy-dispersive X-ray spectroscopy (SEM-EDS) to clarify the retained crystalline structure well as the homogeneous dispersion of Al-MOF (MIL-53) in polymer monolith. The developed Al-MOF-polymer (MIL-53) monolithic column was evaluated according to its extraction recovery of penicillins. Several parameters affecting the extraction recoveries of penicillins using fabricated Al-MOF-polymer (MIL-53) monolithic column including different MIL-53 weight percentages, column length, pH, desorption solvent, and mobile phase flow rate were investigated. For comparison, different Al-based MOFs (MIL-68, CYCU-4 and DUT-5) were fabricated using the optimized condition for MIL-53-polymer (sample matrix at pH 3, 200μL desorption volume using methanol, 37.5% of MOF, 4-cm column length at 0.100mLmin(-1) flow rate). Among all the Al-MOF-polymers, MIL-53(Al)-polymer still afforded the best extraction recovery for penicillins ranging from 90.5 to 95.7% for intra-day with less than 3.5% relative standard deviations (RSDs) and inter-day precision were in the range of 90.7-97.6% with less than 4.2% RSDs. Meanwhile, the recoveries for column-to-column were in the range of 89.5-93.5% (polymer was applied for the extraction of penicillin in river water and milk by spiking trace-level penicillin for as low as 50μgL(-1) and 100μgL(-1) with recoveries ranging from 80.8% to 90.9% (<6.7% RSDs) in

  13. Comparison of fully wettable RPLC stationary phases for LC-MS-based cellular metabolomics.

    Science.gov (United States)

    Si-Hung, Le; Causon, Tim J; Hann, Stephan

    2017-07-10

    Reversed-phase LC combined with high-resolution mass spectrometry (HRMS) is one of the most popular methods for cellular metabolomics studies. Due to the difficulties in analyzing a wide range of polarities encountered in the metabolome, 100%-wettable reversed-phase materials are frequently used to maximize metabolome coverage within a single analysis. Packed with silica-based sub-3 μm diameter particles, these columns allow high separation efficiency and offer a reasonable compromise for metabolome coverage within a single analysis. While direct performance comparison can be made using classical chromatographic characterization approaches, a comprehensive assessment of the column's performance for cellular metabolomics requires use of a full LC-HRMS workflow in order to reflect realistic study conditions used for cellular metabolomics. In this study, a comparison of several reversed-phase LC columns for metabolome analysis using such a dedicated workflow is presented. All columns were tested under the same analytical conditions on an LC-TOF-MS platform using a variety of authentic metabolite standards and biotechnologically relevant yeast cell extracts. Data on total workflow performance including retention behavior, peak capacity, coverage, and molecular feature extraction repeatability from these columns are presented with consideration for both nontargeted screening and differential metabolomics workflows using authentic standards and Pichia pastoris cell extract samples. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. The primary transcriptome of Salmonella enterica Serovar Typhimurium and its dependence on ppGpp during late stationary phase.

    Directory of Open Access Journals (Sweden)

    Vinoy K Ramachandran

    Full Text Available We have used differential RNA-seq (dRNA-seq to characterise the transcriptomic architecture of S. Typhimurium SL1344, and its dependence on the bacterial alarmone, guanosine tetraphosphate (ppGpp during late stationary phase, (LSP. Under LSP conditions we were able to identify the transcriptional start sites (TSSs for 53% of the S. Typhimurium open reading frames (ORFs and discovered 282 candidate non-coding RNAs (ncRNAs. The mapping of LSP TSSs enabled a detailed comparison with a previous dRNA-seq study of the early stationary phase (ESP transcriptional architecture of S. Typhimurium SL1344 and its dependence on ppGpp. For the purposes of this study, LSP was defined as an aerobic LB culture grown to a later optical density reading (OD600 = 3.6 compared to ESP (OD600 = 2.3. The precise nucleotide positions of the majority of S. Typhimurium TSSs at LSP agreed closely with those identified at ESP. However, the identification of TSSs at different positions, or where additional or fewer TSSs were found at LSP compared to ESP enabled the genome-wide categorisation of growth phase dependent changes in promoter structure, the first time such an analysis has been done on this scale. Comparison of the ppGpp-dependency LSP and ESP TSSs for mRNAs and ncRNAs revealed a similar breadth of ppGpp-activation and repression. However, we note several ncRNAs previously shown to be involved in virulence were highly ppGpp-dependent at LSP. Finally, although SPI1 was expressed at ESP, we found SPI1 was not as highly expressed at LSP, instead we observed elevated expression of SPI2 encoded genes. We therefore also report an analysis of SPI2 transcriptional architecture at LSP resulting in localisation of SsrB binding sites and identification of a previously unreported SPI2 TSS. We also show that ppGpp is required for nearly all of SPI2 expression at LSP as well as for expression of SPI1 at ESP.

  15. The primary transcriptome of Salmonella enterica Serovar Typhimurium and its dependence on ppGpp during late stationary phase.

    Science.gov (United States)

    Ramachandran, Vinoy K; Shearer, Neil; Thompson, Arthur

    2014-01-01

    We have used differential RNA-seq (dRNA-seq) to characterise the transcriptomic architecture of S. Typhimurium SL1344, and its dependence on the bacterial alarmone, guanosine tetraphosphate (ppGpp) during late stationary phase, (LSP). Under LSP conditions we were able to identify the transcriptional start sites (TSSs) for 53% of the S. Typhimurium open reading frames (ORFs) and discovered 282 candidate non-coding RNAs (ncRNAs). The mapping of LSP TSSs enabled a detailed comparison with a previous dRNA-seq study of the early stationary phase (ESP) transcriptional architecture of S. Typhimurium SL1344 and its dependence on ppGpp. For the purposes of this study, LSP was defined as an aerobic LB culture grown to a later optical density reading (OD600 = 3.6) compared to ESP (OD600 = 2.3). The precise nucleotide positions of the majority of S. Typhimurium TSSs at LSP agreed closely with those identified at ESP. However, the identification of TSSs at different positions, or where additional or fewer TSSs were found at LSP compared to ESP enabled the genome-wide categorisation of growth phase dependent changes in promoter structure, the first time such an analysis has been done on this scale. Comparison of the ppGpp-dependency LSP and ESP TSSs for mRNAs and ncRNAs revealed a similar breadth of ppGpp-activation and repression. However, we note several ncRNAs previously shown to be involved in virulence were highly ppGpp-dependent at LSP. Finally, although SPI1 was expressed at ESP, we found SPI1 was not as highly expressed at LSP, instead we observed elevated expression of SPI2 encoded genes. We therefore also report an analysis of SPI2 transcriptional architecture at LSP resulting in localisation of SsrB binding sites and identification of a previously unreported SPI2 TSS. We also show that ppGpp is required for nearly all of SPI2 expression at LSP as well as for expression of SPI1 at ESP.

  16. Determination of imidazole derivatives by micellar electrokinetic chromatography combined with solid-phase microextraction using activated carbon-polymer monolith as adsorbent.

    Science.gov (United States)

    Shih, Yung-Han; Lirio, Stephen; Li, Chih-Keng; Liu, Wan-Ling; Huang, Hsi-Ya

    2016-01-08

    In this study, an effective method for the separation of imidazole derivatives 2-methylimidazole (2-MEI), 4- methylimidazole (4-MEI) and 2-acetyl-4-tetrahydroxybutylimidazole (THI) in caramel colors using cation-selective exhaustive injection and sweeping micellar electrokinetic chromatography (CSEI-sweeping-MEKC) was developed. The limits of detection (LOD) and quantitation (LOQ) for the CSEI-sweeping-MEKC method were in the range of 4.3-80μgL(-1) and 14-270μgL(-1), respectively. Meanwhile, a rapid fabrication activated carbon-polymer (AC-polymer) monolithic column as adsorbent for solid-phase microextraction (SPME) of imidazole colors was developed. Under the optimized SPME condition, the extraction recoveries for intra-day, inter-day and column-to-column were in the range of 84.5-95.1% (<6.3% RSDs), 85.6-96.1% (<4.9% RSDs), and 81.3-96.1% (<7.1% RSDs), respectively. The LODs and LOQs of AC-polymer monolithic column combined with CSEI-sweeping-MEKC method were in the range of 33.4-60.4μgL(-1) and 111.7-201.2μgL(-1), respectively. The use of AC-polymer as SPME adsorbent demonstrated the reduction of matrix effect in food samples such as soft drink and alcoholic beverage thereby benefiting successful determination of trace-level caramel colors residues using CSEI-sweeping-MEKC method. The developed AC-polymer monolithic column can be reused for more than 30 times without any significant loss in the extraction recovery for imidazole derivatives.

  17. Effective extraction of triazines from environmental water samples using magnetism-enhanced monolith-based in-tube solid phase microextraction.

    Science.gov (United States)

    Mei, Meng; Huang, Xiaojia; Yang, Xiaodong; Luo, Qing

    2016-09-21

    This article reports on the effective extraction of triazines from environmental water samples using magnetism-enhanced monolith-based in-tube solid phase microextraction (ME-MB/IT-SPME). Firstly, monolithic poly (octyl methacrylate-co-ethyleneglycol dimethacrylate) capillary column doped with magnetic nanoparticles was synthesized inside a fused silica. After that, the monolithic capillary column was placed inside a magnetic coil that allowed the exertion of a variable magnetic field during adsorption and desorption steps. The effects of intensity of magnetic field, adsorption and desorption flow rate, volume of sample and desorption solvent, pH value and ionic strength in sample matrix on the performance of ME-MB/IT-SPME for triazines were investigated in details. Under the optimized conditions, the developed ME-MB/IT-SPME showed satisfactory quantitative extraction efficiencies of the target analytes between 64.8% and 99.7%. At the same time, the ME-MB/IT-SPME was combined with high-performance liquid chromatography with diode array detection to detect six triazines in water samples. The limits of detection (S/N = 3) and limits of quantification (S/N = 10) were in the ranges of 0.074-0.23 μg/L and 0.24-0.68 μg/L, respectively. The precision of the proposed method was evaluated in terms of intra- and inter-assay variability calculated as relative standard deviation, and it was found that the values were all below 10%. Finally, the developed method was successfully applied for environmental water samples such as farmland, lake and river water with spiked recoveries in the range of 70.7-119%.

  18. A novel ionic liquid-modified organic-polymer monolith as the sorbent for in-tube solid-phase microextraction of acidic food additives.

    Science.gov (United States)

    Wang, Ting-Ting; Chen, Yi-Hui; Ma, Jun-Feng; Hu, Min-Jie; Li, Ying; Fang, Jiang-Hua; Gao, Hao-Qi

    2014-08-01

    A novel ionic liquid-modified organic-polymer monolithic capillary column was prepared and used for in-tube solid-phase microextraction (SPME) of acidic food additives. The primary amino group of 1-aminopropyl-3-methylimidazolium chloride was reacted with the epoxide group of glycidyl methacrylate. The as-prepared new monomer was then copolymerized in situ with acrylamide and N,N'-methylenebisacrylamide in the presence of polyethylene glycol (PEG)-8000 and PEG-10,000 as porogens. The extraction performance of the developed monolithic sorbent was evaluated for benzoic acid, 3-hydroxybenzoic acid, cinnamic acid, 2,4-dichlorophenoxyacetic acid, and 3-(trifluoromethyl)-cinnamic acid. Such a sorbent, bearing hydrophobic and anion-exchange groups, had high extraction efficiency towards the test compounds. The adsorption capacities for the analytes dissolved in water ranged from 0.18 to 1.74 μg cm(-1). Good linear calibration curves (R(2) > 0.99) were obtained, and the limits of detection (S/N = 3) for the analytes were found to be in the range 1.2-13.5 ng mL(-1). The recoveries of five acidic food additives spiked in Coca-Cola beverage samples ranged from 85.4 % to 98.3 %, with RSD less than 6.9 %. The excellent applicability of the ionic liquid (IL)-modified monolithic column was further tested by the determination of benzoic acid content in Sprite samples, further illustrating its good potential for analyzing food additives in complex samples.

  19. Preparation and evaluation of open tubular C18-silica monolithic microcartridges for preconcentration of peptides by on-line solid phase extraction capillary electrophoresis.

    Science.gov (United States)

    Ortiz-Villanueva, Elena; Benavente, Fernando; Giménez, Estela; Yilmaz, Fatma; Sanz-Nebot, Victoria

    2014-10-10

    In this study, C18-silica monoliths were synthesized as a porous layer in open tubular capillary columns, to be cut later into microcartridges for the analysis of neuropeptides by on-line solid-phase extraction capillary electrophoresis with UV and MS detection (SPE-CE-UV and SPE-CE-MS). First, several types of C18-silica monolithic (MtC18) microcartridges were used to analyse standard solutions of five neuropeptides (i.e. dynorphin A (1-7), substance P (7-11), endomorphin 1, methionine enkephalin and [Ala]-methionine enkephalin). The MtC18 sorbents were especially selective against endomorphin 1 and substance P (7-11)). The best results in terms of sensitivity and inter-microcartridge reproducibility were achieved with the microcartridges obtained from a 10-cm open tubular capillary column with a thin monolithic coating with large through-pores (1-5μm). Run-to-run repeatability, microcartridge durability, linearity ranges and LODs were studied by MtC18-SPE-CE-MS. As expected due to their greater selectivity, the best LOD enhancement was obtained for End1 and SP (7-11) (50 times with regard to CE-MS). Finally, the suitability of the methodology for analysing biological fluids was tested with plasma samples spiked with End1 and SP (7-11). Results obtained were promising because both neuropeptides could be detected at 0.05μgmL(-1), which was almost the same concentration level as for the standard solutions (0.01μgmL(-1)).

  20. Enhanced resolution of Mentha piperita volatile fraction using a novel medium-polarity ionic liquid gas chromatography stationary phase.

    Science.gov (United States)

    Ragonese, Carla; Sciarrone, Danilo; Grasso, Elisa; Dugo, Paola; Mondello, Luigi

    2016-02-01

    The evaluation of a novel medium-polarity ionic-liquid-based gas chromatography column, SLB-IL60, towards the analysis of a complex essential oil, namely, a peppermint essential oil sample, is reported. The SLB-IL60 30 m column was subjected to bleeding measurements, by means of conventional gas chromatography with mass spectrometry. The SLB-IL60 column was then evaluated in the analysis of pure standard compounds, chosen as typical constituents of peppermint essential oil. Resolution and peak symmetry (expressed as tailing factors at 10% of peak height) were measured and the results were compared to those obtained on the most widely used columns in such an application, namely a medium-polarity [100% poly(ethyleneglycol)] stationary phase, and an apolar 5% diphenyl/95% dimethyl siloxane. The final part of the evaluation was dedicated to the gas chromatography with mass spectrometry analysis of a peppermint essential oil sample and again the data were compared to those obtained on the 100% poly(ethyleneglycol) and the 5% diphenyl/95% dimethyl siloxane phase. Linear retention indices were determined for all the identified components on the ionic liquid capillary. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Enantioseparation Using Cellulose Tris(3,5-dimethylphenylcarbamate as Chiral Stationary Phase for HPLC: Influence of Molecular Weight of Cellulose

    Directory of Open Access Journals (Sweden)

    Yuji Okada

    2016-11-01

    Full Text Available The cellulose oligomers with different degrees of polymerization (DP, 7, 11, 18, 24, 26, 40 and 52, were prepared by hydrolysis of microcrystalline cellulose with phosphoric acid. These oligomers including the starting microcrystalline cellulose (DP 124 were converted to tris(3,5-dimethylphenylcarbamate (CDMPC derivatives by the reaction with an excess of 3,5-dimethylphenyl isocyanate to be used as the chiral stationary phase (CSP in high-performance liquid chromatography (HPLC. The structures of the CDMPC derivatives were investigated by infrared spectroscopy (IR, 1H-NMR, circular dichroism (CD and size exclusion chromatography (SEC, and the DPs of the derivatives estimated by SEC agreed with those estimated by 1H-NMR. After coating the derivatives on silica gel, their chiral recognition abilities were evaluated using eight racemates under a normal phase condition with a hexane-2-propanol (99/1 mixture as an eluent. The chiral recognition abilities of 7- and 11-mers, particularly the former, were lower than those of the higher oligomers from DP 18 to 52, which had rather similar abilities to that of 124-mer, although the abilities depended on the racemates. DP 18 seems to be sufficient for CDMPC to exhibit chiral recognition similar to that of the CDMPC with larger DPs.

  2. Modeling, analysis, and design of stationary reference frame droop controlled parallel three-phase voltage source inverters

    DEFF Research Database (Denmark)

    Vasquez, Juan Carlos; Guerrero, Josep M.; Savaghebi, Mehdi;

    2013-01-01

    Power electronics based MicroGrids consist of a number of voltage source inverters (VSIs) operating in parallel. In this paper, the modeling, control design, and stability analysis of parallel connected three-phase VSIs are derived. The proposed voltage and current inner control loops and the mat......Power electronics based MicroGrids consist of a number of voltage source inverters (VSIs) operating in parallel. In this paper, the modeling, control design, and stability analysis of parallel connected three-phase VSIs are derived. The proposed voltage and current inner control loops...... and the mathematical models of the VSIs are based on the stationary reference frame. A hierarchical control scheme for the paralleled VSI system is developed comprising two levels. The primary control includes the droop method and the virtual impedance loops, in order to share active and reactive power. The secondary...... control restores the frequency and amplitude deviations produced by the primary control. Also, a synchronization algorithm is presented in order to connect the MicroGrid to the grid. Experimental results are provided to validate the performance and robustness of the parallel VSI system control...

  3. The effect of acidic and basic additives on the enantioseparation of basic drugs using polysaccharide-based chiral stationary phases.

    Science.gov (United States)

    Ye, Yun K; Stringham, Rodger W

    2006-08-01

    The enantioseparation of nine commercially available basic drugs was achieved on polysaccharide-based chiral stationary phases with the acidic additive ethanesulfonic acid and the basic additive butylamine. Seven different commercially available CSPs were used for the study (AD, AS, OD, OJ, OG, OB, and OC). Mobile phase additives have been proven to be essential in obtaining satisfactory enantio-resolution in terms of both efficiency and selectivity. Significantly improved selectivities were obtained for the basic probe drugs with the acidic additive, ethanesulfonic acid, rather than the basic additive, butylamine. This is best seen with Chiralpak AS CSP. No enantioseparation for the nine drugs was observed when butylamine was used as an additive; however, satisfactory enantioseparation for the nine drugs was achieved using ethanesulfonic acid. Higher column efficiencies were observed with the acidic additive, especially when isopropanol was used as a modifier. Higher sensitivity was also achieved with ethanesulfonic acid because of the significantly lower background at the UV detection wavelength. The acidic additive was demonstrated to be superior to the basic additive for the enantioseparation of basic drugs using seven different polysaccharide-based CSPs. These results are counterintuitive to the common "rule of thumb" in enantioseparation that states acidic additives work best for acidic analytes and basic additives work best for basic analytes. The beneficial effects of acidic additive in enantioseparations observed in this study could significantly improve the applicability of polysaccharide-based CSPs for the enantioseparation of basic analytes.

  4. Time-Frequency Integrals and the Stationary Phase Method in Problems of Waves Propagation from Moving Sources

    Directory of Open Access Journals (Sweden)

    Gennadiy Burlak

    2012-12-01

    Full Text Available The time-frequency integrals and the two-dimensional stationary phase method are applied to study the electromagnetic waves radiated by moving modulated sources in dispersive media. We show that such unified approach leads to explicit expressions for the field amplitudes and simple relations for the field eigenfrequencies and the retardation time that become the coupled variables. The main features of the technique are illustrated by examples of the moving source fields in the plasma and the Cherenkov radiation. It is emphasized that the deeper insight to the wave effects in dispersive case already requires the explicit formulation of the dispersive material model. As the advanced application we have considered the Doppler frequency shift in a complex single-resonant dispersive metamaterial (Lorenz model where in some frequency ranges the negativity of the real part of the refraction index can be reached. We have demonstrated that in dispersive case the Doppler frequency shift acquires a nonlinear dependence on the modulating frequency of the radiated particle. The detailed frequency dependence of such a shift and spectral behavior of phase and group velocities (that have the opposite directions are studied numerically.

  5. Enantioselective determination of protein amino acids in fertilizers by liquid chromatography-tandem mass spectrometry on chiral teicoplanin stationary phase.

    Science.gov (United States)

    Taujenis, Lukas; Olšauskaitė, Vilma; Padarauskas, Audrius

    2014-11-19

    High-performance liquid chromatography on a glycopeptide antibiotic teicoplanin-based chiral stationary phase coupled with tandem mass spectrometry was developed for fast and reliable enantioseparation and determination of protein amino acids in hydrolyzed fertilizer samples. The effect of the mobile phase parameters (type and content of organic modifier and pH) and the column temperature on the enantioselectivity was investigated. Under optimized conditions, the majority (15 of 19) of d/l-amino acid pairs were resolved with a resolution factor (Rs) higher than 1.5 with a run time of 15 min. A triple quadrupole tandem mass spectrometer operating in multiple reaction monitoring mode with an electrospray ionization (ESI) ion source was employed for detection. The method was validated in terms of linearity, limits of detection, limits of quantitation, precision, and accuracy. Linear responses were obtained with determination coefficients higher than 0.998 for all analytes, and limits of detection were from 0.04 to 0.24 μg/mL. Sample spike/recovery experiments gave recovery values ranging from 73% for d-threonine to 116% for L-tryptophan. Relative standard deviations for inter- and intraday precision experiments were lower than 21.7%. The developed method was successfully applied for determination of the free amino acid enantiomers in five commercially available hydrolyzed protein fertilizer samples.

  6. Staphylococcus aureus aconitase inactivation unexpectedly inhibits post-exponential-phase growth and enhances stationary-phase survival

    OpenAIRE

    Somerville, Greg A.; Chaussee, Michael S.; Morgan, Carrie I; Fitzgerald, Ross; Dorward, D W; Reitzer, Lawrence J.; Musser, James M.

    2002-01-01

    Staphylococcus aureus preferentially catabolizes glucose, generating pyruvate, which is subsequently oxidized to acetate under aerobic growth conditions. Catabolite repression of the tricarboxylic acid (TCA) cycle results in the accumulation of acetate. TCA cycle derepression coincides with exit from the exponential growth phase, the onset of acetate catabolism, and the maximal expression of secreted virulence factors. These data suggest that carbon and energy for post-exponential-phase growt...

  7. Study of Some Alkanes Thermodynamic Parameters Using New Liquid Crystals Containing Sulfur as Stationary Phases

    Directory of Open Access Journals (Sweden)

    Mebrouk Djedid

    2015-06-01

    Full Text Available Most of the synthesized compounds which have liquid crystalline character in their composition comprise aromatic molecules. Furthermore there are few jobs that replace this type of molecules by inhomogeneous molecules that have LC character. We will replace the aromatic rings by units of 1,3,4-oxadiazole and study the effects of these new components of the transition temperatures and the Thermodynamic characteristics of n- alkanes in these two LC's phases. have been investigated by inverse gas chromatography. The transition temperatures obtained by GC are in good agreement with those found by DSC. The results are interpreted in terms of parameters “b” and related thermodynamic quantities

  8. Phenyl Functionalized Sol-gel Silica Sorbent for Capillary Microextraction and Chromia-Based Sol-gel Ucon Stationary Phase for Capillary Gas Chromatography

    Science.gov (United States)

    McLean, Michael M.

    The first chapter of this thesis presents an introduction to sol-gel methodology whose usefulness as a synthetic route will be demonstrated with two applications in chromatography. The first application involves the fabrication of a capillary micro-extraction (CME) device by coating a phenyl functionalized extracting phase on the inner surface of a fused silica capillary for analyte pre-concentration. The device was coupled on-line to a RP-HPLC system and practicality was demonstrated using allergens as target analytes. The allergens chosen as model analytes are typically found in fragrance products and food. Most of the 26 fragrance allergens that are monitored by various government authorities have a phenyl organic moiety (a strong chromophore), thus making them appropriate probes for exploring the extraction efficiency of the coating using a UV detector. The CME device showed ppt level limit of detection which makes it suitable for trace analyses of allergens and similar compounds in a variety of matrices. The second application explores the feasibility of using sol-gel derived chromia-based stationary phase in gas chromatographic columns. The organic moiety of the stationary phase was derived from Ucon 75-H-90,000 while the inorganic backbone was prepared using chromium(III) dichloride hydroxide - methacrylic acid - aqua complex, 40% in isopropanol/acetone . Usefulness of prepared chromia-based GC stationary phase was examined for petrochemical application. Promising results were obtained using aliphatic-aromatics, polyaromatic hydrocarbons, BTEX test mixture, cycloalkanes, branched alkanes and akylbenzenes. The column was able to perform without degradation despite being rinsed multiples times sequentially with the following solvents: dichloromethane, methanol, water and finally methanol again. Maximum theoretical plate number calculated is around 2,400 plates/m. The plate number clearly needs improvement but is a promising result for the newly explored

  9. Ionic liquid-based zwitterionic organic polymer monolithic column for capillary hydrophilic interaction chromatography.

    Science.gov (United States)

    Wang, Tingting; Chen, Yihui; Ma, Junfeng; Zhang, Xiaodan; Zhang, Lihua; Zhang, Yukui

    2015-08-21

    In the current study, a novel ionic liquid-based zwitterionic organic polymer monolithic column was developed by copolymerizing 1-vinyl-3-(butyl-4-sulfonate) imidazolium, acrylamide and N,N'-methylenebisacrylamide in a quaternary porogenic solvent consisting of formamide, dimethyl sulphoxide, polyethylene glycol 8000 and polyethylene glycol 10,000 for capillary hydrophilic interaction chromatography. The monolithic stationary phase was optimized by adjusting the amount of monomer in the polymerization solution along with the composition of porogenic solvent. The optimized monolith exhibited excellent selectivity and favorable retention for nucleosides and benzoic acid derivatives. The primary factors affecting the separation efficiency of the monolithic column (including acetonitrile content, pH, and buffer salt concentration in the mobile phase) have been thoroughly evaluated. Excellent reproducibility of the retention times for five nucleosides was achieved, with relative standard deviations of run-to-run (n = 3), column-to-column (n = 3) and batch-to-batch (n = 3) in the range of 0.18-0.48%, 2.33-4.20% and 3.07-6.50%, respectively.

  10. Suitability of teicoplanin-aglycone bonded stationary phase for simulated moving bed enantio separation of racemic amino acids employing composition-constrained eluents

    NARCIS (Netherlands)

    Bechtold, M; Heinemann, M; Panke, S

    2006-01-01

    The suitability of a teicoplanin-aglycone based chiral stationary phase for the simulated moving bed (SMB) enantioseparation of amino acids under enzyme-compatible conditions was shown following a procedure that is based solely on model-based simulations and HPLC experiments. A set of eight amino ac

  11. Identifying important structural features of ionic liquid stationary phases for the selective separation of nonpolar analytes by comprehensive two-dimensional gas chromatography.

    Science.gov (United States)

    Zhang, Cheng; Ingram, Isaiah C; Hantao, Leandro W; Anderson, Jared L

    2015-03-20

    A series of dicationic ionic liquid (IL)-based stationary phases were evaluated as secondary columns in comprehensive two-dimensional gas chromatography (GC×GC) for the separation of aliphatic hydrocarbons from kerosene. In order to understand the role that structural features of ILs play on the selectivity of nonpolar analytes, the solvation parameter model was used to probe the solvation properties of the IL-based stationary phases. It was observed that room temperature ILs containing long free alkyl side chain substituents and long linker chains between the two cations possess less cohesive forces and exhibited the highest resolution of aliphatic hydrocarbons. The anion component of the IL did not contribute significantly to the overall separation, as similar selectivities toward aliphatic hydrocarbons were observed when examining ILs with identical cations and different anions. In an attempt to further examine the separation capabilities of the IL-based GC stationary phases, columns of the best performing stationary phases were prepared with higher film thickness and resulted in enhanced selectivity of aliphatic hydrocarbons.

  12. Thermodynamics of the adsorption of volatile organic compounds in a binary polydimethylsiloxane-permethylated β-cyclodextrin stationary phase, as measured by gas chromatography

    Science.gov (United States)

    Onuchak, L. A.; Platonov, V. I.; Kuraeva, Yu. G.

    2013-05-01

    The sorption behavior of 29 organic compounds in a binary polydimethylsiloxane-permethylated β-cyclodextrin stationary phase is investigated using gas chromatography. The effect of the sorbates' polarity, shape, and chirality on the formation of sorbate-cyclodextrin inclusion complexes is studied.

  13. The Enantiomer Separations of Allethrone and Propargyllone Using Two Long Chain Acylated β-Cyclodextrin Derivatives as CGC Capillary Stationary Phases

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Using two β-cyclodextrin derivatives (CDs) with long chain of acyl groups as chiral stationary phases (CSPs) of capillary gas chromatography (CGC), the enantiomers of racemic allethrone and propargyllone were well resolved after derived with acetyl chloride. The enantiomer excess values (e.e.%) of 1S-allethrone and 1S-propargyllone were also determined successfully using these CDs.

  14. Temperature effects on the chromatographic behaviour of racemic 2-amidotetralins on a Whelk-O 1 stationary phase in super- and subcritical fluid chromatography

    NARCIS (Netherlands)

    Selditz, U; Copinga, S; Grol, CJ; Franke, JP; de Zeeuw, RA; Wikstrom, H; Gyllenhaal, O

    1999-01-01

    The chromatographic behaviour of thirty structurally related racemic 2-amidotetralins on a cis-(3R, 4S)-Whelk-O 1 stationary phase was investigated at five different temperatures between 50 and -15 degrees C using super- or subscritical fluid chromatography (SFC/SubFC). Generally, low selectivity fa

  15. Increments to chiral recognition facilitating enantiomer separations of chiral acids, bases, and ampholytes using Cinchona-based zwitterion exchanger chiral stationary phases.

    Science.gov (United States)

    Wernisch, Stefanie; Pell, Reinhard; Lindner, Wolfgang

    2012-07-01

    The intramolecular distances of anion and cation exchanger sites of zwitterionic chiral stationary phases represent potential tuning sites for enantiomer selectivity. In this contribution, we investigate the influence of alkanesulfonic acid chain length and flexibility on enantiomer separations of chiral acids, bases, and amphoteric molecules for six Cinchona alkaloid-based chiral stationary phases in comparison with structurally related anion and cation exchangers. Employing polar-organic elution conditions, we observed an intramolecular counterion effect for acidic analytes which led to reduced retention times but did not impair enantiomer selectivities. Retention of amphoteric analytes is based on simultaneous double ion pairing of their charged functional groups with the acidic and basic sites of the zwitterionic selectors. A chiral center in the vicinity of the strong cation exchanger site is vital for chiral separations of bases. Sterically demanding side chains are beneficial for separations of free amino acids. Enantioseparations of free (un-derivatized) peptides were particularly successful in stationary phases with straight-chain alkanesulfonic acid sites, pointing to a beneficial influence of more flexible moieties. In addition, we observed pseudo-enantiomeric behavior of quinine and quinidine-derived chiral stationary phases facilitating reversal of elution orders for all analytes. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Mass-transfer properties of insulin on core-shell and fully porous stationary phases.

    Science.gov (United States)

    Lambert, Nándor; Kiss, Ibolya; Felinger, Attila

    2014-10-31

    The mass-transfer properties of three superficially-porous packing materials, with 2.6 and 3.6μm particle and 1.9, 2.6, and 3.2μm inner core diameter, respectively, were investigated and compared with those of fully porous packings with similar particle properties. Several sources of band spreading in the chromatographic bed have been identified and studied according to the general rate model of chromatography. Besides the axial dispersion in the stream of the mobile phase, and the external mass transfer resistance, the intraparticle diffusion was studied in depth. The first absolute and the second central moments of the peaks of human insulin, over a wide range of mobile phase velocities were measured and used for the calculation of the mass-transfer coefficients. The experimental data were also analyzed using the stochastic or molecular dynamic model of Giddings and Eyring. The dissimilarities of the mass-transfer observed in the different columns were identified and evaluated. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Direct high-performance liquid chromatography enantioseparation of terazosin on an immobilised polysaccharide-based chiral stationary phase under polar organic and reversed-phase conditions.

    Science.gov (United States)

    Ferretti, Rosella; Gallinella, Bruno; La Torre, Francesco; Zanitti, Leo; Turchetto, Luciana; Mosca, Antonina; Cirilli, Roberto

    2009-07-10

    High-performance liquid chromatography (HPLC) enantioseparation of terazosin (TER) was accomplished on the immobilised-type Chiralpak IC chiral stationary phase (CSP) under both polar organic and reversed-phase modes. A simple analytical method was validated using a mixture of methanol-water-DEA 95:5:0.1 (v/v/v) as a mobile phase. Under reversed-phase conditions good linearities were obtained over the concentration range 8.76-26.28 microg mL(-1) for both enantiomers. The limits of detection and quantification were 10 and 30 ng mL(-1), respectively. The intra- and inter-day assay precision was less than 1.66% (RSD%). The optimised conditions also allowed to resolve chiral and achiral impurities from the enantiomers of TER. The proposed HPLC method supports pharmacological studies on the biological effects of the both forms of TER and analytical investigations of potential drug formulations based on a single enantiomer. At the semipreparative scale, 5.3 mg of racemic sample were resolved with elution times less than 12 min using a mobile phase consisting of methanol-DEA 100:0.1 (v/v) and both enantiomers were isolated with a purity of > or = 99% enantiomeric excess (ee). The absolute configuration of TER enantiomers was assigned by comparison of the measured specific rotations with those reported in the literature.

  18. (Global and local) fluctuations of phase space contraction in deterministic stationary nonequilibrium.

    Science.gov (United States)

    Bonetto, F.; Chernov, N. I.; Lebowitz, J. L.

    1998-12-01

    We studied numerically the validity of the fluctuation relation introduced in Evans et al. [Phys. Rev. Lett. 71, 2401-2404 (1993)] and proved under suitable conditions by Gallavotti and Cohen [J. Stat. Phys. 80, 931-970 (1995)] for a two-dimensional system of particles maintained in a steady shear flow by Maxwell demon boundary conditions [Chernov and Lebowitz, J. Stat. Phys. 86, 953-990 (1997)]. The theorem was found to hold if one considers the total phase space contraction sigma occurring at collisions with both walls: sigma=sigma( upward arrow )+sigma( downward arrow ). An attempt to extend it to more local quantities sigma( upward arrow ) and sigma( downward arrow ), corresponding to the collisions with the top or bottom wall only, gave negative results. The time decay of the correlations in sigma( upward arrow, downward arrow ) was very slow compared to that of sigma. (c) 1998 American Institute of Physics.

  19. Sinorhizobium meliloti sigma factors RpoE1 and RpoE4 are activated in stationary phase in response to sulfite.

    Science.gov (United States)

    Bastiat, Bénédicte; Sauviac, Laurent; Picheraux, Carole; Rossignol, Michel; Bruand, Claude

    2012-01-01

    Rhizobia are soil bacteria able to establish a nitrogen-fixing symbiosis with legume plants. Both in soil and in planta, rhizobia spend non-growing periods resembling the stationary phase of in vitro-cultured bacteria. The primary objective of this work was to better characterize gene regulation in this biologically relevant growth stage in Sinorhizobium meliloti. By a tap-tag/mass spectrometry approach, we identified five sigma factors co-purifying with the RNA polymerase in stationary phase: the general stress response regulator RpoE2, the heat shock sigma factor RpoH2, and three extra-cytoplasmic function sigma factors (RpoE1, RpoE3 and RpoE4) belonging to the poorly characterized ECF26 subgroup. We then showed that RpoE1 and RpoE4 i) are activated upon metabolism of sulfite-generating compounds (thiosulfate and taurine), ii) display overlapping regulatory activities, iii) govern a dedicated sulfite response by controlling expression of the sulfite dehydrogenase SorT, iv) are activated in stationary phase, likely as a result of endogenous sulfite generation during bacterial growth. We showed that SorT is required for optimal growth of S. meliloti in the presence of sulfite, suggesting that the response governed by RpoE1 and RpoE4 may be advantageous for bacteria in stationary phase either by providing a sulfite detoxification function or by contributing to energy production through sulfite respiration. This paper therefore reports the first characterization of ECF26 sigma factors, the first description of sigma factors involved in control of sulphur metabolism, and the first indication that endogenous sulfite may act as a signal for regulation of gene expression upon entry of bacteria in stationary phase.

  20. Sinorhizobium meliloti sigma factors RpoE1 and RpoE4 are activated in stationary phase in response to sulfite.

    Directory of Open Access Journals (Sweden)

    Bénédicte Bastiat

    Full Text Available Rhizobia are soil bacteria able to establish a nitrogen-fixing symbiosis with legume plants. Both in soil and in planta, rhizobia spend non-growing periods resembling the stationary phase of in vitro-cultured bacteria. The primary objective of this work was to better characterize gene regulation in this biologically relevant growth stage in Sinorhizobium meliloti. By a tap-tag/mass spectrometry approach, we identified five sigma factors co-purifying with the RNA polymerase in stationary phase: the general stress response regulator RpoE2, the heat shock sigma factor RpoH2, and three extra-cytoplasmic function sigma factors (RpoE1, RpoE3 and RpoE4 belonging to the poorly characterized ECF26 subgroup. We then showed that RpoE1 and RpoE4 i are activated upon metabolism of sulfite-generating compounds (thiosulfate and taurine, ii display overlapping regulatory activities, iii govern a dedicated sulfite response by controlling expression of the sulfite dehydrogenase SorT, iv are activated in stationary phase, likely as a result of endogenous sulfite generation during bacterial growth. We showed that SorT is required for optimal growth of S. meliloti in the presence of sulfite, suggesting that the response governed by RpoE1 and RpoE4 may be advantageous for bacteria in stationary phase either by providing a sulfite detoxification function or by contributing to energy production through sulfite respiration. This paper therefore reports the first characterization of ECF26 sigma factors, the first description of sigma factors involved in control of sulphur metabolism, and the first indication that endogenous sulfite may act as a signal for regulation of gene expression upon entry of bacteria in stationary phase.

  1. [Determination of five avermectins in bovine liver by on-line solid-phase extraction with hydrophobic monolithic column coupled with high performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Li, Xin; Zhang, Yaoqin; Ai, Lianfeng; Wang, Xuesheng; Wang, Manman; Xu, Houjun; Hao, Yulan

    2015-06-01

    A method based on on-line solid-phase extraction (SPE) with hydrophobic monolithic column coupled with high performance liquid chromatography-tandem mass spectrometry was developed for the simultaneous determination of five avermectins in bovine liver. A poly(butyl methacrylate-co-ethylene dimethacrylate) monolithic column was used as the sorbent. The parameters influenced on on-line SPE and separation process such as the loading mobile phase, the eluting flow rate and the solvent for the separation were investigated in detail. Blank samples, spiked samples, matrix effect and recovery experiments were investigated to evaluate the extraction efficiency and potential interfering compounds originating from the matrix. Under the optimized conditions, the method showed a linear range of 1-100 µg/L and the quantification limit of 5 µg/kg for each analyte. The presented method gave recoveries of 77.4%-98.4%. The relative standard deviations of intra-day and inter-day were 4.46%-8.03% and 4.79%-8.68%, respectively. Moreover, no significant changes were found in the extraction performance after more than 400 usages on one monolithic column, and even on the monoliths with various batches. The feasibility of the developed poly (butyl methacrylate-coethylene dimethacrylate) monolithic column based on the on-line SPE method for the determination of avermectins was further demonstrated by the analysis of real samples.

  2. Sensitive Monitoring of Fluoroquinolones in Milk and Honey Using Multiple Monolithic Fiber Solid-Phase Microextraction Coupled to Liquid Chromatography Tandem Mass Spectrometry.

    Science.gov (United States)

    Chen, Lei; Huang, Xiaojia

    2016-11-16

    In the present study, a new multiple monolithic fiber solid-phase microextraction (MMF-SPME) based on poly(apronal-co-divinylbenzene/ethylenedimethacrylate) monolith (APDE) was synthesized. The effect of the preparation parameters of APED on extraction efficiency was studied thoroughly. The combination of APDE/MMF-SPME with high-performance liquid chromatography tandem mass spectrometry detection (HPLC/MS-MS) was developed for sensitive monitoring of ultratrace fluoroquinolones (FQs) in foodstuffs, including milk and honey samples. Under the optimized experimental conditions, the limits of detection (S/N = 3) for the targeted FQs ranged from 0.0019 to 0.018 μg/kg in milk and 0.0010 to 0.0028 μg/kg in honey. The relative standard deviations (RSDs) for method reproducibility were less than 9% in all samples. The established method was successfully applied for the monitoring of FQs residues in milk and honey samples with the recoveries between 74.5% and 116% (RSDs were in the range 0.9-9.5%). In comparison to previous methods, the developed APDE/MMF-SPME-HPLC/MS-MS showed some merits, including satisfactory sensitivity, simplicity, high cost-effectiveness, and low consumption of organic solvent.

  3. Sensitive determination of parabens in human urine and serum using methacrylate monoliths and reversed-phase capillary liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Carrasco-Correa, Enrique Javier; Vela-Soria, Fernando; Ballesteros, Oscar; Ramis-Ramos, Guillermo; Herrero-Martínez, José Manuel

    2015-01-30

    A method for the determination of parabens in human urine and serum by capillary liquid chromatography (cLC) with UV-Vis and mass spectrometry (MS) detection using methacrylate ester-based monolithic columns has been developed. The influence of composition of polymerization mixture was studied. The optimum monolith was obtained with butyl methacrylate monomer at 60/40% (wt/wt) butyl methacrylate/ethylene dimethacrylate ratio and 50wt% porogens (composed of 36wt% of 1,4-butanediol, 54wt% 1-propanol and 10wt% water). Baseline resolution of analytes was achieved through a mobile phase of acetonitrile/water in gradient elution mode. Additionally, dispersive liquid-liquid microextraction (DLLME) was combined with both cLC-UV-Vis and cLC-MS to achieve the determination of parabens in human urine and serum samples with very low limits of detection. Satisfactory intra- and inter-day repeatabilities were obtained in UV-Vis and MS detection, although the latter provided lower detection limits (up to 300-fold) than the UV-Vis detection. Recoveries for the target analytes from spiked biological samples ranged from 95.2% to 106.7%. The proposed methodology for the ultra-low determination of parabens in human urine and serum samples is simple and fast, the consumption of reagents is very low, and very small samples can be analyzed. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Graphene oxide reinforced polymeric ionic liquid monolith solid-phase microextraction sorbent for high-performance liquid chromatography analysis of phenolic compounds in aqueous environmental samples.

    Science.gov (United States)

    Sun, Min; Bu, Yanan; Feng, Juanjuan; Luo, Chuannan

    2016-01-01

    A graphene oxide reinforced polymeric ionic liquids monolith was obtained by copolymerization of graphene oxide doped 1-(3-aminopropyl)-3-(4-vinylbenzyl)imidazolium 4-styrenesulfonate monomer and 1,6-di-(3-vinylimidazolium) hexane bihexafluorophosphate cross-linking agent. Coupled to high-performance liquid chromatography, the monolith was used as a solid-phase microextraction sorbent to analyze several phenolic compounds in aqueous samples. Under the optimized extraction and desorption conditions, linear ranges were 5-400 μg/L for 3-nitrophenol, 2-nitrophenol, and 2,5-dichlorophenol and 2-400 μg/L for 4-chlorophenol, 2-methylphenol, and 2,4,6-trichlorophenol (R(2) = 0.9973-0.9988). The limits of detection were 0.5 μg/L for 3-nitrophenol and 2-nitrophenol and 0.2 μg/L for the rest of the analytes. The proposed method was used to determine target analytes in groundwater from an industrial park and river water. None of the analytes was detected. Relative recoveries were in the range of 75.5-113%.

  5. High-performance liquid chromatography analysis of polyacetylenes and polyenes in Echinacea pallida by using a monolithic reversed-phase silica column.

    Science.gov (United States)

    Pellati, Federica; Calò, Samuele; Benvenuti, Stefania

    2007-05-11

    In this study, a RP-HPLC method for the analysis of polyacetylenes and polyenes in Echinacea pallida roots and phytopharmaceuticals was developed. The reference compounds used for quantification were isolated from the plant material and their structures were determined on the basis of the analysis of UV, IR, NMR and MS data. The complete structure elucidation of three compounds, namely 8-hydroxy-tetradec-(9E)-ene-11,13-diyn-2-one (1), tetradec-(8Z)-ene-11,13-diyn-2-one (6) and pentadec-(8Z)-en-2-one (9) is described. In the analysis of the n-hexane extracts of E. pallida roots, the comparison between conventional and monolithic columns showed that the elution order in both cases is identical and the selectivity is equivalent. However, the retention times achieved by the monolithic column are shorter, resulting in a faster separation (20 min). Therefore, the analyses were carried out on a Chromolith Performance RP-18e (100 mm x 4.6 mm i.d.), with a gradient mobile phase composed by H(2)O and ACN at the flow rate of 2 mL/min. The column was thermostatted at 20 degrees C. The photodiode array detector monitored the eluent at 210 nm. The validation procedure confirmed that this technique affords reliable analysis of these components and is appropriate for the quality control of complex matrices, such as E. pallida roots and phytopharmaceuticals.

  6. Thermodynamic Study of Racemic Ibuprofen Separation by Liquid Chromatography Using Cellulose-Based Stationary Phase

    Directory of Open Access Journals (Sweden)

    Wilson M. Ferrari

    2016-01-01

    Full Text Available Ibuprofen is a nonsteroidal anti-inflammatory drug (NSAID, also known for its significant antipyretic and analgesic properties. This chiral drug is commercialized in racemic form; however, only S-(+-ibuprofen has clinical activities. In this paper the effect of temperature change (from 288.15 to 308.15 K on the ibuprofen resolution was studied. A column (250×4.6 mm packed with tris(3,5-dimethylphenylcarbamate was used to obtain the thermodynamic parameters, such as enthalpy change (ΔH, entropy change (ΔS, variation enthalpy change (ΔΔH, variation entropy change (ΔΔS, and isoenantioselective temperature (Tiso. The mobile phase was a combination of hexane (99%, isopropyl alcohol (1%, and TFA (0.1%, as an additive. The conditions led to a selectivity of 1.20 and resolution of 4.55. The first peak, R-(−-ibuprofen, presented an enthalpy change of 7.21 kJ/mol and entropy change of 42.88 kJ/K·mol; the last peak, S-(+-ibuprofen, has an enthalpy change of 8.76 kJ/mol and 49.40 kJ/K·mol of entropy change.

  7. Synthesis and characterization of nanometric zinc oxide for a stationary phase in liquid chromatography

    Science.gov (United States)

    Gordillo-Delgado, F.; Soto-Barrera, C. C.; Plazas-Saldaña, J.

    2017-01-01

    The increasing demand for equipment to remove organic compounds in industry and research activity has led to evaluate nanometric zinc oxide (ZnO). In this work, we present the ZnO nanoparticles synthesis for reusing of discarded columns, as a low-cost alternative. The compound was obtained by sol-gel technique using zinc chloride and sodium hydroxide as precursors and a drying temperature of 169°C. An X-ray diffractometer was used to estimate the average particle size at 20.3±0.2nm the adsorption capacity was 0.0144L/g and the chemical resistance was tested with HCl and NaOH. The ZnO nanopowder was packed with 100psi pressure in an empty C-18 column cavity. The column packing resolution was evaluated using a high performance liquid chromatographer (HPLC-Thermo Scientific Dionex UltiMate 3000); using a caffeine standard, the following parameters were established: solvent flow: 1.2mL/min, average column temperature: 40°C, running time: 10 minutes, mobile phase acetonitrile-water composition (9:1). These results validate the potential of ZnO nanopowder as a column packing material in HPLC technique.

  8. Nano-electrospray tandem mass spectrometric analysis of the acetylation state of histones H3 and H4 in stationary phase in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Patterton Hugh G

    2011-07-01

    Full Text Available Abstract Background The involvement of histone acetylation in facilitating gene expression is well-established, particularly in the case of histones H3 and H4. It was previously shown in Saccharomyces cerevisiae that gene expression was significantly down-regulated and chromatin more condensed in stationary phase compared to exponential phase. We were therefore interested in establishing the acetylation state of histone H3 and H4 in stationary and in exponential phase, since the regulation of this modification could contribute to transcriptional shut-down and chromatin compaction during semi-quiescence. Results We made use of nano-spray tandem mass spectrometry to perform a precursor ion scan to detect an m/z 126 immonium ion, diagnostic of an Nε-acetylated lysine residue that allowed unambiguous identification of acetylated as opposed to tri-methylated lysine. The fragmentation spectra of peptides thus identified were searched with Mascot against the Swiss-Prot database, and the y-ion and b-ion fragmentation series subsequently analyzed for mass shifts compatible with acetylated lysine residues. We found that K9, K14 and K36 of histone H3 and K12 and K16 of histone H4 were acetylated in exponential phase (bulk histones, but could not detect these modifications in histones isolated from stationary phase cells at the sensitivity level of the mass spectrometer. The corresponding un-acetylated peptides were, however, observed. A significantly higher level of acetylation of these residues in exponential phase was confirmed by immuno-blotting. Conclusion H4K16 acetylation was previously shown to disrupt formation of condensed chromatin in vitro. We propose that de-acetylation of H4K16 allowed formation of condensed chromatin in stationary phase, and that acetylation of H3K9, H3K14, H3K36, and H4K12 reflected the active transcriptional state of the yeast genome in exponential phase.

  9. Polymer-coated fibrous materials as the stationary phase in packed capillary gas chromatography.

    Science.gov (United States)

    Saito, Yoshihiro; Tahara, Ai; Imaizumi, Motohiro; Takeichi, Tsutomu; Wada, Hiroo; Jinno, Kiyokatsu

    2003-10-15

    Synthetic polymer filaments have been introduced as the support material in packed capillary gas chromatography (GC). The filaments of the heat-resistant polymers, Zylon, Kevlar, Nomex, and Technora, were longitudinally packed into a short fused-silica capillary, followed by the conventional coating process for open-tubular GC columns. The separation of several test mixtures such as n-alkylbenzenes and n-alkanes was carried out with these polymer-coated fiber-packed capillary columns. With the coating by various polymeric materials on the surface of these filaments, the retentivity was significantly improved over the parent fiber-packed column (without polymer coating) as well as a conventional open-tubular capillary of the same length. The results demonstrated a good combination of Zylon as the support and poly(dimethylsiloxane)-based materials as the coating liquid-phase for the successful GC separation of n-alkanes and polycyclic aromatic hydrocarbons (PAHs), while successful applications for other separations such as poly(ethylene glycol) coating for the separation of alcohols were also obtained. From the results it has been suggested that the selectivity of the fiber-packed column could be tuned by selecting different coating materials, indicating the promising possibility for a novel usage of fine fibrous polymers as the support material that can be combined with newly synthesized coating materials specially designed for particular separations. Taking advantage of good thermal stability of the fibers, the column temperature could be elevated to higher than 350 degrees C with the combination of a short metallic capillary.

  10. Stationary Frame Current Control Evaluations for Three-Phase Grid-Connected Inverters with PVR-based Active Damped LCL Filters

    DEFF Research Database (Denmark)

    Han, Yang; Shen, Pan; Guerrero, Josep M.

    2016-01-01

    under highly distorted grid conditions. In this paper, a synchronous reference frame equivalent proportional-integral (SRF-EPI) controller in αβ stationary frame using the parallel virtual resistance-based active damping (PVR -AD) strategy for grid-interfaced distributed generation (DG) systems...... to suppress the LCL resonance is proposed. Although both proportional-resonant (PR) controller in αβ stationary frame and PI controller in dq synchronous frame achieve zero steady-state error, the amplitude- and phase-frequency characteristics differ greatly from each other except for the reference tracking...... at fundamental frequency. Therefore, an accurate SRF-EPI controller in αβ stationary frame is established to achieve precise tracking accuracy. Moreover, the robustness, harmonic rejection capabilities, and influence of control delay are investigated by the Nyquist stability criterion when the PVR-based AD...

  11. A chromatographic estimate of the degree of surface heterogeneity of reversed-phase liquid chromatography packing materials II-Endcapped monomeric C18-bonded stationary phase

    Energy Technology Data Exchange (ETDEWEB)

    Gritti, Fabrice [University of Tennessee, Knoxville (UTK); Guiochon, Georges A [ORNL

    2006-01-01

    In a previous report, the heterogeneity of a non-endcapped C{sub 30}-bonded stationary phase was investigated, based on the results of the measurements of the adsorption isotherms of two neutral compounds (phenol and caffeine) and two ionizable compounds (sodium naphthalene sulfonate and propranololium chloride) by frontal analysis (FA). The same method is applied here for the characterization of the surface heterogeneity of two new brands of endcapped C{sub 18}-bonded stationary phases (Gemini and Sunfire). The adsorption isotherms of the same four chemicals were measured by FA and the results confirmed by the independent calculation of the adsorption energy distribution (AED), using the expectation-maximization (EM) method. The effect of the length of the bonded alkyl chain was investigated. Shorter alkyl-bonded-chains (C{sub 18} versus C{sub 30}) and the end-capping of the silica surface contribute to decrease the surface heterogeneity under the same experimental conditions (30% methanol, 25 mM NaCl). The AEDs of phenol and caffeine are bimodal with the C{sub 18}-bonded columns while they are trimodal and quadrimodal, respectively, with a non-endcapped C{sub 30}-bonded column. The 'supersites' (adsorption energy >20 kJ/mol) found on the C{sub 30}-Prontosil column and attributed to a cation exchange mechanism completely disappear on the C{sub 18}-Gemini and C{sub 18}-Sunfire, probably because the end-capping of the silica surface eliminates most if not all the ionic interactions.

  12. Separation of monoclonal antibody charge state variants by open tubular capillary electrochromatography with immobilised protein as stationary phase.

    Science.gov (United States)

    Zhang, Yamin; Wang, Wentao; Xiao, Xue; Jia, Li

    2016-09-30

    Monoclonal antibodies (mAbs) are highly heterogeneous and complex glycoproteins requiring powerful analytical tools for characterization and quality control. In this work, we utilize adsorbed bovine serum albumin (BSA) as a stationary phase in open tubular (OT) capillary electrochromatography for separation of charge state variants of mAbs. Poly(diallydimethylammonium chloride) (PDDA) was used to assist fabrication of BSA coated OT column by electrostatic self-assembly. Scanning electron microscopy and electroosmotic flow measurement were carried out to characterize the as-prepared BSA coated PDDA OT columns. The electrochromatographic performance of the OT columns was evaluated by separation of basic proteins and different charge state variants of mAbs. The effects of background solution pH and concentration on separation were investigated. A rapid separation of charge state variants of mAbs was successfully achieved in the BSA coated PDDA OT column. Separation of seven variants of the mAb cetuximab was achieved using the prepared column. Two basic variants and one acidic variant of rituximab, and two basic variants and four acidic variants of trastuximab were successfully distinguished from the main forms. In addition, the columns demonstrated good repeatability and stability with the run-to-run, day-to-day and batch-to-batch relative standard deviations of migration times less than 3.7%.

  13. Interaction between Antibacterial Peptide Apep10 and Escherichia coli Membrane Lipids Evaluated Using Liposome as Pseudo-Stationary Phase

    Science.gov (United States)

    Li, Man

    2017-01-01

    Liposomes constructed from Escherichia coli membrane lipids were used as a pseudo-stationary phase in capillary electrophoresis and immobilised liposome chromatography to evaluate the interaction between antibacterial peptide (ABP) Apep10 and bacterial membrane lipids. The peptide mobility decreased as the concentration of liposomes increased, providing evidence for the existence of this interaction. The binding constant between Apep10 and the Escherichia coli membranes lipid liposome was higher than that of Apep10 with a mixed phospholipids liposome at the same temperature. The capillary electrophoresis results indicate that the binding ability of Apep10 with a liposome was dependent on the liposome’s lipid compositions. Thermodynamic analysis by immobilised liposome chromatography indicated that hydrophobic and electrostatic effects contributed to the partitioning of Apep10 in the membrane lipids. The liposomes constructed from bacterial membrane lipid were more suitable as the model membranes used to study dynamic ABP/membrane interactions than those constructed from specific ratios of particular phospholipids, with its more biomimetic phospholipid composition and contents. This study provides an appropriate model for the evaluation of ABP-membrane interactions. PMID:28052090

  14. High enantioselective Novozym 435-catalyzed esterification of (R,S)-flurbiprofen monitored with a chiral stationary phase.

    Science.gov (United States)

    Siódmiak, Tomasz; Mangelings, Debby; Vander Heyden, Yvan; Ziegler-Borowska, Marta; Marszałł, Michał Piotr

    2015-03-01

    Lipases form Candida rugosa and Candida antarctica were tested for their application in the enzymatic kinetic resolution of (R,S)-flurbiprofen by enantioselective esterification. Successful chromatographic separation with well-resolved peaks of (R)- and (S)-flurbiprofen and their esters was achieved in one run on chiral stationary phases by high-performance liquid chromatography (HPLC). In this study screening of enzymes was performed, and Novozym 435 was selected as an optimal catalyst for obtaining products with high enantiopurity. Additionally, the influence of organic solvents (dichloromethane, dichloroethane, dichloropropane, and methyl tert-butyl ether), primary alcohols (methanol, ethanol, n-propanol, and n-butanol), reaction time, and temperature on the enantiomeric ratio and conversion was tested. The high values of enantiomeric ratio (E in the range of 51.3-90.5) of the esterification of (R,S)-flurbiprofen were obtained for all tested alcohols using Novozym 435, which have a great significance in the field of biotechnological synthesis of drugs. The optimal temperature range for the performed reactions was from 37 to 45 °C. As a result of the optimization, (R)-flurbiprofen methyl ester was obtained with a high optical purity, eep = 96.3 %, after 96 h of incubation. The enantiomeric ratio of the reaction was E = 90.5 and conversion was C = 35.7 %.

  15. Enantiomeric Separation of 1-(Benzofuran-2-yl)alkylamines on Chiral Stationary Phases Based on Chiral Crown Ethers

    Energy Technology Data Exchange (ETDEWEB)

    Park, Soohyun; Kim, Sang Jun; Hyun, Myung Ho [Pusan National Univ., Busan (Korea, Republic of)

    2012-10-15

    Optically active chiral amines are important as building blocks for pharmaceuticals and as scaffolds for chiral ligands and, consequently, many efforts have been devoted to the development of efficient methods for their preparation. For example, reduction of amine precursors with chiral catalysts, enzymatic kinetic resolution or dynamic kinetic resolution of racemic amines and the direct amination of ketones with transaminases have been developed as the efficient methods for the preparation of optically active chiral amines. During the process of developing or utilizing optically active chiral amines, the methods for the determination of their enantiomeric composition are essential. Among various methods, liquid chromatographic resolution of enantiomers on chiral stationary phases (CSPs) have been known to be one of the most accurate and economic means for the determination of the enantiomeric composition of optically active chiral compounds. Especially, CSPs based on chiral crown ethers have been successfully used for the resolution of racemic primary amines. For example, CSPs based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid (CSP 1, Figure 1) or (3,3'-diphenyl-1,1'-binaphthyl)-20-crown-6 (CSP 2 and CSP 3, Figure 1) have been known to be quite effective for the resolution of cyclic and non-cyclic amines, various fluoroquinolone antibacterials containing a primary amino group, tocainide (antiarrhythmic agent) and its analogues, aryl-a-amino ketones and 3-amino-1,4-benzodiazepin-2-ones.

  16. X-Ray Solution Scattering Study of Four Escherichia coli Enzymes Involved in Stationary-Phase Metabolism.

    Science.gov (United States)

    Dadinova, Liubov A; Shtykova, Eleonora V; Konarev, Petr V; Rodina, Elena V; Snalina, Natalia E; Vorobyeva, Natalia N; Kurilova, Svetlana A; Nazarova, Tatyana I; Jeffries, Cy M; Svergun, Dmitri I

    2016-01-01

    The structural analyses of four metabolic enzymes that maintain and regulate the stationary growth phase of Escherichia coli have been performed primarily drawing on the results obtained from solution small angle X-ray scattering (SAXS) and other structural techniques. The proteins are (i) class I fructose-1,6-bisphosphate aldolase (FbaB); (ii) inorganic pyrophosphatase (PPase); (iii) 5-keto-4-deoxyuronate isomerase (KduI); and (iv) glutamate decarboxylase (GadA). The enzyme FbaB, that until now had an unknown structure, is predicted to fold into a TIM-barrel motif that form globular protomers which SAXS experiments show associate into decameric assemblies. In agreement with previously reported crystal structures, PPase forms hexamers in solution that are similar to the previously reported X-ray crystal structure. Both KduI and GadA that are responsible for carbohydrate (pectin) metabolism and acid stress responses, respectively, form polydisperse mixtures consisting of different oligomeric states. Overall the SAXS experiments yield additional insights into shape and organization of these metabolic enzymes and further demonstrate the utility of hybrid methods, i.e., solution SAXS combined with X-ray crystallography, bioinformatics and predictive 3D-structural modeling, as tools to enrich structural studies. The results highlight the structural complexity that the protein components of metabolic networks may adopt which cannot be fully captured using individual structural biology techniques.

  17. Preparation of chitosan-graft-(β-cyclodextrin) based sol-gel stationary phase for open-tubular capillary electrochromatography.

    Science.gov (United States)

    Lü, Haixia; Li, Qingyin; Yu, Xiaowei; Yi, Jiaojiao; Xie, Zenghong

    2013-07-01

    A novel open-tubular CEC column coated with chitosan-graft-(β-CD) (CDCS) was prepared using sol-gel technique. In the sol-gel approach, owing to the 3D network of sol-gel and the strong chemical bond between the stationary phase and the surface of capillary columns, good chromatographic characteristics and unique selectivity in separating isomers were shown. The column efficiencies of 55,000∼163,000 plates/m for the isomeric xanthopterin and phenoxy acid herbicides using the sol-gel-derived CDCS columns were achieved. Good stabilities were demonstrated that the RSD values for the retention time of thiourea and isoxanthopterin were 1.3 and 1.4% (run to run, n = 5), 1.6 and 2.0% (day to day, n = 3), 2.9 and 3.1% (column to column, n = 3), respectively. The sol-gel-coated CDCS columns have shown improved separations of isomeric xanthopterin in comparison with CDCS-bonded capillary column.

  18. Selective high performance liquid chromatography imprinted-stationary phases for the screening of phenylurea herbicides in vegetable samples.

    Science.gov (United States)

    Tamayo, F G; Martin-Esteban, A

    2005-12-09

    Molecularly imprinted polymers (MIPs) for the determination of phenylurea herbicides have been synthesized by polymerisation of the appropriated reagents mixture within the pores of preformed spherical silica particles leading to a silica-MIP composite material. Subsequently, the silica matrix was etched away resulting in MIP beads which can be considered the "mirror image" of the original silica mold. The MIP particles were packed in stainless steal HPLC columns (125mmx4.6mm I.D.) and the materials were evaluated as imprinted-stationary phases for phenylurea herbicides. The imprinting effect of the originated specific binding sites for the selective recognition of phenylurea herbicides was clearly demonstrated. An efficient separation of a mixture of phenylurea herbicides in two groups, with or without a methoxy group in the chemical structure, was achieved and well shaped and defined peaks were obtained. Finally, the optimum imprinted column (prepared using linuron as template, 2-(trifluoromethyl)-acrylic acid as monomer, 72h of polymerisation time and the subsequent dissolution of silica matrix) was used for the LC-UV screening of phenylurea herbicides directly from vegetable sample extracts without any previous clean-up step at low concentration level in less than 10min.

  19. Application of Homochiral Alkylated Organic Cages as Chiral Stationary Phases for Molecular Separations by Capillary Gas Chromatography

    Directory of Open Access Journals (Sweden)

    Shengming Xie

    2016-11-01

    Full Text Available Molecular organic cage compounds have attracted considerable attention due to their potential applications in gas storage, catalysis, chemical sensing, molecular separations, etc. In this study, a homochiral pentyl cage compound was synthesized from a condensation reaction of (S,S-1,2-pentyl-1,2-diaminoethane and 1,3,5-triformylbenzene. The imine-linked pentyl cage diluted with a polysiloxane (OV-1701 was explored as a novel stationary phase for high-resolution gas chromatographic separation of organic compounds. Some positional isomers were baseline separated on the pentyl cage-coated capillary column. In particular, various types of enantiomers including chiral alcohols, esters, ethers and epoxides can be resolved without derivatization on the pentyl cage-coated capillary column. The reproducibility of the pentyl cage-coated capillary column for separation was investigated using nitrochlorobenzene and styrene oxide as analytes. The results indicate that the column has good stability and separation reproducibility after being repeatedly used. This work demonstrates that molecular organic cage compounds could become a novel class of chiral separation media in the near future.

  20. Application of Homochiral Alkylated Organic Cages as Chiral Stationary Phases for Molecular Separations by Capillary Gas Chromatography.

    Science.gov (United States)

    Xie, Shengming; Zhang, Junhui; Fu, Nan; Wang, Bangjin; Hu, Cong; Yuan, Liming

    2016-11-08

    Molecular organic cage compounds have attracted considerable attention due to their potential applications in gas storage, catalysis, chemical sensing, molecular separations, etc. In this study, a homochiral pentyl cage compound was synthesized from a condensation reaction of (S,S)-1,2-pentyl-1,2-diaminoethane and 1,3,5-triformylbenzene. The imine-linked pentyl cage diluted with a polysiloxane (OV-1701) was explored as a novel stationary phase for high-resolution gas chromatographic separation of organic compounds. Some positional isomers were baseline separated on the pentyl cage-coated capillary column. In particular, various types of enantiomers including chiral alcohols, esters, ethers and epoxides can be resolved without derivatization on the pentyl cage-coated capillary column. The reproducibility of the pentyl cage-coated capillary column for separation was investigated using nitrochlorobenzene and styrene oxide as analytes. The results indicate that the column has good stability and separation reproducibility after being repeatedly used. This work demonstrates that molecular organic cage compounds could become a novel class of chiral separation media in the near future.

  1. X-Ray Solution Scattering Study of Four Escherichia coli Enzymes Involved in Stationary-Phase Metabolism.

    Directory of Open Access Journals (Sweden)

    Liubov A Dadinova

    Full Text Available The structural analyses of four metabolic enzymes that maintain and regulate the stationary growth phase of Escherichia coli have been performed primarily drawing on the results obtained from solution small angle X-ray scattering (SAXS and other structural techniques. The proteins are (i class I fructose-1,6-bisphosphate aldolase (FbaB; (ii inorganic pyrophosphatase (PPase; (iii 5-keto-4-deoxyuronate isomerase (KduI; and (iv glutamate decarboxylase (GadA. The enzyme FbaB, that until now had an unknown structure, is predicted to fold into a TIM-barrel motif that form globular protomers which SAXS experiments show associate into decameric assemblies. In agreement with previously reported crystal structures, PPase forms hexamers in solution that are similar to the previously reported X-ray crystal structure. Both KduI and GadA that are responsible for carbohydrate (pectin metabolism and acid stress responses, respectively, form polydisperse mixtures consisting of different oligomeric states. Overall the SAXS experiments yield additional insights into shape and organization of these metabolic enzymes and further demonstrate the utility of hybrid methods, i.e., solution SAXS combined with X-ray crystallography, bioinformatics and predictive 3D-structural modeling, as tools to enrich structural studies. The results highlight the structural complexity that the protein components of metabolic networks may adopt which cannot be fully captured using individual structural biology techniques.

  2. Chiral Separation of Novel a-Aminophosphonates Containing a Benzothiazole Moiety by Liquid Chromatography Using an Amylose Stationary Phase

    Institute of Scientific and Technical Information of China (English)

    LU Ping; ZHANG Yu-Ping; SONG Bao-An; YANG Song; BHADURY Shankar Pinaki; HU De-Yu; XUE Wei; CHEN Zhuo; JIN Lin-Hong

    2008-01-01

    The present report describes a chiral HPLC method for the enantiomeric separation of a-aminophosphonate derivatives using two new coated and immobilized amylose-based chiral stationary phases (CSP,Chiralpak IA and Chiralpak AD-H).The chromatographic parameters such as retention factor (k),separation factor (a),and resolution (Rs) of the solutes were investigated on these two CSPs.Reasonably good baseline separation for these compounds was achieved using Chiralpak IA column.The influences of temperature,content of ethanol modifier and the structure of analyte were also studied.THF,EtOAc and CH2Cl2 were used as ehients on analytical and semi-preparative columns.Highly enriched enantiomers with purities of up to 96.4%--100% (ee) and yields of 90.2%--95.5% were obtained,respectively.The proposed methods were found to be suitable and accurate for rapid separation and semi-preparation of enantiomeric a-aminophosphonate derivatives available.

  3. Synergistic effects on enantioselectivity of zwitterionic chiral stationary phases for separations of chiral acids, bases, and amino acids by HPLC.

    Science.gov (United States)

    Hoffmann, Christian V; Pell, Reinhard; Lämmerhofer, Michael; Lindner, Wolfgang

    2008-11-15

    In an attempt to overcome the limited applicability scope of earlier proposed Cinchona alkaloid-based chiral weak anion exchangers (WAX) and recently reported aminosulfonic acid-based chiral strong cation exchangers (SCX), which are conceptionally restricted to oppositely charged solutes, their individual chiral selector (SO) subunits have been fused in a combinatorial synthesis approach into single, now zwitterionic, chiral SO motifs. The corresponding zwitterionic ion-exchange-type chiral stationary phases (CSPs) in fact combined the applicability spectra of the parent chiral ion exchangers allowing for enantioseparations of chiral acids and amine-type solutes in liquid chromatography using polar organic mode with largely rivaling separation factors as compared to the parent WAX and SCX CSPs. Furthermore, the application spectrum could be remarkably expanded to various zwitterionic analytes such as alpha- and beta-amino acids and peptides. A set of structurally related yet different CSPs consisting of either a quinine or quinidine alkaloid moiety as anion-exchange subunit and various chiral or achiral amino acids as cation-exchange subunits enabled us to derive structure-enantioselectivity relationships, which clearly provided strong unequivocal evidence for synergistic effects of the two oppositely charged ion-exchange subunits being involved in molecular recognition of zwitterionic analytes by zwitterionic SOs driven by double ionic coordination.

  4. Single-walled carbon nanotubes as stationary phase in gas chromatographic separation and determination of argon, carbon dioxide and hydrogen.

    Science.gov (United States)

    Safavi, Afsaneh; Maleki, Norooz; Doroodmand, Mohammad Mahdi

    2010-08-24

    A chromatographic technique is introduced based on single-walled carbon nanotubes (SWCNTs) as stationary phase for separation of Ar, CO(2) and H(2) at parts per million (ppm) levels. The efficiency of SWCNTs was compared with solid materials such as molecular sieve, charcoal, multi-walled carbon nanotubes and carbon nanofibers. The morphology of SWCNTs was optimized for maximum adsorption of H(2), CO(2) and Ar and minimum adsorption of gases such as N(2), O(2), CO and H(2)O vapour. To control temperature of the gas chromatography column, peltier cooler was used. Mixtures of Ar, CO(2) and H(2) were separated according to column temperature program. Relative standard deviation for nine replicate analyses of 0.2 mL H(2) containing 10 microL of each Ar or CO(2) was 2.5% for Ar, 2.8% for CO(2) and 3.6% for H(2). The interfering effects of CO, and O(2) were investigated. Working ranges were evaluated as 40-600 ppm for Ar, 30-850 ppm for CO(2) and 10-1200 ppm for H(2). Significant sensitivity, small relative standard deviation (RSD) and acceptable limit of detection (LOD) were obtained for each analyte, showing capability of SWCNTs for gas separation and determination processes. Finally, the method was used to evaluate the contents of CO(2) in air sample.

  5. Numerical determination of competitive adsorption isotherm of mandelic acid enantiomers on cellulose-based chiral stationary phase.

    Science.gov (United States)

    Zhang, Yan; Rohani, Sohrab; Ray, Ajay K

    2008-08-15

    The use of inverse method for the determination of competitive adsorption isotherm of mandelic acid enantiomers on cellulose tris(3,5-diethylphenyl carbamate) stationary phase is proposed in this work. Non-dominated sorting genetic algorithm with jumping genes (NSGA-II-JG) was applied to acquire the isotherm parameters by minimizing the sum of square deviations of the model predictions from the measured elution profiles. Three different competitive isotherm models, i.e., Langmuir, biLangmuir and Tóth, combined with transport-dispersive chromatographic model were used in predicting the elution profiles. Orthogonal collocation on finite element (OCFE) method was applied to obtain the calculated elution profiles. Results indicate that biLangmuir isotherm and Tóth isotherm give remarkably similar equilibrium isotherms within the investigated liquid concentration range. Band profiles calculated from both isotherm models are in good agreement with the experimental data. The validity of the determined parameters was verified by comparing the model predictions with experimental elution profiles at various experimental conditions.

  6. Methacrylate-based monolithic layers for planar chromatography of polymers.

    Science.gov (United States)

    Maksimova, E F; Vlakh, E G; Tennikova, T B

    2011-04-29

    A series of macroporous monolithic methacrylate-based materials was synthesized by in situ free radical UV-initiated copolymerization of functional monomers, such as glycidyl methacrylate (GMA), butyl methacrylate (BuMA), 2-aminoethyl methacrylate (AEMA), 2-hydroxyethyl methacrylate (HEMA) and 2-cyanoethyl methacrylate (CEMA), with crosslinking agent, namely, ethylene glycol dimethacrylate (EDMA). The materials obtained were applied as the stationary phases in simple and robust technique - planar chromatography (PLC). The method of separation layer fabrication representing macroporous polymer monolith bound to the specially prepared glass surface was developed and optimized. The GMA-EDMA and BuMA-EDMA matrixes were successfully applied for the separation of low molecular weight compounds (the mixture of several dies), as well as poly(vinylpyrrolidone) and polystyrene homopolymers of different molecular weights using reversed-phase mechanism. The materials based on copolymers AEMA-HEMA-EDMA and CEMA-HEMA-EDMA were used for normal-phase PLC separation of 2,4-dinitrophenyl amino acids and polystyrene standards.

  7. Formate hydrogen lyase mediates stationary-phase deacidification and increases survival during sugar fermentation in acetoin-producing enterobacteria

    Directory of Open Access Journals (Sweden)

    Bram eVivijs

    2015-02-01

    Full Text Available Two fermentation types exist in the Enterobacteriaceae family. Mixed-acid fermenters produce substantial amounts of lactate, formate, acetate and succinate, resulting in lethal medium acidification. On the other hand, 2,3-butanediol fermenters switch to the production of the neutral compounds acetoin and 2,3-butanediol and even deacidify the environment after an initial acidification phase, thereby avoiding cell death. We equipped three mixed-acid fermenters (Salmonella Typhimurium, S. Enteritidis and Shigella flexneri with the acetoin pathway from Serratia plymuthica to investigate the mechanisms of deacidification. Acetoin production caused attenuated acidification during exponential growth in all three bacteria, but stationary-phase deacidification was only observed in Escherichia coli and Salmonella, suggesting that it was not due to the consumption of protons accompanying acetoin production. To identify the mechanism, 34 transposon mutants of acetoin-producing E. coli that no longer deacidified the culture medium were isolated. The mutations mapped to 16 genes, all involved in formate metabolism. Formate is an end product of mixed-acid fermentation that can be converted to H2 and CO2 by the formate hydrogen lyase (FHL complex, a reaction that consumes protons and thus can explain medium deacidification. When hycE, encoding the large subunit of hydrogenase 3 that is part of the FHL complex, was deleted in acetoin-producing E. coli, deacidification capacity was lost. Metabolite analysis in E. coli showed that introduction of the acetoin pathway reduced lactate and acetate production, but increased glucose consumption and formate and ethanol production. Analysis of a hycE mutant in S. plymuthica confirmed that medium deacidification in this organism is also mediated by FHL. These findings improve our understanding of the physiology and function of fermentation pathways in Enterobacteriaceae.

  8. Chromatographic performance of synthetic polycrystalline diamond as a stationary phase in normal phase high performance liquid chromatography.

    Science.gov (United States)

    Peristyy, Anton; Paull, Brett; Nesterenko, Pavel N

    2015-04-24

    The chromatographic properties of high pressure high temperature synthesised diamond (HPHT) are investigated in normal phase mode of high performance liquid chromatography. Purified nonporous irregular shape particles of average particles size 1.2 μm and specific surface area 5.1 m(2) g(-1) were used for packing 100×4.6 mm ID or 50×4.6 mm ID stainless steel columns. The retention behaviour of several classes of compounds including alkyl benzenes, polyaromatic hydrocarbons (PAH), alkylphenylketones, phenols, aromatic acids and bases were studied using n-hexane-2-propanol mixtures as mobile phase. The results are compared with those observed for microdispersed sintered detonation nanodiamond (MSDN) and porous graphitic carbon (PGC). HPHT diamond revealed distinctive separation selectivity, which is orthogonal to that observed for porous graphitic carbon; while selectivities of HPHT diamond and microdispersed sintered detonation nanodiamonds are similar. Owing to non-porous particle nature, columns packed with high pressure high temperature diamond exhibited excellent mass transfer and produce separations with maximum column efficiency of 128,200 theoretical plates per meter.

  9. Synthesis and characterization of a novel stationary phase, Si-Zr/Ti(PMTDS), based upon ternary oxide support for high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Amparo, Maura R.; Marques, Fabiana A.; Faria, Anizio M., E-mail: anizio@pontal.ufu.br [Universidade Federal de Uberlandia (FACIP/UFU), Ituiutaba, MG (Brazil). Faculdade de Ciencias Integradas do Pontal

    2013-09-15

    A new stationary phase based on the thermal immobilization of poly(methyltetradecylsiloxane) (PMTDS) on silica particles coated with a mixture of zirconia and titania was prepared and evaluated for the chromatographic separation of test mixtures. The spherical particles were characterized by elemental analysis, SEM, FTIR and {sup 29}Si NMR. The physicochemical properties of PMTDS phase supported on Si-Zr/Ti were intermediate between PMTDS phases supported on titanized silica and zirconized silica. The chromatographic performance of Si-Zr/Ti(PMTDS) phase was similar to PMTDS phases based on metal oxide coated silica having only one metal oxide and the preparation of a Si-Zr/Ti(PMTDS) phase allowed evaluation of the effect of each oxide, zirconia and titania, on the separation process and on the stability of the immobilized polymer phase. The hydrolytic stability of Si-Zr/Ti(PMTDS) stationary phase was similar to the Si-Ti(PMTDS) phase, improving the chemical stability of the silica-based PMTDS phase by about 100%. (author)

  10. Preparation of a weak anion exchange/hydrophobic interaction dual-function mixed-mode chromatography stationary phase for protein separation using click chemistry.

    Science.gov (United States)

    Zhao, Kailou; Yang, Fan; Xia, Hongjun; Wang, Fei; Song, Qingguo; Bai, Quan

    2015-03-01

    In this study, 3-diethylamino-1-propyne was covalently bonded to the azide-silica by a click reaction to obtain a novel dual-function mixed-mode chromatography stationary phase for protein separation with a ligand containing tertiary amine and two ethyl groups capable of electrostatic and hydrophobic interaction functionalities, which can display hydrophobic interaction chromatography character in a high-salt-concentration mobile phase and weak anion exchange character in a low-salt-concentration mobile phase employed for protein separation. As a result, it can be employed to separate proteins with weak anion exchange and hydrophobic interaction modes, respectively. The resolution and selectivity of the stationary phase were evaluated in both hydrophobic interaction and ion exchange modes with standard proteins, respectively, which can be comparable to that of conventional weak anion exchange and hydrophobic interaction chromatography columns. Therefore, the synthesized weak anion exchange/hydrophobic interaction dual-function mixed-mode chromatography column can be used to replace two corresponding conventional weak anion exchange and hydrophobic interaction chromatography columns to separate proteins. Based on this mixed-mode chromatography stationary phase, a new off-line two-dimensional liquid chromatography technology using only a single dual-function mixed-mode chromatography column was developed. Nine kinds of tested proteins can be separated completely using the developed method within 2.0 h.

  11. Effect of anaerobic and stationary phase growth conditions on the heat shock and oxidative stress responses in Escherichia coli K-12.

    Science.gov (United States)

    Díaz-Acosta, Alondra; Sandoval, María L; Delgado-Olivares, Luis; Membrillo-Hernández, Jorge

    2006-06-01

    The natural living style of Escherichia coli occurs in the gastrointestinal tract, where most of its existence is spent under anaerobic conditions and in stationary phase of growth. Here we report on the heat shock response of E. coli K-12 cells growing in the presence or absence of oxygen. An rpoH mutant (impaired in the synthesis of the sigma(32) transcriptional factor) exhibited an increased sensitivity to heat shock but only in the exponential phase of aerobic growth, suggesting that in anaerobic growth conditions, and in aerobic stationary phase, sigma(32)-independent mechanisms are playing a prime role in protecting cells from heat stress. Our results demonstrated that sigma(S) is not involved in this protection system. Studies on the kinetics of synthesis of Heat shock proteins (Hsp) after an abrupt rise in temperature demonstrated that in the absence of oxygen, the synthesis of Hsp is triggered faster and is sustained for a longer period of time compared to aerobic growth conditions. Finally, the heated cells in the exponential phase of aerobic growth displayed a high concentration of oxidatively damaged proteins in the presence of 4 mM H(2)O(2), in sharp contrast to cultures of stationary phase or anaerobic growth.

  12. Growth phase-dependent switch in osmolyte strategy in a moderate halophile: ectoine is a minor osmolyte but major stationary phase solute in Halobacillus halophilus.

    Science.gov (United States)

    Saum, Stephan H; Müller, Volker

    2008-03-01

    The moderately halophilic, chloride-dependent bacterium Halobacillus halophilus switches its osmolyte strategy with the salinity in its environment by the production of different compatible solutes. Ectoine is produced predominantly at very high salinities, along with proline. Interestingly, ectoine production is growth phase dependent which led to a more than 1000-fold change in the ectoine : proline ratio from 0.04 in exponential to 27.4 in late stationary phase cultures. The genes encoding the ectoine biosynthesis pathway were identified on the chromosome in the order ectABC. They form an operon that is expressed in a salinity-dependent manner with low-level expression below 1.5 M NaCl but 10-fold and 23-fold increased expression at 2.5 and 3.0 M NaCl respectively. The temporal expression of genes involved in osmoresponse is different with gdh/gln and pro genes being first, followed by ect genes. Chloride had no effect on expression of ect genes, but stimulated cellular EctC synthesis as well as ectoine production. These data demonstrate, for the first time, a growth-phase dependent switch in osmolyte strategy in a moderate halophile and, additionally, represent another piece of the chloride regulon of H. halophilus.

  13. 2-DE based proteomic analysis of Saccharomyces cerevisiae wild and K+ transport-affected mutant (trk1,2) strains at the growth exponential and stationary phases.

    Science.gov (United States)

    Curto, Miguel; Valledor, Luis; Navarrete, Clara; Gutiérrez, Dolores; Sychrova, Hana; Ramos, José; Jorrin, Jesús

    2010-11-10

    By using a 2-DE based workflow, the proteome of wild and potassium transport mutant trk1,2 under optimal growth potassium concentration (50mM) has been analyzed. At the exponential and stationary phases, both strains showed similar growth, morphology potassium content, and Vmax of rubidium transport, the only difference found being the Km values for this potassium analogue transport, higher for the mutant (20mM) than for the wild (3-6mM) cells. Proteins were buffer-extracted, precipitated, solubilized, quantified, and subjected to 2-DE analysis in the 5-8 pH range. More differences in protein content (37-64mgg(-1) cell dry weight) and number of resolved spots (178-307) were found between growth phases than between strains. In all, 164 spots showed no differences between samples and a total of 105 were considered to be differential after ANOVA test. 171 proteins, corresponding to 71 unique gene products have been identified, this set being dominated by cytosolic species and glycolitic enzymes. The ranking of the more abundant spots revealed no differences between samples and indicated fermentative metabolism, and active cell wall biosynthesis, redox homeostasis, biosynthesis of amino acids, coenzymes, nucleotides, and RNA, and protein turnover, apart from cell division and growth. PCA analysis allowed the separation of growth phases (PC1 and 2) and strains at the stationary phase (PC3 and 4), but not at the exponential one. These results are also supported by clustering analysis. As a general tendency, a number of spots newly appeared at the stationary phase in wild type, and to a lesser extent, in the mutant. These up-accumulated spots corresponded to glycolitic enzymes, indicating a more active glucose catabolism, accompanied by an accumulation of methylglyoxal detoxification, and redox-homeostasis enzymes. Also, more extensive proteolysis was observed at the stationary phase with this resulting in an accumulation of low Mr protein species.

  14. Metal Organic Vapor Phase Epitaxy of Monolithic Two-Color Light-Emitting Diodes Using an InGaN-Based Light Converter

    Science.gov (United States)

    Damilano, Benjamin; Kim-Chauveau, Hyonju; Frayssinet, Eric; Brault, Julien; Hussain, Sakhawat; Lekhal, Kaddour; Vennéguès, Philippe; De Mierry, Philippe; Massies, Jean

    2013-09-01

    Monolithic InGaN-based light-emitting diodes (LEDs) using a light converter fully grown by metal organic vapor phase epitaxy are demonstrated. The light converter, consisting of 10-40 InGaN/GaN quantum wells, is grown first, followed by a violet pump LED. The structure and growth conditions of the pump LED are specifically adapted to avoid thermal degradation of the light converter. Electroluminescence analysis shows that part of the pump light is absorbed by the light converter and reemitted at longer wavelength. Depending on the emission wavelength of the light converter, different LED colors are achieved. In particular, for red-emitting light converters, a color temperature of 2100 K corresponding to a tint between warm white and candle light is demonstrated.

  15. Extent of the influence of phosphate buffer and ionic liquids on the reduction of the silanol effect in a C18 stationary phase.

    Science.gov (United States)

    Carda-Broch, S; García-Alvarez-Coque, M C; Ruiz-Angel, M J

    2017-06-01

    The presence of anionic free silanols in the silica-based stationary phases gives rise to broad and asymmetrical peaks when cationic basic compounds are chromatographed using hydro-organic mobile phases. The addition to the mobile phase of a reagent with ionic character prevents the access of analytes to the free silanols, improving the peak shape. The silanol activity can be affected by the buffer concentration and mobile phase pH, factors that are not always considered sufficiently in the literature. In this work, the chromatographic behaviour of three basic β-adrenoceptor antagonists (acebutolol, nadolol and timolol), using mobile phases containing acetonitrile, was examined at different phosphate buffer concentrations (5-50mM) and mobile phase pH (2-8), in the absence and presence of three imidazolium-based ionic liquids (1-ethyl-, 1-butyl- and 1-hexyl-3-methylimidazolium chloride). All factors were evaluated through both the retention and peak shape. The imidazolium cations can block the access of cationic analytes through electrostatic interaction with the anionic silanols, or association with the alkyl chains bound to the stationary phase. In previous reports, the protection mechanism was demonstrated to be directly related to the cation size. The studies in this work reveal that the effectiveness of the mobile phase additive as silanol blocker also depends on the concentration of the buffer anion and the protonation degree of the silanols on the stationary phase. Increasing amounts of phosphate at low pH give rise to increasing retention times. Also, the peak shape is improved, which indicates the influence of phosphate on blocking the activity of free silanols. However, the benefits obtained by the combined effect of buffering the mobile phase at low pH and the use of a bulky additive are lost at pH>6. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Comparison of three development approaches for Stationary Phase Optimised Selectivity Liquid Chromatography based screening methods Part II: A group of structural analogues (PDE-5 inhibitors in food supplements).

    Science.gov (United States)

    Deconinck, E; Ghijs, L; Kamugisha, A; Courselle, P

    2016-02-01

    Three approaches for the development of a screening method to detect adulterated dietary supplements, based on Stationary Phase Optimised Selectivity Liquid Chromatography were compared for their easiness/speed of development and the performance of the optimal method obtained. This comparison was performed for a heterogeneous group of molecules, i.e. slimming agents (Part I) and a group of structural analogues, i.e. PDE-5 inhibitors (Part II). The first approach makes use of primary runs at one isocratic level, the second of primary runs in gradient mode and the third of primary runs at three isocratic levels to calculate the optimal combination of segments of stationary phases. In each approach the selection of the stationary phase was followed by a gradient optimisation. For the PDE-5 inhibitors, the group of structural analogues, only the method obtained with the third approach was able to differentiate between all the molecules in the development set. Although not all molecules are baseline separated, the method allows the identification of the selected adulterants in dietary supplements using only diode array detection. Though, due to the mobile phases used, the method could also be coupled to mass spectrometry. The method was validated for its selectivity following the guidelines as described for the screening of pesticide residues and residues of veterinary medicines in food.

  17. Long-term high-temperature and pH stability assessment of modern commercially available stationary phases by using retention factor analysis.

    Science.gov (United States)

    Haun, Jakob; Oeste, Katja; Teutenberg, Thorsten; Schmidt, Torsten C

    2012-11-09

    High-temperature liquid chromatography provides several advantages concerning HPLC method development and is needed to realize special hyphenation techniques. However, hardware limitations and stationary phase degradation can prevent the successful application of this technique. Therefore, column stability is of major importance. The presented study contains results of long-term high-temperature stability tests for eight modern commercially available HPLC stationary phases. Six of them were silica-based reversed-phase C18 columns tested according to an earlier reported procedure (Teutenberg, 2009 [12]). On the basis of the extended data set that covers a wide range of column technologies, a comparative approach using retention factor analysis was evaluated in order to categorize the columns according to their long-term high-temperature stability at 150 °C and changing pH value. This approach offers a more objective alternative to conventional listings in which temperature maxima are compared for columns tested under different conditions. Additionally, customized tests are presented for one promising polymer-based C18 stationary phase and an amide column for hydrophilic interaction liquid chromatography.

  18. Preparation of a novel weak cation exchange/hydrophobic interaction chromatography dual-function polymer-based stationary phase for protein separation using "thiol-ene click chemistry".

    Science.gov (United States)

    Yang, Fan; Bai, Quan; Zhao, Kailou; Gao, Dong; Tian, Lei

    2015-02-01

    A novel dual-function mixed-mode stationary phase based on poly(glycidyl methacrylate-co-ethylene dimethacrylate) microspheres was synthesized by thiol-ene click chemistry and characterized by infrared spectroscopy and elemental analysis. The new system displays both hydrophobic interaction chromatography (HIC) character in a high salt concentration mobile phase, and weak cation exchange (WCX) chromatography character in a low salt concentration mobile phase. It can be used to separate proteins in both ion-exchange chromatography (IEC) mode and HIC mode. The resolution and selectivity of the stationary phase were evaluated in both HIC mode and IEC mode using protein standards. In comparison with the conventional WCX and HIC columns, the results were satisfactory and acceptable. Protein mass and bioactivity recoveries of more than 96% can be achieved in both HIC mode and IEC mode using this column. The results indicate that the novel dual-function mixed-mode column in many cases can replace the use of two individual WCX and HIC columns. In addition, the effects on protein separation of different ligand structures in the dual-function stationary phase and the pH of the mobile phase used were also investigated in detail. The results show that electrostatic interaction of the ligand with proteins must match the hydrophobicity of the ligand, which is an important factor to prepare the dual-function stationary phase. On the basis of this dual-function mixed-mode chromatography column, a new two-dimensional liquid chromatography technology with a single column system was also developed in this study, and was used to renature and purify recombinant human interferon-γ from inclusion bodies. The mass recovery, purity, and specific bioactivity obtained for the purified recombinant human interferon-γ were 87.2%, 92.4%, and 2.8 × 10(7) IU/mg, respectively, in IEC mode, and 83.4%, 95.2%, and 4.3 × 10(7) IU/mg, respectively, in HIC mode. The results indicate that the

  19. Monoliths in Bioprocess Technology

    Directory of Open Access Journals (Sweden)

    Vignesh Rajamanickam

    2015-04-01

    Full Text Available Monolithic columns are a special type of chromatography column, which can be used for the purification of different biomolecules. They have become popular due to their high mass transfer properties and short purification times. Several articles have already discussed monolith manufacturing, as well as monolith characteristics. In contrast, this review focuses on the applied aspect of monoliths and discusses the most relevant biomolecules that can be successfully purified by them. We describe success stories for viruses, nucleic acids and proteins and compare them to conventional purification methods. Furthermore, the advantages of monolithic columns over particle-based resins, as well as the limitations of monoliths are discussed. With a compilation of commercially available monolithic columns, this review aims at serving as a ‘yellow pages’ for bioprocess engineers who face the challenge of purifying a certain biomolecule using monoliths.

  20. A novel chiral stationary phase LC-MS/MS method to evaluate oxidation mechanisms of edible oils.

    Science.gov (United States)

    Ito, Junya; Shimizu, Naoki; Kobayashi, Eri; Hanzawa, Yasuhiko; Otoki, Yurika; Kato, Shunji; Hirokawa, Takafumi; Kuwahara, Shigefumi; Miyazawa, Teruo; Nakagawa, Kiyotaka

    2017-08-30

    The elucidation of lipid oxidation mechanisms of food is vital. In certain lipids, characteristic lipid hydroperoxide isomers are formed by different oxidation mechanisms (i.e., photo-oxidation or auto-oxidation). For example, linoleic acid is photo-oxidized to 13-9Z, 11E-hydroperoxyoctadecadienoic acid (HPODE), 12-9Z,13E-HPODE, 10-8E,12Z-HPODE and 9-10E,12Z-HPODE, whereas 13-9Z, 11E-HPODE, 13-9E,11E-HPODE, 9-10E,12Z-HPODE and 9-10E,12E-HPODE are formed by auto-oxidation. Therefore, we considered that oxidation mechanisms could be evaluated by analyzing these characteristic positional and cis/trans lipid hydroperoxide isomers. In this study, we developed a novel chiral stationary phase LC-MS/MS (CSP-LC-MS/MS) method to analyze the positional and cis/trans isomers of HPODE, with the use of a chiral column and sodium ion. Also, as an application of the method, either light-exposed or heated edible oils were treated with lipase to hydrolyze triacylglycerols. The resultant fatty acids including HPODE isomers were analyzed with the developed method. As a result, HPODE isomers characteristic to photo-oxidation were certainly detected in light-exposed edible oils. On the other hand, in heated edible oils, the HPODE isomers characteristic to auto-oxidation were largely increased. Thus, the combination of the developed CSP-LC-MS/MS method with lipase proves to be a powerful tool to evaluate the involvement and mechanisms of lipid oxidation in the process of food deterioration.

  1. Separation of opiate alkaloids by electrokinetic chromatography with sulfated-cyclodextrin as a pseudo-stationary phase.

    Science.gov (United States)

    Zakaria, Philip; Macka, Miroslav; Haddad, Paul R

    2003-01-24

    The separation of six related opiate alkaloids (morphine, thebaine, 10-hydroxythebaine, codeine, oripavine and laudanine) was studied using sulfated-cyclodextrin (s-CD) as a cation-exchange pseudo-stationary phase. Cation-exchange interactions between the cationic analytes and the anionic s-CD (7-11 mol of sulfate groups per mole CD) were found to bethe predominant mechanism, allowing the separations to be performed at low pH where the opiates are protonated and exhibit very similar mobilities. The concentrations of the s-CD and the competing ion (Na+ or Mg2+) in the electrolyte were used to govern the extent of the ion-exchange interactions. Interactions with the sulfated-cyclodextrin differed for each analyte, with oripavine exhibiting the strongest interaction and 10-thebaine and laudanine showing the weakest interactions. Despite the very similar structures of the analytes, these differences resulted in significant changes in separation selectivity. The separation was modelled using a migration equation derived from first principles and based on ion-exchange interactions between the s-CD and the opiates. Constants within the model were obtained by non-linear regression using a small subset of experimentally determined migration times. These constants related to the ion-exchange affinities of the s-CD for the various opiates. When the model was used to predict migration times under other experimental conditions, a very good correlation was obtained between observed and predicted mobilities (r2=0.996). Optimisation of the system was performed using the normalised resolution product and minimum resolution criteria and this process provided two optimised separations, each exhibiting a different separation selectivity.

  2. The CGC enantiomer separation of 2-arylcarboxylic acid esters by using β-cyclodextrin derivatives as chiral stationary phases.

    Science.gov (United States)

    Shi, Xueyan; Liu, Feipeng; Mao, Jianyou

    2016-03-17

    Chiral 2-arylcarboxylic acid esters are important intermediates in preparation of enantioenriched 2-arylpropionic acids type Non-steroidal anti-inflammatory drugs (NSAIDs). Enantiomer separation of 2-arylcarboxylic acid esters is crucial for evaluation of the asymmetric synthesis efficiency and the enantiomer excess of chiral 2-arylcarboxylic acid derivatives. The capillary gas chromatography (CGC) enantiomer separation of 17 pairs of 2-arylcarboxylic acid esters enantiomers was conducted by using seven different β-cyclodextrin derivatives (CDs) as chiral stationary phases. It was found that for the 7 pairs of 2-phenylpropionates enantiomers, CDs with both alkyl and acyl substituents especially 2,6-di-O-pentyl-3-O-butyryl-β-cyclodextrin exhibited better enantiomer separation abilities than the other CDs examined. For the 7 pairs of 2-(4-substituted phenyl)propionates enantiomers, 2,3,6-tri-O-methyl-β-cyclodextrin possessed better enantiomer separation abilities than the other CDs. Among the 3 pairs of 2-phenylbutyrates enantiomers examined, only methyl 2-phenylbutyrate enantiomers could be separated by three CDs among the 7 CDs tested, while enantiomers of ethyl 2-phenylbutyrate and isopropyl 2-phenylbutyrate couldn't be separated by any of the 7 CDs tested. Besides the structures of CDs, the structures of 2-arylcarboxylic acid esters including different ester moieties, substituents of phenyl, and different carboxylic acids moieties in 2-arylcarboxylic acid esters also affected the enantiomer separation results greatly. The CGC enantiomer separation results of 2-arylcarboxylic acid esters on different CDs are useful for solving the enantiomer separation problem of 2-arylcarboxylic acid esters.

  3. Synthesis of a new cyclosporine-based stationary phase and separation behaviors toward aromatic positional isomers by high-performance liquid chromatography.

    Science.gov (United States)

    Liu, Wen-na; Fan, Jun; Lin, Chun; Zheng, Sheng-run; Zhang, Wei-guang

    2015-04-01

    A new cyclosporine-bonded stationary phase has been synthesized through the Staudinger reaction between azido cyclosporine A (CsA) and aminopropyl silica gel and applied in separations of six disubstituted aromatic analytes by high-performance liquid chromatography. For dimethyl phthalate, nitroaniline and chloronitrobenzene, their positional isomers were well-separated on this CsA stationary phase, in which the separation factor for m-/o-dimethyl phthalates was the biggest. For nitrotoluene, dichlorobenzene and benzenediol, the m-/o-isomers were co-eluted. Then, effects of chromatographic conditions (such as types and content of alcoholic modifiers) on separation of nitroaniline positional isomers have been investigated. Retention behaviors of nitroaniline isomers on the column exhibited the strengthening trend along with increasing carbon number of alcohols, from ethanol to 1-propanol, and to 1-butanol. A similar phenomenon was observed by lowering the content of alcohol.

  4. L—PROLINE—FUNCTIONALIZED POLYSTYRENE WITH 1—METHYLDECYLENE SPACER AS A CHIRAL STATIONARY PHASE FOR LIGAND—EXCHANGE CHROMATOGRAPHIC RESOLUTION OF AMINO ACID RECEMATES

    Institute of Scientific and Technical Information of China (English)

    MaJianbiao; ChenLi; 等

    1995-01-01

    The L-proline-functionalized polystyrene with 1-methyldecylene spacer was synthesized from 2% divinylbenzene-crosslinked polystyrene gel via 10-bromo-1-methyldecyl polystyrene intermediate.After complexed with copper(Ⅱ) ion,the polymer with L-proline ligand was used as the chiral stationary phase (CSP) for ligand exchange chromatography of amino acid racemates.The results showed that the CSP possessed powerful enantioselectivity and all racemates of the fifteen tried amino acids were completely separated.

  5. Perpentylated (2, 3, 6-Tri-O-pentyl)-β-cyclodextrin Used as Capillary Gas Chromatographic Stationary Phase Prepared by Sol-gel Technology

    Institute of Scientific and Technical Information of China (English)

    Min Min LIANG; Mei Ling QI; Yu LENG; Ruo Nong FU

    2005-01-01

    Capillary column preparation using perpentylated (2,3,6-tri-O-pentyl)-β-cyclodextrin as stationary phase by sol-gel technology with simplicity and rapidity is described. Multiple preparation steps in conventional column technology were avoided. The prepared columns exhibit satisfactory chromatographic performances and pronounced selectivity for a wide range of test solutes, and have been successfully used for the separation of nitrotoluene, dimethoxybenzene,alcohols, alkanes, dimethylphenol and cresol isomers.

  6. Stationary phase induction of dnaN and recF, two genes of Escherichia coli involved in DNA replication and repair.

    Science.gov (United States)

    Villarroya, M; Pérez-Roger, I; Macián, F; Armengod, M E

    1998-03-16

    The beta subunit of DNA polymerase III holoenzyme, the Escherichia coli chromosomal replicase, is a sliding DNA clamp responsible for tethering the polymerase to DNA and endowing it with high processivity. The gene encoding beta, dnaN, maps between dnaA and recF, which are involved in initiation of DNA replication at oriC and resumption of DNA replication at disrupted replication forks, respectively. In exponentially growing cells, dnaN and recF are expressed predominantly from the dnaA promoters. However, we have found that stationary phase induction of the dnaN promoters drastically changes the expression pattern of the dnaA operon genes. As a striking consequence, synthesis of the beta subunit and RecF protein increases when cell metabolism is slowing down. Such an induction is dependent on the stationary phase sigma factor, RpoS, although the accumulation of this factor alone is not sufficient to activate the dnaN promoters. These promoters are located in DNA regions without static bending, and the -35 hexamer element is essential for their RpoS-dependent induction. Our results suggest that stationary phase-dependent mechanisms have evolved in order to coordinate expression of dnaN and recF independently of the dnaA regulatory region. These mechanisms might be part of a developmental programme aimed at maintaining DNA integrity under stress conditions.

  7. Involvement of GDH3-encoded NADP+-dependent glutamate dehydrogenase in yeast cell resistance to stress-induced apoptosis in stationary phase cells.

    Science.gov (United States)

    Lee, Yong Joo; Kim, Kyung Jin; Kang, Hong Yong; Kim, Hye-Rim; Maeng, Pil Jae

    2012-12-28

    Glutamate metabolism is linked to a number of fundamental metabolic pathways such as amino acid metabolism, the TCA cycle, and glutathione (GSH) synthesis. In the yeast Saccharomyces cerevisiae, glutamate is synthesized from α-ketoglutarate by two NADP(+)-dependent glutamate dehydrogenases (NADP-GDH) encoded by GDH1 and GDH3. Here, we report the relationship between the function of the NADP-GDH and stress-induced apoptosis. Gdh3-null cells showed accelerated chronological aging and hypersusceptibility to thermal and oxidative stress during stationary phase. Upon exposure to oxidative stress, Gdh3-null strains displayed a rapid loss in viability associated with typical apoptotic hallmarks, i.e. reactive oxygen species accumulation, nuclear fragmentation, DNA breakage, and phosphatidylserine translocation. In addition, Gdh3-null cells, but not Gdh1-null cells, had a higher tendency toward GSH depletion and subsequent reactive oxygen species accumulation than did WT cells. GSH depletion was rescued by exogenous GSH or glutamate. The hypersusceptibility of stationary phase Gdh3-null cells to stress-induced apoptosis was suppressed by deletion of GDH2. Promoter swapping and site-directed mutagenesis of GDH1 and GDH3 indicated that the necessity of GDH3 for the resistance to stress-induced apoptosis and chronological aging is due to the stationary phase-specific expression of GDH3 and concurrent degradation of Gdh1 in which the Lys-426 residue plays an essential role.

  8. Involvement of GDH3-encoded NADP+-dependent Glutamate Dehydrogenase in Yeast Cell Resistance to Stress-induced Apoptosis in Stationary Phase Cells*

    Science.gov (United States)

    Lee, Yong Joo; Kim, Kyung Jin; Kang, Hong Yong; Kim, Hye-Rim; Maeng, Pil Jae

    2012-01-01

    Glutamate metabolism is linked to a number of fundamental metabolic pathways such as amino acid metabolism, the TCA cycle, and glutathione (GSH) synthesis. In the yeast Saccharomyces cerevisiae, glutamate is synthesized from α-ketoglutarate by two NADP+-dependent glutamate dehydrogenases (NADP-GDH) encoded by GDH1 and GDH3. Here, we report the relationship between the function of the NADP-GDH and stress-induced apoptosis. Gdh3-null cells showed accelerated chronological aging and hypersusceptibility to thermal and oxidative stress during stationary phase. Upon exposure to oxidative stress, Gdh3-null strains displayed a rapid loss in viability associated with typical apoptotic hallmarks, i.e. reactive oxygen species accumulation, nuclear fragmentation, DNA breakage, and phosphatidylserine translocation. In addition, Gdh3-null cells, but not Gdh1-null cells, had a higher tendency toward GSH depletion and subsequent reactive oxygen species accumulation than did WT cells. GSH depletion was rescued by exogenous GSH or glutamate. The hypersusceptibility of stationary phase Gdh3-null cells to stress-induced apoptosis was suppressed by deletion of GDH2. Promoter swapping and site-directed mutagenesis of GDH1 and GDH3 indicated that the necessity of GDH3 for the resistance to stress-induced apoptosis and chronological aging is due to the stationary phase-specific expression of GDH3 and concurrent degradation of Gdh1 in which the Lys-426 residue plays an essential role. PMID:23105103

  9. Validated method for bioactive lignans in Schisandra chinensis in vitro cultures using a solid phase extraction and a monolithic column application.

    Science.gov (United States)

    Brezinová, Lenka; Vlasínová, Helena; Havel, Ladislav; Humpa, Otakar; Slanina, Jirí

    2010-09-01

    A simple and rapid method for determination of six lignans found in plant cell cultures of Schisandra chinensis was developed and validated. The lignans were extracted from plant samples with methanol and the extracts were effectively cleaned by solid-phase extraction using Strata C18-E (Phenomenex) cartridges. Chromatographic separation was carried out on a Chromolith Performance RP-18e monolithic column (100 x 4.6 mm, Merck) using an isocratic mobile phase of acetonitrile and water in a 50:50 (v/v) ratio. The eluent was monitored at 220 nm. The baseline separation of schizandrin, gomisin A, deoxyschizandrin, gamma-schizandrin, gomisin N and wuweizisu C was achieved in a relatively short time period (20 min), which was made possible by the relatively high flow rate of the mobile phase (2 mL/min). The lower limit of quantitation was 0.1 mg/L for schizandrin and gomisin A, 0.3 mg/L for deoxyschizandrin, gamma-schizandrin, and gomisin N and 1 mg/L for wuweizisu C. The analysis of spiked samples containing six lignans provided absolute recoveries between 93 and 101% in all cases. The validated method was successfully applied to the determination of lignans in embryogenic plant cell cultures of Schisandra chinensis.

  10. Adsorption mechanism in reversed-phase liquid chromatography. Effect of the surface coverage of a monomeric C18-silica stationary phase

    Energy Technology Data Exchange (ETDEWEB)

    Gritti, Fabrice [University of Tennessee, Knoxville (UTK); Guiochon, Georges A [ORNL

    2006-04-01

    The effect of the bonding density of the octadecyl chains onto the same silica on the adsorption and retention properties of low molecular weight compounds (phenol, caffeine, and sodium 2-naphthalene sulfonate) was investigated. The same mobile phase (methanol:water, 20:80, v/v) and temperature (T = 298 K) were applied and two duplicate columns (A and B) from each batch of packing material (neat silica, simply endcapped or C{sub 1} phase, 0.42, 1.01, 2.03, and 3.15 {micro}mol/m{sup 2} of C{sub 18} alkyl chains) were tested. Adsorption data of the three compounds were acquired by frontal analysis (FA) and the adsorption energy distributions (AEDs) were calculated using the expectation-maximization method. Results confirmed earlier findings in linear chromatography of a retention maximum at an intermediate bonding density. From a general point of view, the saturation capacity of the adsorbent tends to decrease with increasing bonding density, due to the vanishing space intercalated between the C{sub 18} bonded chains and to the decrease of the specific surface area of the stationary phase. The equilibrium constants are maximum for an intermediary bonding density ({approx}2 {micro}mol/m{sup 2}). An enthalpy-entropy compensation was found for the thermodynamic parameters of the isotherm data. Weak equilibrium constants (small {Delta}H) and high saturation capacities (large {Delta}S) were observed at low bonding densities, higher equilibrium constants and lower saturation capacities at high bonding densities, the combinations leading to similar apparent retention in RPLC. The use of a low surface coverage column is recommended for preparative purposes.

  11. pi-Selective stationary phases: (I) Influence of the spacer chain length of phenyl type phases on the aromatic and methylene selectivity of aromatic compounds in reversed phase high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Mayfield, Kirsty [University of Western Sydney, Australia; Dennis, Gary [University of Western Sydney, Australia; Gritti, Fabrice [University of Tennessee, Knoxville (UTK); Guiochon, Georges A [ORNL; Shalliker, R. Andrew [University of Western Sydney, Australia

    2010-01-01

    Phenyl type stationary phases of increasing spacer chain length (phenyl, methyl phenyl, ethyl phenyl, propyl phenyl and butyl phenyl, with 0-4 carbon atoms in the spacer chain, respectively) were synthesized and packed in house to determine the impact that the spacer chain length has on the retention process. Two trends in the aromatic selectivity, q{sub aromatic}, were observed, depending on whether the number of carbon atoms in the spacer chain is even or odd. Linear log k' vs {phi} plots were obtained for each stationary phase and the S coefficient was determined from the gradient of these plots. For the phenyl type phases, the S vs n{sub c} plots of the retention factors of linear polycyclic aromatic hydrocarbons vs the number of rings exhibit a distinct discontinuity that between 3 and 4 rings, which increases with increasing spacer chain length for even phases but decreases for odd phases. Accordingly, we suggest that the retention factors depend differently on the number of carbon atoms in the spacer chain depending on whether this number is even or odd and that this effect is caused by different orientations of the aromatic ring relative to the silica surface.

  12. Ionic liquids as porogens in the microwave-assisted synthesis of methacrylate monoliths for chromatographic application

    Energy Technology Data Exchange (ETDEWEB)

    Singco, Brenda; Lin, Chen-Lan; Cheng, Yi-Jie; Shih, Yung-Han [Department of Chemistry and Center for Nanotechnology, CYCU (Chung Yuan Christian University), 200 Chung Pei Road, Chung-Li 320, Taiwan (China); Huang, Hsi-Ya, E-mail: hyhuang@cycu.edu.tw [Department of Chemistry and Center for Nanotechnology, CYCU (Chung Yuan Christian University), 200 Chung Pei Road, Chung-Li 320, Taiwan (China)

    2012-10-09

    Highlights: Black-Right-Pointing-Pointer An efficient and cleaner monoliths preparation utilizing ionic liquids in conjunction with microwave Black-Right-Pointing-Pointer These ionic liquids with varied cation alkyl chain and anion type successfully tuned the morphology of different alkyl methacrylates Black-Right-Pointing-Pointer Small molecules and peptide digests separated well in these monoliths. - Abstract: Several imidazolium-based ionic liquids (ILs) with varying cation alkyl chain length (C{sub 4}-C{sub 10}) and anion type (tetrafluoroborate ([BF{sub 4}]{sup -}), hexafluorophosphate ([PF{sub 6}]{sup -}) and bis(trifluoromethylsulfonyl)imide ([Tf{sub 2}N]{sup -})) were used as reaction media in the microwave polymerization of methacrylate-based stationary phases. Scanning electron micrographs and backpressures of poly(butyl methacrylate-ethylene dimethacrylate) (poly(BMA-EDMA)) monoliths synthesized in the presence of these ionic liquids demonstrated that porosity and permeability decreased when cation alkyl chain length and anion hydrophobicity were increased. Performance of these monoliths was assessed for their ability to separate parabens by capillary electrochromatography (CEC). Intra-batch precision (n = 3 columns) for retention time and peak area ranged was 0.80-1.13% and 3.71-4.58%, respectively. In addition, a good repeatability of RSD{sub Retentiontime} = <0.30% and {approx}1.0%, RSD{sub Peakarea} = <1.30% and <4.3%, and RSD{sub Efficiency} = <0.6% and <11.5% for intra-day and inter-day, respectively exemplify monolith performance reliability for poly(BMA-EDMA) fabricated using 1-hexyl-3-methylimidazolium tetrafluoroborate ([C{sub 6}mim][BF{sub 4}]) porogen. This monolith was also tested for its potential in nanoLC to separate protein digests in gradient mode. ILs as porogens also fabricated different alkyl methacrylate (AMA) (C4-C18) monoliths. Furthermore, employing binary IL porogen mixture such as 1-butyl-3-methylimidazolium tetrafluoroborate

  13. Hydrophobic-hydrophilic monolithic dual-phase layer for two-dimensional thin-layer chromatography coupled with surface-enhanced Raman spectroscopy detection.

    Science.gov (United States)

    Zheng, Binxing; Liu, Yanhua; Li, Dan; Chai, Yifeng; Lu, Feng; Xu, Jiyang

    2015-08-01

    Hydrophobic-hydrophilic monolithic dual-phase plates have been prepared by a two-step polymerization method for two-dimensional thin-layer chromatography of low-molecular-weight compounds, namely, several dyes. The thin 200 μm poly(glycidyl methacrylate-co-ethylene dimethacrylate) layers attached to microscope glass plates were prepared using a UV-initiated polymerization method within a simple glass mold. After cutting and cleaning the specific area of the layer, the reassembled mold was filled with a polymerization mixture of butyl methacrylate and ethylene dimethacrylate and subsequently irradiated with UV light. During the second polymerization process, the former layer was protected from the UV light with a UV mask. After extracting the porogens and hydrolyzing the poly(glycidyl methacrylate-co-ethylene dimethacrylate) area, these two-dimensional layers were used to separate a mixture of dyes with great difference in their polarity using reversed-phase chromatography mode within the hydrophobic layer and then hydrophilic interaction chromatography mode along the hydrophilic area. In the latter dimension only the specific spot was developed further. Detection of the separated dyes could be achieved with surface-enhanced Raman spectroscopy.

  14. Application of a cholesterol stationary phase in the analysis of phosphorothioate oligonucleotides by means of ion pair chromatography coupled with tandem mass spectrometry.

    Science.gov (United States)

    Studzińska, Sylwia; Krzemińska, Katarzyna; Szumski, Michał; Buszewski, Bogusław

    2016-07-01

    The main aim of this study was the investigation of the influence of several ion pair reagents towards both the retention and the mass spectrometry sensitivity of phosphorothioate oligonucleotides. A cholesterol stationary phase was applied for the first time in the analysis of this group of compounds. The mobile phase composition was modified by changing the concentration and the type of amines and acetates or 1,1,1,3,3,3-hexafluoroisopropanol. It has been shown that the increase of amines concentration results in the retention factor increase for each oligonucleotide, on each adsorbent. The only exception was the mobile phase composed of triethylamine and 1,1,1,3,3,3-hexafluoroisopropanol. This is a consequence of interactions taking place between a cholesterol molecule and an alcohol. This effect was convenient when the mass spectrometry detection was applied, since it allowed an increase in the sensitivity. Moreover, optimization of the mobile phase composition and its impact on the efficiency of ionization process and on the sensitivity in mass spectrometry were also presented. The optimization of this new method, based on cholesterol stationary phase coupled with mass spectrometry detection, was finally applied for the determination of phosphorothioate oligonucleotides impurity in a real sample. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Ion-exchange and hydrophobic interactions affecting selectivity for neutral and charged solutes on three structurally similar agglomerated ion-exchange and mixed-mode stationary phases.

    Science.gov (United States)

    Kazarian, Artaches A; Taylor, Mark R; Haddad, Paul R; Nesterenko, Pavel N; Paull, Brett

    2013-11-25

    The nature and extent of mixed-mode retention mechanisms evident for three structurally related, agglomerated, particle-based stationary phases were evaluated. These three agglomerated phases were Thermo Fisher ScientificIon PacAS11-HC - strong anion exchange, Thermo Fisher Scientific IonPac CS10--strong cation-exchange PS-DVB, and the Thermo Fisher Scientific Acclaim Trinity P1silica-based substrate, which is commercially marketed as a mixed-mode stationary phase. All studied phases can exhibit zwitterionic and hydrophobic properties, which contribute to the retention of charged organic analytes. A systematic approach was devised to investigate the relative ion-exchange capacities and hydrophobicities for each of the three phases, together with the effect of eluent pH upon selectivity, using a specifically selected range of anionic, cationic and neutral aromatic compounds. Investigation of the strong anion-exchange column and the Trinity P1 mixed-mode substrate, in relation to ion-exchange capacity and pH effects, demonstrated similar retention behaviour for both the anionic and ampholytic solutes, as expected from the structurally related phases. Further evaluation revealed that the ion-exchange selectivity of the mixed-mode phase exhibited properties similar to that of the strong anion-exchange column, with secondary cation-exchange selectivity, albeit with medium to high anion-exchange and cation-exchange capacities, allowing selective retention for each of the anionic, cationic and ampholytic solutes. Observed mixed-mode retention upon the examined phases was found to be a sum of anion- and cation-exchange interactions, secondary ion-exchange and hydrophobic interactions, with possible additional hydrogen bonding. Hydrophobic evaluation of the three phases revealed logP values of 0.38-0.48, suggesting low to medium hydrophobicity. These stationary phases were also benchmarked against traditional reversed-phase substrates namely, octadecylsilica YMC-Pac Pro C18

  16. Discriminação quiral por CLAE em carbamatos de polissacarídeos: desenvolvimento, aplicações e perspectivas Polysaccharide carbamate as chiral stationary phases for HPLC: development, applications and perspectives

    Directory of Open Access Journals (Sweden)

    Quezia B. Cass

    1997-02-01

    Full Text Available The importance of chromatographic separation of enantiomers has been acknowledged. This review surveys the development, application and perspectives of polysaccharide carbamates as chiral stationary phase for HPLC.

  17. Effect of chromatographic conditions on retention behavior and system efficiency for HPTLC of selected psychotropic drugs on chemically bonded stationary phases.

    Science.gov (United States)

    Petruczynik, Anna; Wróblewski, Karol; Waksmundzka-Hajnos, Monika

    2015-01-01

    Selected psychotropic drug standards have been chromatographed on RP18, CN and diol layers with a variety of aqueous and nonaqueous mobile phases. The effect of buffers at acidic or basic pH, acetic acid, ammonia and diethylamine (DEA) in aqueous mobile phases on retention, efficiency and peak symmetry was examined. Improved peak symmetry and separation selectivity for investigated compounds were observed when ammonia or DEA were used as mobile phase additives. The effect of diethylamine concentration in aqueous eluents on retention, peak symmetry and theoretical plate number obtained on CN plates was also investigated. Because of the strong retention of these basic drugs on stationary phases bonded on silica matrix, nonaqueous eluents containing medium polar diluents, strongly polar modifiers and silanol blockers (ammonia or diethylamine) were applied. Aqueous and nonaqueous eluent systems with the best selectivity and efficiency were used for separate psychotropic drug standards' mixture on CN layer by 2D TLC.

  18. Effects of the dynamic modification of stationary phases by sorbates in gas chromatography: The possibility of separating enantiomers in achiral systems

    Science.gov (United States)

    Zenkevich, I. G.; Pavlovskii, A. A.

    2016-10-01

    It is shown that the gas chromatographic separation of enantiomers on columns with achiral nonpolar stationary phases is principally possible as a result of the dynamic modification of stationary phases by sorbates under analysis. It is found that a number of key characteristic features is intrinsic to such separation: it can be only partial, it does not occur for all chromatographic columns, and it is observed only for some compounds and only within narrow ranges of quantities of sorbates that are close to the limits of mass overload of chromatographic systems. These characteristic features are illustrated by the examples of separating (1 R,5 R)-(+)- and (1 S,5 S)-(-)-α-pinenes on a WCOT column with an RTX-5 phase. The main characteristic feature of the separation of enantiomers as a result of the dynamic modification of stationary phases is the nonconformity of peaks in chromatograms with two individual enantiomers, compared to other ways and means for their separation; the first eluting peak belongs to the enantiomer that predominates in a mixture irrespective of its configuration, while the second peak corresponds to the racemic mixture of enantiomers; i.e., the ratio of peak areas in chromatograms does not correspond to the actual ratio of enantiomers in samples under analysis and is strongly distorted as a result of their incomplete separation. It is concluded that the separation of racemic mixtures in achiral systems is fundamentally impossible under any conditions, and this is one of the key criteria of the validity of the considered concept as a whole.

  19. Preparation and retention mechanism study of graphene and graphene oxide bonded silica microspheres as stationary phases for high performance liquid chromatography.

    Science.gov (United States)

    Zhang, Xiaoqiong; Chen, Sha; Han, Qiang; Ding, Mingyu

    2013-09-13

    Graphene oxide (GO) bonded stationary phase for high performance liquid chromatography (HPLC) was fabricated by coating GO sheets onto aminosilica microspheres via covalent coupling. Graphene (G) functionalized HPLC stationary phase was then prepared through hydrazine reduction of GO bonded silica (GO@SiO2) composite, which was the first example of using graphene as stationary-phase component for HPLC. Effective separations of the tested neutral and polar compounds on both GO@SiO2 and graphene bonded silica (G@SiO2) columns were achieved under the optimal experimental conditions. Compared with commercial C18 column, the different chromatographic performances of GO and graphene bonded columns were ascribed to their unique retention mechanisms. The polyaromatic scaffold of GO and graphene gives π-π stacking property and hydrophobic effect, and other retention mechanisms, such as π-π electron-donor-acceptor (EDA) interaction for the separation of nitroaromatic compounds and hydrogen bonding for hydroxyl and amino compounds, may also be taken into consideration. Experimental results indicated that the mixed-mode retention mechanism can facilitate the separation of analytes with similar hydrophobicity, which is a unique property compared with C18 column. Additionally, G@SiO2 showed higher affinity to aromatic analytes in contrast with GO@SiO2 and its retention mechanism was not consistent with the typical reversed phase behavior. The separation of aromatic compounds on G@SiO2 column relies primarily on the π-π stacking interaction and then the hydrophobicity, while the two interactions have equal shares on GO@SiO2 column.

  20. Monolithic Fuel Fabrication Process Development

    Energy Technology Data Exchange (ETDEWEB)

    C. R. Clark; N. P. Hallinan; J. F. Jue; D. D. Keiser; J. M. Wight

    2006-05-01

    The pursuit of a high uranium density research reactor fuel plate has led to monolithic fuel, which possesses the greatest possible uranium density in the fuel region. Process developments in fabrication development include friction stir welding tool geometry and cooling improvements and a reduction in the length of time required to complete the transient liquid phase bonding process. Annealing effects on the microstructures of the U-10Mo foil and friction stir welded aluminum 6061 cladding are also examined.

  1. Monolithic molecular imprinted polymer fiber for recognition and solid phase microextraction of ephedrine and pseudoephedrine in biological samples prior to capillary electrophoresis analysis.

    Science.gov (United States)

    Deng, Dong-Li; Zhang, Ji-You; Chen, Chen; Hou, Xiao-Ling; Su, Ying-Ying; Wu, Lan

    2012-01-01

    A novel capillary electrophoresis (CE) method coupled with monolithic molecular imprinted polymer (MIP) fiber based solid phase microextraction (SPME) was developed for selective and sensitive determination of ephedrine (E) and pseudoephedrine (PE). With in situ polymerization in a silica capillary mold and E as template, the MIP fibers could be produced in batch reproducibly and each fiber was available for 50 extraction cycles without significant decrease in extraction ability. Using the MIP fiber under optimized extraction conditions, CE detection limits of E and PE were greatly lowered from 0.20 to 0.00096 μg/mL and 0.12 to 0.0011 μg/mL, respectively. Analysis of urine and serum samples by the MIP-SPME-CE method was also performed, with results indicating that E and PE could be selectively extracted. The recoveries and relative standard deviations (RSDs) for sample analysis were found in the range of 91-104% and 3.8-9.1%, respectively.

  2. In Bacillus subtilis LutR is part of the global complex regulatory network governing the adaptation to the transition from exponential growth to stationary phase.

    Science.gov (United States)

    Irigül-Sönmez, Öykü; Köroğlu, Türkan E; Öztürk, Büşra; Kovács, Ákos T; Kuipers, Oscar P; Yazgan-Karataş, Ayten

    2014-02-01

    The lutR gene, encoding a product resembling a GntR-family transcriptional regulator, has previously been identified as a gene required for the production of the dipeptide antibiotic bacilysin in Bacillus subtilis. To understand the broader regulatory roles of LutR in B. subtilis, we studied the genome-wide effects of a lutR null mutation by combining transcriptional profiling studies using DNA microarrays, reverse transcription quantitative PCR, lacZ fusion analyses and gel mobility shift assays. We report that 65 transcriptional units corresponding to 23 mono-cistronic units and 42 operons show altered expression levels in lutR mutant cells, as compared with lutR(+) wild-type cells in early stationary phase. Among these, 11 single genes and 25 operons are likely to be under direct control of LutR. The products of these genes are involved in a variety of physiological processes associated with the onset of stationary phase in B. subtilis, including degradative enzyme production, antibiotic production and resistance, carbohydrate utilization and transport, nitrogen metabolism, phosphate uptake, fatty acid and phospholipid biosynthesis, protein synthesis and translocation, cell-wall metabolism, energy production, transfer of mobile genetic elements, induction of phage-related genes, sporulation, delay of sporulation and cannibalism, and biofilm formation. Furthermore, an electrophoretic mobility shift assay performed in the presence of both SinR and LutR revealed a close overlap between the LutR and SinR targets. Our data also revealed a significant overlap with the AbrB regulon. Together, these findings reveal that LutR is part of the global complex, interconnected regulatory systems governing adaptation of bacteria to the transition from exponential growth to stationary phase.

  3. Common screening approaches for efficient analytical method development in LC and SFC on columns packed with immobilized polysaccharide-derived chiral stationary phases.

    Science.gov (United States)

    Franco, Pilar; Zhang, Tong

    2013-01-01

    Owing to their remarkable enantioselectivity, versatility, and stability, immobilized polysaccharide-based chiral stationary phases (CSPs) have been successfully integrated into the tool box of many research and industry groups for the separation of enantiomers or stereoisomers by liquid and supercritical fluid chromatography. Due to the structurally diverse range of compounds available, efficient method development for chiral separations utilizing such CSPs is a challenging subject. In this chapter, we will discuss simplified screening protocols and straightforward approaches to achieve chiral separations in HPLC and SFC using the column series CHIRALPAK™ IA, IB, IC, and ID in reasonable time frame and with limited experimental work and a high success rate.

  4. Effect of ligand structure of stationary phase of high per-formance hydrophobic interaction chromatography on re-naturation efficiency of GuHCl-denatured α-chymotrypsin

    Institute of Scientific and Technical Information of China (English)

    SHEN; Yehua; WANG; Haibo; BAI; Quan; GENG; Xindu

    2005-01-01

    The renaturation of the denatured α-chymotrypsin (α-Chy) with 1.7 mol · L-1 guanidine hydrochloride (GuHCI) by three kinds of stationary phase of high performance hydrophobic interaction chromatography (STHIC) with a comparable hydrophobicity but different ligand structures was investigated. The obtained result indicates that the ligand structures of the three STHIC contribute to the renaturation efficiency of α-Chy in the order of the end ligands PEG-600< phenyl group < tetrahydrofurfuryl alcohol (THFA).

  5. One-Pot Click Access to a Cyclodextrin Dimer-Based Novel Aggregation Induced Emission Sensor and Monomer-Based Chiral Stationary Phase

    Directory of Open Access Journals (Sweden)

    Xiaoli Li

    2016-11-01

    Full Text Available A ‘two birds, one stone’ strategy was developed via a one-pot click reaction to simultaneously prepare a novel cyclodextrin (CD dimer based aggregation induced emission (AIE sensor (AIE-DCD and a monomer based chiral stationary phase (CSP-MCD for chiral high performance liquid chromatography (CHPLC. AIE-DCD was found to afford satisfactory AIE response for specific detection of Zn2+ with a detection limit of 50 nM. CSP-MCD exhibits excellent enantioseparation ability toward dansyl amino acids, where the resolution of dansyl amino leucine reaches 5.43.

  6. Analysis of dynamic changes in the proteome of a Bcl-XL overexpressing Chinese hamster ovary cell culture during exponential and stationary phases.

    Science.gov (United States)

    Carlage, Tyler; Kshirsagar, Rashmi; Zang, Li; Janakiraman, Vijay; Hincapie, Marina; Lyubarskaya, Yelena; Weiskopf, Andy; Hancock, William S

    2012-01-01

    Mammalian cell cultures used for biopharmaceutical production undergo various dynamic biological changes over time, including the transition of cells from an exponential growth phase to a stationary phase during cell culture. To better understand the dynamic aspects of cell culture, a quantitative proteomics approach was used to identify dynamic trends in protein expression over the course of a Chinese hamster ovary (CHO) cell culture for the production of a recombinant monoclonal antibody and overexpressing the antiapoptotic gene Bcl-xl. Samples were analyzed using a method incorporating iTRAQ labeling, two-dimensional LC/MS, and linear regression calculations to identify significant dynamic trends in protein abundance. Using this approach, 59 proteins were identified with significant temporal changes in expression. Pathway analysis tools were used to identify a putative network of proteins associated with cell growth and apoptosis. Among the differentially expressed proteins were molecular chaperones and isomerases, such as GRP78 and PDI, and reported cell growth markers MCM2 and MCM5. In addition, two proteins with growth-regulating properties, transglutaminase-2 and clusterin, were identified. These proteins are associated with tumor proliferation and apoptosis and were observed to be expressed at relatively high levels during stationary phase, which was confirmed by western blotting. The proteomic methodology described here provides a dynamic view of protein expression throughout a CHO fed-batch cell culture, which may be useful for further elucidating the biological processes driving mammalian cell culture performance. Copyright © 2012 American Institute of Chemical Engineers (AIChE).

  7. Comparison of the Separation Performances of Cinchona Alkaloid-Based Zwitterionic Stationary Phases in the Enantioseparation of β2- and β3-Amino Acids

    Directory of Open Access Journals (Sweden)

    István Ilisz

    2014-12-01

    Full Text Available The enantiomers of twelve unusual β2- and β3-homoamino acids containing the same side-chains were separated on chiral stationary phases containing a quinine- or quinidine-based zwitterionic ion-exchanger as chiral selector. The effects of the mobile phase composition, the nature and concentration of the acid and base additives and temperature on the separations were investigated. The changes in standard enthalpy, ∆(∆H°, entropy, ∆(∆S°, and free energy, ∆(∆G°, were calculated from the linear van’t Hoff plots derived from the ln α vs. 1/T curves in the studied temperature range (10–50 °C. The values of the thermodynamic parameters depended on the nature of the selectors, the structures of the analytes, and the positions of the substituents on the analytes. A comparison of the zwitterionic stationary phases revealed that the quinidine-based ZWIX(−™ column exhibited much better selectivity for both β2- and β3-amino acids than the quinine-based ZWIX(+™ column, and the separation performances of both the ZWIX(+™ and ZWIX(−™ columns were better for β2-amino acids. The elution sequence was determined in some cases and was observed to be R < S and S < R on the ZWIX(+™ and ZWIX(−™ columns, respectively.

  8. Core shell stationary phases for a novel separation of triglycerides in plant oils by high performance liquid chromatography with electrospray-quadrupole-time of flight mass spectrometer.

    Science.gov (United States)

    La Nasa, Jacopo; Ghelardi, Elisa; Degano, Ilaria; Modugno, Francesca; Colombini, Maria Perla

    2013-09-20

    A new method for the analysis of triglycerides (TAGs) in vegetable oils was developed using a Poroshell 120 EC-C18 column (3.0 mm×50 mm, 2.7 μm) with a high resolution ESI-Q-ToF tandem mass spectrometer as detection system. We used an Agilent Poroshell column, which is characterized by a recently developed stationary phase based on non-porous core particles. The results highlighted the advantages of this column in terms of the dramatic improvement in the number of theoretical plates and in low column backpressure. The developed method enabled us to analyze complex mixtures of more than 40 TAGs within less than 25 min and with a low backpressure (lower than 100 bar), and represents the first application of a core-shell stationary phase in reverse phase HPLC using an ESI-Q-ToF as detection system. The method was optimized on standards of TAGs, validated and applied to several plant oils. By a quantitative point of view, the method showed a very good linearity (r(2)>0.999) in the range 0.1-2.4 μg/g; high intra- and inter-day precision both in terms of retention times (RSD%<0.04%) and peak areas (RSD%<0.3%). Limits of detection and quantitation were lower than 0.03 μg/g and 0.10 μg/g, respectively.

  9. Investigation on enantiomeric separations of fluorenylmethoxycarbonyl amino acids and peptides by high-performance liquid chromatography using native cyclodextrins as chiral stationary phases.

    Science.gov (United States)

    Tang, Y; Zukowski, J; Armstrong, D W

    1996-09-06

    A systematic study was carried out to investigate enantiomeric separations of fluorenylmethoxycarbonyl (FMOC) amino acids and their peptides. Twenty amino acids were derivatized by 9-fluorenylmethyl chloroformate (FMOC-Cl) and its analogues, FMOC-glycyl-Cl and FMOC-beta-alanyl-Cl. All derivatives were chromatographed on native beta- and gamma-cyclodextrin columns using acetonitrile as the main mobile phase component. The results indicated that glycyl and beta-alanyl groups between FMOC and amino acid moieties enhanced chiral selectivities of amino acid derivatives. The addition of modifiers, triethylamine, acetic acid and methanol, into the mobile phase caused alterations in retention, enantiorecognition and elution order. The structures of amino acids and the type of chiral stationary phase employed exhibited significant impacts on chiral resolutions. It is also found that the number and position of glycyl moieties affect the retentions and enantioselectivities of FMOC derivatized glycyl containing peptides.

  10. A comparative quantitative structure-retention relationships study for lipophilicity determination of compounds with a phenanthrene skeleton on cyano-, reversed phase-, and normal phase-thin layer chromatography stationary phases.

    Science.gov (United States)

    Ciura, Krzesimir; Nowakowska, Joanna; Pikul, Piotr; Struck-Lewicka, Wiktoria; Markuszewski, Michał J

    2015-01-01

    The phenanthrene skeleton is an important moiety in medical chemistry as it is present in steroidal drugs used as anti-inflammatory and anti-asthmatic agents as well as synthetic hormones or potassium sparing diuretics. Chromatographic properties of 14 derivatives containing the phenanthrene skeleton in their structure with known lipophilicity have been studied. NP, RP, and cyano-bonded silica stationary phases with three binary mobile phases (acetonitrile-water, acetone-water, and acetone-petroleum ether) were tested. Obtained chromatographic data were correlated with the lipophilicity expressed as values of log partition coefficient (P). The presented study was undertaken to find the best TLC system and chromatographic data processing method in order to predict log P values. Correlations between chromatographic data and measurements of lipophilicity of compounds were presented as results of established quantitative structure-retention relationships. Principal component analysis and cluster analysis were used to investigate the similarities among chromatographic systems.

  11. Strong cation-exchange chromatography of proteins on a sulfoalkylated monolithic cryogel.

    Science.gov (United States)

    Perçin, Işık; Khalaf, Rushd; Brand, Bastian; Morbidelli, Massimo; Gezici, Orhan

    2015-03-20

    A new strong cation exchanger (SCX) monolithic column was synthesized by at-line surface modification of a cryogel prepared by copolymerization of 2-hydroxyethylmethacrylate (HEMA) and glycidylmethacrylate (GMA). Sodium salt of 3-Mercaptopropane sulfonic acid (3-MPS) was used as the ligand to transform the surface of the monolith into a strong cation exchanger. The obtained material was characterized in terms of elemental analysis, infrared spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Brunauer-Emmett-Teller (BET) N2 adsorption, and used as a stationary phase for strong-cation exchange chromatography of some proteins, such as α-chymotrypsinogen, cytochrome c and lysozyme. Water permeability of the column was calculated according to Darcy's law (2.66×10(-13)m(2)). The performance of the monolithic cryogel column was evaluated on the basis of Height Equivalent to a Theoretical Plate (HETP). Retention behavior of the studied proteins was modeled on the basis of Yamamoto model to understand the role of the ion-exchange mechanism in retention behaviors. The considered proteins were successfully separated, and the obtained chromatogram was compared with that obtained with a non-functionalized cryogel column.

  12. A Ka-band monolithic low phase noise coplanar waveguide oscillator using InAlAs/InGaAs HBT

    Science.gov (United States)

    Cui, Delong; Hsu, Shawn; Pavlidis, Dimitris; Chin, Patrick; Block, Tom

    2002-02-01

    A Ka-band oscillator has been designed, fabricated and tested using InAlAs/InGaAs HBTs. Coplanar waveguide technology has been employed to improve the Q-factor of the circuit. An output power of 2.6 dBm with DC to RF conversion efficiency of 7.8% was measured at 31.7 GHz. Low phase noise of -87 and -112 dBc/Hz were achieved at an offset frequency of 100 kHz and 1 MHz respectively. These low phase noise values can be attributed to the low 1/ f noise of the InAlAs/InGaAs HBT devices and the coplanar design used for the circuit.

  13. One-step synthesis of mussel-inspired molecularly imprinted magnetic polymer as stationary phase for chip-based open tubular capillary electrochromatography enantioseparation.

    Science.gov (United States)

    Wang, Xiao-Ni; Liang, Ru-Ping; Meng, Xiang-Ying; Qiu, Jian-Ding

    2014-10-03

    A facile approach for preparation of molecularly imprinted polymers was developed and successfully used as chiral stationary phase for rapid enantioseparation by open tubular capillary electrochromatography (OT-CEC). In this work, molecularly imprinted polymers were one-step prepared employing Fe3O4 nanoparticles (NPs) as the supporting substrate and dopamine as the functional monomer. By simply mixing Fe3O4 NPs with template molecules in a weak alkaline solution of dopamine, a thin adherent polydopamine (PDA) film imprinted with template molecules was formed by the self-polymerization of dopamine on the surface of Fe3O4 NPs. After extracting the embedded template molecules, the produced imprinted Fe3O4@PDA NPs are of three dimensional shape of template molecules favoring high binding capacity and magnetism property for easy manipulation. The imprinted Fe3O4@PDA NPs prepared with l-tryptophan, l-tyrosine, Gly-l-Phe or s-ofloxacin as template molecules were packed in the PDMS microchannel via magnetic field as novel stationary phase for the successful enantioseparation of corresponding target analysts. In addition, the imprinted Fe3O4@PDA NPs-based OT-CEC system exhibited excellent reproducibility, stability and repeatability, which provides a powerful protocol for separation enantiomers within a short analytical time and opens up a promising avenue for high-throughput screening of chiral compounds.

  14. Fragment-based approach to calculate hydrophobicity of anionic and nonionic surfactants derived from chromatographic retention on a C18 stationary phase.

    Science.gov (United States)

    Hammer, Jort; Haftka, Joris J-H; Scherpenisse, Peter; Hermens, Joop L M; de Voogt, Pim W P

    2017-02-01

    To predict the fate and potential effects of organic contaminants, information about their hydrophobicity is required. However, common parameters to describe the hydrophobicity of organic compounds (e.g., octanol-water partition constant [KOW ]) proved to be inadequate for ionic and nonionic surfactants because of their surface-active properties. As an alternative approach to determine their hydrophobicity, the aim of the present study was therefore to measure the retention of a wide range of surfactants on a C18 stationary phase. Capacity factors in pure water (k'0 ) increased linearly with increasing number of carbon atoms in the surfactant structure. Fragment contribution values were determined for each structural unit with multilinear regression, and the results were consistent with the expected influence of these fragments on the hydrophobicity of surfactants. Capacity factors of reference compounds and log KOW values from the literature were used to estimate log KOW values for surfactants (log KOWHPLC). These log KOWHPLC values were also compared to log KOW values calculated with 4 computational programs: KOWWIN, Marvin calculator, SPARC, and COSMOThermX. In conclusion, capacity factors from a C18 stationary phase are found to better reflect hydrophobicity of surfactants than their KOW values. Environ Toxicol Chem 2017;36:329-336. © 2016 The Authors. Environmental Toxicology and Chemistry Published by Wiley Periodicals, Inc. on behalf of SETAC.

  15. Exploring chiral separation of 3-carboxamido-5-aryl isoxazole derivatives by supercritical fluid chromatography on amylose and cellulose tris dimethyl- and chloromethyl phenylcarbamate polysaccharide based stationary phases.

    Science.gov (United States)

    Zehani, Yasmine; Lemaire, Lucas; Ghinet, Alina; Millet, Régis; Chavatte, Philippe; Vaccher, Claude; Lipka, Emmanuelle

    2016-10-07

    Four polysaccharide based chiral stationary phases were chosen, two chlorinated: Lux™ Amylose-2 (tris-5-chloro-2-methylphenylcarbamate of amylose) and Lux™ Cellulose-2 (tris-3-chloro-4-methylphenylcarbamate of cellulose) and two methylated: Chiralpak(®) AD-H (tris-3,5-dimethylphenylcarbamate of amylose) and Chiralcel(®) OD-H (tris-3,5-dimethylphenylcarbamate of cellulose) to separate four 3-carboxamido-5-aryl isoxazole derivatives by supercritical fluid chromatography. The effect of chiral stationary phase, co-solvent nature (MeOH, EtOH, 2-PrOH and ACN) and percentage (10-20%), temperature (20-45°C) and chemical structure of the compounds on retention, resolution and elution order were thoroughly studied. In addition, thermodynamic parameters were determined from the linear portion of the Van't Hoff plots. For all the derivatives, the Lux™ Cellulose-2 and Chiralpak(®) AD-H provided excellent resolutions (Rs=9.78) in short run time (under 6min). The preparation of about 10mg of each of the eight enantiomers was achieved successfully on a Chiralpak(®) AD-H with various percentages of ethanol as a co-solvent. Lastly, the enantiomeric purity of each of the eight individual enantiomer generated was determined and found higher than 98%.

  16. Graphene-ZIF8 composite material as stationary phase for high-resolution gas chromatographic separations of aliphatic and aromatic isomers.

    Science.gov (United States)

    Yang, Xiaohong; Li, Changxia; Qi, Meiling; Qu, Liangti

    2016-08-19

    This work presents the separation performance of graphene-ZIF8 (G-Z) composite material as stationary phase for capillary gas chromatography (GC). The G-Z stationary phase achieved high column efficiency of 5000 plates/m determined by n-dodecane (k=1.22) at 120°C and showed weakly polar nature. Importantly, it exhibited high selectivity and resolving capability for branched alkane isomers and aromatic positional isomers, showing clear advantages over the reported neat graphene and ZIF8. In addition, it attained high resolution for geometric cis-/trans-isomers. The G-Z column exhibited good column thermal stability up to 300°C and column repeatability with RSD values of retention times in the range of 0.01-0.19% for intra-day, 0.05-0.88% for inter-day and 0.66-5.6% for between-column, respectively, Moreover, the G-Z column was employed for the determination of minor impurity isomers in real reagent samples, which demonstrates its promising potential in GC applications.

  17. The use of Stationary Phase Optimized Selectivity Liquid Chromatography for the development of herbal fingerprints to detect targeted plants in plant food supplements.

    Science.gov (United States)

    Deconinck, E; Djiogo, C A Sokeng; Kamugisha, A; Courselle, P

    2017-08-01

    The consumption of plant food supplements is increasing steadily and more and more, these products are bought through internet. Often the products sold through internet are not registered or declared with a national authority, meaning that no or minimal quality control is performed and that they could contain herbs or plants that are regulated. Stationary Phase Optimized Selectivity Liquid Chromatography (SOS-LC) was evaluated for the development of specific fingerprints, to be used for the detection of targeted plants in plant food supplements. Three commonly used plants in plant food supplements and two regulated plants were used to develop fingerprints with SOS-LC. It was shown that for all plants specific fingerprints could be obtained, allowing the detection of these targeted plants in triturations with different herbal matrices as well as in real samples of suspicious supplements seized by the authorities. For three of the five plants a more specific fingerprint was obtained, compared to the ones developed on traditional columns described in literature. It could therefore be concluded that the combination of segments of different types of stationary phases, as used in SOS-LC, has the potential of becoming a valuable tool in the quality control and the identification of crude herbal or plant material and in the detection of regulated plants in plant food supplements or other herbal preparations. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Synthesis of Cellulose-2,3-bis(3,5-dimethylphenylcarbamate) in an Ionic Liquid and Its Chiral Separation Efficiency as Stationary Phase

    Science.gov (United States)

    Liu, Runqiang; Zhang, Yijun; Bai, Lianyang; Huang, Mingxian; Chen, Jun; Zhang, Yuping

    2014-01-01

    A chiral selector of cellulose-2,3-bis(3,5-dimethylphenylcarbamate) (CBDMPC) was synthesized by reacting 3,5-dimethylphenyl isocyanate with microcrystalline cellulose dissolved in an ionic liquid of 1-allyl-3-methyl-imidazolium chloride (AMIMCl). The obtained chiral selector was effectively characterized by infrared spectroscopy, elemental analysis and 1H NMR. The selector was reacted with 3-aminopropylsilanized silica gel and the CBDMPC bonded chiral stationary phase (CSP) was obtained. Chromatographic evaluation of the prepared CSPs was conducted by high performance liquid chromatographic (HPLC) and baseline separation of three typical fungicides including hexaconazole, metalaxyl and myclobutanil was achieved using n-hexane/isopropanol as the mobile phase with a flow rate 1.0 mL/min. Experimental results also showed that AMIMCl could be recycled easily and reused in the preparation of CSPs as an effective reaction media. PMID:24733066

  19. Synthesis of Cellulose-2,3-bis(3,5-dimethylphenylcarbamate in an Ionic Liquid and Its Chiral Separation Efficiency as Stationary Phase

    Directory of Open Access Journals (Sweden)

    Runqiang Liu

    2014-04-01

    Full Text Available A chiral selector of cellulose-2,3-bis(3,5-dimethylphenylcarbamate (CBDMPC was synthesized by reacting 3,5-dimethylphenyl isocyanate with microcrystalline cellulose dissolved in an ionic liquid of 1-allyl-3-methyl-imidazolium chloride (AMIMCl. The obtained chiral selector was effectively characterized by infrared spectroscopy, elemental analysis and 1H NMR. The selector was reacted with 3-aminopropylsilanized silica gel and the CBDMPC bonded chiral stationary phase (CSP was obtained. Chromatographic evaluation of the prepared CSPs was conducted by high performance liquid chromatographic (HPLC and baseline separation of three typical fungicides including hexaconazole, metalaxyl and myclobutanil was achieved using n-hexane/isopropanol as the mobile phase with a flow rate 1.0 mL/min. Experimental results also showed that AMIMCl could be recycled easily and reused in the preparation of CSPs as an effective reaction media.

  20. Fabrication of a polymeric ionic liquid-based adsorbent for multiple monolithic fiber solid-phase microextraction of endocrine disrupting chemicals in complicated samples.

    Science.gov (United States)

    Pei, Miao; Zhang, Zirui; Huang, Xiaojia; Wu, Yuanfei

    2017-04-01

    A multiple monolithic fiber solid-phase microextraction (MMF-SPME) utilizing polymeric ionic liquid-based adsorbent was prepared. The adsorbent was obtained by in situ copolymerization of an ionic liquid, 1-trimethyl-(4-vinylbenzyl) aminium chloride and dual cross-linkers (divinylbenzene and ethylenedimethacrylate). The effect of preparation conditions including the content of ionic liquid and porogen in the polymerization mixture on extraction performance was studied in detail. Infrared spectroscopy, elemental analysis, scanning electron microscopy and mercury intrusion porosimetry were used to inspect the physicochemical properties of the new adsorbent. The applicability of the new MMF-SPME was demonstrated by the extraction of trace endocrine disrupting chemicals (EDCs). Results indicated that the prepared MMF-SPME could extract EDCs effectively through multi-interactions such as ion-exchange, π-π and hydrophobic interactions. After optimization of extraction parameters, a method of MMF-SPME coupled to high performance liquid chromatography/diode array detection was conducted to detect trace EDCs in complicated samples including environmental water and human urine. The limits of detection (S/N=3) and quantification (S/N=10) for targeted compounds were 0.011-0.065μg/L and 0.036-0.21μg/L, respectively. Satisfactory precision was also achieved by evaluating the repeatability and intermediate precision with relative standard deviations (RSDs) of less than 9% and 10%, respectively. At the same time, the proposed method was successfully applied for the determination of EDCs in water and human urine with spiking recoveries ranged from 70.6% to 119%.

  1. 混合色谱填料的研究进展%State of art of mixed-mode stationary phase

    Institute of Scientific and Technical Information of China (English)

    徐基伟; 孙元社; 唐涛; 夏明珠; 雷武; 王风云; 李彤

    2015-01-01

    随着科学技术的发展,人们需要分离分析的样品越来越复杂,尤其是多肽、蛋白质类生物样品的复杂性使得单一模式色谱难以满足分离分析的要求。混合模式色谱因其独特的分离性能,可以在一次分离中获得与多维色谱相当的分离效果,而且可以避免多维色谱系统结构复杂、流动相兼容性差、分析时间长等问题,成为近年来的研究热点之一。混合模式色谱的研究重点是色谱固定相的设计与开发。混合模式色谱固定相包括反相/离子交换混合固定相、反相/亲水混合固定相、亲水/离子交换混合固定相、两性离子交换混合固定相及三相混合固定相。本文综述了近年来混合模式色谱填料的研究及应用进展,并对混合模式色谱及固定相的发展前景进行了展望。%With the development of science and technology,more and more complex samples like peptides and proteins need to be separated. It is difficult to separate them by single mode chromatography. Due to the unique separation character of mixed-mode chromatography,the same separation performance can be obtained in mixed-mode chromatography in one separation as multidimensional chromatography. And some disadvantages of multidimensional chromatog-raphy can be avoided in mixed-mode chromatography,such as the complexity of the system, the poor compatibility of mobile phases and the long analytical time. More and more attention is devoted to the mixed-mode chromatography in recent years. The focus on the mixed-mode chromatography is to design new structures of mixed-mode stationary phase. At present, mixed-mode stationary phases included reversed-phase/ion-exchange,reversed-phase/hydro-philic,hydrophilic/ion-exchange,zwitterionic and trimode mixed-mode stationary phases. The kinds of mixed-mode stationary phases are summarized and their features and applications in different fields are discussed in this review.

  2. A chromatographic estimate of the degree of heterogeneity of RPLC packing materials. 1. Non-endcapped polymeric C30-bonded stationary phase

    Energy Technology Data Exchange (ETDEWEB)

    Gritti, Fabrice [University of Tennessee, Knoxville (UTK); Guiochon, Georges A [ORNL

    2006-01-01

    A new chromatographic method estimating the degree of heterogeneity of RPLC packing materials is based on the results of systematic measurements of the adsorption data in a wide concentration range for selected probe compounds. These data are acquired by frontal analysis (FA), modeled, and used for the calculation of the adsorption energy distribution (AED). Four compounds were used, two neutral compounds of different molecular sizes (caffeine and phenol) and two ionizable compounds of opposite charges, 2-naphthalene sulfonate, an anion, and propranololium, a cation. This work was done on a C{sub 30}-bonded silica stationary phase (Prontosil-C{sub 30}), using the same aqueous mobile phase (30% methanol, v/v) for all compounds, except that sodium chloride (25 mM) was added to elute the ionizable compounds. All four adsorption isotherms have Langmuirian behavior. The AEDs are tri-modal for phenol, quadri-modal for caffeine. The total saturation capacity of the stationary phase is four-fold lower for caffeine than for phenol, due in part to its larger molecular size. The equilibrium constants on the low-energy sites of types 1 and 2 are eight-fold larger. These two types of sites characterize the heterogeneity of the bonded layer itself. The density of the high-energy sites of types 3 and 4 is higher for caffeine, suggesting that caffeine molecules can be accommodated in some hydrophobic cages into which smaller molecules like phenol cannot. These high-energy types of sites characterize the heterogeneity of the whole stationary phase (silica support included). The ionizable compounds have larger molecules than the neutral ones and, accordingly, a lower relative density of sites of type 2 to sites of type 1. A tri-modal and a quadri-modal energy distributions were observed for the 2-naphthalene sulfonate anion and the propranololium cation, respectively. The fourth types of sites measured and its unusually high equilibrium constant are most probably due to ion

  3. Influence of the polymerization-mixture composition for monolithic methacrylate-based columns on the electrochromatographic performance of drug molecules.

    Science.gov (United States)

    Tanret, Indiana; Mangelings, Debby; Vander Heyden, Yvan

    2008-09-29

    Methacrylate-based monolithic stationary phases were evaluated for the analysis of drug molecules using capillary electrochromatography (CEC) as separation technique. The effect of the polymerization-mixture composition on the retention behavior of a small test set of mainly drug molecules was studied. Two factors were varied in a central-composite design-based approach: the ratio between the pore-forming solvents and the monomers on one hand, and the ratio within the pore-forming solvents on the other hand, resulting in nine different stationary phases. The central point of the design was chosen at 70% (m/m) pore-forming solvents (PFS) of which 30% (m/m) is 1,4-butanediol, i.e. 21% of the total polymerization mixture. Experiments were conducted using both a basic (pH 11.5) and an acidic (pH 3) mobile phase. Retention times, retention factors, peak asymmetry and number of theoretical plates are the responses used to evaluate the performance of the resulting monoliths. The best compromise between the different responses was found around 67% PFS and 18% 1,4-butanediol (relative to the total mass), i.e. rather close to the center point. At these conditions, retention times were generally below 15min and retention factors below 5. Asymmetry values between close to 1 were found, and theoretical plate numbers up to 10,900, which were improvements compared to the central point of the design.

  4. Monolithically integrated multi-wavelength MQW-DBR laser diodes fabricated by selective metalorganic vapor phase epitaxy

    Science.gov (United States)

    Sasaki, Tatsuya; Yamaguchi, Masayuki; Kitamura, Mitsuhiro

    1994-12-01

    Selective metalorganic vapor phase epitaxy (MOVPE) was used to grow InGaAsP/InP layers for fabricating multi-wavelength laser diodes. Multiple quantum well (MQW) active and passive waveguides were simultaneously grown by one step selective growth. The selectively grown layer thickness increases with the mask stripe width. This growth enhancement can be used to control the lasing wavelength of distributed Bragg reflector (DBR) laser diodes, because the effective refractive index of the MQW passive waveguide at the DBR region can be controlled by the mask stripe width. This simple technique was used to fabricate multi-wavelength MQW-DBR laser diodes. In the selective growth, the MQW structure was grown under 150 Torr to obtain large bandgap energy shift for the MQW passive waveguides compared to the active waveguide, which was effective for wide wavelength tuning range. On the contrary, a bulk InGaAsP guide layer was grown under 35 Torr to prevent too much composition shift and maintain high crystalline quality of the MQW passive waveguide. For 10 consecutive laser diodes, a wavelength span of over 20 nm with accurate wavelength control was achieved.

  5. Temperature-induced inversion of elution order in the chromatographic enantioseparation of 1,1'-bi-2-naphthol on an immobilized polysaccharide-based chiral stationary phase.

    Science.gov (United States)

    Yao, Bixia; Zhan, Fengping; Yu, Guangyan; Chen, Zhifen; Fan, Wenjing; Zeng, Xiongping; Zeng, Qingle; Weng, Wen

    2009-07-10

    In this work, the enantioseparations of 1,1'-bi-2-naphthol (BINOL) and its three derivatives were performed on an immobilized polysaccharide-based chiral stationary phase, Chiralpak IA, under normal-phase mode. The effects of the content of polar modifier in the mobile phase and the column temperature on the retention and enantioseparation were investigated in detail. Temperature-induced inversion of elution order for BINOL was observed directly when n-hexane/2-propanol (92/8, v/v) was used as mobile phase. The isoenantioselective temperature (T(iso)) was calculated to be 31.4 degrees C. When n-hexane/2-propanol/THF (93/2/5, v/v/v) was used as mobile phase, the T(iso) value decreased to -8.2 degrees C. Entropically driven enantioseparation which had practical application was obtained successfully (separation factor being 1.189 and 1.332 at 25 degrees C and 50 degrees C, respectively). The corresponding thermodynamic parameters for other three binaphthyl compounds were compared with that for BINOL. Some inferences about chiral recognition mechanism were stressed.

  6. Construction and Evaluation of Continuous Two-dimensional Open-tubular Ion Exchange/Reversed Phase Monolithic Column Capillary Electrochromatography System%连续二维开管离子交换/反相整体柱毛细管电色谱的构建与评价

    Institute of Scientific and Technical Information of China (English)

    朱亚仙; 张凌怡; 雷雯; 张维冰

    2011-01-01

    在毛细管中原位合成反相整体色谱柱,并在同一根毛细管柱中的其余部分通过在内表面涂覆N-[3-(三甲氧基硅烷)-丙基)]-乙二胺(PEDA)使其具有离子交换功能,制备成连续二维开管离子交换/反相整体柱毛细管电色谱柱.通过对7种有机酸的分离探讨了开管柱中离子交换对分离的影响,进一步以天麻提取物为样品,对二维分离系统加以评价,出峰数量达到34个.在构建的新型电色谱柱系统中,可以通过电渗流、电泳流以及溶质与固定相的作用同时对分离选择性加以调节,适用于强疏水性和强亲水性同时存在的复杂样品的分析方法的建立.%A continuous two-dimensional open-tubular ion exchange/reversed phase monolithic column capillary electrochromatography system was developed. Reversed phase monolithic column was firstly synthesized in the end part of the capillary, then N-[3-(trimethoxysilyl)-propyl]-ethlenediamine was surface coated to the other part of the capillary. The separation of seven organic acids verified the effect on the separation of ion exchange in the open-tubular capillary. Rhizoma gastrodiae was separated to evaluate the system, and 34 peaks were recognized. This system is designed for complex samples with highly hydrophobicity components and highly hydrophilicity components simultaneous. This new column system combining advantages of multidimensional separation and adjusted stationary phase selectivity is suitable for the analysis method development of complicated samples.

  7. Normal and polar-organic-phase high-performance liquid chromatographic enantioresolution of omeprazole, rabeprazole, lansoprazole and pantoprazole using monochloro-methylated cellulose-based chiral stationary phase and determination of dexrabeprazole.

    Science.gov (United States)

    Dixit, Shuchi; Dubey, Rituraj; Bhushan, Ravi

    2014-01-01

    Enantioresolution of four anti-ulcer drugs (chiral sulfoxides), namely, omeprazole, rabeprazole, lansoprazole and pantoprazole, was carried out by high-performance liquid chromatography using a polysaccharide-based chiral stationary phase consisting of monochloromethylated cellulose (Lux cellulose-2) under normal and polar-organic-phase conditions with ultraviolet detection at 285 nm. The method was validated for linearity, accuracy, precision, robustness and limit of detection. The optimized enantioresolution method was compared for both the elution modes. The optimized method was further utilized to check the enantiomeric purity of dexrabeprazole.

  8. Comparison between polymerized ionic liquids synthesized using chain-growth and step-growth mechanisms used as stationary phase in gas chromatography.

    Science.gov (United States)

    Roeleveld, Kevin; David, Frank; Lynen, Frédéric

    2016-06-17

    In this study the merits of polymerized imidazolium based ionic liquid (PIL) stationary phases obtained via condensation and free radical polymerizations are compared as stationary phases in gas chromatography (GC). Poly(1-vinyl-3-butyl-imidazolium - bis(trifluoromethane)sulfonamide) (poly(ViC4Im(+) NTf2(-))) was obtained via a chain-growth mechanism while poly(propylimidazolium-NTf2) (poly(C3Im(+) NTf2(-))) was synthesized via a step-growth polymerization. The thermal stability of both polymers was assessed using thermal gravimetric analysis and compared with bleeding profiles obtained from the statically coated GC columns (30m×0.25mm×0.25μm). The performance was compared to what could be obtained on commercially available 1,5-di(2,3-dimethylimidazolium)pentane(2+) 2NTf2(-) (SLB-IL111) ionic liquid based columns. It was observed that the step-growth polymer was more thermally stable, up to 325°C, while the chain-growth polymer showed initial degradation at 250°C. Both polymers allowed reaching minimal plate heights of 0.400-0.500mm for retained solutes such as benzaldehyde, acetophenone, 1-methylnaphthalene and aniline. Assessment of the McReynolds constants illustrated that the polarity of the step-growth polymer was similar to the SLB-IL111 column, while displaying improved column stability. The PIL phases and particularly the so far little studied condensation based polymer shows particular retention and satisfactory column performance for polar moieties such as esters, amine and carbonyl functionalities.

  9. Detailed insights into the retention mechanism of caffeine metabolites on the amide stationary phase in hydrophilic interaction chromatography.

    Science.gov (United States)

    Guo, Yong; Shah, Rajan

    2016-09-09

    The amide phase was investigated using a wide range of acetonitrile content in the mobile phase in both the HILIC and RPLC modes. Using caffeine metabolites as the model compounds, the retention, thermodynamic and kinetic data was obtained under various mobile phase conditions and supported the previous postulation that there might be a transition of the predominant retention mechanism in relation to the acetonitrile content in HILIC. On the amide phase, hydrophilic partitioning seemed to be the predominant retention mechanism below 85% acetonitrile; and a different retention mechanism (presumably surface adsorption) made more and more significant contributions to the overall retention when the acetonitrile content reached above 85%. This study also provided more direct evidences to explain the effect of salt concentration on the retention of non-charged solutes in HILIC. In addition, the retention, thermodynamic and kinetic data suggest that the amide phase behaved very differently from the conventional C18 phase in the RPLC mode.

  10. Carbon Fiber Composite Monoliths as Catalyst Supports

    Energy Technology Data Exchange (ETDEWEB)

    Contescu, Cristian I [ORNL; Gallego, Nidia C [ORNL; Pickel, Joseph M [ORNL; Blom, Douglas Allen [ORNL; Burchell, Timothy D [ORNL

    2006-01-01

    Carbon fiber composite monoliths are rigid bodies that can be activated to a large surface area, have tunable porosity, and proven performance in gas separation and storage. They are ideal as catalyst supports in applications where a rigid support, with open structure and easy fluid access is desired. We developed a procedure for depositing a dispersed nanoparticulate phase of molybdenum carbide (Mo2C) on carbon composite monoliths in the concentration range of 3 to 15 wt% Mo. The composition and morphology of this phase was characterized using X-ray diffraction and electron microscopy, and a mechanism was suggested for its formation. Molybdenum carbide is known for its catalytic properties that resemble those of platinum group metals, but at a lower cost. The materials obtained are expected to demonstrate catalytic activity in a series of hydrocarbon reactions involving hydrogen transfer. This project demonstrates the potential of carbon fiber composite monoliths as catalyst supports.

  11. Carbon Fiber Composite Monoliths for Catalyst Supports

    Energy Technology Data Exchange (ETDEWEB)

    Contescu, Cristian I [ORNL; Gallego, Nidia C [ORNL; Pickel, Joseph M [ORNL; Blom, Douglas Allen [ORNL; Burchell, Timothy D [ORNL

    2006-01-01

    Carbon fiber composite monoliths are rigid bodies that can be activated to a large surface area, have tunable porosity, and proven performance in gas separation and storage. They are ideal as catalyst supports in applications where a rigid support, with open structure and easy fluid access is desired. We developed a procedure for depositing a dispersed nanoparticulate phase of molybdenum carbide (Mo2C) on carbon composite monoliths in the concentration range of 3 to 15 wt% Mo. The composition and morphology of this phase was characterized using X-ray diffraction and electron microscopy, and a mechanism was suggested for its formation. Molybdenum carbide is known for its catalytic properties that resemble those of platinum group metals, but at a lower cost. The materials obtained are expected to demonstrate catalytic activity in a series of hydrocarbon reactions involving hydrogen transfer. This project demonstrates the potential of carbon fiber composite monoliths as catalyst supports.

  12. Comparison of the peak resolution and the stationary phase retention between the satellite and the planetary motions using the coil satellite centrifuge with counter-current chromatographic separation of 4-methylumbelliferyl sugar derivatives.

    Science.gov (United States)

    Shinomiya, Kazufusa; Zaima, Kazumasa; Harada, Yukina; Yasue, Miho; Harikai, Naoki; Tokura, Koji; Ito, Yoichiro

    2017-01-20

    Coil satellite centrifuge (CSC) produces the complex satellite motion consisting of the triplicate rotation of the coiled column around three axes including the sun axis (the angular velocity, ω1), the planet axis (ω2) and the satellite axis (the central axis of the column) (ω3) according to the following formula: ω1=ω2+ω3. Improved peak resolution in the separation of 4-methylumbelliferyl sugar derivatives was achieved using the conventional multilayer coiled columns with ethyl acetate/1-butanol/water (3: 2: 5, v/v) for the lower mobile phase at the combination of the rotation speeds (ω1, ω2, ω3)=(300, 150, 150rpm), and (1:4:5, v/v) for the upper mobile phase at (300:100:200rpm). The effect of the satellite motion on the peak resolution and the stationary phase retention was evaluated by each CSC separation with the different rotation speeds of ω2 and ω3 under the constant revolution speed at ω1=300rpm. With the lower mobile phase, almost constant peak resolution and stationary phase retention were yielded regardless of the change of ω2 and ω3, while with the upper mobile phase these two values were sensitively varied according to the different combination of ω2 and ω3. For example, when ω2=147 or 200rpm is used, no stationary phase was retained in the coiled column while ω2=150rpm could retain enough volume of stationary phase for separation. On the other hand, the combined rotation speeds at (ω1, ω2, ω3)=(300, 300, 0rpm) or (300, 0, 300rpm) produced insufficient peak resolution regardless of the choice of the mobile phase apparently due to the lack of rotation speed except at (300, 0, 300rpm) with the upper mobile phase. At lower rotation speed of ω1=300rpm, better peak resolution and stationary phase retention were obtained by the satellite motion (ω3) than by the planetary motion (ω2), or ω3>ω2. The effect of the hydrophobicity of the two-phase solvent systems on the stationary phase retention was further examined using the n

  13. The PaPsr1 and PaWhi2 genes are members of the regulatory network that connect stationary phase to mycelium differentiation and reproduction in Podospora anserina.

    Science.gov (United States)

    Timpano, Hélène; Chan Ho Tong, Laetitia; Gautier, Valérie; Lalucque, Hervé; Silar, Philippe

    2016-09-01

    In filamentous fungi, entrance into stationary phase is complex as it is accompanied by several differentiation and developmental processes, including the synthesis of pigments, aerial hyphae, anastomoses and sporophores. The regulatory networks that control these processes are still incompletely known. The analysis of the "Impaired in the development of Crippled Growth (IDC)" mutants of the model filamentous ascomycete Podospora anserina has already yielded important information regarding the pathway regulating entrance into stationary phase. Here, the genes affected in two additional IDC mutants are identified as orthologues of the Saccharomyces cerevisiae WHI2 and PSR1 genes, known to regulate stationary phase in this yeast, arguing for a conserved role of these proteins throughout the evolution of ascomycetes.

  14. Development of a Stationary Phase Optimised Selectivity Liquid Chromatography based screening method for adulterations of food supplements for the treatment of pain.

    Science.gov (United States)

    Deconinck, E; Kamugisha, A; Van Campenhout, P; Courselle, P; De Beer, J O

    2015-06-01

    Illegally adulterated dietary supplements are an increasing problem worldwide. One of the important groups of often adulterated products are the dietary supplements, sold for the treatment of pain. These often contain analgesics, a heterogeneous group of molecules, containing both hydrophilic and hydrophobic compounds. The development of a screening method for these components, especially when mass spectrometric detection is not available, necessitates chromatographic separation, difficult to achieve with traditional chromatographic columns. In this paper Stationary Phase Optimised Selectivity Liquid Chromatography was used for the development of a screening method for nine analgesics, codeine and caffeine, often present in this type of dietary supplements. The method shows a good separation of all the compounds, allowing the screening to be performed with diode array detection and is fully compatible with mass spectrometry. The method was validated for its selectivity following the guidelines as described for the screening of pesticide residues and residues of veterinary medicines in food.

  15. Synthesis and Evaluation of Molecularly Imprinted Polymeric Microspheres for Chloramphenicol by Aqueous Suspension Polymerization as a High Performance Liquid Chromatography Stationary Phase

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yan [Beijing Forestry Univ., Beijing (China); Lei, Jiandu [Univ., of Science and Technology, Beijing (Korea, Republic of)

    2013-06-15

    Molecularly imprinted microsphere for chloramphenicol (CAP) with high adsorption capacity and excellent selectivity is prepared by aqueous suspension polymerization, in which chloramphenicol is used as template molecule and ethyl acetate as porogen. The CAP-imprinted microspheres are used as high performance liquid chromatography (HPLC) stationary phase and packed into stainless steel column (150 mm Χ 4.6 mm i. d.) for selective separation of chloramphenicol. HPLC analysis suggests that chloramphenicol can be distinguished from not only its structural analogs but also other broad-spectrum antibiotic such as erythromycin and tetracycline. In addition, the binding experiments of CAP-imprinted microspheres are carried out in ethanol/water (1:4, V:V), the results indicate that the maximum apparent static binding capacity of molecularly imprinted microspheres is up to 66.64 mg g{sup -1} according to scatchard model.

  16. Gas chromatographic-mass spectrometric characterization of all acyclic C5-C7 alkenes from fluid catalytic cracked gasoline using polydimethylsiloxane and squalane stationary phases.

    Science.gov (United States)

    Soják, Ladislav; Addová, Gabriela; Kubinec, Róbert; Kraus, Angelika; Hu, Gengyuan

    2002-02-15

    Published retention indices of acyclic alkenes C5-C7 on squalane and polydimethylsiloxane as stationary phases were investigated, and reliable retention indices of alkenes from various sources were converted to separation systems used in a laboratory. Retention indices measured on available authentic commercial alkenes and on alkenic fraction of gasoline, published retention indices as well as means of GC-MS were used for verification of calculated retention indices. Retention of some gas chromatographic unseparated isomer pairs was obtained by mass spectrometric deconvolution using a specific single-ion monitoring. On the basis of these retention data, C5-C7 alkenes were identified and analyzed in the gasoline from fluid catalytic cracking. In the gasoline all 59 acyclic C5-C7 isomeric alkenes were determined at significantly different concentration levels.

  17. Separation of alkanes and aromatic compounds by packed column gas chromatography using functionalized multi-walled carbon nanotubes as stationary phases.

    Science.gov (United States)

    Speltini, Andrea; Merli, Daniele; Quartarone, Eliana; Profumo, Antonella

    2010-04-23

    In the present work, we show a novel application of pristine and functionalized Multi-Walled Carbon Nanotubes (MWCNTs) as stationary phase in low-cost packed columns for the gas chromatographic separation of alkanes and aromatic hydrocarbons. The MWCNTs were deeply investigated by means of physical and chemical methods, like thermal analysis, IR and atomic force microscopy, and Inverse Gas Chromatography (IGC) in order to correlate the adsorption process and surface properties with the material purity level and functionalization degree. The derivatization process of the pristine nanotubes was a key factor to achieve a successful separation of both the light n-alkanes (C3-C5) and the related isomers (C4-C5 branched alkanes). Satisfactory results were similarly obtained in the case of separation of aromatic hydrocarbons (BTX).