Development and Validation of a Precise and Stability Indicating LC Method for the Determination of Benzalkonium Chloride in Pharmaceutical Formulation Using an Experimental Design

Directory of Open Access Journals (Sweden)

Harshal K. Trivedi

2010-01-01

Full Text Available A simple, precise, shorter runtime and stability indicating reverse-phase high performance liquid chromatographic method has been developed and validated for the quantification of benzalkonium chloride (BKC preservative in pharmaceutical formulation of sparfloxacin eye drop. The method was successfully applied for determination of benzalkonium chloride in various ophthalmic formulations like latanoprost, timolol, dexametasone, gatifloxacin, norfloxacin, combination of moxifloxacin and dexamethasone, combination of nepthazoline HCl, zinc sulphate and chlorpheniramine maleate, combination of tobaramycin and dexamethasone, combination of phenylephrine HCl, naphazoline HCl, menthol and camphor. The RP-LC separation was achieved on an Purospher Star RP-18e 75 mm × 4.0 mm, 3.0 μ in the isocratic mode using buffer: acetonitrile (35: 65, v/v, as the mobile phase at a flow rate of 1.8 mL/min. The methods were performed at 215 nm; in LC method, quantification was achieved with PDA detection over the concentration range of 50 to 150 μg/mL. The method is effective to separate four homologs with good resolution in presence of excipients, sparfloxacin and degradable compound due to sparfloxacin and BKC within five minutes. The method was validated and the results were compared statistically. They were found to be simple, accurate, precise and specific. The proposed method was validated in terms of specificity, precision, recovery, solution stability, linearity and range. All the validation parameters were within the acceptance range and concordant to ICH guidelines.

N-linked glycoprotein analysis using dual-extraction ultrahigh-performance liquid chromatography and electrospray tandem mass spectrometry.

Science.gov (United States)

Siu, S O; Lam, Maggie P Y; Lau, Edward; Yeung, William S B; Cox, David M; Chu, Ivan K

2010-01-01

Although reverse-phase liquid chromatography (RP-LC) is a common technique for peptide separation in shotgun proteomics and glycoproteomics, it often provides unsatisfactory results for the analysis of glycopeptides and glycans. This bias against glycopeptides makes it difficult to study glycoproteins. By coupling mass spectrometry (MS) with a combination of RP-LC and normal-phase (NP)-LC as an integrated front-end separation system, we demonstrate that effective identification and characterization of both peptides and glycopeptides mixtures, and their constituent glycan structures, can be achieved from a single sample injection event.

Isocratic Solid Phase Extraction-Liquid Chromatography (SPE-LC) Interfaced to High-Performance Tandem Mass Spectrometry for Rapid Protein Identification

DEFF Research Database (Denmark)

Hørning, Ole B; Kjeldsen, Frank; Theodorsen, Søren

2008-01-01

the isocratic solid phase extraction-liquid chromatography (SPE-LC) technology for rapid separation ( approximately 8 min) of simple peptide samples. We now extend these studies to demonstrate the potential of SPE-LC separation in combination with a hybrid linear ion trap-Orbitrap tandem mass spectrometer...

Isotope ratio monitoring LC/MS (IRM-LC/MS): new applications

International Nuclear Information System (INIS)

Juchelka, D; Hilkert, A.; Krummen, M.

2005-01-01

With the introduction of compound specific isotope analysis by isotope ratio monitoring GC/ MS (IRM-GC/MS) the immediate demand for similar applications using HPLC was created. Many compounds of biological, medical, pharmaceutical and environmental interest are not volatile or too polar. Consequently, they cannot be directly analyzed by gas chromatography. In IRM-GC/MS the carrier is helium, which does not interfere with the essential combust ion step prior to isotope ratio mass spectrometry (IRMS). In opposite the LC mobile phase has inhibited a similar direct conversion up to now. All earlier IRM-LC/MS approaches were based on the removal of the liquid phase prior to combustion risking fractionation of the isotope ratios of the eluted compounds. To avoid such restrictions we developed a new continuous flow concept for the coupling of an HPLC system to the isotope ratio MS for 13 C/ 12C isotope ratio analysis. In the Finnigan LC IsoLink, the liquid phase is not removed from the sample prior to oxidation. The sample is oxidized still in the mobile phase followed by on-line separation of the CO 2 from the liquid phase and transfer into the isotope ratio MS. Therefore, this strategy is based on water and inorganic buffers as mobile phase. The new approach opens up a whole new world in the application of gas isotope ratio mass spectrometry. The 13 C/ 12 C ratio s of organic acids, amino acids, carbohydrates and nucleotides can now be measured. These components typically within a complex matrix are separated by liquid chromatography followed by on-line determination of the isotope ratios. The draw backs of using derivatization and off-line preparation procedures can now be over come. This new technique allow s studying biochemical cycles, running tracer experiments and determining the origin of components. Applications from different scientific areas such as biogeochemistry, molecular biology, and pharmacy as well as authenticity control o f foods will be presented

[Study on the phase II metabolites of phenoprolamine hydrochloride in rat bile by LC/DAD/MSD].

Science.gov (United States)

Ding, L; Zhang, Z X; Ni, P Z; Wang, G J; An, D K

2001-06-01

To study the phase II metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. DDPH was administered by i.p. to bile duct-cannulated rats. Bile samples were collected before drug administration and up to 12 h after drug administration. After being purified and enriched with C-18 SPE columns the rat bile samples were analyzed by LC/DAD/MSD to identify the peaks of phase II metabolites. The fractions of phase II metabolites were prepared by HPLC and treated with beta-glucuronidase, and then were purified and enriched with C-18 SPE columns and analyzed by LC/DAD/MSD. The corresponding reference standards of DDPH phase I metabolites were analyzed by LC/DAD/MSD under identical conditions. The peaks M7, M8 and M9 in the chromatograms of rat bile samples were the phase II metabolites of DDPH and the enzymatic hydrolysates of M7, M8 and M9 were 1-(2, 6-dimethyl-4-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M3), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxyphenylethylamino)-propane (M2) and 1-(2,6-dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethyl-amino)-propane (M1) respectively. beta-1-O-[3,5-dimethyl-4-[-2-methyl-2-(3,4-dimethoxy-phenylethylamino)- ethoxy]-phenyl]-glucuronic acid (M7, glucuronide of M3), beta-1-O-[2, 4-dimethyl-3-[2-methyl-2-(3, 4-dimethoxy-phenylethylamino)-ethoxy]-phenyl]-glucuronic acid (M8, glucuronide of M2) and beta-1-O-[2-methoxy-4-[1-methyl-2-(2, 6-dimethylphenoxy)-ethylamino-ethyl]-phenyl]-glucuronic acid (M9, glucuronide of M1) were the phase II metabolites of DDPH in rat bile.

Identification of Forced Degradation Products of Itopride by LC-PDA and LC-MS

OpenAIRE

Joshi, Payal; Bhoir, Suvarna; Bhagwat, A. M.; Vishwanath, K.; Jadhav, R. K.

2011-01-01

Degradation products of itopride formed under different forced conditions have been identified using LC-PDA and LC-MS techniques. Itopride was subjected to forced degradation under the conditions of hydrolysis, photolysis, oxidation, dry and wet heat, in accordance with the International Conference on Harmonization. The stress solutions were chromatographed on reversed phase C18 (250×4.6 mm, 5 μm) column with a mobile phase methanol:water (55:45, v/v) at a detection wavelength of 215 nm. Itop...

Expression of wild-type Rp1 protein in Rp1 knock-in mice rescues the retinal degeneration phenotype.

Directory of Open Access Journals (Sweden)

Qin Liu

Full Text Available Mutations in the retinitis pigmentosa 1 (RP1 gene are a common cause of autosomal dominant retinitis pigmentosa (adRP, and have also been found to cause autosomal recessive RP (arRP in a few families. The 33 dominant mutations and 6 recessive RP1 mutations identified to date are all nonsense or frameshift mutations, and almost exclusively (38 out of 39 are located in the 4(th and final exon of RP1. To better understand the underlying disease mechanisms of and help develop therapeutic strategies for RP1 disease, we performed a series of human genetic and animal studies using gene targeted and transgenic mice. Here we report that a frameshift mutation in the 3(rd exon of RP1 (c.686delC; p.P229QfsX35 found in a patient with recessive RP1 disease causes RP in the homozygous state, whereas the heterozygous carriers are unaffected, confirming that haploinsufficiency is not the causative mechanism for RP1 disease. We then generated Rp1 knock-in mice with a nonsense Q662X mutation in exon 4, as well as Rp1 transgenic mice carrying a wild-type BAC Rp1 transgene. The Rp1-Q662X allele produces a truncated Rp1 protein, and homozygous Rp1-Q662X mice experience a progressive photoreceptor degeneration characterized disorganization of photoreceptor outer segments. This phenotype could be prevented by expression of a normal amount of Rp1 protein from the BAC transgene without removal of the mutant Rp1-Q662X protein. Over-expression of Rp1 protein in additional BAC Rp1 transgenic lines resulted in retinal degeneration. These findings suggest that the truncated Rp1-Q662X protein does not exert a toxic gain-of-function effect. These results also imply that in principle gene augmentation therapy could be beneficial for both recessive and dominant RP1 patients, but the levels of RP1 protein delivered for therapy will have to be carefully controlled.

Validated RP-HPLC Method for Quantification of Phenolic ...

African Journals Online (AJOL)

Purpose: To evaluate the total phenolic content and antioxidant potential of the methanol extracts of aerial parts and roots of Thymus sipyleus Boiss and also to determine some phenolic compounds using a newly developed and validated reversed phase high performance liquid chromatography (RP-HPLC) method.

Accessible silanol sites - beneficial for the RP-HPLC separation of constitutional and diastereomeric azaspirovesamicol isomers.

Science.gov (United States)

Wenzel, Barbara; Fischer, Steffen; Brust, Peter; Steinbach, Jörg

2010-12-10

Different RP-HPLC columns (phenyl, conventional ODS, cross-linked C(18) and special end-capped C(8) and C(18) phases) were used to investigate the separation of four basic ionizable isomers. Using ACN/20mM NH(4)OAc aq., a separation was observed exclusively on RP columns with higher silanol activity at unusual high ACN concentration, indicating cation-exchange as main retention mechanism. Using MeOH/20mM NH(4)OAc aq., another separation at low MeOH concentrations was observed on both, RP columns with higher as well as RP columns with lower silanol activity, which is mainly based on hydrophobic interactions. The isomers were also separated on a bare silica column at higher MeOH content using NH(4)OAc. Since cation-exchange governs this retention, the elution order was different compared to the RP phases. A strong retention on the silica column was observed in ACN, which could be attributed to partition processes as additional retention mechanism. Copyright © 2010 Elsevier B.V. All rights reserved.

Effect of TiO2 on conjugative transfer of RP4 plasmid

International Nuclear Information System (INIS)

Qian Di; Zhang Buchang; Yang Dong; Chen Zhaoli; Jin Min; Qiu Zhigang; Li Junwen

2013-01-01

Objective: To explore the effect and law of nano-titanium dioxide on the conjugative transfer of RP4 plasmid. Methods: Mating was conducted between Escherichia coli HB101 (RP4) and E. coli K12Rif in saline without stirring under certain conditions and the donor per recipient ratio was 1:1 constantly. The selective LB agar medium plates containing appropriate antibiotics were used to count the number of transconjugants and the conjugative transfer frequency. Results: Nano-titanium dioxide could promote the conjugative transfer of RP4. The nano-titanium dioxide concentration, bacterial concentration, mating temperature and mating time could affect the conjugative transfer of RP4. Conclusion: Nano-titanium dioxide can promote plasmid conjugal transfer in the liquid phase under certain conditions, which may pose a potential hazard to environmental and human health. (authors)

Integrated Solid-Phase Extraction-Capillary Liquid Chromatography (speLC) Interfaced to ESI-MS/MS for Fast Characterization and Quantification of Protein and Proteomes

DEFF Research Database (Denmark)

Falkenby, Lasse Gaarde; Such-Sanmartín, Gerard; Larsen, Martin Røssel

2014-01-01

min speLC-MS/MS experiment. Analysis by selected reaction monitoring by speLC-SRM-MS/MS of distinct peptides derived from the blood proteins IGF1, IGF2, IBP2, and IBP3 demonstrated protein quantification with CV values below 10% across 96 replicates. The speLC-MS/MS system is ideally suited for fast......The high peptide sequencing speed provided by modern hybrid tandem mass spectrometers enables the utilization of fast liquid chromatographic (LC) separation techniques. We present a robust solid-phase extraction/capillary LC system (speLC) for 5-10 min separation of semicomplex peptide mixtures...

Determination of the working voltage of the RP-0 and RP-10 nuclear reactors starting chains

International Nuclear Information System (INIS)

Zuniga, Agustin; Veramendi, Emilio; Salazar, Alberto

2014-01-01

This report presents the determination of the voltage curve of the RP-0 and RP-10 nuclear reactor starting chains, as an approach for determining the working voltages. The method consisted in changing the voltage and recording the corresponding counts. Then, the area of the best response was determined according to the detector characteristics, and also was found that for the RP-0, the response was in the proportional region, meanwhile for the RP-10, the response was in the region of ionization chambers. The values obtained were: BF3-A3 (RP-0) and 1150 V for RP-10, CF-A1 = 400 V, CF-A2 = 400 V, CF-A3 = 400 V. (authors).

A time-variant analysis of the 1/f^(2) phase noise in CMOS parallel LC-Tank quadrature oscillators

DEFF Research Database (Denmark)

Andreani, Pietro

2006-01-01

This paper presents a study of 1/f2 phase noise in quadrature oscillators built by connecting two differential LC-tank oscillators in a parallel fashion. The analysis clearly demonstrates the necessity of adopting a time-variant theory of phase noise, where a more simplistic, time...

Detecting pM concentrations of prostaglandins in cell culture supernatants by capillary SCX-LC-MS/MS

DEFF Research Database (Denmark)

Dahl, Sandra Rinne; Kleiveland, Charlotte Ramstad; Kassem, Moustapha

2008-01-01

A highly sensitive, improved online strong cation exchange (SCX)--RP capillary liquid chromatographic (cLC) method with IT mass spectrometric (IT-MS/MS) detection for the simultaneous determination of prostaglandin (PG)A(1), PGD(2), PGE(1), PGE(2), PGF(2alpha), 8-iso-(8i)PGF(2alpha), 6-keto-(6k...

Determination of Critical Properties of Endothermic Hydrocarbon Fuel RP-3 Based on Flow Visualization

Science.gov (United States)

Wang, Ning; Zhou, Jin; Pan, Yu; Wang, Hui

2014-01-01

The critical pressure and temperature of an endothermic hydrocarbon fuel RP-3 were determined by flow visualization. The flow pattern images of RP-3 at different pressures and temperatures were obtained. The critical pressure is identified by disappearance of the phase change while the critical temperature is determined by appearance of the opalescence phenomenon under the critical pressure. The opalescence phenomenon is unique to the critical point. The critical pressure and temperature of RP-3 are determined to be 2.3 MPa and 646 K, respectively.

Numerical study of the enlarged O(5) symmetry of the 3D antiferromagnetic RP2 spin model

International Nuclear Information System (INIS)

Fernandez, L.A.; Martin-Mayor, V.; Sciretti, D.; Tarancon, A.; Velasco, J.L.

2005-01-01

We investigate by means of Monte Carlo simulation and finite-size scaling analysis the critical properties of the three-dimensional O(5) non-linear σ model and of the antiferromagnetic RP 2 model, both of them regularized on a lattice. High accuracy estimates are obtained for the critical exponents, universal dimensionless quantities and critical couplings. It is concluded that both models belong to the same universality class, provided that rather non-standard identifications are made for the momentum-space propagator of the RP 2 model. We have also investigated the phase diagram of the RP 2 model extended by a second-neighbor interaction. A rich phase diagram is found, where most of the phase transitions are of the first order

An automated online turboflow cleanup LC/MS/MS method for the determination of 11 plasticizers in beverages and milk.

Science.gov (United States)

Ates, Ebru; Mittendorf, Klaus; Senyuva, Hamide

2013-01-01

An automated sample preparation technique involving cleanup and analytical separation in a single operation using an online coupled TurboFlow (RP-LC system) is reported. This method eliminates time-consuming sample preparation steps that can be potential sources for cross-contamination in the analysis of plasticizers. Using TurboFlow chromatography, liquid samples were injected directly into the automated system without previous extraction or cleanup. Special cleanup columns enabled specific binding of target compounds; higher MW compounds, i.e., fats and proteins, and other matrix interferences with different chemical properties were removed to waste, prior to LC/MS/MS. Systematic stepwise method development using this new technology in the food safety area is described. Selection of optimum columns and mobile phases for loading onto the cleanup column followed by transfer onto the analytical column and MS detection are critical method parameters. The method was optimized for the assay of 10 phthalates (dimethyl, diethyl, dipropyl, butyl benzyl, diisobutyl, dicyclohexyl, dihexyl, diethylhexyl, diisononyl, and diisododecyl) and one adipate (diethylhexyl) in beverages and milk.

Identification of Forced Degradation Products of Itopride by LC-PDA and LC-MS.

Science.gov (United States)

Joshi, Payal; Bhoir, Suvarna; Bhagwat, A M; Vishwanath, K; Jadhav, R K

2011-05-01

Degradation products of itopride formed under different forced conditions have been identified using LC-PDA and LC-MS techniques. Itopride was subjected to forced degradation under the conditions of hydrolysis, photolysis, oxidation, dry and wet heat, in accordance with the International Conference on Harmonization. The stress solutions were chromatographed on reversed phase C18 (250×4.6 mm, 5 μm) column with a mobile phase methanol:water (55:45, v/v) at a detection wavelength of 215 nm. Itopride degraded in acid, alkali and oxidative stress conditions. The stability indicating method was developed and validated. The degradation pathway of the drug to products II-VIII is proposed.

STABILITY-INDICATING RP-HPLC METHOD FOR SIMULTANEOUS ...

African Journals Online (AJOL)

B. S. Chandravanshi

pharmacopoeia, a careful and thorough literature review also showed absence of any reversed phase LC .... cetostearyl alcohol, petroleum jelly) of the ointment and human serum. Synthetic ... acid/base hydrolysis and oxidation. Acid-stress ...

Characterization of Isomeric Glycans by Reversed Phase Liquid Chromatography-Electronic Excitation Dissociation Tandem Mass Spectrometry

Science.gov (United States)

Tang, Yang; Wei, Juan; Costello, Catherine E.; Lin, Cheng

2018-04-01

The occurrence of numerous structural isomers in glycans from biological sources presents a severe challenge for structural glycomics. The subtle differences among isomeric structures demand analytical methods that can provide structural details while working efficiently with on-line glycan separation methods. Although liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a powerful tool for mixture analysis, the commonly utilized collision-induced dissociation (CID) method often does not generate a sufficient number of fragments at the MS2 level for comprehensive structural characterization. Here, we studied the electronic excitation dissociation (EED) behaviors of metal-adducted, permethylated glycans, and identified key spectral features that could facilitate both topology and linkage determinations. We developed an EED-based, nanoscale, reversed phase (RP)LC-MS/MS platform, and demonstrated its ability to achieve complete structural elucidation of up to five structural isomers in a single LC-MS/MS analysis. [Figure not available: see fulltext.

A new acetonitrile-free mobile phase method for LC-ELSD quantification of fructooligosaccharides in onion (Allium cepa L.).

Science.gov (United States)

Downes, Katherine; Terry, Leon A

2010-06-30

Onion soluble non-structural carbohydrates consist of fructose, glucose and sucrose plus fructooligosaccharides (FOS) with degrees of polymerisation (DP) in the range of 3-19. In onion, sugars and FOS are typically separated using liquid chromatography (LC) with acetonitrile (ACN) as a mobile phase. In recent times, however, the production of ACN has diminished due, in part, to the current worldwide economic recession. A study was therefore undertaken, to find an alternative LC method to quantify sugars and FOS from onion without the need for ACN. Two mobile phases were compared; the first taken from a paper by Vågen and Slimestad (2008) using ACN mobile phase, the second, a newly reported method using ethanol (EtOH). The EtOH mobile phase eluted similar concentrations of all FOS compared to the ACN mobile phase. In addition, limit of detection, limit of quantification and relative standard deviation values were sufficiently and consistently lower for all FOS using the EtOH mobile phase. The drawback of the EtOH mobile phase was mainly the inability to separate all individual sugar peaks, yet FOS could be successfully separated. However, using the same onion extract, a previously established LC method based on an isocratic water mobile phase could be used in a second run to separate sugars. Although the ACN mobile phase method is more convenient, in the current economic climate a method based on inexpensive and plentiful ethanol is a valid alternative and could potentially be applied to other fresh produce types. In addition to the mobile phase solvent, the effect of extraction solvents on sugar and FOS concentration was also investigated. EtOH is still widely used to extract sugars from onion although previous literature has concluded that MeOH is a superior solvent. For this reason, an EtOH-based extraction method was compared with a MeOH-based method to extract both sugars and FOS. The MeOH-based extraction method was more efficacious at extracting sugars and

Stationary phases for superheated water chromatography

International Nuclear Information System (INIS)

Saha, Shikha

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 18 (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 system. Although high efficiencies were not achieved with superheated water on PS-DVB and Xterra RP 18 columns, a higher efficiency was achieved on a PBD-zirconia column with superheated water than with 25-35% ACN at room temperature. The proposed theoretical increases in u opt were measured on three columns using superheated water as the mobile phase. The application of the superheated water chromatographic method to the separation of the pungent constituents of ginger by superheated water chromatography-NMR coupling system was studied. The coupling of superheated water chromatography using deuterium oxide to NMR spectroscopy for the separation of dry ginger extract was successful, although the NMR sensitivity in on-line mode coupling system was low. However, four compounds were identified in the ginger extract by stop-flow mode on superheated water chromatography-UV-NMR detection system. (author)

Development and Validation of a RP-HPLC Method for the ...

African Journals Online (AJOL)

Erah

Methods: Reverse phase (RP) chromatographic separation and estimation was achieved ... Recently, the use of natural products in ... one of the major consequences of herb-drug ... assurance and routine analysis of the blend. .... The robustness of the method was evaluated .... compliance with regulatory requirements) of.

Development and validation of a dissolution test with reversed-phase liquid chromatography analysis for rupatadine in tablet dosage forms

Directory of Open Access Journals (Sweden)

Sérgio Luiz Dalmora

2010-01-01

Full Text Available A dissolution test for in vitro evaluation of tablet dosage forms containing 10 mg of rupatadine was developed and validated by RP-LC. A discriminatory dissolution method was established using apparatus paddle at a stirring rate of 50 rpm with 900 mL of deaerated 0.01 M hydrochloric acid. The proposed method was validated yielding acceptable results for the parameters evaluated, and was applied for the quality control analysis of rupatadine tablets, and to evaluate the formulation during an accelerated stability study. Moreover, quantitative analyses were also performed, to compare the applicability of the RP-LC and the LC-MS/MS methods.

METHOD 544. DETERMINATION OF MICROCYSTINS AND NODULARIN IN DRINKING WATER BY SOLID PHASE EXTRACTION AND LIQUID CHROMATOGRAPHY/TANDEM MASS SPECTROMETRY (LC/MS/MS)

Science.gov (United States)

Method 544 is an accurate and precise analytical method to determine six microcystins (including MC-LR) and nodularin in drinking water using solid phase extraction and liquid chromatography tandem mass spectrometry (SPE-LC/MS/MS). The advantage of this SPE-LC/MS/MS is its sensi...

LC-MS/MS assay for the quantitation of the tyrosine kinase inhibitor neratinib in human plasma.

Science.gov (United States)

Kiesel, Brian F; Parise, Robert A; Wong, Alvin; Keyvanjah, Kiana; Jacobs, Samuel; Beumer, Jan H

2017-02-05

Neratinib is an orally available tyrosine kinase inhibitor targeting HER2 (ERBB2) and EGFR (ERBB). It is being clinically evaluated for the treatment of breast and other solid tumors types as a single agent or in combination with other chemotherapies. In support of several phase I/II clinical trials investigating neratinib combinations, we developed and validated a novel LC-MS/MS assay for the quantification of neratinib in 100μL of human plasma with a stable isotopic internal standard. Analytes were extracted from plasma using protein precipitation and evaporation of the resulting supernatant followed by resuspension. Chromatographic separation was achieved using an Acquity UPLC BEH Shield RP18 column and a gradient methanol-water mobile phase containing 10% ammonium acetate. An ABI 4000 mass spectrometer and electrospray positive mode ionization were used for detection. The assay was linear from 2 to 1,000ng/mL and proved to be accurate (98.9-106.5%) and precise (neratinib. Copyright © 2016 Elsevier B.V. All rights reserved.

Metabolite Profiling of Human Amniotic Fluid by Hyphenated Nuclear Magnetic Resonance Spectroscopy

OpenAIRE

Graça, Gonçalo; Duarte, Iola F.; Goodfellow, Brian J.; Carreira, Isabel M.; Couceiro, Ana Bela; Domingues, Maria do Rosário; Spraul, Manfred; Tseng, Li-Hong; Gil, Ana M.

2008-01-01

The metabolic profiling of human amniotic fluid (HAF) is of potential interest for the diagnosis of disorders in the mother or the fetus. In order to build a comprehensive metabolite database for HAF, hyphenated NMR has been used, for the first time, for systematic HAF profiling. Experiments were carried out using reverse-phase (RP) and ion-exchange liquid chromatography (LC), in order to detect less and more polar compounds, respectively. RP-LC conditions achieved good separation of amino ac...

The simultaneous detection and quantification of p-aminobenzoic acid and its phase 2 biotransformation metabolites in human urine using LC-MS/MS.

Science.gov (United States)

Nortje, Carla; Jansen van Rensburg, Peet; Cooke, Cecile; Erasmus, Elardus

2015-01-01

p-Aminobenzoic acid (PABA) can be used as a probe substance to investigate glycine conjugation, a reaction of phase 2 biotransformation. An LC-MS/MS method for simultaneous quantification of PABA and its metabolites from human urine was developed and validated. The metabolites can be quantified with acceptable precision and accuracy directly from human urine samples after ingestion of 550 mg PABA. The developed LC-MS/MS assay is to our knowledge the first method available for the simultaneous quantification of PABA and its glycine conjugation metabolites in human urine and provides important quantitative data for studies of this phase 2 biotransformation pathway.

A Novel Neural Network Vector Control for Single-Phase Grid-Connected Converters with L, LC and LCL Filters

Directory of Open Access Journals (Sweden)

Xingang Fu

2016-04-01

Full Text Available This paper investigates a novel recurrent neural network (NN-based vector control approach for single-phase grid-connected converters (GCCs with L (inductor, LC (inductor-capacitor and LCL (inductor-capacitor-inductor filters and provides their comparison study with the conventional standard vector control method. A single neural network controller replaces two current-loop PI controllers, and the NN training approximates the optimal control for the single-phase GCC system. The Levenberg–Marquardt (LM algorithm was used to train the NN controller based on the complete system equations without any decoupling policies. The proposed NN approach can solve the decoupling problem associated with the conventional vector control methods for L, LC and LCL-filter-based single-phase GCCs. Both simulation study and hardware experiments demonstrate that the neural network vector controller shows much more improved performance than that of conventional vector controllers, including faster response speed and lower overshoot. Especially, NN vector control could achieve very good performance using low switch frequency. More importantly, the neural network vector controller is a damping free controller, which is generally required by a conventional vector controller for an LCL-filter-based single-phase grid-connected converter and, therefore, can overcome the inefficiency problem caused by damping policies.

Influence of variation in mobile phase pH and solute pK(a) with the change of organic modifier fraction on QSRRs of hydrophobicity and RP-HPLC retention of weakly acidic compounds.

Science.gov (United States)

Han, Shu-ying; Liang, Chao; Zou, Kuan; Qiao, Jun-qin; Lian, Hong-zhen; Ge, Xin

2012-11-15

The variation in mobile phase pH and ionizable solute dissociation constant (pK(a)) with the change of organic modifier fraction in hydroorganic mobile phase has seemingly been a troublesome problem in studies and applications of reversed phase high performance liquid chromatography (RP-HPLC). Most of the early studies regarding the RP-HPLC of acid-base compounds have to measure the actual pH of the mixed mobile phase rigorously, sometimes bringing difficulties in the practices of liquid chromatographic separation. In this paper, the effect of this variation on the apparent n-octanol/water partition coefficient (K(ow)″) and the related quantitative structure-retention relationship (QSRR) of logK(ow)″ vs. logk(w), the logarithm of retention factor of analytes in neat aqueous mobile phases, was investigated for weakly acidic compounds. This QSRR is commonly used as a classical method for K(ow) measurement by RP-HPLC. The theoretical and experimental derivation revealed that the variation in mobile phase pH and solute pK(a) will not affect the QSRRs of acidic compounds. This conclusion is proved to be suitable for various types of ion-suppressors, i.e., strong acid (perchloric acid), weak acid (acetic acid) and buffer salt (potassium dihydrogen phosphate/phosphoric acid, PBS). The QSRRs of logK(ow)″ vs. logk(w) were modeled by 11 substituted benzoic acids using different types of ion-suppressors in a binary methanol-water mobile phase to confirm our deduction. Although different types of ion-suppressor all can be used as mobile phase pH modifiers, the QSRR model obtained by using perchloric acid as the ion-suppressor was found to have the best result, and the slightly inferior QSRRs were obtained by using acetic acid or PBS as the ion-suppressor. Copyright © 2012 Elsevier B.V. All rights reserved.

An Analysis of 1/f2 Phase Noise in Bipolar Colpitts Oscillators (With a Digression on Bipolar Differential-Pair LC Oscillators)

DEFF Research Database (Denmark)

Fard, A.; Andreani, Pietro

2007-01-01

This work presents an analysis of phase noise in the $1/f^{2}$ region displayed by both single-ended and differential bipolar Colpitts oscillators. Very accurate and rigorous symbolic phase noise expressions are derived, enabling a deeper insight into the major mechanisms of phase noise generatio......, and providing new tools for design optimization. Phase noise expressions for the cross-coupled differential-pair LC-tank oscillator are derived as well....

Identification of alkyl dimethylbenzylammonium surfactants in water samples by solid-phase extraction followed by ion trap LC/MS and LC/MS/MS

Science.gov (United States)

Ferrer, I.; Furlong, E.T.

2001-01-01

A novel methodology was developed for the determination of alkyl (C12, C14, and C16) dimethylbenzylammonium chloride (benzalkonium chloride or BAC, Chemical Abstract Service number: 8001-54-5) in water samples. This method is based on solid-phase extraction (SPE) using polymeric cartridges, followed by high-performance liquid chromatography/ion trap mass spectrometry (LC/MS) and tandem mass spectrometry(MS/MS) detection, equipped with an electrospray interface in positive ion mode. Chromatographic separation was achieved for three BAC homologues by using a C18 column and a gradient of acetonitrile/10 millimolar aqueous ammonium formate. Total method recoveries were higher than 71% in different water matrices. The main ions observed by LC/MS were at mass-to-charge ratios (m/z) of 304, 332, and 360, which correspond to the molecular ions of the C12, C14, and C16 alkyl BAC, respectively. The unequivocal structural identification of these compounds in water samples was performed by LC/MS/MS after isolation and subsequent fragmentation of each molecular ion. The main fragmentation observed for the three different homologues corresponded to the loss of the toluyl group in the chemical structure, which leads to the fragment ions at m/z 212, 240, and 268 and a tropylium ion, characteristic of all homologues, at m/z 91. Detection limits for the methodology developed in this work were in the low nanogram-per-liter range. Concentration levels of BAC - ranging from 1.2 to 36.6 micrograms per liter - were found in surface-water samples collected downstream from different wastewater-treatment discharges, thus indicating its input and persistence through the wastewater-treatment process.

TLR accessory molecule RP105 (CD180 is involved in post-interventional vascular remodeling and soluble RP105 modulates neointima formation.

Directory of Open Access Journals (Sweden)

Jacco C Karper

Full Text Available BACKGROUND: RP105 (CD180 is TLR4 homologue lacking the intracellular TLR4 signaling domain and acts a TLR accessory molecule and physiological inhibitor of TLR4-signaling. The role of RP105 in vascular remodeling, in particular post-interventional remodeling is unknown. METHODS AND RESULTS: TLR4 and RP105 are expressed on vascular smooth muscle cells (VSMC as well as in the media of murine femoral artery segments as detected by qPCR and immunohistochemistry. Furthermore, the response to the TLR4 ligand LPS was stronger in VSMC from RP105(-/- mice resulting in a higher proliferation rate. In RP105(-/- mice femoral artery cuff placement resulted in an increase in neointima formation as compared to WT mice (4982 ± 974 µm(2 vs.1947 ± 278 µm(2,p = 0.0014. Local LPS application augmented neointima formation in both groups, but in RP105(-/- mice this effect was more pronounced (10316±1243 µm(2 vs.4208 ± 555 µm(2,p = 0.0002, suggesting a functional role for RP105. For additional functional studies, the extracellular domain of murine RP105 was expressed with or without its adaptor protein MD1 and purified. SEC-MALSanalysis showed a functional 2∶2 homodimer formation of the RP105-MD1 complex. This protein complex was able to block the TLR4 response in whole blood ex-vivo. In vivo gene transfer of plasmid vectors encoding the extracellular part of RP105 and its adaptor protein MD1 were performed to initiate a stable endogenous soluble protein production. Expression of soluble RP105-MD1 resulted in a significant reduction in neointima formation in hypercholesterolemic mice (2500 ± 573 vs.6581 ± 1894 µm(2,p<0.05, whereas expression of the single factors RP105 or MD1 had no effect. CONCLUSION: RP105 is a potent inhibitor of post-interventional neointima formation.

On the interaction of the morphological structure and the LC behaviour of LC side chain block copolymers

NARCIS (Netherlands)

Fischer, H.R.; Poser, S.; Arnold, M.

1995-01-01

The interaction between morphological structure and phase behaviour of a LC side group block copolymer has been investigated using DSC, TEM and small angle X-ray diffraction. All samples of Polystyrene-block-2-(3-cholesteryloxycarbonyloxy)ethyl methacrylate (PS-b-PChEMA) show a phase separation

SIMULTANEOUS ESTIMATION OF MOXIFLOXACIN HYDROCHLORIDE AND DEXAMETHASONE SODIUM PHOSPHATE IN BULK AND IN OPHTHALMIC SOLUTION BY RP- HPLC

OpenAIRE

DHUMAL, D. M; SHIRKHEDKAR, A. A; NERKAR, P. P; SURANA, S. J

2012-01-01

A new simple, precise, accurate and selective RP-HPLC method has been developed and validated for simultaneous estimation of Moxifloxacin Hydrochloride (MOX) and Dexamethasone Sodium Phosphate (DSP) in Ophthalmic Solution. The method was carried out on a Qualisil RP C-8 (250 mm x 4.6 mm, 5 µm) column with a mobile phase consisting of Methanol: Water (75:25 v/v) pH adjusted to 3.0 with ortho-phosphoric acid of aqueous phase and flow rate of 1.0 mL min¹. Detection was carried out at 240 nm. The...

New Stability Indicating RP-HPLC Method for the Estimation of Cefpirome Sulphate in Bulk and Pharmaceutical Dosage Forms

OpenAIRE

Rao, Kareti Srinivasa; Kumar, Keshar Nargesh; Joydeep, Datta

2011-01-01

A simple stability indicating reversed-phase HPLC method was developed and subsequently validated for estimation of Cefpirome sulphate (CPS) present in pharmaceutical dosage forms. The proposed RP-HPLC method utilizes a LiChroCART-Lichrosphere100, C18 RP column (250 mm ? 4mm ? 5 ?m) in an isocratic separation mode with mobile phase consisting of methanol and water in the proportion of 50:50 % (v/v), at a flow rate 1ml/min, and the effluent was monitored at 270 nm. The retention time of CPS wa...

A size exclusion-reversed phase two dimensional-liquid chromatography methodology for stability and small molecule related species in antibody drug conjugates.

Science.gov (United States)

Li, Yi; Gu, Christine; Gruenhagen, Jason; Zhang, Kelly; Yehl, Peter; Chetwyn, Nik P; Medley, Colin D

2015-05-08

Antibody drug conjugates (ADCs) are complex therapeutic agents combining the specific targeting properties of antibodies and highly potent cytotoxic small molecule drugs to selectively eliminate tumor cells while limiting the toxicity to normal healthy tissues. One unique critical quality attribute of ADCs is the content of unconjugated small molecule drug present from either incomplete conjugation or degradation of the ADC. In this work, size exclusion chromatography (SEC) was coupled with reversed-phase (RP) HPLC in an online 2-dimensional chromatography format for identification and quantitation of unconjugated small molecule drugs and related small molecule impurities in ADC samples directly without sample preparation. The SEC method in the 1st dimension not only separated the small molecule impurities from the intact ADC, but also provided information about the size variants (monomer, dimer, aggregates, etc.) of the ADC. The small molecule peak from the SEC was trapped and sent to a RP-HPLC in the 2nd dimension to further separate and quantify the different small molecule impurities present in the ADC sample. This SEC-RP 2D-LC method demonstrated excellent precision (%RSDmolecule degradation products and aggregation of the conjugate were observed in the stability samples and the degradation pathways of the ADC were investigated. This 2D-LC method offers a powerful tool for ADC characterization and provides valuable information for conjugation and formulation development. Copyright © 2015 Elsevier B.V. All rights reserved.

Development of a New RP-UPLC Method for the Determination of ...

African Journals Online (AJOL)

Development of a New RP-UPLC Method for the Determination of Rabeprazole Sodium in Pharmaceutical Formulation and Application in Dissolution Studies. ... particle size 1.7 μm) column using an isocratic method with mobile phase composed of acetonitrile and phosphate buffer (pH 7.4) in the ratio of 35:65 (v/v).

Fast and simple online sample preparation coupled with capillary LC-MS/MS for determination of prostaglandins in cell culture supernatants

DEFF Research Database (Denmark)

Rinne, Sandra; Ramstad Kleiveland, Charlotte; Kassem, Moustapha

2007-01-01

An online 2-D strong cation exchange (SCX)-RP capillary liquid chromatographic (cLC) method with IT mass spectrometric (IT-MS/MS) detection for the simultaneous determination of prostaglandin (PG) A(1), PGD(2), PGE(1), PGE(2), PGF(2a), 6-keto-(6k)PGF(1a), and 15-Delta(12,14)-deoxy-PGJ(2) (15dPGJ(2...

On the interaction of the morphological structure and the LC behaviour of LC side chain block copolymers

NARCIS (Netherlands)

Fischer, H.R.; Arnold, M.

1995-01-01

The interaction between morphological structure and phase behaviour of a group of LC side group block copolymers have been investigated using DSC, TEM and small angle X-ray diffraction. Generally, phase separation between the two blocks was observed. It was found that in the case of those samples,

Cyclic life of superalloy IN738LC under in-phase and out-of-phase thermo-mechanical fatigue loading

International Nuclear Information System (INIS)

Chen Hongjun; Wahi, R.P.; Wever, H.

1995-01-01

The cyclic life of IN738LC, a widely used nickel base superalloy for blades in stationary gas turbines, was investigated under thermo-mechanical fatigue loading using a temperature variation range of 1023 to 1223 K, with temperature variation rate in the range of 6 to 15 K/min. Simple thermo-mechanical cycles with linear sequences corresponding to in-phase (IP) and out-of-phase (OP) tests were performed. Both the IP and OP tests were carried out at different constant mechanical strain ranges varied between 0.8 to 2.0% and at a constant mechanical strain rate of 10 -5 s -1 . Thermo-mechanical fatigue lives under both test conditions were compared with each other and with those of isothermal LCF tests at a temperature of 1223 K. The results show that the life under thermo-mechanical fatigue is strongly dependent on the nature of the test, i.e. stress controlled or strain controlled. (orig.)

Autosomal recessive retinitis pigmentosa with RP1 mutations is associated with myopia.

Science.gov (United States)

Chassine, Thomas; Bocquet, Béatrice; Daien, Vincent; Avila-Fernandez, Almudena; Ayuso, Carmen; Collin, Rob Wj; Corton, Marta; Hejtmancik, J Fielding; van den Born, L Ingeborgh; Klevering, B Jeroen; Riazuddin, S Amer; Sendon, Nathacha; Lacroux, Annie; Meunier, Isabelle; Hamel, Christian P

2015-10-01

To determine the refractive error in patients with autosomal recessive retinitis pigmentosa (arRP) caused by RP1 mutations and to compare it with that of other genetic subtypes of RP. Twenty-six individuals had arRP with RP1 mutations, 25 had autosomal dominant RP (adRP) with RP1 mutation, 8 and 33 had X-linked RP (xlRP) with RP2 and RPGR mutations, respectively, 198 and 93 had Usher syndrome and arRP without RP1 mutations, respectively. The median of the spherical equivalent (SE) and the IQR (Q25-Q75) was determined and multiple comparisons were performed. arRP patients with RP1 mutations had SE median at -4.0 dioptres (D) OD (Ocula Dextra); -3.88 D OS (Ocula Sinistra), whereas arRP patients without RP1 mutations (-0.50 D OD; -0.75 D OS) and Usher syndrome patients (-0.50 D OD; -0.38 D OS) were significantly less myopic (pUsher syndrome and adRP with RP1 mutation had a narrow IQR (-9.06 to -1.13 D), whereas arRP with RP1 mutations and xlRP with RP2 or RPGR mutations had a larger range (-9.06; -1.13 D). arRP patients with RP1 mutations have myopia not different from patients with xlRP with RP2 or RPGR mutations, while RP patients from other genetic subgroups were emmetropic or mildly myopic. We suggest that arRP patients with high myopic refractive error should be preferentially analysed for RP1 mutations. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Generation of accurate peptide retention data for targeted and data independent quantitative LC-MS analysis: Chromatographic lessons in proteomics.

Science.gov (United States)

Krokhin, Oleg V; Spicer, Vic

2016-12-01

The emergence of data-independent quantitative LC-MS/MS analysis protocols further highlights the importance of high-quality reproducible chromatographic procedures. Knowing, controlling and being able to predict the effect of multiple factors that alter peptide RP-HPLC separation selectivity is critical for successful data collection for the construction of ion libraries. Proteomic researchers have often regarded RP-HPLC as a "black box", while vast amount of research on peptide separation is readily available. In addition to obvious parameters, such as the type of ion-pairing modifier, stationary phase and column temperature, we describe the "mysterious" effects of gradient slope, column size and flow rate on peptide separation selectivity. Retention time variations due to these parameters are governed by the linear solvent strength (LSS) theory on a peptide level by the value of its slope S in the basic LSS equation-a parameter that can be accurately predicted. Thus, the application of shallower gradients, higher flow rates, or smaller columns will each increases the relative retention of peptides with higher S-values (long species with multiple positively charged groups). Simultaneous changes to these parameters that each drive shifts in separation selectivity in the same direction should be avoided. The unification of terminology represents another pressing issue in this field of applied proteomics that should be addressed to facilitate further progress. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Analysis of iridoids, secoiridoids and xanthones in Centaurium erythraea, Frasera caroliniensis and Gentiana lutea using LC-MS and RP-HPLC.

Science.gov (United States)

Aberham, Anita; Pieri, Valerio; Croom, Edward M; Ellmerer, Ernst; Stuppner, Hermann

2011-02-20

This study presents a new and validated HPLC method for the simultaneous determination of bioactive compounds in Centaurium erythraea, Frasera caroliniensis and Gentiana lutea. The iridoid loganic acid, four secoiridoids and 29 xanthones were separated on a RP-18 column, using aqueous o-phosphoric acid (0.085%, v/v) and acetonitrile as mobile phase. Phytochemical investigation of C. erythraea herb and F. caroliniensis roots resulted into isolation of 25 xanthones and three secoiridoids the structure of which was elucidated by spectroscopic means (NMR, MS and UV). 1,3,8-Trihydroxy-5,6-dimethoxyxanthone, isolated from C. erythraea, turned out to be a novel xanthone. The stability of the analytes was tested by subjecting samples to light, moisture and different temperatures. After six months of storage, decomposition of gentiopicroside and sweroside was observed. The swertiamarin content was nearly unchanged when stored at room temperature or in the refrigerator, but high temperature conditions reduced the content to 85%. In contrast, xanthones were stable under long-term, refrigerated and accelerated conditions. The established chromatographic method has been successfully applied for the quantification of the bioactive compounds in the three plants. The presence and distribution of polyoxygenated xanthones within the three members of the Gentianaceae family and their significance as analytical markers are discussed. Copyright © 2010 Elsevier B.V. All rights reserved.

Radiation Protection Section (SC/SL/RP)

CERN Document Server

2006-01-01

We should like to inform you that the Radiation Protection Section (SC/SL/RP) located on the Prévessin site has moved from Building 865 (ground floor) to new premises in Wing A of Building 892 (second floor). Telephone numbers remain the same. SC/SL/RP section

Polymerization speed and diffractive experiments in polymer network LC test cells

Science.gov (United States)

Braun, Larissa; Gong, Zhen; Habibpourmoghadam, Atefeh; Schafforz, Samuel L.; Wolfram, Lukas; Lorenz, Alexander

2018-02-01

Polymer-network liquid crystals (LCs), where the response properties of a LC can be enhanced by the presence of a porous polymer network, are investigated. In the reported experiments, liquid crystals were doped with a small amount (situ generated polymer network, the electro-optic response properties of photo cured samples were enhanced. For example, their continuous phase modulation properties led to more localized responses in samples with interdigitated electrodes, which caused suppression of selected diffraction orders in the diffraction patterns recorded in polymer network LC samples. Moreover, capacitance changes were investigated during photopolymerization of a blue phase LC.

Development and validation of simple RP-HPLC-PDA analytical protocol for zileuton assisted with Design of Experiments for robustness determination

OpenAIRE

Saurabh B. Ganorkar; Dinesh M. Dhumal; Atul A. Shirkhedkar

2017-01-01

A simple, rapid, sensitive, robust, stability-indicating RP-HPLC-PDA analytical protocol was developed and validated for the analysis of zileuton racemate in bulk and in tablet formulation. Development of method and resolution of degradation products from forced; hydrolytic (acidic, basic, neutral), oxidative, photolytic (acidic, basic, neutral, solid state) and thermal (dry heat) degradation was achieved on a LC – GC Qualisil BDS C18 column (250 mm × 4.6 mm × 5 μm) by isocratic mode at ambie...

Autosomal recessive retinitis pigmentosa with RP1 mutations is associated with myopia

NARCIS (Netherlands)

Chassine, T.; Bocquet, B.; Daien, V.; Avila-Fernandez, A.; Ayuso, C.; Collin, R.W.J.; Corton, M.; Hejtmancik, J.F.; Born, L.I. van den; Klevering, B.J.; Riazuddin, S.A.; Sendon, N.; Lacroux, A.; Meunier, I.; Hamel, C.P.

2015-01-01

OBJECTIVE: To determine the refractive error in patients with autosomal recessive retinitis pigmentosa (arRP) caused by RP1 mutations and to compare it with that of other genetic subtypes of RP. METHODS: Twenty-six individuals had arRP with RP1 mutations, 25 had autosomal dominant RP (adRP) with RP1

Improvement of Nicotinic Acid and Nicotinamide Analysis in Meats and Meat Products by HPLC and LC-MS/MS with Solid-Phase Extraction.

Science.gov (United States)

Hiki, Asako; Yamajima, Yukiko; Uematsu, Yoko

2016-01-01

A method for nicotinic acid (NA) and nicotinamide (NAA) analysis in meats was developed. NA and NAA were extracted from meats or meat products with metaphosphate aqueous solution. The extract was cleaned up with an Oasis MCX cartridge. The cartridge was washed with 2% acetic acid (v/v) and acetic acid-methanol solution. NA and NAA were eluted with ammonia-methanol solution. NA and NAA in the eluate were chromatographed on a Scherzo SM-C18 (3.0×150 mm, 3.0 μm) column with 20 mmol/L ammonium acetate containing 0.1% acetic acid-acetonitrile (97 : 3) as a mobile phase and were monitored at 261 nm. Quantification was performed by LC and LC-MS/MS. Calibration curves showed high linearity (correlation coefficient>0.998) between 1-25 μg/mL for LC and LC-MS/MS. Recoveries were 84-108% (CV≦5.8%) by HPLC and 79-105% (CV≦9.0%) by LC-MS/MS. The limit of quantitation for NA was 0.005-0.01 g/kg and that for NAA was 0.01-0.02 g/kg.

The mitochondrial SIR2 related protein 2 (SIR2RP2 impacts Leishmania donovani growth and infectivity.

Directory of Open Access Journals (Sweden)

Nimisha Mittal

2017-05-01

Full Text Available Leishmania donovani, a protozoan parasite is the major causative agent of visceral leishmaniasis. Increased toxicity and resistance to the existing repertoire of drugs has been reported. Hence, an urgent need exists for identifying newer drugs and drug targets. Previous reports have shown sirtuins (Silent Information Regulator from kinetoplastids as promising drug targets. Leishmania species code for three SIR2 (Silent Information Regulator related proteins. Here, we for the first time report the functional characterization of SIR2 related protein 2 (SIR2RP2 of L. donovani.Recombinant L. donovani SIR2RP2 was expressed in E. coli and purified. The enzymatic functions of SIR2RP2 were determined. The subcellular localization of LdSIR2RP2 was done by constructing C-terminal GFP-tagged full-length LdSIR2RP2. Deletion mutants of LdSIR2RP2 were generated in Leishmania by double targeted gene replacement methodology. These null mutants were tested for their proliferation, virulence, cell cycle defects, mitochondrial functioning and sensitivity to known SIR2 inhibitors.Our data suggests that LdSIR2RP2 possesses NAD+-dependent ADP-ribosyltransferase activity. However, NAD+-dependent deacetylase and desuccinylase activities were not detected. The protein localises to the mitochondrion of the promastigotes. Gene deletion studies showed that ΔLdSIR2RP2 null mutants had restrictive growth phenotype associated with accumulation of cells in the G2/M phase and compromised mitochondrial functioning. The null mutants had attenuated infectivity. Deletion of LdSIR2RP2 resulted in increased sensitivity of the parasites to the known SIR2 inhibitors. The sirtuin inhibitors inhibited the ADP-ribosyltransferase activity of recombinant LdSIR2RP2. In conclusion, sirtuins could be used as potential new drug targets for visceral leishmaniasis.

Determination of Scopolamine in Human Saliva Using Solid Phase Extraction and LC/MS/MS

Science.gov (United States)

Wang, Zuwei; Vaksman, Zalman; Boyd, Jason; Putcha, Lakshmi

2007-01-01

Purpose: Scopolamine is the preferred treatment for motion sickness during space flight because of its quick onset of action, short half-life and favorable side-effect profile. The dose administered depends on the mode of administration and usually ranges between 0.1 and 0.8 mg. Such small doses make it difficult to detect concentrations of scopolamine in biological fluids by using conventional HPLC methods. To measure scopolamine in saliva and thereby to evaluate the pharmacokinetics of scopolamine, we developed an LC/MS/MS method using off-line solid phase extraction. Method: Samples (0.5mL) were loaded onto Waters Oasis HLB co-polymer cartridges (10 mg, 1 mL) and eluted with 0.5 mL methanol without evaporation and reconstitution. HPLC separation of the eluted sample was performed using an Agilent Zorbax SB-CN column (50 x 2.1 mm) at a flow rate of 0.2 mL/min for 4 minutes. The mobile phase for separation was 90:10 (v/v) methanol: ammonium acetate (2 mM) in water, pH 5.0 +/- 0.1. Concentrations of scopolamine were determined using a Micromass Quattro Micro(TM) mass spectrometer with electrospray ionization (ESI). ESI mass spectra were acquired in positive ion mode with multiple reaction monitoring for the determination of scopolamine m/z = 304.2 yields 138.1 and internal standard (IS) hyoscyamine m/z = 290.2 yields 124.1. Results: The method is rapid, reproducible, specific and has the following parameters: scopolamine and the IS are eluted at 1.7 and 3.2 min respectively. The linear range is 50-5000 pg/mL for scopolamine in saliva with correlation coefficients > 0.99 with a CV < 0.5 %. The intra-day and inter-day CVs are < 15 % for quality control samples with concentrations of 75, 300, 750 and 3000 pg/mL of scopolamine in human saliva. Conclusion: Solid phase extraction allows more rapid sample preparation and greater precision than liquid extraction. Furthermore, we increased the sensitivity and specificity by adjusting the LC mobile phase and using an MS

Systematic Assessment of Seven Solvent and Solid-Phase Extraction Methods for Metabolomics Analysis of Human Plasma by LC-MS

Science.gov (United States)

Sitnikov, Dmitri G.; Monnin, Cian S.; Vuckovic, Dajana

2016-12-01

The comparison of extraction methods for global metabolomics is usually executed in biofluids only and focuses on metabolite coverage and method repeatability. This limits our detailed understanding of extraction parameters such as recovery and matrix effects and prevents side-by-side comparison of different sample preparation strategies. To address this gap in knowledge, seven solvent-based and solid-phase extraction methods were systematically evaluated using standard analytes spiked into both buffer and human plasma. We compared recovery, coverage, repeatability, matrix effects, selectivity and orthogonality of all methods tested for non-lipid metabolome in combination with reversed-phased and mixed-mode liquid chromatography mass spectrometry analysis (LC-MS). Our results confirmed wide selectivity and excellent precision of solvent precipitations, but revealed their high susceptibility to matrix effects. The use of all seven methods showed high overlap and redundancy which resulted in metabolite coverage increases of 34-80% depending on LC-MS method employed as compared to the best single extraction protocol (methanol/ethanol precipitation) despite 7x increase in MS analysis time and sample consumption. The most orthogonal methods to methanol-based precipitation were ion-exchange solid-phase extraction and liquid-liquid extraction using methyl-tertbutyl ether. Our results help facilitate rational design and selection of sample preparation methods and internal standards for global metabolomics.

LC-MS/MS signal suppression effects in the analysis of pesticides in complex environmental matrices.

Science.gov (United States)

Choi, B K; Hercules, D M; Gusev, A I

2001-02-01

The application of LC separation and mobile phase additives in addressing LC-MS/MS matrix signal suppression effects for the analysis of pesticides in a complex environmental matrix was investigated. It was shown that signal suppression is most significant for analytes eluting early in the LC-MS analysis. Introduction of different buffers (e.g. ammonium formate, ammonium hydroxide, formic acid) into the LC mobile phase was effective in improving signal correlation between the matrix and standard samples. The signal improvement is dependent on buffer concentration as well as LC separation of the matrix components. The application of LC separation alone was not effective in addressing suppression effects when characterizing complex matrix samples. Overloading of the LC column by matrix components was found to significantly contribute to analyte-matrix co-elution and suppression of signal. This signal suppression effect can be efficiently compensated by 2D LC (LC-LC) separation techniques. The effectiveness of buffers and LC separation in improving signal correlation between standard and matrix samples is discussed.

Experimental investigation on flow patterns of RP-3 kerosene under sub-critical and supercritical pressures

Science.gov (United States)

Wang, Ning; Zhou, Jin; Pan, Yu; Wang, Hui

2014-02-01

Active cooling with endothermic hydrocarbon fuel is proved to be one of the most promising approaches to solve the thermal problem for hypersonic aircraft such as scramjet. The flow patterns of two-phase flow inside the cooling channels have a great influence on the heat transfer characteristics. In this study, phase transition processes of RP-3 kerosene flowing inside a square quartz-glass tube were experimentally investigated. Three distinct phase transition phenomena (liquid-gas two phase flow under sub-critical pressures, critical opalescence under critical pressure, and corrugation under supercritical pressures) were identified. The conventional flow patterns of liquid-gas two phase flow, namely bubble flow, slug flow, churn flow and annular flow are observed under sub-critical pressures. Dense bubble flow and dispersed flow are recognized when pressure is increased towards the critical pressure whilst slug flow, churn flow and annular flow disappear. Under critical pressure, the opalescence phenomenon is observed. Under supercritical pressures, no conventional phase transition characteristics, such as bubbles are observed. But some kind of corrugation appears when RP-3 transfers from liquid to supercritical. The refraction index variation caused by sharp density gradient near the critical temperature is thought to be responsible for this corrugation.

Focusing and non-focusing modulation strategies for the improvement of on-line two-dimensional hydrophilic interaction chromatography × reversed phase profiling of complex food samples.

Science.gov (United States)

Montero, Lidia; Ibáñez, Elena; Russo, Mariateresa; Rastrelli, Luca; Cifuentes, Alejandro; Herrero, Miguel

2017-09-08

Comprehensive two-dimensional liquid chromatography (LC × LC) is ever gaining interest in food analysis, as often, food-related samples are too complex to be analyzed through one-dimensional approaches. The use of hydrophilic interaction chromatography (HILIC) combined with reversed phase (RP) separations has already been demonstrated as a very orthogonal combination, which allows attaining increased resolving power. However, this coupling encompasses different analytical challenges, mainly related to the important solvent strength mismatch between the two dimensions, besides those common to every LC × LC method. In the present contribution, different strategies are proposed and compared to further increase HILIC × RP method performance for the analysis of complex food samples, using licorice as a model sample. The influence of different parameters in non-focusing modulation methods based on sampling loops, as well as under focusing modulation, through the use of trapping columns in the interface and through active modulation procedures are studied in order to produce resolving power and sensitivity gains. Although the use of a dilution strategy using sampling loops as well as the highest possible first dimension sampling rate allowed significant improvements on resolution, focusing modulation produced significant gains also in peak capacity and sensitivity. Overall, the obtained results demonstrate the great applicability and potential that active modulation may have for the analysis of complex food samples, such as licorice, by HILIC × RP. Copyright © 2017 Elsevier B.V. All rights reserved.

Can LC and LC-MS ever replace immunoassays?

Directory of Open Access Journals (Sweden)

Timothy G. Cross

2016-10-01

Full Text Available Immunoassays have been the technology of choice for the analysis of biomolecules for many decades across a wide range of applications in research, diagnostics and infectious disease monitoring. There are good reasons for the wide adoption of immunoassays but even such a well established and characterised technique has limitations and as such investigators are looking at alternative technologies. One such alternative is liquid chromatography (LC and, more specifically, liquid chromatography coupled with mass spectrometry (LC-MS. This article will review both immunoassay and LC and LC-MS technologies and methodologies and discuss the advantages and limitations of both approaches. In addition, the next developments that will need to occur before there is widespread adoption of LC and LC-MS technology preferentially over immunoassays will be examined.

Synthesis of RP 48497, an Impurity of Eszopiclone

Directory of Open Access Journals (Sweden)

Mao-Sheng Cheng

2008-08-01

Full Text Available Abstract: RP 48497 is a photodegradation product of eszopiclone, a non-benzodiazepine sedative-hypnotic used in the treatment of insomnia. We report herein the first synthesis of RP 48497 via reduction, chlorination, and recyclization of 6-(5-chloropyridin-2-yl-7-hydroxy-6,7-dihydropyrrolo[3,4-b]pyrazin-5-one (3, a key intermediate in the synthesis of eszopiclone. The structure of RP 48497 was confirmed by its 1H-NMR and MS data. The mechanism of the reduction step in the synthesis of RP 48497 was also studied and the key parameters were determined. These findings should be important for quality control purposes in the manufacture of eszopiclone.

Enrichment and identification of integral membrane proteins from barley aleurone layers by reversed-phase chromatography, SDS-PAGE and LC-MS/MS

DEFF Research Database (Denmark)

Hynek, Radovan; Svensson, Birte; Nørregaard Jensen, Ole

2006-01-01

was developed, comprising batch reversed-phase chromatography with stepwise elution of hydrophobic proteins by 2-propanol. Proteins in the most hydrophobic fraction were separated by SDS-PAGE and identified by LC-MS/MS and barley EST sequence database search. The method was efficient for enrichment of integral...

[Study on the phase I metabolites of phenoprolamine hydrochloride in rat bile by LC/DAD/MSD].

Science.gov (United States)

Ding, L; Zhang, Z X; Ni, P Z; Wang, G J; An, D K

2001-03-01

To study the phase I metabolites of phenoprolamine hydrochloride (DDPH) in rat bile. DDPH was administered i.p. to bile duct-cannulated rats. Bile samples were collected before administration and up to 12 h after administration. After being treated with beta-glucuronidase, the bile samples were purified and enriched with C-18 SPE columns, and then were analyzed by LC/DAD/MSD. The samples containing synthesized reference standards of DDPH metabolite 1-(2, 6-dimethylphenoxy)-2-(3-methoxy-4-hydroxyphenylethylamino)-propane (M1), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3, 4-methoxy-phenylethylamino)-propane (M2), 1-(2,6-dimethyl-4-hydroxyphenoxy)-2-(3,4- methoxyphenylethylamino)-propane (M3), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2-(3-hydroxy-4- methoxyphenylethylamino)-propane (M4), 1-(2, 6-dimethyl-3-hydroxyphenoxy)-2- (3-hydroxy-4-methoxyphenylethylamino)-propane (M5) and 1-(2,6-dimethyl-4-hydroxyphenoxy)-2-(3-methoxy-4- hydroxyphenylethylamino)-propane (M6) were analyzed by LC/DAD/MSD under identical conditions. The retention times, UV spectra, molecular weights and production spectra (obtained by collision-induced dissociation) of the apparent ions of peak A, B, C, D, E and F in the total ion chromatogram of DDPH treated rat bile sample were consistent with those of M1, M2, M3, M5, M4 and M6, respectively. M1, M2, M3, M4, M5 and M6 were identified as the phase I metabolites of DDPH in the rat.

Annotation of the human serum metabolome by coupling three liquid chromatography methods to high-resolution mass spectrometry.

Science.gov (United States)

Boudah, Samia; Olivier, Marie-Françoise; Aros-Calt, Sandrine; Oliveira, Lydie; Fenaille, François; Tabet, Jean-Claude; Junot, Christophe

2014-09-01

This work aims at evaluating the relevance and versatility of liquid chromatography coupled to high resolution mass spectrometry (LC/HRMS) for performing a qualitative and comprehensive study of the human serum metabolome. To this end, three different chromatographic systems based on a reversed phase (RP), hydrophilic interaction chromatography (HILIC) and a pentafluorophenylpropyl (PFPP) stationary phase were used, with detection in both positive and negative electrospray modes. LC/HRMS platforms were first assessed for their ability to detect, retain and separate 657 metabolite standards representative of the chemical families occurring in biological fluids. More than 75% were efficiently retained in either one LC-condition and less than 5% were exclusively retained by the RP column. These three LC/HRMS systems were then evaluated for their coverage of serum metabolome. The combination of RP, HILIC and PFPP based LC/HRMS methods resulted in the annotation of about 1328 features in the negative ionization mode, and 1358 in the positive ionization mode on the basis of their accurate mass and precise retention time in at least one chromatographic condition. Less than 12% of these annotations were shared by the three LC systems, which highlights their complementarity. HILIC column ensured the greatest metabolome coverage in the negative ionization mode, whereas PFPP column was the most effective in the positive ionization mode. Altogether, 192 annotations were confirmed using our spectral database and 74 others by performing MS/MS experiments. This resulted in the formal or putative identification of 266 metabolites, among which 59 are reported for the first time in human serum. Copyright © 2014 Elsevier B.V. All rights reserved.

Positive inotropic effects of RP 62719, a new pure class III antiarrhythmic agent, on guinea pig myocardium.

Science.gov (United States)

Beregi, J P; Escande, D; Coudray, N; Mery, P; Mestre, M; Chemla, D; Lecarpentier, Y

1992-12-01

The mechanical effects of RP 62719 [(-)1-[-2-(3,4-dihydro-2H-1- benzopyran-4-yl)ethyl]-4-(3,4-dimethoxyphenyl)-piperidine] were tested in vitro on guinea pig left ventricular papillary muscle. RP 62719 is a novel pure class III antiarrhythmic agent known to prolong the cardiac action potential duration by selectively blocking the inward rectifying K+ current. Mechanical parameters were determined from contraction and relaxation phases under isotonic and isometric conditions. At a concentration of 0.02 microM, RP 62719 did not produce significant effects on inotropy or lusitropy. At 0.2 and 2 microM, the drug improved contraction under both heavy and low loading conditions, as evidenced by a 30% increase in maximum unloaded shortening velocity (Vmax, P delta L, P force normalized per cross-sectional area (AF/s, P force derivative per mm2 (+dF/s, P force derivative per mm2 (-dF/s, P < .001). At a higher concentration (20 microM), effects of RP 62719 on inotropy and lusitropy were less marked, thus accounting for the bell-shaped form of the dose-response curve. An increase in the extracellular Ca++ concentration from 2.5 to 3.75 mM improved inotropy to a similar extent (+30-50%) as did 2 microM RP 62719. However, lusitropy and mechanical coupling between contraction and relaxation were not modified in the same proportion under RP 62719 and under 3.75 mM Ca++.(ABSTRACT TRUNCATED AT 250 WORDS)

An enhanced close-in phase noise LC-VCO using parasitic V-NPN transistors in a CMOS process

International Nuclear Information System (INIS)

Gao Peijun; Min Hao; Oh, N J

2009-01-01

A differential LC voltage controlled oscillator (VCO) employing parasitic vertical-NPN (V-NPN) transistors as a negative g m -cell is presented to improve the close-in phase noise. The V-NPN transistors have lower flicker noise compared to MOS transistors. DC and AC characteristics of the V-NPN transistors are measured to facilitate the VCO design. The proposed VCO is implemented in a 0.18 μm CMOS RF/mixed signal process, and the measurement results show the close-in phase noise is improved by 3.5-9.1 dB from 100 Hz to 10 kHz offset compared to that of a similar CMOS VCO. The proposed VCO consumes only 0.41 mA from a 1.5 V power supply. (semiconductor integrated circuits)

RP-HPLCdetermination of ginsenosides RG1 and RB1 in Panax ...

African Journals Online (AJOL)

Y Liu, Q He, D Liu, W Wan, X Liu, C Wen, W Fang, W Zhou, Z Xia ... RP-HPLC method is adopted, column used is a ZOBAX SB-C18 column (4.6 mm × 250 mnl, 5 μm), mobile phase A is water, and B is acetonitrile, gradient elution conditions are: 0~20 min (A:B 20:80), 20~60 min (A:B 20~35: 80~65); and detection ...

Arcuate AgRP neurons mediate orexigenic and glucoregulatory actions of ghrelin★

Science.gov (United States)

Wang, Qian; Liu, Chen; Uchida, Aki; Chuang, Jen-Chieh; Walker, Angela; Liu, Tiemin; Osborne-Lawrence, Sherri; Mason, Brittany L.; Mosher, Christina; Berglund, Eric D.; Elmquist, Joel K.; Zigman, Jeffrey M.

2013-01-01

The hormone ghrelin stimulates eating and helps maintain blood glucose upon caloric restriction. While previous studies have demonstrated that hypothalamic arcuate AgRP neurons are targets of ghrelin, the overall relevance of ghrelin signaling within intact AgRP neurons is unclear. Here, we tested the functional significance of ghrelin action on AgRP neurons using a new, tamoxifen-inducible AgRP-CreERT2 transgenic mouse model that allows spatiotemporally-controlled re-expression of physiological levels of ghrelin receptors (GHSRs) specifically in AgRP neurons of adult GHSR-null mice that otherwise lack GHSR expression. AgRP neuron-selective GHSR re-expression partially restored the orexigenic response to administered ghrelin and fully restored the lowered blood glucose levels observed upon caloric restriction. The normalizing glucoregulatory effect of AgRP neuron-selective GHSR expression was linked to glucagon rises and hepatic gluconeogenesis induction. Thus, our data indicate that GHSR-containing AgRP neurons are not solely responsible for ghrelin's orexigenic effects but are sufficient to mediate ghrelin's effects on glycemia. PMID:24567905

Arcuate AgRP neurons mediate orexigenic and glucoregulatory actions of ghrelin.

Science.gov (United States)

Wang, Qian; Liu, Chen; Uchida, Aki; Chuang, Jen-Chieh; Walker, Angela; Liu, Tiemin; Osborne-Lawrence, Sherri; Mason, Brittany L; Mosher, Christina; Berglund, Eric D; Elmquist, Joel K; Zigman, Jeffrey M

2014-02-01

The hormone ghrelin stimulates eating and helps maintain blood glucose upon caloric restriction. While previous studies have demonstrated that hypothalamic arcuate AgRP neurons are targets of ghrelin, the overall relevance of ghrelin signaling within intact AgRP neurons is unclear. Here, we tested the functional significance of ghrelin action on AgRP neurons using a new, tamoxifen-inducible AgRP-CreER(T2) transgenic mouse model that allows spatiotemporally-controlled re-expression of physiological levels of ghrelin receptors (GHSRs) specifically in AgRP neurons of adult GHSR-null mice that otherwise lack GHSR expression. AgRP neuron-selective GHSR re-expression partially restored the orexigenic response to administered ghrelin and fully restored the lowered blood glucose levels observed upon caloric restriction. The normalizing glucoregulatory effect of AgRP neuron-selective GHSR expression was linked to glucagon rises and hepatic gluconeogenesis induction. Thus, our data indicate that GHSR-containing AgRP neurons are not solely responsible for ghrelin's orexigenic effects but are sufficient to mediate ghrelin's effects on glycemia.

Nuclear structure and weak rates of heavy waiting point nuclei under rp-process conditions

Science.gov (United States)

Nabi, Jameel-Un; Böyükata, Mahmut

2017-01-01

The structure and the weak interaction mediated rates of the heavy waiting point (WP) nuclei 80Zr, 84Mo, 88Ru, 92Pd and 96Cd along N = Z line were studied within the interacting boson model-1 (IBM-1) and the proton-neutron quasi-particle random phase approximation (pn-QRPA). The energy levels of the N = Z WP nuclei were calculated by fitting the essential parameters of IBM-1 Hamiltonian and their geometric shapes were predicted by plotting potential energy surfaces (PESs). Half-lives, continuum electron capture rates, positron decay rates, electron capture cross sections of WP nuclei, energy rates of β-delayed protons and their emission probabilities were later calculated using the pn-QRPA. The calculated Gamow-Teller strength distributions were compared with previous calculation. We present positron decay and continuum electron capture rates on these WP nuclei under rp-process conditions using the same model. For the rp-process conditions, the calculated total weak rates are twice the Skyrme HF+BCS+QRPA rates for 80Zr. For remaining nuclei the two calculations compare well. The electron capture rates are significant and compete well with the corresponding positron decay rates under rp-process conditions. The finding of the present study supports that electron capture rates form an integral part of the weak rates under rp-process conditions and has an important role for the nuclear model calculations.

Assessment of the chromatographic lipophilicity of eight cephalosporins on different stationary phases.

Science.gov (United States)

Dąbrowska, Monika; Starek, Małgorzata; Komsta, Łukasz; Szafrański, Przemysław; Stasiewicz-Urban, Anna; Opoka, Włodzimierz

2017-04-01

The retention behaviors were investigated for a series of eight cephalosporins in thin-layer chromatography (TLC) using stationary phases of RP-2, RP-8, RP-18, NH 2 , DIOL, and CN chemically bonded silica gel. Additionally, various binary mobile phases (water/methanol and water/acetone) were used in different volume proportions. The retention behavior of the analyzed molecules was defined by R M0 constant. In addition, reversed phase high performance liquid chromatography (RP-HPLC) was performed in lipophilicity studies by using immobilized artificial membrane (IAM) stationary phase. Obtained chromatographic data (R M0 and logk' IAM ) were correlated with the lipophilicity, expressed as values of the log calculated (logP calc ) and experimental (logP exp(shake-flask) ) partition coefficient. Principal component analysis (PCA) was applied in order to obtain an overview of similarity or dissimilarity among the analyzed compounds. Hierarchical cluster analysis (HCA) was performed to compare the separation characteristics of the applied stationary phases. This study was undertaken to identify the best chromatographic system and chromatographic data processing method to enable the prediction of logP values. A comprehensive chromatographic investigation into the retention of the analyzed cephalosporins revealed a similar behavior on RP-18, RP-8 and CN stationary phases. The weak correlations obtained between experimental and certain computed lipophilicity indices revealed that R M0 and PC1/RM are relevant lipophilicity parameters and the RP-8, CN and RP-18 plates are appropriate stationary phases for lipophilicity investigation, whereas computational approaches still cannot fully replace experimentation. Copyright © 2017 Elsevier B.V. All rights reserved.

Development and validation of RP-UHPLC procedure for estimation of 5-amino salicyclic acid in 5-amino salicyclic acid rectal suppositories

Science.gov (United States)

Balaji, Jayagopal; Shivashankar, Murugesh

2017-11-01

The present study describes a simple and robust reverse phase ultra performance liquid chromatography (RP-UPLC) method for the quantification of 5-amino salicyclic acid in 5-amino salicyclic acid rectal capsules. Successful separation of Mesalamine peak from excipient peaks and diluent were achieved on a Acquity C8 (50 × 2.1 mm, 1.7 μm) and UV detector at 254 nm, 0.3 mL/min as a flow rate, and 3 μL as an injection volume. For the RP-UPLC method, phosphate buffer and methanol was used as mobile phases at ratio of 83:17 and the column temperature was 25 °C. Percentage recovery obtained in the range of 98.7 - 99.7 % and the method is linear for Mesalamine for specified concentration range with coefficient of variation (r) not less than 0.99. The proposed RP-UPLC method was found to be specific, linear, precise, accurate and robust.

Development, validation and comparison of two stability-indicating RP-LC methods using charged aerosol and UV detectors for analysis of lisdexamfetamine dimesylate in capsules

Directory of Open Access Journals (Sweden)

Graciela Carlos

2016-11-01

Full Text Available Two new stability-indicating liquid chromatographic methods using two detectors, an ultraviolet (UV and a charged aerosol detector (CAD simultaneously connected in series were validated for the assessment of lisdexamfetamine dimesylate (LDX in capsule. The method was optimized and the influence of individual parameters on UV and CAD response and sensitivity was studied. Chromatography was performed on a Zorbax CN column (250 mm × 4.6 mm, 5 μm in an isocratic elution mode, using acetonitrile and 20 mM ammonium formate at pH 4.0 (50:50, v/v as mobile phase and UV detection at 207 nm. The developed method was validated according to ICH guidelines and the parameters’ specificity, limit of detection, limit of quantitation, linearity, accuracy, precision and robustness were evaluated. CAD is designated to be a non-linear detector in a wide dynamic range, however, the method was linear over the concentration range of 70–130 μg mL−1 in both detectors. The method was precise and accurate. Robustness study was performed by a Plackett–Burman design, delivering results within the acceptable range. Neither the excipients nor the degradation products showed interference in the method after studies of specificity as well as under stress conditions. The results of the LC-UV and LC-CAD methods were statistically compared through ANOVA and showed no significant difference (p > 0.05. Both proposed methods could be considered interchangeable and stability-indicating, and can be applied as an appropriate quality control tool for routine analysis of LDX in capsule.

Short vowel placements in RP past and present

DEFF Research Database (Denmark)

Fabricius, Anne

This study addresses diachronic change in the short vowel system of RP. While TRAP lowering and backing in RP has been reported previously, the movements STRUT has undergone have proven more difficult to determine. This study identifies a TRAP/STRUT 'rotation' using acoustic measurements...

CHARACTERIZATION OF DANSYLATED CYSTEINE, GLUTATHIONE DISULFIDE, CYSTEINE AND CYSTINE BY NARROW BORE LIQUID CHROMATOGRAPHY/ELECTROSPRAY IONIZATION MASS SPECTROMETRY

Science.gov (United States)

A method using reversed phase high performance liquid chromatography/electrospray ionization-mass spectrometric (RP-LC/ESI-MS) method has been developed to confirm the identity of dansylated derivatives of cysteine and glutathione, and their respective dimers. Cysteine, GSH, CSSC...

Swirl Coaxial Injector Testing with LOX/RP-J

Science.gov (United States)

Greene, Sandra Elam; Casiano, Matt

2013-01-01

Testing was conducted at NASA fs Marshall Space Flight Center (MSFC) in the fall of 2012 to evaluate the operation and performance of liquid oxygen (LOX) and kerosene (RP ]1) in an existing swirl coaxial injector. While selected Russian engines use variations of swirl coaxial injectors, component level performance data has not been readily available, and all previously documented component testing at MSFC with LOX/RP ]1 had been performed using a variety of impinging injector designs. Impinging injectors have been adequate for specific LOX/RP ]1 engine applications, yet swirl coaxial injectors offer easier fabrication efforts, providing cost and schedule savings for hardware development. Swirl coaxial elements also offer more flexibility for design changes. Furthermore, testing with LOX and liquid methane propellants at MSFC showed that a swirl coaxial injector offered improved performance compared to an impinging injector. So, technical interest was generated to see if similar performance gains could be achieved with LOX/RP ]1 using a swirl coaxial injector. Results would allow such injectors to be considered for future engine concepts that require LOX/RP ]1 propellants. Existing injector and chamber hardware was used in the test assemblies. The injector had been tested in previous programs at MSFC using LOX/methane and LOX/hydrogen propellants. Minor modifications were made to the injector to accommodate the required LOX/RP ]1 flows. Mainstage tests were performed over a range of chamber pressures and mixture ratios. Additional testing included detonated gbombs h for stability data. Test results suggested characteristic velocity, C*, efficiencies for the injector were 95 ]97%. The injector also appeared dynamically stable with quick recovery from the pressure perturbations generated in the bomb tests.

Arcuate AgRP neurons mediate orexigenic and glucoregulatory actions of ghrelin

OpenAIRE

Wang, Qian; Liu, Chen; Uchida, Aki; Chuang, Jen-Chieh; Walker, Angela; Liu, Tiemin; Osborne-Lawrence, Sherri; Mason, Brittany L.; Mosher, Christina; Berglund, Eric D.; Elmquist, Joel K.; Zigman, Jeffrey M.

2014-01-01

The hormone ghrelin stimulates eating and helps maintain blood glucose upon caloric restriction. While previous studies have demonstrated that hypothalamic arcuate AgRP neurons are targets of ghrelin, the overall relevance of ghrelin signaling within intact AgRP neurons is unclear. Here, we tested the functional significance of ghrelin action on AgRP neurons using a new, tamoxifen-inducible AgRP-CreERT2 transgenic mouse model that allows spatiotemporally-controlled re-expression of physiologi...

CHARACTERIZATION OF DANSYLATED CYSTEINE, CYSTINE, GLUTATHIONE, AND GLUTATHIONE DISULFIDE BY NARROW BORE LIQUID CHROMATOGRAPHY - ELECTROSPRAY IONIZATION MASS SPECTROMETRY

Science.gov (United States)

A method using reversed phase high performance liquid chromtography/electrospray ionization-mass spectrometry (RP-LC/ESI-MS) has been developed to confirm the dientity of dansylated derivatives of cysteine (C) and glutathione (GSH), and their respective dimers, cystine (CSSC) and...

Development of Chromatographic Fingerprints of Eurycoma longifolia (Tongkat Ali Roots Using Online Solid Phase Extraction-Liquid Chromatography (SPE-LC

Directory of Open Access Journals (Sweden)

Nor Nasriah Zaini

2016-04-01

Full Text Available E. longifolia is attracting interest due to its pharmacological properties and pro-vitality effects. In this study, an online SPE-LC approach using polystyrene divinyl benzene (PSDVB and C18 columns was developed in obtaining chromatographic fingerprints of E. longifolia. E. longifolia root samples were extracted using pressurized liquid extraction (PLE technique prior to online SPE-LC. The effects of mobile phase compositions and column switching time on the chromatographic fingerprint were optimized. Validation of the developed method was studied based on eurycomanone. Linearity was in the range of 5 to 50 µg∙mL−1 (r2 = 0.997 with 3.2% relative standard deviation of peak area. The developed method was used to analyze 14 E. longifolia root samples and 10 products (capsules. Selected chemometric techniques: cluster analysis (CA, discriminant analysis (DA, and principal component analysis (PCA were applied to the fingerprint datasets of 37 selected peaks to evaluate the ability of the chromatographic fingerprint in classifying quality of E. longifolia. Three groups were obtained using CA. DA yielded 100% correlation coefficient with 19 discriminant compounds. Using PCA, E. longifolia root samples were clearly discriminated from the products. This study showed that the developed online SPE-LC method was able to provide comprehensive evaluation of E. longifolia samples for quality control purposes.

LC-ESI-MS/MS determination of 4-methylpyrazole in dog plasma and its application to a pharmacokinetic study in dogs.

Science.gov (United States)

Chandrasekhar, Devaraj V; Suresh, Ponnayyan Sulochana; Dittakavi, Sreekanth; Hiremath, Rakesh A; Bhamidipati, Ravi Kanth; Richter, Wolfgang; Srinivas, Nuggehally R; Mullangi, Ramesh

2018-02-01

A simple, specific, sensitive and rapid LC-ESI-MS/MS method has been developed and validated for the quantification of 4-methylpyrazole in dog plasma using N-methylnicotinamide-d 4 as an internal standard (IS) as per regulatory guidelines. Sample preparation was accomplished through a simple protein precipitation. Chromatographic separation of 4-methylpyrazole and the IS was performed on a monolithic (Chromolith RP 18e ) column using an isocratic mobile phase comprising 0.2% formic acid in water and acetonitrile (20:80, v/v) at a flow rate of 1.0 mL/min. Elution of 4-methylpyrazole and the IS occurred at ~1.60 and 1.56 min, respectively. The total chromatographic run time was 3.2 min. A linear response function was established in the concentration range of 4.96-4955 ng/mL. The intra- and inter-day accuracy and precision were in the ranges 1.81-12.9 and 3.80-11.1%, respectively. This novel method has been applied to a pharmacokinetic study in dogs. Copyright © 2017 John Wiley & Sons, Ltd.

RP TLC-based lipophilicity assessment of some natural and synthetic coumarins

Energy Technology Data Exchange (ETDEWEB)

Rabtti, El Hadi M.A.; Natic, Maja M.; Milojkovic-Opsenica, Dusanka M.; Trifkovic, Jelena D.; Vuckovic, Ivan M.; Tesic, Zivoslav Lj., E-mail: ztesic@chem.bg.ac.rs [Faculty of Chemistry, University of Belgrade, Belgrade (Serbia); Vajs, Vlatka E. [Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Belgrade (Serbia)

2012-03-15

The lipophilic character of twelve coumarins was investigated by reversed-phase thin-layer chromatography (RP TLC) on RP-18 silica. The three different binary solvent systems composed of water and organic modifier (methanol, tetrahydrofuran or acetonitrile) were used in order to determine retention parameter (R{sub M}{sup 0}) and octanol-water partition coefficient (log P{sub OW})as a measure of the lipophilicity of the tested compounds. Lipophilicity parameter (log P{sub OW}) was experimentally determined using eight standard solutes with known log P{sub OW} values which were analyzed under the same chromatographic conditions as the target substances. Lipophilicity parameters together with 2D molecular descriptors were subjected to the multivariate statistical analysis (principal component analysis (PCA) and partial least square (PLS) regression) in order to reveal the most influential factors governing the retention, i.e., lipophilicity of the investigated compounds. The quantitative structure-retention relationship models reveal the importance of descriptors which describe the size and the shape of the molecule as well as their polar properties. (author)

RpA ratio: total shadowing due to running coupling

OpenAIRE

Iancu, E.; Triantafyllopoulos, D. N.

2007-01-01

We predict that the RpA ratio at the most forward rapidities to be measured at LHC should be strongly suppressed, close to "total shadowing'' (RpA = A^(-1/3)), as a consequence of running coupling effects in the nonlinear QCD evolution.

An LC-MS Assay with Isocratic Separation and On-Line Solid Phase Extraction to Improve the Routine Therapeutic Drug Monitoring of Busulfan in Plasma

Directory of Open Access Journals (Sweden)

Ialongo Cristiano

2017-04-01

Full Text Available Background: Busulfan (Bu requires therapeutic drug monitoring (TDM in subjects undergoing a conditioning regimen for hematopoietic stem cell transplantation (HSCT. To speed up the procedure and increase reproducibility, we improved our routine LC-MS/MS assay using the on-line solid-phase extraction (SPE of samples.

Exploring Phonetic and Phonological Variation: RP and the Nigerian ...

African Journals Online (AJOL)

... Nigerian university, a hierarchy of the intelligibility of RP vowel phonemes is established. This not only provides evidence that intelligibility is a phenomenon which may be examined from a non-native speaker perspective, it also identifies specific features of RP segmental phonology which presents problems to Nigerians.

Complex mixture analysis of peptides using LC/LC-MS/MS and data-dependent protein identification

International Nuclear Information System (INIS)

Wasinger, V.; Corthals, G.

2001-01-01

The comprehensive identification of proteins within complex solutions by mass-spectrometry largely depends on the sensitivity, resolving power and sampling efficiency of the technology. An integrated orthogonal approach using Strong Cation Exchange-Reverse Phase-MS/MS (SCX-RP-MS/MS) was used to evaluate the data-dependent Collision Induced Dissociation (CID) of yeast peptides. Reverse phase gradient times of 4, 10. 30, 90, and 180 minutes allowed the identification of hundreds of proteins in a nearly automated fashion from nuclear, membrane, and cytosolic distributions. Many proteins from typically difficult to resolve regions of two-dimensional gels, such as >100kDa, > pI 9.0 and Codon Adaptation Index < 0.2, were also identified using this multi-dimensional separation technology. Few low mass proteins (<10kDa) were identified. The impact of scan-range and duty-cycle on CID of peptides will be discussed

Calibration of "Babyline" RP instruments

CERN Multimedia

2015-01-01

  If you have old RP instrumentation of the “Babyline” type, as shown in the photo, please contact the Radiation Protection Group (Joffrey Germa, 73171) to have the instrument checked and calibrated. Thank you. Radiation Protection Group

Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) in toxicological analysis. Studies on the detection of clobenzorex and its metabolites within a systematic toxicological analysis procedure by GC-MS and by immunoassay and studies on the detection of alpha- and beta-amanitin in urine by atmospheric pressure ionization electrospray LC-MS.

Science.gov (United States)

Maurer, H H; Kraemer, T; Ledvinka, O; Schmitt, C J; Weber, A A

1997-02-07

GC-MS is the method of choice for toxicological analysis of toxicants volatile in GC while non-volatile and/or thermally labile toxicants need LC-MS for their determination. Studies are presented on the toxicological detection of the amphetamine-like anorectic clobenzorex in urine by GC-MS after acid hydrolysis, extraction and acetylation and by fluorescence polarization immunoassay (FPIA, TDx (meth)amphetamine II). After ingestion of 60 mg of clobenzorex, the parent compound and/or its metabolites could be detected by GC-MS for up to 84 h or by FPIA for up to 60 h. Since clobenzorex shows no cross-reactivity with the used immunoassay, the N-dealkylated metabolite amphetamine is responsible for the positive TDx results. The intake of clobenzorex instead of amphetamine can be differentiated by GC-MS detection of hydroxyclobenzorex which is detectable for at least as long as amphetamine. In addition, the described GC-MS procedure allows the simultaneous detection of most of the toxicologically relevant drugs. Furthermore, studies are described on the atmospheric pressure ionization electrospray LC-MS detection of alpha- and beta-amanitin, toxic peptides of amanita mushrooms, in urine after solid-phase extraction on RP-18 columns. Using the single ion monitoring mode with the ions m/z 919 and 920 the amanitins could be detected down to 10 ng/ml of urine which allows us to diagnose intoxications with amanita mushrooms.

Ferro-based derivatizing agents for LC/MS an LC/EC/MS

NARCIS (Netherlands)

Seiwert, Bettina

2007-01-01

Within this thesis, the development and application of ferrocene-based derivatizing agents for LC/MS and LC/EC/MS is presented. The advantages of derivatization by ferrocenes are the similtaneous introduction of a mass tag and an electroactive group, which make them ideally suited for LC/MS and

Radiation protection Group (SC/RP) desperately seeking...

CERN Document Server

A. Hervé, Y. Donjoux / SC

2006-01-01

We are trying to trace two transit permits (passavants), which constitute the customs clearance documentation for two 'AD6'portable radiation detectors. The two permits (No. 1308 and No. 1309) were sent in the same internal mail envelope towards the middle of March 2006 but never reached their final destination. After weeks of searching in vain, we are now appealing for your help. If you have these two permits in your possession, please get in touch with us. Many thanks in advance. A.HERVE - SC/RP- ( 163168 / 70927) Y.DONJOUX - SC/RP - (160105 / 73171)

Comparative evaluation of electrostatic repulsion-hydrophilic interaction chromatography (ERLIC) and high-pH reversed phase (Hp-RP) chromatography in profiling of rat kidney proteome.

Science.gov (United States)

Hao, Piliang; Ren, Yan; Dutta, Bamaprasad; Sze, Siu Kwan

2013-04-26

ERLIC and high-pH RP (Hp-RP) have been reported to be promising alternatives to strong cation exchange (SCX) in proteome fractionation. Here we compared the performance of ERLIC, concatenated ERLIC and concatenated Hp-RP in proteome profiling. The protein identification is comparable in these three strategies, but significantly more unique peptides are identified by the two concatenation methods, resulting in a significant increase of the average protein sequence coverage. The pooling of fractions from spaced intervals results in more uniform distribution of peptides in each fraction compared with the chromatogram-based pooling of adjacent fractions. ERLIC fractionates peptides according to their pI and GRAVY values. These properties remains but becomes less remarkable in concatenated ERLIC. In contrast, the average pI and GRAVY values of the peptides are comparable in each fraction in concatenated Hp-RP. ERLIC performs the best in identifying peptides with pI>9 among the three strategies, while concatenated Hp-RP is good at identifying peptides with pI<4. These advantages are useful when either basic or acidic peptides/proteins are analytical targets. The power of ERLIC in identification of basic peptides seems to be due to their efficient separation from acidic peptides. This study facilitates the choice of proper fractionation strategies based on specific objectives. For in-depth proteomic analysis of a cell, tissue and plasma, multidimensional liquid chromatography (MDLC) is still necessary to reduce sample complexity for improving analytical dynamic range and proteome coverage. This work conducts a direct comparison of three promising first-dimensional proteome fractionation methods. They are comparable in identifying proteins, but concatenated ERLIC and concatenated Hp-RP identify significantly more unique peptides than ERLIC. ERLIC is good at analyzing basic peptides, while concatenated Hp-RP performs the best in analyzing acidic peptides with pI<4. This

Measurement of the Diffusion Coefficient of Water in RP-3 and RP-5 Jet Fuels Using Digital Holography Interferometry

Science.gov (United States)

Li, Chaoyue; Feng, Shiyu; Shao, Lei; Pan, Jun; Liu, Weihua

2018-04-01

The diffusion coefficient of water in jet fuel was measured employing double-exposure digital holographic interferometry to clarify the diffusion process and make the aircraft fuel system safe. The experimental method and apparatus are introduced in detail, and the digital image processing program is coded in MATLAB according to the theory of the Fourier transform. At temperatures ranging from 278.15 K to 333.15 K in intervals of 5 K, the diffusion coefficient of water in RP-3 and RP-5 jet fuels ranges from 2.6967 × 10 -10 m2·s-1 to 8.7332 × 10 -10 m2·s-1 and from 2.3517 × 10 -10 m2·s-1 to 8.0099 × 10-10 m2·s-1, respectively. The relationship between the measured diffusion coefficient and temperature can be well fitted by the Arrhenius law. The diffusion coefficient of water in RP-3 jet fuel is higher than that of water in RP-5 jet fuel at the same temperature. Furthermore, the viscosities of the two jet fuels were measured and found to be expressible in the form of the Arrhenius equation. The relationship among the diffusion coefficient, viscosity and temperature is analyzed according to the classic prediction model, namely the Stokes-Einstein correlation, and this correlation is further revised via experimental data to obtain a more accurate predication result.

Resource Prospector (RP) - Early Prototyping and Development

Science.gov (United States)

Andrews, D.; Colaprete, A.; Quinn, J.; Bluethmann, B.; Trimble, J.

2015-01-01

The Resource Prospector (RP) is an In-Situ Resource Utilization (ISRU) technology demonstration mission under study by the NASA Human Exploration and Operations Mission Directorate's (HEOMD) Advanced Exploration Systems (AES) Division. The mission, currently planned to launch in 2020, will demonstrate extraction of oxygen from lunar regolith to validate ISRU capability. The mission will address key Strategic Knowledge Gaps (SKGs) for robotic and human exploration to the Moon, Near Earth Asteroids (NEAs), and ultimately Mars, as well as meet the strategic goals of the Global Exploration Roadmap (GER), offered by the International Space Exploration Coordination Group (ISECG). In this roadmap, the use of local resources is specifically addressed relating to human exploration. RP will provide knowledge to inform the selection of future mission destinations, support the development of exploration systems, and reduce the risk associated with human exploration. Expanding human presence beyond low-Earth orbit to asteroids and Mars will require the maximum possible use of local materials, so-called in-situ resources. The moon presents a unique destination to conduct robotic investigations that advance ISRU capabilities, as well as providing significant exploration and science value. Lunar regolith contains useful resources such as oxygen, water, silicon, and light metals, like aluminum and titanium. Oxygen can be separated from the regolith for life support (breathable air), or used to create rocket propellant (oxidizer). Regolith can be used to protect against radiation exposure, be processed into solar cells, or used to manufacture construction materials such as bricks and glass. RP will characterize the constituents and distribution of water and other volatiles at the poles of the Moon, enabling innovative uses of local resources, in addition to validating ISRU capabilities. This capability, as well as a deeper understanding of regolith, will be valuable in the

Improved methods for urinary atrazine mercapturate analysis-Assessment of an enzyme-linked immunosorbent assay (ELISA) and a novel liquid chromatography-mass spectrometry (LC-MS) method utilizing online solid phase extraction (SPE)

International Nuclear Information System (INIS)

Koivunen, Marja E.; Dettmer, Katja; Vermeulen, Roel; Bakke, Berit; Gee, Shirley J.; Hammock, Bruce D.

2006-01-01

Elimination of interfering substances in urine by solid phase extraction (SPE) prior to analysis resulted in 10-fold improvement in the sensitivity of atrazine mercapturate (AM) enzyme-linked immunosorbent assay (ELISA) compared to previous reports. Of the two tested SPE systems, Oasis[reg] HLB and MCX, the mixed-mode MCX gave good recoveries (82%) of AM in spiked samples measured by ELISA, whereas the reverse-phase HLB phase was not compatible with the immunochemical method. At relatively high concentrations of urinary AM (>20 ng mL -1 ), sample dilution was effective enough for the elimination of interfering substances. The new liquid chromatography-mass spectrometry (LC-MS) method developed for AM utilizes online-SPE with Oasis[reg] HLB, column switching and a stable-isotope internal standard. The limit of quantification (0.05 ng mL -1 ) indicates improved sensitivity compared with most previously published LC-MS methods for AM. Validation of all three methods, LC-MS, ELISA + SPE and ELISA + dilution with spiked urine samples showed good correlation between the known and measured concentrations with R 2 values of 0.996, 0.957 and 0.961, respectively. When a set (n = 70 plus 12 blind duplicates) of urine samples from farmers exposed to atrazine was analyzed, there was a good agreement (R 2 = 0.917) between the log normalized data obtained by ELISA + SPE and LC-MS. High correlation among the data obtained by the two tested methods and the LC-MS method by the Center of Disease Control and Prevention (CDC), together with low variability among the blind duplicates, suggests that both methods reported here would be suitable for the analysis of urinary AM as a biomarker for human exposure of atrazine

DNV RP-F116 integrity management of submarine pipeline system

Energy Technology Data Exchange (ETDEWEB)

Leinum, B.H.; Etterdal, B. [Det Norske Veritas (DNV), Hoevik (Norway); Augustinovic, Z. [Dong Energy, Fredericia (Denmark); Paula, A. [Det Norske Veritas (DNV), Rio de Janeiro, RJ (Brazil)

2009-12-19

The need to keep pipelines operating safely and efficiently is paramount for any operator. Additionally, there is an increasing awareness at both regulatory and public level. Authorities around the world are adopting a more proactive approach as they seek to minimize the risk of human injuries or harm to the environment resulting from pipeline leaks. A joint industry project (JIP) led by DNV Energy is formulating guidelines for how to manage the integrity of submarine pipeline system. The resultant document of recommended practice, DNV RP-F116, will provide the oil and gas industry with a useful tool in an area where no such formal guidance currently exists. This paper gives an overview of the RP and demonstrates the use with a case study for the Siri field. The Siri field is operated by DONG Energy, Denmark and all infield pipelines were subjected to an integrity assessment using the Integrity Management process as outlined in the upcoming DNV RP-116. In parallel with establishing the RP, DNV has further developed the web-based software system, Orbit+, which is aligned with the RP. The software system is implemented and used by Dong Energy. (author)

LC-UV-solid-phase extraction-NMR-MS combined with a cryogenic flow probe and its application to the identification of compounds present in Greek oregano

NARCIS (Netherlands)

Exarchou, V.; Godejohann, M.; Beek, van T.A.; Gerothanassis, I.P.; Vervoort, J.J.M.

2003-01-01

Structure elucidation of natural products usually relies on a combination of NMR spectroscopy with mass spectrometry whereby NMR trails MS in terms of the minimum sample amount required. In the present study, the usefulness of on-line solid-phase extraction (SPE) in LC-NMR for peak storage after the

Hypothalamic AgRP-neurons control peripheral substrate utilization and nutrient partitioning

Science.gov (United States)

Joly-Amado, Aurélie; Denis, Raphaël G P; Castel, Julien; Lacombe, Amélie; Cansell, Céline; Rouch, Claude; Kassis, Nadim; Dairou, Julien; Cani, Patrice D; Ventura-Clapier, Renée; Prola, Alexandre; Flamment, Melissa; Foufelle, Fabienne; Magnan, Christophe; Luquet, Serge

2012-01-01

Obesity-related diseases such as diabetes and dyslipidemia result from metabolic alterations including the defective conversion, storage and utilization of nutrients, but the central mechanisms that regulate this process of nutrient partitioning remain elusive. As positive regulators of feeding behaviour, agouti-related protein (AgRP) producing neurons are indispensible for the hypothalamic integration of energy balance. Here, we demonstrate a role for AgRP-neurons in the control of nutrient partitioning. We report that ablation of AgRP-neurons leads to a change in autonomic output onto liver, muscle and pancreas affecting the relative balance between lipids and carbohydrates metabolism. As a consequence, mice lacking AgRP-neurons become obese and hyperinsulinemic on regular chow but display reduced body weight gain and paradoxical improvement in glucose tolerance on high-fat diet. These results provide a direct demonstration of a role for AgRP-neurons in the coordination of efferent organ activity and nutrient partitioning, providing a mechanistic link between obesity and obesity-related disorders. PMID:22990237

Determination of etoricoxib in human plasma using automated on-line solid-phase extraction coupled with LC-APCI/MS/MS

Directory of Open Access Journals (Sweden)

Sérgio Luiz Dalmora

2008-01-01

Full Text Available A liquid chromatography-tandem mass spectrometry method with atmospheric pressure chemical ionization (LC-APCI/MS/MS was validated for the determination of etoricoxib in human plasma using antipyrin as internal standard, followed by on-line solid-phase extraction. The method was performed on a Luna C18 column and the mobile phase consisted of acetonitrile:water (95:5, v/v/ammonium acetate (pH 4.0; 10 mM, run at a flow rate of 0.6 mL/min. The method was linear in the range of 1-5000 ng/mL (r²>0.99. The lower limit of quantitation was 1 ng/mL. The recoveries were within 93.72-96.18%. Moreover, method validation demonstrated acceptable results for the precision, accuracy and stability studies.

Autophagosome Proteins LC3A, LC3B and LC3C Have Distinct Subcellular Distribution Kinetics and Expression in Cancer Cell Lines.

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Michael I Koukourakis

Full Text Available LC3s (MAP1-LC3A, B and C are structural proteins of autophagosomal membranes, widely used as biomarkers of autophagy. Whether these three LC3 proteins have a similar biological role in autophagy remains obscure. We examine in parallel the subcellular expression patterns of the three LC3 proteins in a panel of human cancer cell lines, as well as in normal MRC5 fibroblasts and HUVEC, using confocal microscopy and western blot analysis of cell fractions. In the cytoplasm, there was a minimal co-localization between LC3A, B and C staining, suggesting that the relevant autophagosomes are formed by only one out of the three LC3 proteins. LC3A showed a perinuclear and nuclear localization, while LC3B was equally distributed throughout the cytoplasm and localized in the nucleolar regions. LC3C was located in the cytoplasm and strongly in the nuclei (excluding nucleoli, where it extensively co-localized with the LC3A and the Beclin-1 autophagy initiating protein. Beclin 1 is known to contain a nuclear trafficking signal. Blocking nuclear export function by Leptomycin B resulted in nuclear accumulation of all LC3 and Beclin-1 proteins, while Ivermectin that blocks nuclear import showed reduction of accumulation, but not in all cell lines. Since endogenous LC3 proteins are used as major markers of autophagy in clinical studies and cell lines, it is essential to check the specificity of the antibodies used, as the kinetics of these molecules are not identical and may have distinct biological roles. The distinct subcellular expression patterns of LC3s provide a basis for further studies.

Supercritical fluid chromatography-A Hybrid of GC and LC

Directory of Open Access Journals (Sweden)

Neha Sethi

2010-01-01

Full Text Available High performance specifications and unique functionality of chromatographic techniques is a demand of pharmaceutical industry and research. This leads to the origin of Supercritical Fluid Chromatography (SFC. It is a rapidly expanding analytical technique. The main feature that differentiates SFC from other chromatographic techniques is the replacement of either the liquid or gas mobile phase with a supercritical fluid mobile phase. It is considered a hybrid of GC and LC technique. High diffusion coefficient and low viscosity of supercritical fluids is responsible for high speed analysis, high efficiency and high sensitivity. Low mobile-phase flow rate, density programming and compatability with GC and LC detectors make SFC a versatile chromatographic technique in analytical re-search and development. It has a unique characteristic of analyzing thermo labile or non-volatile substances. This review highlights the role of supercritical fluid chromatography in the separation of polymers, thermally labile pesticides, fatty acids, metal chelates and organometallic compounds, chiral and achiral molecules, identification and analysis of polar samples, explosives, drugs of abuse and application of SFC in forensic science (fingerprint-ing.

Stability-indicating RP-HPLC method for the simultaneous determination of escitalopram oxalate and clonazepam.

Science.gov (United States)

Kakde, Rajendra B; Satone, Dinesh D; Gadapayale, Kamalesh K; Kakde, Megha G

2013-07-01

The objective of the current study was to develop a validated, specific stability-indicating reversed-phase liquid chromatographic (LC) method for the quantitative determination of escitalopram oxalate and clonazepam and their related substances in bulk drugs and pharmaceutical dosage forms in the presence of degradation products. Forced degradation studies were performed on the pure drugs of escitalopram oxalate and clonazepam, as per the stress conditions prescribed by the International Conference on Harmonization (ICH) using acid, base, oxidation, thermal stress and photolytic degradation to show the stability-indicating power of the method. Significant degradation was observed during acid and alkaline hydrolysis and no degradation was observed in other stress conditions. The chromatographic method was optimized using the samples generated from forced degradation studies. Good resolution between the peaks corresponded to the active pharmaceutical ingredients, escitalopram oxalate and clonazepam, and degradation products from the analyte were achieved on an ODS Hypersil C18 column (250 × 4.6 mm) using a mobile phase consisting of a mixture of acetonitrile-50 mM phosphate buffer + 10 mM triethylamine (70:30, v/v). The detection was conducted at 268 nm. The limit of detection and the limit of quantitation for escitalopram oxalate and clonazepam were established. The stress test solutions were assayed against the qualified working standards of escitalopram oxalate and clonazepam, which indicated that the developed LC method was stability-indicating. Validation of the developed LC method was conducted as per ICH requirements. The developed LC method was found to be suitable to check the quality of bulk samples of escitalopram oxalate and clonazepam.

A UKSeRP for SAIL: striking a balance

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Kerina Jones

2017-04-01

The SAIL UKSeRP represents a powerful analytical environment within a privacy-protecting safe haven and secure remote access system which has been designed to be scalable and adaptable to meet the needs of the rapidly growing data linkage community. Further challenges lie ahead as the landscape develops and emerging data types become more available. UKSeRP technology is available and customisable for other use cases within the UK and international jurisdictions, to operate within their respective governance frameworks.

A Simple and Validated Reverse Phase HPLC Methodfor the Determination of Rabeprazole inPharmaceutical Dosage Forms

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Uma Mahesh Karra

2010-01-01

Full Text Available A simple and rapid reverse phase high performance liquid chromatography (RP-HPLC method was developed and validated for quantitative determination of rabeprazole in bulk drug samples and formulations. Rabeprazole was analyzed by using reverse phase LC-GC column (Inertsil ODS, 4.6 mm x 25 cm, 5 microns, with mobile phase consisting of methanol: water (78:22 v/v. The flow rate was set 1.0 mL/min and analysis was performed at wavelength 288 nm using Photo Diode Array (PDA detector at ambient temperature. The method was validated and stability studies were conducted under different conditions. The retention time for rabeprazole was around 4.12 minutes. The calibration curves were linear (r≥0.9998 over a concentration range from 20.0 to 80.0 μg/mL. Limit of detection (LOD and Limit of quantitation (LOQ were 8 ng/mL and 24 ng/mL respectively. The developed method was successfully applied to estimate the amount of rabeprazole in tablet formulations.

Experience in initial training required for the recognition of the qualified RP expert in Spain

International Nuclear Information System (INIS)

Rodriguez Suarez, M.; Marco Arboli, M.; Menarguez, J.

2003-01-01

An important point of the actions inside the European framework to achieve the harmonisation of the training programmes and recognition was included in the European directive 96/29/Euratom which includes definition and specific tasks of the European Qualified Expert on Radiation Protection (RP). Basic syllabus for training of those experts was developed in the communication 98/C 133/03 concerning BSS applications. Although, in the Spanish education system, the training and recognition requirements of the high level qualified experts on RP was defined since 1977, until 2001, the figure of the Technical Qualified Expert on RP does not appear in the legal framework. In December 2002, a new regulation of the Spanish Regulatory Body, CSN, about qualifications to obtain the recognition of RP Expert in Spain (both high qualified and technical RP experts) was published. Concerning the qualified expert on RP, (RP Officer), responsible of the RP Service, which takes charge of the effective protection and advise radioactive and nuclear facilities in Rp aspects,has to be authorised by the regulatory body. to obtain the RP officer diploma, conceded by the CSN, an initial training of 300 hours and a three-year minimum experience are required (for X-ray installation a 6-month experience is enough). The technical qualified expert on RP is the worker who carried out the tasks in the a RP Service under the supervision of the RP officer. A Technician Qualified Expert on RP does not need an specific accreditation of the Regulatory Body, but an initial RP training and a three-month minimum experience are required and has hold a certificate by the RP officer. Continuous training is also required and as well has to receive a certificate from the RP officer. Since 1977, The Institute for energy Studies has been implementing specific training courses for those professionals who want to obtain the diploma of RP officer (high degree qualified RP expert), conceded by the CSN. Since then

Effects of phase transformation of steam-water relative permeabilities

Energy Technology Data Exchange (ETDEWEB)

Verma, A.K.

1986-03-01

A combined theoretical and experimental study of steam-water relative permeabilities (RPs) was carried out. First, an experimental study of two-phase concurrent flow of steam and water was conducted and a set of RP curves was obtained. These curves were compared with semi-empirical and experimental results obtained by other investigators for two-phase, two-component flow (oil/gas; gas/water; gas/oil). It was found that while the wetting phase RPs were in good agreement, RPs for the steam phase were considerably higher than the non-wetting phase RPs in two-component systems. This enhancement of steam RP is attributed to phase transformation effects at the pore level in flow channels. The effects of phase transformation were studied theoretically. This study indicates that there are two separate mechanisms by which phase transformation affects RP curves: (1) Phase transformation is converging-diverging flow channels can cause an enhancement of steam phase RP. In a channel dominated by steam a fraction of the flowing steam condenses upstream from the constriction, depositing its latent heat of condensation. This heat is conducted through the solid grains around the pore throat, and evaporation takes place downstream from it. Therefore, for a given bulk flow quality; a smaller fraction of steam actually flows through the throat segments. This pore-level effect manifests itself as relative permeability enhancement on a macroscopic level; and (2) phase transformation along the interface of a stagnant phase and the phase flowing around it controls the irreducible phase saturation. Therefore, the irreducible phase saturation in steam-water flow will depend, among other factors, on the boundary conditions of the flow.

Development and validation of simple RP-HPLC-PDA analytical protocol for zileuton assisted with Design of Experiments for robustness determination

Directory of Open Access Journals (Sweden)

Saurabh B. Ganorkar

2017-02-01

Full Text Available A simple, rapid, sensitive, robust, stability-indicating RP-HPLC-PDA analytical protocol was developed and validated for the analysis of zileuton racemate in bulk and in tablet formulation. Development of method and resolution of degradation products from forced; hydrolytic (acidic, basic, neutral, oxidative, photolytic (acidic, basic, neutral, solid state and thermal (dry heat degradation was achieved on a LC – GC Qualisil BDS C18 column (250 mm × 4.6 mm × 5 μm by isocratic mode at ambient temperature, employing a mobile phase methanol and (0.2%, v/v orthophosphoric acid in ratio of (80:20, v/v at a flow rate of 1.0 mL min−1 and detection at 260 nm. ‘Design of Experiments’ (DOE employing ‘Central Composite Design’ (CCD and ‘Response Surface Methodology’ (RSM were applied as an advancement to traditional ‘One Variable at Time’ (OVAT approach to evaluate the effects of variations in selected factors (methanol content, flow rate, concentration of orthophosphoric acid as graphical interpretation for robustness and statistical interpretation was achieved with Multiple Linear Regression (MLR and ANOVA. The method succeeded over the validation parameters: linearity, precision, accuracy, limit of detection and limit of quantitation, and robustness. The method was applied effectively for analysis of in-house zileuton tablets.

[Positive inotropic and lusitropic effect of RP 62719, a new class III antiarrhythmia agent].

Science.gov (United States)

Beregi, J P; Escande, D; Coudray, N; Chemla, D; Mestre, M; Péry, N; Lecarpentier, Y

1994-02-01

Antiarrhythmic drugs, especially the Class I family, exert a negative inotropic effect on the myocardium which is particularly undesirable in patients with depressed left ventricular function. Therefore, research has been directed to the development of new, more specific molecules of the Class III family. The authors studies the mechanical effects of RP 62719 on guinea pig left ventricular papillary muscle. This new molecule is a pure Class III antiarrhythmic, known to lengthen the duration of the cardiac action potential by selectively blocking the potassium current iK1 (inward rectifier K+ current). The mechanical parameters were determined during the phases of contraction and relaxation under isotonic and isometric conditions. At 0.2 and 2 microM concentrations, RP 62719 improved cardiac contraction under both isotonic and isometric conditions with an increase of about 30% of Vmax (p delta 1 (p force normalized per cross-sectional area [AF/S (p actions could provide significant therapeutic advantages especially in patients cardiac failure.

AgRP Neurons Control Systemic Insulin Sensitivity via Myostatin Expression in Brown Adipose Tissue.

Science.gov (United States)

Steculorum, Sophie M; Ruud, Johan; Karakasilioti, Ismene; Backes, Heiko; Engström Ruud, Linda; Timper, Katharina; Hess, Martin E; Tsaousidou, Eva; Mauer, Jan; Vogt, Merly C; Paeger, Lars; Bremser, Stephan; Klein, Andreas C; Morgan, Donald A; Frommolt, Peter; Brinkkötter, Paul T; Hammerschmidt, Philipp; Benzing, Thomas; Rahmouni, Kamal; Wunderlich, F Thomas; Kloppenburg, Peter; Brüning, Jens C

2016-03-24

Activation of Agouti-related peptide (AgRP) neurons potently promotes feeding, and chronically altering their activity also affects peripheral glucose homeostasis. We demonstrate that acute activation of AgRP neurons causes insulin resistance through impairment of insulin-stimulated glucose uptake into brown adipose tissue (BAT). AgRP neuron activation acutely reprograms gene expression in BAT toward a myogenic signature, including increased expression of myostatin. Interference with myostatin activity improves insulin sensitivity that was impaired by AgRP neurons activation. Optogenetic circuitry mapping reveals that feeding and insulin sensitivity are controlled by both distinct and overlapping projections. Stimulation of AgRP → LHA projections impairs insulin sensitivity and promotes feeding while activation of AgRP → anterior bed nucleus of the stria terminalis (aBNST)vl projections, distinct from AgRP → aBNSTdm projections controlling feeding, mediate the effect of AgRP neuron activation on BAT-myostatin expression and insulin sensitivity. Collectively, our results suggest that AgRP neurons in mice induce not only eating, but also insulin resistance by stimulating expression of muscle-related genes in BAT, revealing a mechanism by which these neurons rapidly coordinate hunger states with glucose homeostasis. Copyright © 2016 Elsevier Inc. All rights reserved.

Temperature Jump Pyrolysis Studies of RP 2 Fuel

Science.gov (United States)

2017-01-09

Briefing Charts 3. DATES COVERED (From - To) 15 December 2016 – 11 January 2017 4. TITLE AND SUBTITLE Temperature Jump Pyrolysis Studies of RP-2 Fuel...Rev. 8- 98) Prescribed by ANSI Std. 239.18 1 TEMPERATURE JUMP PYROLYSIS STUDIES OF RP-2 FUEL Owen Pryor1, Steven D. Chambreau2, Ghanshyam L...17026 7 Temperature Jump Pyrolysis at AFRL Edwards Rapid heating of a metal filament at a rate of 600 – 800 K/s, and the set temperature is held for

Analytical methodologies based on LC-MS/MS for monitoring selected emerging compounds in liquid and solid phases of the sewage sludge.

Science.gov (United States)

Boix, C; Ibáñez, M; Fabregat-Safont, D; Morales, E; Pastor, L; Sancho, J V; Sánchez-Ramírez, J E; Hernández, F

2016-01-01

In this work, two analytical methodologies based on liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) were developed for quantification of emerging pollutants identified in sewage sludge after a previous wide-scope screening. The target list included 13 emerging contaminants (EC): thiabendazole, acesulfame, fenofibric acid, valsartan, irbesartan, salicylic acid, diclofenac, carbamazepine, 4-aminoantipyrine (4-AA), 4-acetyl aminoantipyrine (4-AAA), 4-formyl aminoantipyrine (4-FAA), venlafaxine and benzoylecgonine. The aqueous and solid phases of the sewage sludge were analyzed making use of Solid-Phase Extraction (SPE) and UltraSonic Extraction (USE) for sample treatment, respectively. The methods were validated at three concentration levels: 0.2, 2 and 20 μg L(-1) for the aqueous phase, and 50, 500 and 2000 μg kg(-1) for the solid phase of the sludge. In general, the method was satisfactorily validated, showing good recoveries (70-120%) and precision (RSD metabolites of dipyrone had not been studied before in sewage sludge.

Simultaneous analysis of 17 diuretics in dietary supplements by HPLC and LC-MS/MS.

Science.gov (United States)

Woo, H; Kim, J W; Han, K M; Lee, J H; Hwang, I S; Lee, J H; Kim, J; Kweon, S J; Cho, S; Chae, K R; Han, S Y; Kim, J

2013-01-01

In order to test health foods for illegally added diuretics for weight loss, we developed simple, rapid, selective, and sensitive methods using HPLC and LC-MS/MS for the simultaneous analysis of 17 diuretics in dietary supplements. HPLC conditions were set with a Capcell-pak C18, using a mobile phase consisting of gradient conditions, UV detection at 254 nm and validated for linearity (r(2)> 0.999), precision (CV ≤ 3%), recoveries (90.4-102.8%) and reproducibility. Identification and quantification of 17 diuretics were accomplished by ion-spray LC-MS/MS using multiple reaction monitoring (MRM). The chromatographic separation was carried out under the reversed-phase mechanism on an HSS-T3 column. The LC-MS/MS method was validated for linearity (r(2)> 0.99) and precision (CV Diuretics were not detected in all samples. Extraction recovery was also investigated and the extraction recoveries in different formulations were from 88% to 110% and from 81% to 116% using HPLC and LC-MS/MS, respectively. There was no significant difference in recoveries in the type of dietary supplements. Based on this result, the developed methods to monitor illegal drug adulterations in dietary supplements using HPLC and LC-MS/MS are simple, fast and reliable. Therefore, it is applicable to routine drug-adulteration screening.

[Separation and identification of bovine lactoferricin by high performance liquid chromatography-matrix-assisted laser desorption/ionization time of flight/ time of flight mass spectrometry].

Science.gov (United States)

An, Meichen; Liu, Ning

2010-02-01

A high performance liquid chromatography-matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (HPLC-MALDI-TOF/TOF MS) method was developed for the separation and identification of bovine lactoferricin (LfcinB). Bovine lactoferrin was hydrolyzed by pepsin and then separated by ion exchange chromatography and reversed-phase liquid chromatography (RP-LC). The antibacterial activities of the fractions from RP-LC separation were determined and the protein concentration of the fraction with the highest activity was measured, whose sequence was indentified by MALDI-TOF/TOF MS. The relative molecular mass of LfcinB was 3 124.89 and the protein concentration was 18.20 microg/mL. The method of producing LfcinB proposed in this study has fast speed, high accuracy and high resolution.

RP1 is a phosphorylation target of CK2 and is involved in cell adhesion.

Science.gov (United States)

Stenner, Frank; Liewen, Heike; Göttig, Stephan; Henschler, Reinhard; Markuly, Norbert; Kleber, Sascha; Faust, Michael; Mischo, Axel; Bauer, Stefan; Zweifel, Martin; Knuth, Alexander; Renner, Christoph; Wadle, Andreas

2013-01-01

RP1 (synonym: MAPRE2, EB2) is a member of the microtubule binding EB1 protein family, which interacts with APC, a key regulatory molecule in the Wnt signalling pathway. While the other EB1 proteins are well characterized the cellular function and regulation of RP1 remain speculative to date. However, recently RP1 has been implicated in pancreatic cancerogenesis. CK2 is a pleiotropic kinase involved in adhesion, proliferation and anti-apoptosis. Overexpression of protein kinase CK2 is a hallmark of many cancers and supports the malignant phenotype of tumor cells. In this study we investigate the interaction of protein kinase CK2 with RP1 and demonstrate that CK2 phosphorylates RP1 at Ser(236) in vitro. Stable RP1 expression in cell lines leads to a significant cleavage and down-regulation of N-cadherin and impaired adhesion. Cells expressing a Phospho-mimicking point mutant RP1-ASP(236) show a marked decrease of adhesion to endothelial cells under shear stress. Inversely, we found that the cells under shear stress downregulate endogenous RP1, most likely to improve cellular adhesion. Accordingly, when RP1 expression is suppressed by shRNA, cells lacking RP1 display significantly increased cell adherence to surfaces. In summary, RP1 phosphorylation at Ser(236) by CK2 seems to play a significant role in cell adhesion and might initiate new insights in the CK2 and EB1 family protein association.

RP1 is a phosphorylation target of CK2 and is involved in cell adhesion.

Directory of Open Access Journals (Sweden)

Frank Stenner

Full Text Available RP1 (synonym: MAPRE2, EB2 is a member of the microtubule binding EB1 protein family, which interacts with APC, a key regulatory molecule in the Wnt signalling pathway. While the other EB1 proteins are well characterized the cellular function and regulation of RP1 remain speculative to date. However, recently RP1 has been implicated in pancreatic cancerogenesis. CK2 is a pleiotropic kinase involved in adhesion, proliferation and anti-apoptosis. Overexpression of protein kinase CK2 is a hallmark of many cancers and supports the malignant phenotype of tumor cells. In this study we investigate the interaction of protein kinase CK2 with RP1 and demonstrate that CK2 phosphorylates RP1 at Ser(236 in vitro. Stable RP1 expression in cell lines leads to a significant cleavage and down-regulation of N-cadherin and impaired adhesion. Cells expressing a Phospho-mimicking point mutant RP1-ASP(236 show a marked decrease of adhesion to endothelial cells under shear stress. Inversely, we found that the cells under shear stress downregulate endogenous RP1, most likely to improve cellular adhesion. Accordingly, when RP1 expression is suppressed by shRNA, cells lacking RP1 display significantly increased cell adherence to surfaces. In summary, RP1 phosphorylation at Ser(236 by CK2 seems to play a significant role in cell adhesion and might initiate new insights in the CK2 and EB1 family protein association.

Nutritive, Post-ingestive Signals Are the Primary Regulators of AgRP Neuron Activity

Directory of Open Access Journals (Sweden)

Zhenwei Su

2017-12-01

Full Text Available Summary: The brain regulates food intake by processing sensory cues and peripheral physiological signals, but the neural basis of this integration remains unclear. Hypothalamic, agouti-related protein (AgRP-expressing neurons are critical regulators of food intake. AgRP neuron activity is high during hunger and is rapidly reduced by the sight and smell of food. Here, we reveal two distinct components of AgRP neuron activity regulation: a rapid but transient sensory-driven signal and a slower, sustained calorie-dependent signal. We discovered that nutrients are necessary and sufficient for sustained reductions in AgRP neuron activity and that activity reductions are proportional to the calories obtained. This change in activity is recapitulated by exogenous administration of gut-derived satiation signals. Furthermore, we showed that the nutritive value of food trains sensory systems—in a single trial—to drive rapid, anticipatory AgRP neuron activity inhibition. Together, these data demonstrate that nutrients are the primary regulators of AgRP neuron activity. : Su et al. demonstrate that nutrient content in the GI tract is rapidly signaled to hypothalamic neurons activated by hunger. This rapid effect is mediated by three satiation signals that synergistically reduce the activity of AgRP neurons. These findings uncover how hunger circuits in the brain are regulated and raise the possibility that hunger can be pharmacologically controlled. Keywords: calcium imaging, AgRP neurons, calories, satiation signals, sensory regulation, single trial learning, cholecystokinin, CCK, peptide tyrosine tyrosine, PYY, amylin, homeostasis

Verification of propofol sulfate as a further human propofol metabolite using LC-ESI-QQQ-MS and LC-ESI-QTOF-MS analysis.

Science.gov (United States)

Maas, Alexandra; Maier, Christoph; Michel-Lauter, Beate; Broecker, Sebastian; Madea, Burkhard; Hess, Cornelius

2017-03-01

Propofol (2,6-diisopropylphenol) is a water-insoluble, intravenous anesthetic that is widely used for the induction and maintenance of anesthesia as well as for endoscopic and pediatric sedation. After admission, propofol undergoes extensive hepatic and extrahepatic metabolism, including direct conjugation to propofol glucuronide and hydroxylation to 2,6-diisopropyl-1,4-quinol. The latter substance subsequently undergoes phase II metabolism, resulting in the formation of further metabolites (1quinolglucuronide, 4quinolglucuronide and 4quinol-sulfate). Further minor phase I propofol metabolites (2-(ω-propanol)-6-isopropylphenol and 2-(ω-propanol)-6-isopropyl-1,4-quinol)) are also described. Due to its chemical structure with the phenolic hydroxyl group, propofol is also an appropriate substrate for sulfation by sulfotransferases. The existence of propofol sulfate was investigated by liquid chromatography electrospray ionization triple quadrupole mass spectrometry (LCESIQQQ-MS) and liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LCESI-QTOF-MS). A propofol sulfate reference standard was used for identification and method development, yielding a precursor at m/z 257 (deprotonated propofol sulfate) and product ions at m/z 177 (deprotonated propofol) and m/z 80 ([SO3]-). Propofol sulfate - a further phase II metabolite of propofol - was verified in urine samples by LC-ESI-QQQ-MS and LC-ESI-QTOF-MS. Analyses of urine samples from five volunteers collected before and after propofol-induced sedation verified the presence of propofol sulfate in urine following propofol administration, whereas ascertained concentrations of this metabolite were significantly lower compared with detected propofol glucuronide concentrations. The existence of propofol sulfate as a further phase II propofol metabolite in humans could be verified by two different detection techniques (LCESIQQQ-MS and LC-ESI-QTOFMS) on the basis of a propofol sulfate

Determination of thromboxanes, leukotrienes and lipoxins using high-temperature capillary liquid chromatography-tandem mass spectrometry and on-line sample preparation

DEFF Research Database (Denmark)

Dahl, Sandra Rinne; Kleiveland, Charlotte Ramstad; Kassem, Moustapha

2009-01-01

An on-line strong cation-exchange (SCX)-reversed-phase (RP) capillary liquid chromatographic (cLC) method with ion-trap tandem mass spectrometric (IT-MS/MS) detection for the simultaneous determination of thromboxane (TX) B(2), TXB(3), leukotriene (LT) B(4), LTD(4) and lipoxin (LX) A(4) in cell...

Pharmacokinetics and safety of DTS-108, a human oligopeptide bound to SN-38 with an esterase-sensitive cross-linker in patients with advanced malignancies: a Phase I study

Directory of Open Access Journals (Sweden)

Coriat R

2016-11-01

Full Text Available Romain Coriat,1 Sandrine J Faivre,2 Olivier Mir,3 Chantal Dreyer,2 Stanislas Ropert,3 Mohammed Bouattour,2 Robert Desjardins,4 François Goldwasser,3 Eric Raymond5 1Gastroenterology and Digestive Oncology Unit, Cochin Teaching Hospital, Université Paris Descartes Sorbonne Paris Cité, Paris, 2Department of Medical Oncology, Beaujon Teaching Hospital, Université Paris Diderot, Paris 7, Clichy, 3Department of Medical Oncology, Cochin Teaching Hospital, Université Paris Descartes Sorbonne Paris Cité, Paris, France; 4Drais Pharmaceuticals, Bridgewater, NJ, USA; 5Groupe Hospitalier Paris Saint-Joseph, Paris, France Background: DTS-108 is a hydrosoluble prodrug, where the SN-38 moiety is covalently linked to a 20-amino acid vector peptide by a specific esterase-sensitive cross-linker, releasing 7-ethyl-10-hydroxycampthotecin (SN-38 by esterase bond cleavage. Methods: The pharmacokinetics of DTS-108, adverse events graded according to NCI-CTCv3.1, dose-limiting toxicities at cycle 1, the maximum tolerated dose (MTD, and the recommended Phase II dose (RP2D of intravenous DTS-108 (1–2 hours every 2 weeks were evaluated in a first-in-human Phase I study in patients with advanced/metastatic carcinomas, according to an accelerated dose escalation design. SN-38 and SN-38 glucuronide (SN-38G levels were evaluated with fluorescence high-performance liquid chromatography (HPLC test, then liquid chromatography–tandem mass spectrometry (LC/MS/MS methods. Results: Forty-two patients received DTS-108 across 14 dosing cohorts (range 3–416 mg/m2. At 416 mg/m2, three out of six patients had grade 4 neutropenia thereby defining the MTD and the RP2D at 313 mg/m2. Fluorescence HPLC was inaccurate to quantify DTS-108 and its metabolites (SN-38 and SN-38G. New processes and analytical LC/MS/MS methods for testing SN-38 were implemented. At a dose of 313 mg/m2, mean DTS-108, SN-38, and SN-38G area under the plasma concentration–time curve to infinity

An integrated top-down and bottom-up strategy for characterization protein isoforms and modifications

Energy Technology Data Exchange (ETDEWEB)

Wu, Si; Tolic, Nikola; Tian, Zhixin; Robinson, Errol W.; Pasa-Tolic, Ljiljana

2011-04-15

Bottom-up and top-down strategies are two commonly used methods for mass spectrometry (MS) based protein identification; each method has its own advantages and disadvantages. In this chapter, we describe an integrated top-down and bottom-up approach facilitated by concurrent liquid chromatography-mass spectrometry (LC-MS) analysis and fraction collection for comprehensive high-throughput intact protein profiling. The approach employs a high resolution reversed phase (RP) LC separation coupled with LC eluent fraction collection and concurrent on-line MS with a high field (12 Tesla) Fourier-transform ion cyclotron resonance (FTICR) mass spectrometer. Protein elusion profiles and tentative modified protein identification are made using detected intact protein mass in conjunction with bottom-up protein identifications from the enzymatic digestion and analysis of corresponding LC fractions. Specific proteins of biological interest are incorporated into a target ion list for subsequent off-line gas-phase fragmentation that uses an aliquot of the original collected LC fraction, an aliquot of which was also used for bottom-up analysis.

New Stability Indicating RP-HPLC Method for the Estimation of Cefpirome Sulphate in Bulk and Pharmaceutical Dosage Forms.

Science.gov (United States)

Rao, Kareti Srinivasa; Kumar, Keshar Nargesh; Joydeep, Datta

2011-01-01

A simple stability indicating reversed-phase HPLC method was developed and subsequently validated for estimation of Cefpirome sulphate (CPS) present in pharmaceutical dosage forms. The proposed RP-HPLC method utilizes a LiChroCART-Lichrosphere100, C18 RP column (250 mm × 4mm × 5 μm) in an isocratic separation mode with mobile phase consisting of methanol and water in the proportion of 50:50 % (v/v), at a flow rate 1ml/min, and the effluent was monitored at 270 nm. The retention time of CPS was 2.733 min and its formulation was exposed to acidic, alkaline, photolytic, thermal and oxidative stress conditions, and the stressed samples were analyzed by the proposed method. The described method was linear over a range of 0.5-200μg/ml. The percentage recovery was 99.46. F-test and t-test at 95% confidence level were used to check the intermediate precision data obtained under different experimental setups; the calculated value was found to be less than the critical value.

Fabricating Complete Dentures with CAD/CAM and RP Technologies.

Science.gov (United States)

Bilgin, Mehmet Selim; Erdem, Ali; Aglarci, Osman Sami; Dilber, Erhan

2015-06-01

Two techological approaches for fabricating dentures; computer-aided design and computer-aided manufacturing (CAD/CAM) and rapid prototyping (RP), are combined with the conventional techniques of impression and jaw relation recording to determine their feasibility and applicability. Maxillary and mandibular edentulous jaw models were produced using silicone molds. After obtaining a gypsum working model, acrylic bases were crafted, and occlusal rims for each model were fabricated with previously determined standard vertical and centric relationships. The maxillary and mandibular relationships were recorded with guides. The occlusal rims were then scanned with a digital scanner. The alignment of the maxillary and mandibular teeth was verified. The teeth in each arch were fabricated in one piece, or set, either by CAM or RP. Conventional waxing and flasking was then performed for both methods. These techniques obviate a practitioner's need for technicians during design and provide the patient with an opportunity to participate in esthetic design with the dentist. In addition, CAD/CAM and RP reduce chair time; however, the materials and techniques need further improvements. Both CAD/CAM and RP techniques seem promising for reducing chair time and allowing the patient to participate in esthetics design. Furthermore, the one-set aligned artificial tooth design may increase the acrylic's durability. © 2015 by the American College of Prosthodontists.

The Elevation of LC-ESI-Q-TOF-MS Response in the Analysis of Isoquinoline Alkaloids from Some Papaveraceae and Berberidaceae Representatives.

Science.gov (United States)

Kukula-Koch, Wirginia

2017-01-01

Twenty-five methanol extracts obtained from various representatives of Papaveraceae and Berberidaceae botanical families (genera: Papaver , Argemone , Eschscholzia , Chelidonium , Glaucium , and Berberis ) were screened for their alkaloid content in an optimized method suitable for the LC-ESI-Q-TOF-MS analysis. Twelve pharmacologically important isoquinoline alkaloids from four groups, aporphines, benzylisoquinolines, protoberberines, and benzophenanthridines, present in these traditionally used plant species were quantitatively determined in each studied sample, providing their alkaloid profile. A Zorbax Stable Bond RP-18 column and a mobile phase composed of 0.1% formic acid and 0.1% formic acid in acetonitrile (v/v) were used at the flow rate of 0.2 mL/min. A profound study on the optimization of MS response to four groups of isoquinoline alkaloids (validation of capillary voltage, gas flows, nebulizer pressure, skimmer, and fragmentor voltages), repeatability of results, and stability and linearity of measurements were described, showing, among others, 3000 V of capillary voltage, 350°C of gas temperature, 12 L/min of gas flows, nebulizer pressure of 35 psig, 65 V for skimmer voltage, and 30 V for collision energy as the most advantageous operation parameters.

The Elevation of LC-ESI-Q-TOF-MS Response in the Analysis of Isoquinoline Alkaloids from Some Papaveraceae and Berberidaceae Representatives

Directory of Open Access Journals (Sweden)

Wirginia Kukula-Koch

2017-01-01

Full Text Available Twenty-five methanol extracts obtained from various representatives of Papaveraceae and Berberidaceae botanical families (genera: Papaver, Argemone, Eschscholzia, Chelidonium, Glaucium, and Berberis were screened for their alkaloid content in an optimized method suitable for the LC-ESI-Q-TOF-MS analysis. Twelve pharmacologically important isoquinoline alkaloids from four groups, aporphines, benzylisoquinolines, protoberberines, and benzophenanthridines, present in these traditionally used plant species were quantitatively determined in each studied sample, providing their alkaloid profile. A Zorbax Stable Bond RP-18 column and a mobile phase composed of 0.1% formic acid and 0.1% formic acid in acetonitrile (v/v were used at the flow rate of 0.2 mL/min. A profound study on the optimization of MS response to four groups of isoquinoline alkaloids (validation of capillary voltage, gas flows, nebulizer pressure, skimmer, and fragmentor voltages, repeatability of results, and stability and linearity of measurements were described, showing, among others, 3000 V of capillary voltage, 350°C of gas temperature, 12 L/min of gas flows, nebulizer pressure of 35 psig, 65 V for skimmer voltage, and 30 V for collision energy as the most advantageous operation parameters.

Analysis of catecholamines in urine by unique LC/MS suitable ion-pairing chromatography.

Science.gov (United States)

Bergmann, Marianne L; Sadjadi, Seyed; Schmedes, Anne

2017-07-01

The catecholamines, epinephrine (E) and norepinephrine (NE) are small polar, hydrophilic molecules, posing significant challenges to liquid chromatography - tandem mass spectrometry (LC-MS/MS) method development. Specifically, these compounds show little retention on conventional reversed-phase liquid chromatography columns. This work presents development and validation of an LC-MS/MS method for determining catecholamines in urine, based on a new approach to ion-pairing chromatography (IPC), in which the ion-pairing reagent (IPR), 1-Heptane Sulfonic Acid (HSA), is added to the extracted samples instead of the mobile phases. A Hamilton STARlet workstation carried out the solid phase extraction of urine samples. The extracted samples were diluted with 60mmol/L HSA and injected on a Kinetex core-shell biphenyl column with conventional LC-MS/MS suitable mobile phases. Chromatographic separation of E and NE was achieved successfully with very stable retention times (RT). In 484 injections, the RTs were steady with a CV of less than ±4%. Furthermore, HSA was separated from E and NE, allowing HSA to be diverted to waste instead of entering the mass spectrometer ion chamber. The method was validated with good analytical performance, and even though the analysis for urinary catecholamines is increasingly being replaced by plasma free metanephrines in diagnosing pheochromocytomas, this work represents the application of a new analytical technique that can be transferred to other small polar molecules, that are difficult to chromatograph on traditional reversed phase columns. Copyright © 2017 Elsevier B.V. All rights reserved.

RP58 Regulates the Multipolar-Bipolar Transition of Newborn Neurons in the Developing Cerebral Cortex

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Chiaki Ohtaka-Maruyama

2013-02-01

Full Text Available Accumulating evidence suggests that many brain diseases are associated with defects in neuronal migration, suggesting that this step of neurogenesis is critical for brain organization. However, the molecular mechanisms underlying neuronal migration remain largely unknown. Here, we identified the zinc-finger transcriptional repressor RP58 as a key regulator of neuronal migration via multipolar-to-bipolar transition. RP58−/− neurons exhibited severe defects in the formation of leading processes and never shifted to the locomotion mode. Cre-mediated deletion of RP58 using in utero electroporation in RP58flox/flox mice revealed that RP58 functions in cell-autonomous multipolar-to-bipolar transition, independent of cell-cycle exit. Finally, we found that RP58 represses Ngn2 transcription to regulate the Ngn2-Rnd2 pathway; Ngn2 knockdown rescued migration defects of the RP58−/− neurons. Our findings highlight the critical role of RP58 in multipolar-to-bipolar transition via suppression of the Ngn2-Rnd2 pathway in the developing cerebral cortex.

AgRP neurons regulate development of dopamine neuronal plasticity and nonfood-associated behaviors

Science.gov (United States)

Dietrich, Marcelo O; Bober, Jeremy; Ferreira, Jozélia G; Tellez, Luis A; Mineur, Yann S; Souza, Diogo O; Gao, Xiao-Bing; Picciotto, Marina R; Araújo, Ivan; Liu, Zhong-Wu; Horvath, Tamas L

2012-01-01

It is not known whether behaviors unrelated to feeding are affected by hypothalamic regulators of hunger. We found that impairment of Agouti-related protein (AgRP) circuitry by either Sirt1 knockdown in AgRP-expressing neurons or early postnatal ablation of these neurons increased exploratory behavior and enhanced responses to cocaine. In AgRP circuit–impaired mice, ventral tegmental dopamine neurons exhibited enhanced spike timing–dependent long-term potentiation, altered amplitude of miniature postsynaptic currents and elevated dopamine in basal forebrain. Thus, AgRP neurons determine the set point of the reward circuitry and associated behaviors. PMID:22729177

Development and validation of polar RP-HPLC method for screening for ectoine high-yield strains in marine bacteria with green chemistry.

Science.gov (United States)

Chen, Jun; Chen, Jianwei; Wang, Sijia; Zhou, Guangmin; Chen, Danqing; Zhang, Huawei; Wang, Hong

2018-04-02

A novel, green, rapid, and precise polar RP-HPLC method has been successfully developed and screened for ectoine high-yield strain in marine bacteria. Ectoine is a polar and extremely useful solute which allows microorganisms to survive in extreme environmental salinity. This paper describes a polar-HPLC method employed polar RP-C18 (5 μm, 250 × 4.6 mm) using pure water as the mobile phase and a column temperature of 30 °C, coupled with a flow rate at 1.0 mL/min and detected under a UV detector at wavelength of 210 nm. Our method validation demonstrates excellent linearity (R 2  = 0.9993), accuracy (100.55%), and a limit of detection LOQ and LOD of 0.372 and 0.123 μgmL -1 , respectively. These results clearly indicate that the developed polar RP-HPLC method for the separation and determination of ectoine is superior to earlier protocols.

Geometric phases in singlemode fiber lightguides and fiber ring interferometers

International Nuclear Information System (INIS)

Malykin, Grigorii B; Pozdnyakova, Vera I

2004-01-01

We consider various geometric phases (GPs) in singlemode fiber lightguides (SMFs) and in fiber ring interferometers (FRIs): the Pancharatnam phase stemming from the cyclic evolution of the polarization state of radiation (RP state) in SMF, the Rytov-Vladimirskii phase (RV phase) stemming from the Rytov effect (specifically, rotation of the polarization plane due to noncoplanar winding of SMFs), as well as the nonreciprocal phase difference of counterpropagating waves (NPDCW) and nonreciprocal geometric phase of counterpropagating waves (NGPCW), which are caused by polarization nonreciprocity (PN) in FRIs. We show that in the general case, the Pancharatnam phase for an arbitrary RP state is inconsistent with the real phase change of light fluctuations in media that possess not only circular but also linear birefringence. We show that the RV phase, having a geometric origin, can in principle be considered as a dynamic phase (DP). We also show that the NGPCW can be considered as an effect of the evolution of the RP state mapped on the Poincare sphere in Ginzburg's orthogonal screw polarization modes (GSPMs) of SMFs in the FRI contour. We analyze a number of experiments in which geometric phases were detected in FRIs: changing the RV phase and Rytov's angle (RA) in response to change of the pitch of helicoidal winding of SMFs. (methodological notes)

Gs-coupled GPCR signalling in AgRP neurons triggers sustained increase in food intake.

Science.gov (United States)

Nakajima, Ken-ichiro; Cui, Zhenzhong; Li, Chia; Meister, Jaroslawna; Cui, Yinghong; Fu, Ou; Smith, Adam S; Jain, Shalini; Lowell, Bradford B; Krashes, Michael J; Wess, Jürgen

2016-01-08

Agouti-related peptide (AgRP) neurons of the hypothalamus play a key role in regulating food intake and body weight, by releasing three different orexigenic molecules: AgRP; GABA; and neuropeptide Y. AgRP neurons express various G protein-coupled receptors (GPCRs) with different coupling properties, including Gs-linked GPCRs. At present, the potential role of Gs-coupled GPCRs in regulating the activity of AgRP neurons remains unknown. Here we show that the activation of Gs-coupled receptors expressed by AgRP neurons leads to a robust and sustained increase in food intake. We also provide detailed mechanistic data linking the stimulation of this class of receptors to the observed feeding phenotype. Moreover, we show that this pathway is clearly distinct from other GPCR signalling cascades that are operative in AgRP neurons. Our data suggest that drugs able to inhibit this signalling pathway may become useful for the treatment of obesity.

The severity of retinal degeneration in Rp1h gene-targeted mice is dependent on genetic background.

Science.gov (United States)

Liu, Qin; Saveliev, Alexei; Pierce, Eric A

2009-04-01

The severity of disease in patients with retinitis pigmentosa (RP) can vary significantly, even among patients with the same primary mutations. It is hypothesized that modifier genes play important roles in determining the severity of RP, including the retinitis pigmentosa 1 (RP1) form of disease. To investigate the basis of variation in disease expression for RP1 disease, the authors generated congenic mice with a gene-targeted retinitis pigmentosa 1 homolog (Rp1h) allele (Rp1h(tm1Eap)) on several different genetic backgrounds and analyzed their retinal phenotypes. The Rp1h(tm1Eap) allele was placed onto the C57BL/6J, DBA1/J, and A/J backgrounds. Retinal function of the resultant congenic mice was evaluated using electroretinographic analyses. Retinal structure and ultrastructure were evaluated using light and electron microscopy. Rp1h protein location was determined with immunofluorescence microscopy. Analysis of the retinal phenotype of incipient congenic (N6) B6.129S-Rp1h(+/tm1Eap), DBA.129S(B6)-Rp1h(+/tm1Eap), and A.129S(B6)-Rp1h(+/tm1Eap) mice at 1 year of age showed retinal degeneration only in the A.129S(B6)-Rp1h(+/tm1Eap) mice. Further analyses revealed that the photoreceptors of the fully congenic A.129S(B6)-Rp1h(+/tm1Eap) mice show evidence of degeneration at 6 months of age and are almost completely lost by 18 months of age. In contrast, the photoreceptor cells in the fully congenic B6.129S-Rp1h(+/tm1Eap) mice remain healthy up to 18 months. The severity of the retinal degeneration caused by the Rp1h(tm1Eap) allele is notably dependent on genetic background. The development and characterization of the B6.129S-Rp1h(+/tm1Eap) and A.129S(B6)-Rp1h(+/tm1Eap) congenic mouse lines will facilitate identification of sequence alterations in genes that modify the severity of RP1 disease.

Development and validation of RP-HPLC method for analysis of multicomponent cough-cold syrup formulation

OpenAIRE

Ivković, Branka; Marković, Bojan; Vladimirov, Sote

2014-01-01

In this study a reversed phase HPLC method for rapid and simultaneous identification and quantification of doxylamine succinate, ephedrine sulfate, dextrometorphane hydrobromide, paracetamole and sodium benzoate in cough-cold syrup formulation was described. Separation was carried out on XTerraTM RP 18, Waters (150 mm x 4.6 mm column, 5 μm particle size). For the analysis of investigated substances gradient elution was used employing water, pH adjusted at 2.5 with 85 % ortophosphoric acid as ...

Arcuate AgRP neurons and the regulation of energy balance

Directory of Open Access Journals (Sweden)

Céline eCansell

2012-12-01

Full Text Available The arcuate nucleus of the hypothalamus contains at least two crucial populations of neurons that continuously monitor signals reflecting energy status and promote the appropriate behavioral and metabolic responses to changes in energy demand. Neurons making pro-opiomelanocortin (POMC decrease food intake and increase energy expenditure through activation of G protein-coupled receptors melanocortin receptors (MCR via the release of a-melanocyte stimulating hormone. A prevailing idea until recently was that the neighboring neurons expressing the orexigenic neuropeptides, agouti-related protein (AgRP and neuropeptide Y (NPY (AgRP neurons increased feeding by opposing the anorexigenic actions of the POMC neurons. AgRP neurons activation but not POMC neurons inhibition was recently demonstrated to be necessary and sufficient to promote feeding. AgRP expressing axons were identified in mesolimbic, midbrain and pontine structure where they regulate feeding but also feeding-independent functions such as reward or peripheral nutrient partitioning. Post-synaptic Gamma aminobutyric acid (GABA, lasting in a timeline similar to neuromodulation, was identified as the core mechanism by which hunger-activated neurons regulate feeding and non-food related processes in a melanocortin independent manner.

TCPTP Regulates Insulin Signalling in AgRP Neurons to Coordinate Glucose Metabolism with Feeding.

Science.gov (United States)

Dodd, Garron T; Lee-Young, Robert S; Brüning, Jens C; Tiganis, Tony

2018-04-30

Insulin regulates glucose metabolism by eliciting effects on peripheral tissues as well as the brain. Insulin receptor (IR) signalling inhibits AgRP-expressing neurons in the hypothalamus to contribute to the suppression of hepatic glucose production (HGP) by insulin, whereas AgRP neuronal activation attenuates brown adipose tissue (BAT) glucose uptake. The tyrosine phosphatase TCPTP suppresses IR signalling in AgRP neurons. Hypothalamic TCPTP is induced by fasting and degraded after feeding. Here we assessed the influence of TCPTP in AgRP neurons in the control of glucose metabolism. TCPTP deletion in AgRP neurons ( Agrp -Cre; Ptpn2 fl/fl ) enhanced insulin sensitivity as assessed by the increased glucose infusion rates and reduced HGP during hyperinsulinemic-euglycemic clamps, accompanied by increased [ 14 C]-2-deoxy-D-glucose uptake in BAT and browned white adipose tissue. TCPTP deficiency in AgRP neurons promoted the intracerebroventricular insulin-induced repression of hepatic gluconeogenesis in otherwise unresponsive food-restricted mice yet had no effect in fed/satiated mice where hypothalamic TCPTP levels are reduced. The improvement in glucose homeostasis in Agrp -Cre; Ptpn2 fl/fl mice was corrected by IR heterozygosity ( Agrp -Cre; Ptpn2 fl/fl ; Insr fl/+ ), causally linking the effects on glucose metabolism with the IR signalling in AgRP neurons. Our findings demonstrate that TCPTP controls IR signalling in AgRP neurons to coordinate HGP and brown/beige adipocyte glucose uptake in response to feeding/fasting. © 2018 by the American Diabetes Association.

Determination of n-octanol/water partition coefficient for DDT-related compounds by RP-HPLC with a novel dual-point retention time correction.

Science.gov (United States)

Han, Shu-ying; Qiao, Jun-qin; Zhang, Yun-yang; Yang, Li-li; Lian, Hong-zhen; Ge, Xin; Chen, Hong-yuan

2011-03-01

n-Octanol/water partition coefficients (P) for DDTs and dicofol were determined by reversed-phase high performance liquid chromatography (RP-HPLC) on a C(18) column using methanol-water mixture as mobile phase. A dual-point retention time correction (DP-RTC) was proposed to rectify chromatographic retention time (t(R)) shift resulted from stationary phase aging. Based on this correction, the relationship between logP and logk(w), the logarithm of the retention factor extrapolated to pure water, was investigated for a set of 12 benzene homologues and DDT-related compounds with reliable experimental P as model compounds. A linear regression logP=(1.10±0.04) logk(w) - (0.60±0.17) was established with correlation coefficient R(2) of 0.988, cross-validated correlation coefficient R(cv)(2) of 0.983 and standard deviation (SD) of 0.156. This model was further validated using four verification compounds, naphthalene, biphenyl, 2,2-bis(4-chlorophenyl)-1,1-dichloroethane (p,p'-DDD) and 2,2-bis(4-chlorophenyl)-1,1-dichloroethene (p,p'-DDE) with similar structure to DDT. The RP-HPLC-determined P values showed good consistency with shake-flask (SFM) or slow-stirring (SSM) results, especially for highly hydrophobic compounds with logP in the range of 4-7. Then, the P values for five DDT-related compounds, 2-(2-chlorophenyl)-2-(4-chlorophenyl)-1,1,1-trichloroethane (o,p'-DDT), 2-(2-chlorophenyl)-2-(4-chlorophenyl)-1,1-dichloroethane (o,p'-DDD), 2-(2-chlorophenyl)-2-(4-chlorophenyl)-1,1-dichloroethene (o,p'-DDE), and 2,2,2-trichloro-1,1-bis(4-chlorophenyl)ethanol (dicofol) and its main degradation product 4,4'-dichlorobenzophenone (p,p'-DBP) were evaluated by the improved RP-HPLC method for the first time. The excellent precision with SD less than 0.03 proved that the novel DP-RTC protocol can significantly increases the determination accuracy and reliability of P by RP-HPLC. Copyright © 2011 Elsevier Ltd. All rights reserved.

Rapid Screening of Acetylcholinesterase Inhibitors by Effect-Directed Analysis Using LC × LC Fractionation, a High Throughput in Vitro Assay, and Parallel Identification by Time of Flight Mass Spectrometry.

Science.gov (United States)

Ouyang, Xiyu; Leonards, Pim E G; Tousova, Zuzana; Slobodnik, Jaroslav; de Boer, Jacob; Lamoree, Marja H

2016-02-16

Effect-directed analysis (EDA) is a useful tool to identify bioactive compounds in complex samples. However, identification in EDA is usually challenging, mainly due to limited separation power of the liquid chromatography based fractionation. In this study, comprehensive two-dimensional liquid chromatography (LC × LC) based microfractionation combined with parallel high resolution time of flight (HR-ToF) mass spectrometric detection and a high throughput acetylcholinesterase (AChE) assay was developed. The LC × LC fractionation method was validated using analytical standards and a C18 and pentafluorophenyl (PFP) stationary phase combination was selected for the two-dimensional separation and fractionation in four 96-well plates. The method was successfully applied to identify AChE inhibitors in a wastewater treatment plant (WWTP) effluent. Good orthogonality (>0.9) separation was achieved and three AChE inhibitors (tiapride, amisulpride, and lamotrigine), used as antipsychotic medicines, were identified and confirmed by two-dimensional retention alignment as well as their AChE inhibition activity.

RP-UHPLC-UV-ESI-MS/MS analysis of LPMO generated C4-oxidized gluco-oligosaccharides after non-reductive labeling with 2-aminobenzamide.

Science.gov (United States)

Frommhagen, Matthias; van Erven, Gijs; Sanders, Mark; van Berkel, Willem J H; Kabel, Mirjam A; Gruppen, Harry

2017-08-07

Lytic polysaccharide monooxygenases (LPMOs) are able to cleave recalcitrant polysaccharides, such as cellulose, by oxidizing the C1 and/or C4 atoms. The analysis of the resulting products requires a variety of analytical techniques. Up to now, these techniques mainly focused on the identification of non-oxidized and C1-oxidized oligosaccharides. The analysis of C4-oxidized gluco-oligosaccharides is mostly performed by using high pressure anion exchange chromatography (HPAEC). However, the alkaline conditions used during HPAEC analysis lead to tautomerization of C4-oxidized gluco-oligosaccharides, which limits the use of this technique. Here, we describe the use of reverse phase-ultra high performance liquid chromatography (RP-UHPLC) in combination with non-reductive 2-aminobenzamide (2-AB) labeling. Non-reductive 2-AB labeling enabled separation of C4-oxidized gluco-oligosaccharides from their non-oxidized counterparts. Moreover, RP-UHPLC does not require buffered mobile phases, which reduce mass spectrometry (MS) sensitivity. The latter is seen as an advantage over other techniques such as hydrophilic interaction liquid chromatography and porous graphitized carbon coupled to MS. RP-UHPLC coupled to UV detection and mass spectrometry allowed the identification of both labeled non-oxidized and C4-oxidized oligosaccharides. Non-reductive labeling kept the ketone at the C4-position of LPMO oxidized oligosaccharides intact, while selective reducing agents such as sodium triacetoxyborohydride (STAB) reduced this ketone group. Our results show that RP-UHPLC-UV-ESI-MS in combination with non-reductively 2-AB labeling is a suitable technique for the separation and identification of LPMO-generated C4-oxidized gluco-oligosaccharides. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Liquid-chromatography mass spectrometry (LC-MS) of steroid hormone metabolites and its applications

Science.gov (United States)

Penning, Trevor M.; Lee, Seon-Hwa; Jin, Yi; Gutierrez, Alejandro; Blair, Ian A.

2010-01-01

Advances in liquid chromatography-mass spectrometry (LC-MS) can be used to measure steroid hormone metabolites in vitro and in vivo. We find that LC-Electrospray Ionization (ESI)-MS using a LCQ ion trap mass spectrometer in the negative ion mode can be used to monitor the product profile that results from 5α–dihydrotestosterone(DHT)-17β-glucuronide, DHT-17β-sulfate, and tibolone-17β-sulfate reduction catalyzed by human members of the aldo-keto reductase (AKR) 1C subfamily and assign kinetic constants to these reactions. We also developed a stable-isotope dilution LC-electron capture atmospheric pressure chemical ionization (ECAPCI)-MS method for the quantitative analysis of estrone (E1) and its metabolites as pentafluorobenzyl (PFB) derivatives in human plasma in the attomole range. The limit of detection for E1-PFB was 740 attomole on column. Separations can be performed using normal-phase LC because ionization takes place in the gas phase rather than in solution. This permits efficient separation of the regioisomeric 2- and 4-methoxy-E1. The method was validated for the simultaneous analysis of plasma E2 and its metabolites: 2-methoxy-E2, 4-methoxy-E2, 16α-hydroxy-E2, estrone (E1), 2-methoxy-E1, 4-methoxy-EI, and 16α-hydroxy-E1 from 5 pg/mL to 2,000 pg/mL. Our LC-MS methods have sufficient sensitivity to detect steroid hormone levels in prostate and breast tumors and should aid their molecular diagnosis and treatment. PMID:20083198

X-ray phase radiography and tomography with grating interferometry and the reverse projection technique

International Nuclear Information System (INIS)

Wang, Zhili; Gao, Kun; Ge, Xin; Wu, Zhao; Chen, Heng; Wang, Shenghao; Wu, Ziyu; Zhu, Peiping; Yuan, Qingxi; Huang, Wanxia; Zhang, Kai

2013-01-01

X-ray grating interferometry provides substantially increased contrast over conventional absorption-based imaging methods, and therefore new and complementary information. Compared with other phase-contrast imaging techniques, x-ray grating interferometry can overcome some of the problems that have impaired the applications of x-ray phase-contrast radiography and phase tomography. Recently, special attention has been paid to the development of quantitative phase retrieval methods, which is mandatory to perform x-ray phase tomography, to achieve material identification, to differentiate distinct tissues, etc. Typically, the phase-stepping approach has been utilized for phase retrieval in grating interferometry. This method requires a grating scanning and acquisition of multiple radiographic projections, and therefore is disadvantageous in terms of imaging speed and radiation damage. Here we present an innovative, highly sensitive approach, dubbed ‘reverse projection’ (RP), for quantitative phase retrieval. Compared with the phase-stepping approach, the present RP method abandons grating scanning completely, and thus is advantageous due to its much higher efficiency and the reduced radiation dose, without the degradation of reconstruction quality. This review presents a detailed explanation of the principle of the RP method. Both radiography and phase tomography experiments are performed to validate the RP method. We believe that this new technique will find widespread applications in biomedical imaging and in vivo studies. (paper)

Determination of Phthalate Plasticisers in Palm Oil Using Online Solid Phase Extraction-Liquid Chromatography (SPE-LC

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Nazarudin Ibrahim

2014-01-01

Full Text Available Contamination of phthalates plasticisers to food has raised concern as some of the phthalates are suspected to be endocrine disruptors. The phthalates have high affinity with oily environment and analysing these chemicals in such matrices is difficult because of the trace amount of the analyte and interference from matrix. An online solid phase extraction (SPE technique using a large volume (3.5 mL injection was developed for the analysis of 6 common plasticisers in palm oil. A simple sample preparation involving alumina as a fat retainer and methanol : acetonitrile (1 : 1 as the extraction solvent was performed prior to the usage of online SPE-LC system. This system consists of two columns, C16 for the solid phase extraction (SPE and C18 as the analytical column, and a photo diode array detector. The calibration curves were linear from 5 to 1000 μg L−1, with correlation coefficients above 0.99. The instrumental limit of detection was 3 μg L−1 and satisfactory recovery was obtained. A screening on a few samples in the retail market revealed the presence of dibutyl phthalate (DBP and butylbenzylphthalate (BBP in the palm oil, with concentration less than 1 mg L−1.

Characterization of phytoplasmas related to 'Candidatus Phytoplasma asteris' subgroup rpI-L in Iran

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Vali-Sichani Fereshteh

2014-07-01

Full Text Available In two of Iran's central provinces, several herbaceous plants showing phytoplasma disease symptoms were collected to detect 'Canididatus Phytoplasma asteris'-related phytoplasmas. Confirmation of an association of phytoplasmas with diseased plants was done using polymerase chain reaction (PCR assays having the phytoplasma universal primer pairs P1/P7 followed by R16F2n/ R16R2 in nested PCR. Then, for detection of 'Ca. P. asteris', DNA samples were subjected to amplification of rp and tuf genes using specific primer pairs rp(IF1A/rp(IR1A and fTufAy/rTufAy, respectively. Restriction fragment length polymorphism or RFLP analyses of rp gene fragments using Tsp509I restriction enzyme as well as sequence analyses indicated that 'Ca. P. asteris'-related phytoplasmas associated with carrot, niger seed and scallion plants in these regions, belong to the rpI-L subgroup. This research is the first report of carrot, niger seed, and scallion infection with phytoplasmas belonging to the rpI-L subgroup.

Automated Solid Phase Extraction (SPE) LC/NMR Applied to the Structural Analysis of Extractable Compounds from a Pharmaceutical Packaging Material of Construction.

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Norwood, Daniel L; Mullis, James O; Davis, Mark; Pennino, Scott; Egert, Thomas; Gonnella, Nina C

2013-01-01

The structural analysis (i.e., identification) of organic chemical entities leached into drug product formulations has traditionally been accomplished with techniques involving the combination of chromatography with mass spectrometry. These include gas chromatography/mass spectrometry (GC/MS) for volatile and semi-volatile compounds, and various forms of liquid chromatography/mass spectrometry (LC/MS or HPLC/MS) for semi-volatile and relatively non-volatile compounds. GC/MS and LC/MS techniques are complementary for structural analysis of leachables and potentially leachable organic compounds produced via laboratory extraction of pharmaceutical container closure/delivery system components and corresponding materials of construction. Both hyphenated analytical techniques possess the separating capability, compound specific detection attributes, and sensitivity required to effectively analyze complex mixtures of trace level organic compounds. However, hyphenated techniques based on mass spectrometry are limited by the inability to determine complete bond connectivity, the inability to distinguish between many types of structural isomers, and the inability to unambiguously determine aromatic substitution patterns. Nuclear magnetic resonance spectroscopy (NMR) does not have these limitations; hence it can serve as a complement to mass spectrometry. However, NMR technology is inherently insensitive and its ability to interface with chromatography has been historically challenging. This article describes the application of NMR coupled with liquid chromatography and automated solid phase extraction (SPE-LC/NMR) to the structural analysis of extractable organic compounds from a pharmaceutical packaging material of construction. The SPE-LC/NMR technology combined with micro-cryoprobe technology afforded the sensitivity and sample mass required for full structure elucidation. Optimization of the SPE-LC/NMR analytical method was achieved using a series of model compounds

Development and Validation of a UV Spectrophotometric and a RP-HPLC Methods for Moexipril Hydrochloride in Pure Form and Pharmaceutical Dosage Form

International Nuclear Information System (INIS)

Mastiholimath, V.S.; Gupte, P.P.; Mannur, V.S.

2012-01-01

A simple and reliable UV spectrophotometric and high-performance liquid chromatography (HPLC) methods were developed and validated for Moexipril hydrochloride in pure form and pharmaceutical dosage form. The RP-HPLC method was developed on agilant eclipse C 18 , (150 mm x 4.6 mm, 5 μm) with a mobile phase gradient system of 60 % (methanol:acetonitrile (70:30 % v/v)) : 40 % 20 mM ammonium acetate buffer pH 4.5 (v/v) and UV spectrophotometric method was developed in phosphate buffer pH 6.8. The effluent was monitored by SPD-M20A, prominence PDA detector at 210 nm. Calibration curve was linear over the concentration range of 10-35 μg/ml and 1-9 μg/ml for RP-HPLC and UV with a regression coefficient of 0.999. For RP-HPLC method Inter-day and intra-day precision % RSD values were found to be 1.00078 % and 1.49408 % respectively. For UV method 0.73386 % to 1.44111 % for inter day 0.453864 to 1.15542 intra-day precision. Recovery of Moexipril hydrochloride was found to be in the range of 99.8538 % to 101.5614 % and 100.5297586 % to 100.6431587 % for UV and RP-HPLC respectively. The limits of detection (LOD) and quantification (LOQ) for HPLC were 0.98969 and 2.99907 μg/ml, respectively. The developed RP-HPLC and UV spectrophotometric method was successfully applied for the quantitative determination of Moexipril hydrochloride in pharmaceutical dosage. (author)

Simplified RP-HPLC method for multi-residue analysis of abamectin, emamectin benzoate and ivermectin in rice.

Science.gov (United States)

Xie, Xianchuan; Gong, Shu; Wang, Xiaorong; Wu, Yinxing; Zhao, Li

2011-01-01

A rapid, reliable and sensitive reverse-phase high-performance liquid chromatography method with fluorescence detection (RP-FLD-HPLC) was developed and validated for simultaneous analysis of the abamectin (ABA), emamectin (EMA) benzoate and ivermectin (IVM) residues in rice. After extraction with acetonitrile/water (2 : 1) with sonication, the avermectin (AVMs) residues were directly derivatised by N-methylimidazole (N-NMIM) and trifluoroacetic anhydride (TFAA) and then analysed on RP-FLD-HPLC. A good linear relationship (r(2 )> 0.99) was obtained for three AVMs ranging from 0.01 to 5 microg ml(-1), i.e. 0.01-5.0 microg g(-1) in rice matrix. The limit of detection (LOD) and the limit of quantification (LOQ) were between 0.001 and 0.002 microg g(-1) and between 0.004 and 0.006 microg g(-1), respectively. Recoveries were from 81.9% to 105.4% and precision less than 12.4%. The proposed method was successfully applied to routine analysis of the AVMs residues in rice.

Arcuate hypothalamic AgRP and putative POMC neurons show opposite changes in spiking across multiple timescales

Science.gov (United States)

Mandelblat-Cerf, Yael; Ramesh, Rohan N; Burgess, Christian R; Patella, Paola; Yang, Zongfang; Lowell, Bradford B; Andermann, Mark L

2015-01-01

Agouti-related-peptide (AgRP) neurons—interoceptive neurons in the arcuate nucleus of the hypothalamus (ARC)—are both necessary and sufficient for driving feeding behavior. To better understand the functional roles of AgRP neurons, we performed optetrode electrophysiological recordings from AgRP neurons in awake, behaving AgRP-IRES-Cre mice. In free-feeding mice, we observed a fivefold increase in AgRP neuron firing with mounting caloric deficit in afternoon vs morning recordings. In food-restricted mice, as food became available, AgRP neuron firing dropped, yet remained elevated as compared to firing in sated mice. The rapid drop in spiking activity of AgRP neurons at meal onset may reflect a termination of the drive to find food, while residual, persistent spiking may reflect a sustained drive to consume food. Moreover, nearby neurons inhibited by AgRP neuron photostimulation, likely including satiety-promoting pro-opiomelanocortin (POMC) neurons, demonstrated opposite changes in spiking. Finally, firing of ARC neurons was also rapidly modulated within seconds of individual licks for liquid food. These findings suggest novel roles for antagonistic AgRP and POMC neurons in the regulation of feeding behaviors across multiple timescales. DOI: http://dx.doi.org/10.7554/eLife.07122.001 PMID:26159614

Retention prediction of highly polar ionizable solutes under gradient conditions on a mixed-mode reversed-phase and weak anion-exchange stationary phase.

Science.gov (United States)

Balkatzopoulou, P; Fasoula, S; Gika, H; Nikitas, P; Pappa-Louisi, A

2015-05-29

In the present work the retention of three highly polar and ionizable solutes - uric acid, nicotinic acid and ascorbic acid - was investigated on a mixed-mode reversed-phase and weak anion-exchange (RP/WAX) stationary phase in buffered aqueous acetonitrile (ACN) mobile phases. A U-shaped retention behavior was observed for all solutes with respect to the eluent organic modifier content studied in a range of 5-95% (v/v). This retention behavior clearly demonstrates the presence of a HILIC-type retention mechanism at ACN-rich hydro-organic eluents and an RP-like retention at aqueous-rich hydro-organic eluents. Hence, this column should be promising for application under both RP and HILIC gradient elution modes. For this reason, a series of programmed elution runs were carried out with increasing (RP) and decreasing (HILIC) organic solvent concentration in the mobile phase. This dual gradient process was successfully modeled by two retention models exhibiting a quadratic or a cubic dependence of the logarithm of the solute retention factor (lnk) upon the organic modifier volume fraction (φ). It was found that both models produced by gradient retention data allow the prediction of solute retention times for both types of programmed elution on the mixed-mode column. Four, in the case of the quadratic model, or five, in the case of the cubic model, initial HILIC- and RP-type gradient runs gave satisfactory retention predictions of any similar kind elution program, even with different flow rate, with an overall error of only 2.5 or 1.7%, respectively. Copyright © 2015 Elsevier B.V. All rights reserved.

LC-MS n Analysis of Isomeric Chondroitin Sulfate Oligosaccharides Using a Chemical Derivatization Strategy

Science.gov (United States)

Huang, Rongrong; Pomin, Vitor H.; Sharp, Joshua S.

2011-09-01

Improved methods for structural analyses of glycosaminoglycans (GAGs) are required to understand their functional roles in various biological processes. Major challenges in structural characterization of complex GAG oligosaccharides using liquid chromatography-mass spectrometry (LC-MS) include the accurate determination of the patterns of sulfation due to gas-phase losses of the sulfate groups upon collisional activation and inefficient on-line separation of positional sulfation isomers prior to MS/MS analyses. Here, a sequential chemical derivatization procedure including permethylation, desulfation, and acetylation was demonstrated to enable both on-line LC separation of isomeric mixtures of chondroitin sulfate (CS) oligosaccharides and accurate determination of sites of sulfation by MS n . The derivatized oligosaccharides have sulfate groups replaced with acetyl groups, which are sufficiently stable to survive MS n fragmentation and reflect the original sulfation patterns. A standard reversed-phase LC-MS system with a capillary C18 column was used for separation, and MS n experiments using collision-induced dissociation (CID) were performed. Our results indicate that the combination of this derivatization strategy and MS n methodology enables accurate identification of the sulfation isomers of CS hexasaccharides with either saturated or unsaturated nonreducing ends. Moreover, derivatized CS hexasaccharide isomer mixtures become separable by LC-MS method due to different positions of acetyl modifications.

Simultaneous liquid chromatography/mass spectrometry determination of both polar and "multiresidue" pesticides in food using parallel hydrophilic interaction/reversed-phase liquid chromatography and a hybrid sample preparation approach.

Science.gov (United States)

Robles-Molina, José; Gilbert-López, Bienvenida; García-Reyes, Juan F; Molina-Díaz, Antonio

2017-09-29

Pesticide testing of foodstuffs is usually accomplished with generic wide-scope multi-residue methods based on liquid chromatography tandem mass spectrometry (LC-MS/MS). However, this approach does not cover some special pesticides, the so called "single-residue method" compounds, that are hardly compatible with standard reversed-phase (RP) separations due to their specific properties. In this article, we propose a comprehensive strategy for the integration of single residue method compounds and standard multiresidue pesticides within a single run. It is based on the use of a parallel LC column assembly with two different LC gradients performing orthogonal hydrophilic interaction chromatography (HILIC) and reversed-phase (RPLC) chromatography within one analytical run. Two sample aliquots were simultaneously injected on each column, using different gradients, being the eluents merged post-column prior to mass spectrometry detection. The approach was tested with 41 multiclass pesticides covering a wide range of physicochemical properties across several orders of log K ow (from -4 to +5.5). With this assembly, distinct separation from the void was attained for all the pesticides studied, keeping similar performance in terms of sensitivity, peak area reproducibility (pesticides) method was evaluated based on solvent extraction with MeOH and acetonitrile followed by dispersive solid-phase extraction, delivering appropriate recoveries for most of the pesticides included in the study within the log K ow in the range from -4 to +5.5. The proposed strategy may be extended to other fields such as sport drug testing or environmental analysis, where the same type of variety of analytes featuring poor retention within a single chromatographic separation occurs. Copyright © 2017 Elsevier B.V. All rights reserved.

PGC-1α expression in murine AgRP neurons regulates food intake and energy balance

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Jonathan F. Gill

2016-07-01

Full Text Available Objective: Food intake and whole-body energy homeostasis are controlled by agouti-related protein (AgRP and pro-opiomelanocortin (POMC neurons located in the arcuate nucleus of the hypothalamus. Key energy sensors, such as the AMP-activated protein kinase (AMPK or sirtuin 1 (SIRT1, are essential in AgRP and POMC cells to ensure proper energy balance. In peripheral tissues, the transcriptional coactivator PGC-1α closely associates with these sensors to regulate cellular metabolism. The role of PGC-1α in the ARC nucleus, however, remains unknown. Methods: Using AgRP and POMC neurons specific knockout (KO mouse models we studied the consequences of PGC-1α deletion on metabolic parameters during fed and fasted states and on ghrelin and leptin responses. We also took advantage of an immortalized AgRP cell line to assess the impact of PGC-1α modulation on fasting induced AgRP expression. Results: PGC-1α is dispensable for POMC functions in both fed and fasted states. In stark contrast, mice carrying a specific deletion of PGC-1α in AgRP neurons display increased adiposity concomitant with significantly lower body temperature and RER values during nighttime. In addition, the absence of PGC-1α in AgRP neurons reduces food intake in the fed and fasted states and alters the response to leptin. Finally, both in vivo and in an immortalized AgRP cell line, PGC-1α modulates AgRP expression induction upon fasting. Conclusions: Collectively, our results highlight a role for PGC-1α in the regulation of AgRP neuronal functions in the control of food intake and peripheral metabolism. Author Video: Author Video Watch what authors say about their articles Keywords: PGC-1α, Agouti-related protein, Metabolism, Energy homeostasis, Pro-opiomelanocortin, Transcriptional regulation

Argentine Radiation Protection Society Experience in RP education and training

International Nuclear Information System (INIS)

Bomben, A. M.; Ciallella, N. R.; Thomasz, E.; Rudelli, M.; Gisone, P.; Ventura, M.; Gomez Parada, I.; Signoretta, C.

2003-01-01

Since its creation in 1967, the Argentine Radiation Protection Society (SAR) promotes all the radiation protection and nuclear safety aspects not only within the scientific, technical and academic areas, but also to general public. To fulfill this objective, SAR organised training and refresher courses, seminars and workshops on RP subjects. During 2002, SAR organised 7 basic and specialized courses regarding the uses of radioactive materials in industrial applications and the course on medical response in radiological accidents, that was attended by Argentine and other Latin American participants. The programmes of the courses are developed in compliance with the legal requirements and also considering specifics needs. In this paper, the characteristics of the courses are enunciated and basic statistics regarding courses and participants are presented. For the 2003 and 2004, SAR foresees the organisation of 18 courses per year and has the capacity to deliver other courses by request. all the courses are delivered in Spanish language. Based on this educational experience SAR consider a priority the inclusion, of a RP module in all the scientific graduate programmes to generate awareness on the importance of RP. Taking into account the migration of professionals to Europe and North America and the Globalization, SAR advocates the harmonization of RP syllabus to attain an international recognition. (Author)

Development and validation of a highly sensitive LC-ESI-MS/MS method for estimation of IIIM-MCD-211, a novel nitrofuranyl methyl piperazine derivative with potential activity against tuberculosis: Application to drug development.

Science.gov (United States)

Magotra, Asmita; Sharma, Anjna; Gupta, Ajai Prakash; Wazir, Priya; Sharma, Shweta; Singh, Parvinder Pal; Tikoo, Manoj Kumar; Vishwakarma, Ram A; Singh, Gurdarshan; Nandi, Utpal

2017-08-15

In the present study, a simple, sensitive, specific and rapid liquid chromatography (LC) tandem mass spectrometry (MS/MS) method was developed and validated according to the Food and Drug Administration (FDA) guidelines for estimation of IIIM-MCD-211 (a potent oral candidate with promising action against tuberculosis) in mice plasma using carbamazepine as internal standard (IS). Bioanalytical method consisted of one step protein precipitation for sample preparation followed by quantitation in LC-MS/MS using positive electrospray ionization technique (ESI) operating in multiple reaction monitoring (MRM) mode. Elution was achieved in gradient mode on High Resolution Chromolith RP-18e column with mobile phase comprised of acetonitrile and 0.1% (v/v) formic acid in water at the flow rate of 0.4mL/min. Precursor to product ion transitions (m/z 344.5/218.4 and m/z 237.3/194.2) were used to measure analyte and IS, respectively. All validation parameters were well within the limit of acceptance criteria. The method was successfully applied to assess the pharmacokinetics of the candidate in mice following oral (10mg/kg) and intravenous (IV; 2.5mg/kg) administration. It was also effectively used to quantitate metabolic stability of the compound in mouse liver microsomes (MLM) and human liver microsomes (HLM) followed by its in-vitro-in-vivo extrapolation. Copyright © 2017 Elsevier B.V. All rights reserved.

The Determination of Food Dyes in Vitamins by RP-HPLC

Directory of Open Access Journals (Sweden)

Monika Šuleková

2016-10-01

Full Text Available Reversed-phase high performance liquid chromatography (RP-HPLC for the determination of five synthetic food dyes (Quinoline Yellow E104, Sunset Yellow E110, Ponceau 4R E124, Tartrazine E102 and Carmine E120 in vitamins was used. The dyes were analyzed within 10 min using a column with stationary phase C 18 (250 mm × 4.6 mm, 5 μm at 40 °C with isocratic elution, and the mobile phase contained acetonitrile and a mixture of CH3COONa:CH3OH (85:15, v/v in a ratio of 10:90 (v/v for yellow-colored capsules and 20:80 (v/v for red-colored capsules, respectively. A diode-array detector was used to monitor the dyes between 190 and 800 nm. It was established that the analyzed samples contained synthetic dyes in a concentration range from 79.5 ± 0.01 μg/capsule of Ponceau 4R, E124 to 524 ± 0.01 μg/capsule of Tartrazine, E102. The obtained results were compared with existing acceptable daily intakes (ADIs for individual dyes. This paper provides information about the content of dyes in samples of vitamins. This information is not generally available to consumers.

Preparative separation and structural identification of impurities of a new α{sub 2}-Adrenoceptor agonist using stacking injection, LC-MS{sup n} and LC-SPE-NMR

Energy Technology Data Exchange (ETDEWEB)

Cardoso, Josiane O.; Thomasi, Sérgio S.; Venâncio, Tiago; Oliveira, Regina V., E-mail: oliveirarv@ufscar.br [Universidade Federal de São Carlos (UFSCar), SP (Brazil). Departamento de Química; Pitta, Ivan R.; Lima, Maria do Carmo A. de [Universidade Federal de Pernambuco (NUPIT/UFPE), Recife, PE (Brazil). Núcleo de Pesquisa em Inovação Tecnológica

2017-07-01

Identifying impurities in drug substances has become one of the most important issues in pharmaceutical analysis since it can have a significant impact on the efficacy of new pharmaceutical products. Due to the purity requirements, in this paper a new synthetic α{sub 2}-adrenoceptor agonist, called LPSF-PT-31, was purified and its impurities were characterized by liquid chromatography multistage mass spectrometry (LC-MS{sup n}) and liquid chromatography-solid phase extraction-nuclear magnetic resonance (LC-SPE-NMR). The purification step was conducted using a semi-preparative liquid chromatography and stacked injections as a new approach to drug purification. As a result, a total yield of 75% of the pure LPSF-PT-31 and 2.9 L day{sup -}'1 in solvent reduction was obtained. The combination of semi-preparative stacking injection, LC-MS{sup n}, and LC-SPE-NMR, demonstrated to be efficient to purify active drugs and unambiguously identify its impurities. In addition, isolation and identification of drug impurities in the early stages of development can improve the synthetic pathway, preventing the formation of impurities or minimizing this formation to minimum levels. (author)

Collagen films with stabilized liquid crystalline phases and concerns on osteoblast behaviors

Energy Technology Data Exchange (ETDEWEB)

Tang, Minjian; Ding, Shan; Min, Xiang; Jiao, Yanpeng, E-mail: tjiaoyp@jnu.edu.cn; Li, Lihua; Li, Hong; Zhou, Changren, E-mail: tcrz9@jnu.edu.cn

2016-01-01

To duplicate collagen's in vivo liquid crystalline (LC) phase and investigate the relationship between the morphology of LC collagen and osteoblast behavior, a self-assembly method was introduced for preparing collagen films with a stabilized LC phase. The LC texture and topological structure of the films before and after stabilization were observed with polarizing optical microscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM). The relationship between the collagen films and osteoblast behavior was studied with the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide method, proliferation index detection, alkaline phosphatase measurements, osteocalcin assay, inverted microscopy, SEM observation, AFM observation, and cytoskeleton fluorescence staining. The results showed that the LC collagen film had continuously twisting orientations in the cholesteric phase with a typical series of arced patterns. The collagen fibers assembled in a well-organized orientation in the LC film. Compared to the non-LC film, the LC collagen film can promote cell proliferation, and increase ALP and osteocalcin expression, revealing a contact guide effect on osteoblasts. - Highlights: • Collagen film with liquid crystalline (LC) phase was observed by POM, SEM and AFM. • The effect of LC collagen film on osteoblasts behaviors was studied in detail. • LC collagen film promoted osteoblast proliferation and osteogenesis activity.

A Small Potassium Current in AgRP/NPY Neurons Regulates Feeding Behavior and Energy Metabolism.

Science.gov (United States)

He, Yanlin; Shu, Gang; Yang, Yongjie; Xu, Pingwen; Xia, Yan; Wang, Chunmei; Saito, Kenji; Hinton, Antentor; Yan, Xiaofeng; Liu, Chen; Wu, Qi; Tong, Qingchun; Xu, Yong

2016-11-08

Neurons that co-express agouti-related peptide (AgRP) and neuropeptide Y (NPY) are indispensable for normal feeding behavior. Firing activities of AgRP/NPY neurons are dynamically regulated by energy status and coordinate appropriate feeding behavior to meet nutritional demands. However, intrinsic mechanisms that regulate AgRP/NPY neural activities during the fed-to-fasted transition are not fully understood. We found that AgRP/NPY neurons in satiated mice express high levels of the small-conductance calcium-activated potassium channel 3 (SK3) and are inhibited by SK3-mediated potassium currents; on the other hand, food deprivation suppresses SK3 expression in AgRP/NPY neurons, and the decreased SK3-mediated currents contribute to fasting-induced activation of these neurons. Genetic mutation of SK3 specifically in AgRP/NPY neurons leads to increased sensitivity to diet-induced obesity, associated with chronic hyperphagia and decreased energy expenditure. Our results identify SK3 as a key intrinsic mediator that coordinates nutritional status with AgRP/NPY neural activities and animals' feeding behavior and energy metabolism. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

New technologies applied to radiation protection training: Rp course for technical qualified expert

International Nuclear Information System (INIS)

Llorente Herranz, Cristina; Rodriguez, M.; Marco Arboli, Marisa

2008-01-01

This paper shows the development and results of the first Course for technical qualified expert in radiation protection (RP) using the advantages of Communication and Information Technologies (CITs). This project in modality b-learning has born as a result of the RP specialised course demand, the necessity of working-private life balance and working-training time combination as well as the geographic widespread of those interested. The methodology selected has been b-learning (blended learning), which consists of a mixture of e-learning plus face-to-face learning. The RP course for technical qualified RP expert has been designed for 10 weeks using online learning methodology and 2 days of face-to-face learning in a radioactive facility to take account practical sessions and the final knowledge evaluation. A multidisciplinary team of experts has elaborated the RP programme and the docent material, following national normative. The material consists of: a) Multimedia material with theoretical content to be visualised online; b) Additional material to be un-load or printed; c) Practical exercises within multimedia content; d) Practical online and face-to-face sessions. (author)

76 FR 35006 - Recovery Policy RP9523.4, Demolition of Private Structures

Science.gov (United States)

2011-06-15

...] Recovery Policy RP9523.4, Demolition of Private Structures AGENCY: Federal Emergency Management Agency, DHS... (FEMA) is accepting comments on Recovery Policy RP9523.4, Demolition of Private Structures. DATES... guidance in determining the eligibility of demolition of private structures under the provisions of the...

75 FR 67165 - Proposed Collection; Comment Request for Revenue Procedure2007-XX (RP-155430-05)

Science.gov (United States)

2010-11-01

... Revenue Procedure 2007- XX (RP-155430-05) AGENCY: Internal Revenue Service (IRS), Treasury. ACTION: Notice... soliciting comments concerning Revenue Procedure 2007-XX (RP-155430-05), Section 6707/6707A Accelerated... Procedure 2007-XX (RP-155430-05). Abstract: The collection of information this revenue procedure requires is...

Increasing reconstruction quality of diffractive optical elements displayed with LC SLM

Science.gov (United States)

Cheremkhin, Pavel A.; Evtikhiev, Nikolay N.; Krasnov, Vitaly V.; Rodin, Vladislav G.; Starikov, Sergey N.

2015-03-01

Phase liquid crystal (LC) spatial light modulators (SLM) are actively used in various applications. However, majority of scientific applications require stable phase modulation which might be hard to achieve with commercially available SLM due to its consumer origin. The use of digital voltage addressing scheme leads to phase temporal fluctuations, which results in lower diffraction efficiency and reconstruction quality of displayed diffractive optical elements (DOE). Due to high periodicity of fluctuations it should be possible to use knowledge of these fluctuations during DOE synthesis to minimize negative effect. We synthesized DOE using accurately measured phase fluctuations of phase LC SLM "HoloEye PLUTO VIS" to minimize its negative impact on displayed DOE reconstruction. Synthesis was conducted with versatile direct search with random trajectory (DSRT) method in the following way. Before DOE synthesis begun, two-dimensional dependency of SLM phase shift on addressed signal level and time from frame start was obtained. Then synthesis begins. First, initial phase distribution is created. Second, random trajectory of consecutive processing of all DOE elements is generated. Then iterative process begins. Each DOE element sequentially has its value changed to one that provides better value of objective criterion, e.g. lower deviation of reconstructed image from original one. If current element value provides best objective criterion value then it left unchanged. After all elements are processed, iteration repeats until stagnation is reached. It is demonstrated that application of SLM phase fluctuations knowledge in DOE synthesis with DSRT method leads to noticeable increase of DOE reconstruction quality.

Accurate LC peak boundary detection for ¹⁶O/¹⁸O labeled LC-MS data.

Science.gov (United States)

Cui, Jian; Petritis, Konstantinos; Tegeler, Tony; Petritis, Brianne; Ma, Xuepo; Jin, Yufang; Gao, Shou-Jiang S J; Zhang, Jianqiu Michelle

2013-01-01

In liquid chromatography-mass spectrometry (LC-MS), parts of LC peaks are often corrupted by their co-eluting peptides, which results in increased quantification variance. In this paper, we propose to apply accurate LC peak boundary detection to remove the corrupted part of LC peaks. Accurate LC peak boundary detection is achieved by checking the consistency of intensity patterns within peptide elution time ranges. In addition, we remove peptides with erroneous mass assignment through model fitness check, which compares observed intensity patterns to theoretically constructed ones. The proposed algorithm can significantly improve the accuracy and precision of peptide ratio measurements.

Separation and analysis of phenolic acids from Salvia miltiorrhiza and its related preparations by off-line two-dimensional hydrophilic interaction chromatography×reversed-phase liquid chromatography coupled with ion trap time-of-flight mass spectrometry.

Science.gov (United States)

Sun, Wanyang; Tong, Ling; Miao, Jingzhuo; Huang, Jingyi; Li, Dongxiang; Li, Yunfei; Xiao, Hongting; Sun, Henry; Bi, Kaishun

2016-01-29

Salvia miltiorrhiza (SM) is one of the most widely used Traditional Chinese Medicine. Active constituents of SM mainly contain hydrophilic phenolic acids (PAs) and lipophilic tanshinones. However, due to the existing of multiple ester bonds and unsaturated bonds in the structures, PAs have numerous chemical conversion products. Many of them are so low-abundant that hard to be separated using conventional methods. In this study, an off-line two-dimensional liquid chromatography (2D-LC) method was developed to separate PAs in SM and its related preparations. In the first dimension, samples were fractionated by hydrophilic interaction chromatography (HILIC) (Acchrom×Amide, 4.6×250mm, 5μm) mainly based on the hydrogen bonding effects. The fractions were then separated on reversed-phase liquid chromatography (RP-LC) (Acquity HSS T3, 2.1×50mm, 1.7μm) according to hydrophobicity. For the selective identification of PAs, diode array detector (DAD) and electrospray ionization tandem ion trap time-of-flight mass spectrometry (ESI-IT-TOF-MS) were employed. Practical and effective peak capacities of all the samples were greater than 2046 and 1130, respectively, with the orthogonalities ranged from 69.7% to 92.8%, which indicated the high efficiency and versatility of this method. By utilizing the data post-processing techniques, including mass defect filter, neutral loss filter and product ion filter, a total of 265 compounds comprising 196 potentially new PAs were tentatively characterized. Twelve kinds of derivatives, mainly including glycosylated compounds, O-alkylated compounds, condensed compounds and hydrolyzed compounds, constituted the novelty of the newly identified PAs. The HILIC×RP-LC/TOF-MS system expanded our understanding on PAs of S. miltiorrhiza and its related preparations, which could also benefit the separation and characterization of polar constituents in complicated herbal extracts. Copyright © 2016. Published by Elsevier B.V.

75 FR 49508 - Recovery Policy, RP9525.7, Labor Costs-Emergency Work

Science.gov (United States)

2010-08-13

...] Recovery Policy, RP9525.7, Labor Costs--Emergency Work AGENCY: Federal Emergency Management Agency, DHS... (FEMA) is accepting comments on RP9525.7, Labor Costs--Emergency Work. This is an existing policy that is scheduled for review to ensure that Recovery Directorate policies are up to date, incorporate...

Supercritical Fluid Chromatography- A Hybrid of GC and LC

Directory of Open Access Journals (Sweden)

Kaushal K Chandrul

2010-03-01

Full Text Available

High performance specifications and unique functionality of chromatographic techniques is a demand of pharmaceutical industry and research. This leads to the origin of Supercritical Fluid Chromatography (SFC. It is a rapidly expanding analytical technique. The main feature that differentiates SFC from other chromatographic techniques is the replacement of either the liquid or gas mobile phase with a supercritical fluid mobile phase. It is considered a hybrid of GC and LC technique. High diffusion coefficient and low viscosity of supercritical fluids is responsible for high speed analysis, high efficiency and high sensitivity. Low mobile-phase flow rate, density programming and compatability with GC and LC detectors make SFC a versatile chromatographic technique in analytical research and development. It has a unique characteristic of analyzing thermo labile or non-volatile substances. This review highlights the role of supercritical fluid chromatography in the separation of polymers, thermally labile pesticides, fatty acids, metal chelates and organometallic compounds, chiral and achiral molecules, identification and analysis of polar samples, explosives, drugs of abuse and application of SFC in forensic science (fingerprinting. 

LC-MS analysis of the plasma metabolome–a novel sample preparation strategy

DEFF Research Database (Denmark)

Skov, Kasper; Hadrup, Niels; Smedsgaard, Jørn

2015-01-01

Blood plasma is a well-known body fluid often analyzed in studies on the effects of toxic compounds as physiological or chemical induced changes in the mammalian body are reflected in the plasma metabolome. Sample preparation prior to LC-MS based analysis of the plasma metabolome is a challenge...... as plasma contains compounds with very different properties. Besides, proteins, which usually are precipitated with organic solvent, phospholipids, are known to cause ion suppression in electrospray mass spectrometry. We have compared two different sample preparation techniques prior to LC-qTOF analysis...... of plasma samples: The first is protein precipitation; the second is protein precipitation followed by solid phase extraction with sub-fractionation into three sub-samples; a phospholipid, a lipid and a polar sub-fraction. Molecular feature extraction of the data files from LC-qTOF analysis of the samples...

Solid-phase extraction based on hydrophilic interaction liquid chromatography with acetone as eluent for eliminating matrix effects in the analysis of biological fluids by LC-MS.

Science.gov (United States)

Van Damme, T; Lachová, M; Lynen, F; Szucs, R; Sandra, P

2014-01-01

Analysis of drugs and metabolites in biological matrices such as blood or plasma by LC-MS is routinely challenged by the presence of large quantities of competing molecules for ionization in soft ionization sources, such as proteins and phospholipids. While the former can easily be removed by protein precipitation, pre-analytical extraction of the latter is necessary because they show very high retention in reversed-phase LC resulting in long analysis times or in ion suppression effects when not eluted before the next runs. A novel HILIC-based SPE approach, making use of silica cartridges and of acetone as organic solvent, is introduced as a potent alternative to current commercial methods for phospholipid removal. The methodology was developed and tested for a broad polarity range of pharmaceutical solutes (log P from 0 to 6.6) and broad applicability can therefore be envisaged.

Neutronic calculation of safety parameters for the RP-0 and RP-10 nuclear reactors

OpenAIRE

Lázaro, Gerardo; Deen, James R.; Woodruff, William L.

2002-01-01

Theoretical safety calculations were done with proved codes utilized by the staff of the RERTR program in the HEU to LEU core conversions. The studies were designed to evaluate the reactivity coefficients and kinetics parameters of the reactor involved in the evolution of peak power transients by reactivity insertion accidents. It was done to show the trend of these reactivity coefficients as a function of the core size and fuel depletion for RP10 cores. It was useful to get a better underst...

Determination of free formaldehyde in cosmetics containing formaldehyde-releasing preservatives by reversed-phase dispersive liquid-liquid microextraction and liquid chromatography with post-column derivatization.

Science.gov (United States)

Miralles, Pablo; Chisvert, Alberto; Alonso, M José; Hernandorena, Sandra; Salvador, Amparo

2018-03-30

An analytical method for the determination of traces of formaldehyde in cosmetic products containing formaldehyde-releasing preservatives has been developed. The method is based on reversed-phase dispersive liquid-liquid microextraction (RP-DLLME), that allows the extraction of highly polar compounds, followed by liquid chromatography-ultraviolet/visible (LC-UV/vis) determination with post-column derivatization. The variables involved in the RP-DLLME process were studied to provide the best enrichment factors. Under the selected conditions, a mixture of 500 μL of acetonitrile (disperser solvent) and 50 μL of water (extraction solvent) was rapidly injected into 5 mL of toluene sample solution. The extracts were injected into the LC-UV/vis system using phosphate buffer 6 mmol L -1 at pH 2 as mobile phase. After chromatographic separation, the eluate merged with a flow stream of pentane-2,4-dione in ammonium acetate solution as derivatizing reagent and passed throughout a post-column reactor at 85 °C in order to derivatize formaldehyde into 3,5-diacetyl-1,4-dihydrolutidine, according to Hantzsch reaction, which was finally measured spectrophotometrically at 407 nm. The method was successfully validated showing good linearity, an enrichment factor of 86 ± 2, limits of detection and quantification of 0.7 and 2.3 ng mL -1 , respectively, and good repeatability (RSD < 9.2%). Finally, the proposed analytical method was applied to the determination of formaldehyde in different commercial cosmetic samples containing formaldehyde-releasing preservatives, such as bronopol, diazolidinyl urea, imidazolidinyl urea, and DMDM hydantoin, with good relative recovery values (91-113%) thus showing that matrix effects were negligible. The good analytical features of the proposed method besides of its simplicity and affordability, make it useful to carry out the quality control of cosmetic products containing formaldehyde-releasing preservatives. Copyright �

RSAC 6.2 with WinRP 2.0 User Manual

Energy Technology Data Exchange (ETDEWEB)

Bradley Schrader

2005-09-01

The Radiological Safety Analysis Computer Program (RSAC-6.2) calculates the consequences of a release of radionuclides to the atmosphere. Using a personal computer, a user can generate a fission product inventory from either reactor operating history or a nuclear criticality accident. RSAC-6.2 models the effects of high-efficiency particulate air filters or other cleanup systems and calculates decay and ingrowth during transport through processes, facilities, and the environment. Doses are calculated for resuspension, inhalation, immersion, ground surface, and ingestion pathways. WinRP 2.0, a windows based overlay to RSAC-6.2, assists users in creating and running RSAC-6.2 input files. This users manual contains the mathematical models and operating instructions for RSAC-6.2 and WinRP 2.0. Instructions, screens, and examples are provided to guide the user through the functions provided by RSAC-6.2 and WinRP 2.0. These programs are designed for users who are familiar with radiological dose assessment methods.

A recombination outside the BB deletion refines the location of the X-linked retinitis pigmentosa locus RP3

Energy Technology Data Exchange (ETDEWEB)

Fujita, R.; Bingham, E.; Forsythe, P.; McHenry, C. [Univ. of Michigan, Ann Arbor, MI (United States)] [and others

1996-07-01

Genetic loci for X-linked retinitis pigmentosa (XLRP) have been mapped between Xp11.22 and Xp22.13 (RP2, RP3, RP6, and RP15). The RP3 gene, which is responsible for the predominant form of XLRP in most Caucasian populations, has been localized to Xp21.1 by linkage analysis and the map positions of chromosomal deletions associated with the disease. Previous linkage studies have suggested that RP3 is flanked by the markers DXS1110 (distal) and OTC (proximal). Patient BB was though to have RP because of a lesion at the RP3 locus, in addition to chronic granulomatous disease, Duchenne muscular dystrophy (DMD), mild mental retardation, and the McLeod phenotype. This patient carried a deletion extending {approximately}3 Mb from DMD in Xp21.3 to Xp21.1, with the proximal breakpoint located {approximately}40 kb centromeric to DXS1110. The RP3 gene, therefore, is believed to reside between DXS1110 and the proximal breakpoint of the BB deletion. In order to refine the location of RP3 and to ascertain patients with RP3, we have been analyzing several XLRP families for linkage to Xp markers. Linkage analysis in an American family of 27 individuals demonstrates segregation of XLRP with markers in Xp21.1, consistent with the RP3 subtype. One affected male shows a recombination event proximal to DXS1110. Additional markers within the DXS1110-OTC interval show that the crossover is between two novel polymorphic markers, DXS8349 and M6, both of which are present in BB DNA and lie centromeric to the proximal breakpoint. This recombination places the XLRP mutation in this family outside the BB deletion and redefines the location of RP3. 22 refs., 3 figs., 2 tabs.

High sensitivity phase retrieval method in grating-based x-ray phase contrast imaging

Energy Technology Data Exchange (ETDEWEB)

Wu, Zhao; Gao, Kun; Chen, Jian; Wang, Dajiang; Wang, Shenghao; Chen, Heng; Bao, Yuan; Shao, Qigang; Wang, Zhili, E-mail: wangnsrl@ustc.edu.cn [National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei 230029 (China); Zhang, Kai [Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); Zhu, Peiping; Wu, Ziyu, E-mail: wuzy@ustc.edu.cn [National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei 230029, China and Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China)

2015-02-15

Purpose: Grating-based x-ray phase contrast imaging is considered as one of the most promising techniques for future medical imaging. Many different methods have been developed to retrieve phase signal, among which the phase stepping (PS) method is widely used. However, further practical implementations are hindered, due to its complex scanning mode and high radiation dose. In contrast, the reverse projection (RP) method is a novel fast and low dose extraction approach. In this contribution, the authors present a quantitative analysis of the noise properties of the refraction signals retrieved by the two methods and compare their sensitivities. Methods: Using the error propagation formula, the authors analyze theoretically the signal-to-noise ratios (SNRs) of the refraction images retrieved by the two methods. Then, the sensitivities of the two extraction methods are compared under an identical exposure dose. Numerical experiments are performed to validate the theoretical results and provide some quantitative insight. Results: The SNRs of the two methods are both dependent on the system parameters, but in different ways. Comparison between their sensitivities reveals that for the refraction signal, the RP method possesses a higher sensitivity, especially in the case of high visibility and/or at the edge of the object. Conclusions: Compared with the PS method, the RP method has a superior sensitivity and provides refraction images with a higher SNR. Therefore, one can obtain highly sensitive refraction images in grating-based phase contrast imaging. This is very important for future preclinical and clinical implementations.

High sensitivity phase retrieval method in grating-based x-ray phase contrast imaging

International Nuclear Information System (INIS)

Wu, Zhao; Gao, Kun; Chen, Jian; Wang, Dajiang; Wang, Shenghao; Chen, Heng; Bao, Yuan; Shao, Qigang; Wang, Zhili; Zhang, Kai; Zhu, Peiping; Wu, Ziyu

2015-01-01

Purpose: Grating-based x-ray phase contrast imaging is considered as one of the most promising techniques for future medical imaging. Many different methods have been developed to retrieve phase signal, among which the phase stepping (PS) method is widely used. However, further practical implementations are hindered, due to its complex scanning mode and high radiation dose. In contrast, the reverse projection (RP) method is a novel fast and low dose extraction approach. In this contribution, the authors present a quantitative analysis of the noise properties of the refraction signals retrieved by the two methods and compare their sensitivities. Methods: Using the error propagation formula, the authors analyze theoretically the signal-to-noise ratios (SNRs) of the refraction images retrieved by the two methods. Then, the sensitivities of the two extraction methods are compared under an identical exposure dose. Numerical experiments are performed to validate the theoretical results and provide some quantitative insight. Results: The SNRs of the two methods are both dependent on the system parameters, but in different ways. Comparison between their sensitivities reveals that for the refraction signal, the RP method possesses a higher sensitivity, especially in the case of high visibility and/or at the edge of the object. Conclusions: Compared with the PS method, the RP method has a superior sensitivity and provides refraction images with a higher SNR. Therefore, one can obtain highly sensitive refraction images in grating-based phase contrast imaging. This is very important for future preclinical and clinical implementations

Matrix effects break the LC behavior rule for analytes in LC-MS/MS analysis of biological samples.

Science.gov (United States)

Fang, Nianbai; Yu, Shanggong; Ronis, Martin Jj; Badger, Thomas M

2015-04-01

High-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are generally accepted as the preferred techniques for detecting and quantitating analytes of interest in biological matrices on the basis of the rule that one chemical compound yields one LC-peak with reliable retention time (Rt.). However, in the current study, we have found that under the same LC-MS conditions, the Rt. and shape of LC-peaks of bile acids in urine samples from animals fed dissimilar diets differed significantly among each other. To verify this matrix effect, 17 authentic bile acid standards were dissolved in pure methanol or in methanol containing extracts of urine from pigs consuming either breast milk or infant formula and analyzed by LC-MS/MS. The matrix components in urine from piglets fed formula significantly reduced the LC-peak Rt. and areas of bile acids. This is the first characterization of this matrix effect on Rt. in the literature. Moreover, the matrix effect resulted in an unexpected LC behavior: one single compound yielded two LC-peaks, which broke the rule of one LC-peak for one compound. The three bile acid standards which exhibited this unconventional LC behavior were chenodeoxycholic acid, deoxycholic acid, and glycocholic acid. One possible explanation for this effect is that some matrix components may have loosely bonded to analytes, which changed the time analytes were retained on a chromatography column and interfered with the ionization of analytes in the MS ion source to alter the peak area. This study indicates that a comprehensive understanding of matrix effects is needed towards improving the use of HPLC and LC-MS/MS techniques for qualitative and quantitative analyses of analytes in pharmacokinetics, proteomics/metabolomics, drug development, and sports drug testing, especially when LC-MS/MS data are analyzed by automation software where identification of an analyte is based on its exact molecular weight and Rt

Reversed phase parallel artificial membrane permeation assay for log P measurement

Directory of Open Access Journals (Sweden)

Zihao Song

2016-03-01

Full Text Available A reversed phase parallel artificial membrane permeation assay (RP-PAMPA was newly invented for log P measurement. An oil/water/oil sandwich was constructed using a conventional PAMPA instrument. 1 % agarose was used to improve the physical stability of the water phase. A linear correlation between log P and the apparent permeability was observed in the -0.24 < log P < 2.85 region (R2 = 0.98. RP-PAMPA was also applied to pKa measurement.

Development and Validation of a Stability-Indicating LC-UV Method ...

African Journals Online (AJOL)

Development and Validation of a Stability-Indicating LC-UV Method for Simultaneous Determination of Ketotifen and Cetirizine in Pharmaceutical Dosage Forms. ... 5 μm) using an isocratic mobile phase that consisted of acetonitrile and 10 mM disodium hydrogen phosphate buffer (pH 6.5) in a ratio of 45:55 % v/v at a flow ...

77 FR 40626 - RP9580.210, Personal Assistance Services in Shelters Fact Sheet

Science.gov (United States)

2012-07-10

... DEPARTMENT OF HOMELAND SECURITY Federal Emergency Management Agency [Docket ID FEMA-2012-0014] RP9580.210, Personal Assistance Services in Shelters Fact Sheet AGENCY: Federal Emergency Management... Management Agency (FEMA) is accepting comments on Recovery Fact Sheet RP9580.210, Personal Assistance...

The Relevance of AgRP Neuron-Derived GABA Inputs to POMC Neurons Differs for Spontaneous and Evoked Release.

Science.gov (United States)

Rau, Andrew R; Hentges, Shane T

2017-08-02

Hypothalamic agouti-related peptide (AgRP) neurons potently stimulate food intake, whereas proopiomelanocortin (POMC) neurons inhibit feeding. Whether AgRP neurons exert their orexigenic actions, at least in part, by inhibiting anorexigenic POMC neurons remains unclear. Here, the connectivity between GABA-releasing AgRP neurons and POMC neurons was examined in brain slices from male and female mice. GABA-mediated spontaneous IPSCs (sIPSCs) in POMC neurons were unaffected by disturbing GABA release from AgRP neurons either by cell type-specific deletion of the vesicular GABA transporter or by expression of botulinum toxin in AgRP neurons to prevent vesicle-associated membrane protein 2-dependent vesicle fusion. Additionally, there was no difference in the ability of μ-opioid receptor (MOR) agonists to inhibit sIPSCs in POMC neurons when MORs were deleted from AgRP neurons, and activation of the inhibitory designer receptor hM4Di on AgRP neurons did not affect sIPSCs recorded from POMC neurons. These approaches collectively indicate that AgRP neurons do not significantly contribute to the strong spontaneous GABA input to POMC neurons. Despite these observations, optogenetic stimulation of AgRP neurons reliably produced evoked IPSCs in POMC neurons, leading to the inhibition of POMC neuron firing. Thus, AgRP neurons can potently affect POMC neuron function without contributing a significant source of spontaneous GABA input to POMC neurons. Together, these results indicate that the relevance of GABAergic inputs from AgRP to POMC neurons is state dependent and highlight the need to consider different types of transmitter release in circuit mapping and physiologic regulation. SIGNIFICANCE STATEMENT Agouti-related peptide (AgRP) neurons play an important role in driving food intake, while proopiomelanocortin (POMC) neurons inhibit feeding. Despite the importance of these two well characterized neuron types in maintaining metabolic homeostasis, communication between these

Effect of menstrual cycle phase on corticolimbic brain activation by visual food cues.

Science.gov (United States)

Frank, Tamar C; Kim, Ginah L; Krzemien, Alicja; Van Vugt, Dean A

2010-12-02

Food intake is decreased during the late follicular phase and increased in the luteal phase of the menstrual cycle. While a changing ovarian steroid milieu is believed to be responsible for this behavior, the specific mechanisms involved are poorly understood. Brain activity in response to visual food stimuli was compared during the estrogen dominant peri-ovulatory phase and the progesterone dominant luteal phase of the menstrual cycle. Twelve women underwent functional magnetic resonance imaging during the peri-ovulatory and luteal phases of the menstrual cycle in a counterbalanced fashion. Whole brain T2* images were collected while subjects viewed pictures of high calorie (HC) foods, low calorie (LC) foods, and control (C) pictures presented in a block design. Blood oxygen level dependent (BOLD) signal in the late follicular phase and luteal phase was determined for the contrasts HC-C, LC-C, HC-LC, and LC-HC. Both HC and LC stimuli activated numerous corticolimbic brain regions in the follicular phase, whereas only HC stimuli were effective in the luteal phase. Activation of the nucleus accumbens (NAc), amygdala, and hippocampus in response to the HC-C contrast and the hippocampus in response to the LC-C contrast was significantly increased in the late follicular phase compared to the luteal phase. Activation of the orbitofrontal cortex and mid cingulum in response to the HC-LC contrast was greater during the luteal phase. These results demonstrate for the first time that brain responses to visual food cues are influenced by menstrual cycle phase. We postulate that ovarian steroid modulation of the corticolimbic brain contributes to changes in ingestive behavior during the menstrual cycle. Copyright © 2010 Elsevier B.V. All rights reserved.

Application of modern information technologies for monitoring of 600 RP key component performance and lifetime assessment

International Nuclear Information System (INIS)

Karpenko, S.N.; Korotkih, Yu.G.; Levin, A.A.; Sankov, E.I.; Pobedonostzev, A.B.; Shashkin, S.L.

1997-01-01

Safety and reliability of NPP operation in the first turn depend on reliability and quality of the main technological equipment. Diagnostics systems are envisaged for the equipment state inspection to determine its serviceability in NPP designs. An assessment of the equipment residual lifetime at operating plants is calculated as a difference between specified (according to the operation model accepted in the design phase) lifetime and actual time in operation. Real influence of operating modes on the main equipment lifetime, in this case, is not taken into account. Within a design of NPP with VPBER-600 passive safety RP a system for lifetime assessment of RP key component is provided as a part of a process control system. Specialists from OKBM and Scientific Research Institute of Mechanics participate in development of this system. Now the first stage of the work on creation of this system is completed. This paper describes the purpose, the basic configuration principles of the system, its position among other NPP systems, organization of user-system interaction. (author). 5 refs, 1 figs

Application of modern information technologies for monitoring of 600 RP key component performance and lifetime assessment

Energy Technology Data Exchange (ETDEWEB)

Karpenko, S N; Korotkih, Yu G; Levin, A A; Sankov, E I [Research Inst. of Mechanics of Lobachevski State Univ., Nizhny Novgorod (Russian Federation); Pobedonostzev, A B; Shashkin, S L [OKB Mechanical Engineering, Nizhny Novgorod (Russian Federation)

1997-07-01

Safety and reliability of NPP operation in the first turn depend on reliability and quality of the main technological equipment. Diagnostics systems are envisaged for the equipment state inspection to determine its serviceability in NPP designs. An assessment of the equipment residual lifetime at operating plants is calculated as a difference between specified (according to the operation model accepted in the design phase) lifetime and actual time in operation. Real influence of operating modes on the main equipment lifetime, in this case, is not taken into account. Within a design of NPP with VPBER-600 passive safety RP a system for lifetime assessment of RP key component is provided as a part of a process control system. Specialists from OKBM and Scientific Research Institute of Mechanics participate in development of this system. Now the first stage of the work on creation of this system is completed. This paper describes the purpose, the basic configuration principles of the system, its position among other NPP systems, organization of user-system interaction. (author). 5 refs, 1 figs.

An unusual dimeric structure and assembly for TLR4 regulator RP105-MD-1

Energy Technology Data Exchange (ETDEWEB)

Yoon, Sung-il; Hong, Minsun; Wilson, Ian A [Scripps

2011-11-16

RP105-MD-1 modulates the TLR4-MD-2-mediated, innate immune response against bacterial lipopolysaccharide (LPS). The crystal structure of the bovine 1:1 RP105-MD-1 complex bound to a putative endogenous lipid at 2.9 Å resolution shares a similar overall architecture to its homolog TLR4-MD-2 but assembles into an unusual 2:2 homodimer that differs from any other known TLR-ligand assembly. The homodimer is assembled in a head-to-head orientation that juxtaposes the N-terminal leucine-rich repeats (LRRs) of the two RP105 chains, rather than the usual tail-to-tail configuration of C-terminal LRRs in ligand-activated TLR dimers, such as TLR1-TRL2, TLR2-TLR6, TLR3-TLR3 and TLR4-TLR4. Another unusual interaction is mediated by an RP105-specific asparagine-linked glycan, which wedges MD-1 into the co-receptor binding concavity on RP105. This unique mode of assembly represents a new paradigm for TLR complexes and suggests a molecular mechanism for regulating LPS responses.

LcMYB1 is a key determinant of differential anthocyanin accumulation among genotypes, tissues, developmental phases and ABA and light stimuli in Litchi chinensis.

Directory of Open Access Journals (Sweden)

Biao Lai

Full Text Available The red coloration of litchi fruit depends on the accumulation of anthocyanins. The anthocyanins level in litchi fruit varies widely among cultivars, developmental stages and environmental stimuli. Previous studies on various plant species demonstrate that anthocyanin biosynthesis is controlled at the transcriptional level. Here, we describe a litchi R2R3-MYB transcription factor gene, LcMYB1, which demonstrates a similar sequence as other known anthocyanin regulators. The transcription levels of the LcMYB1 and anthocyanin biosynthetic genes were investigated in samples with different anthocyanin levels. The expression of LcMYB1 was strongly associated with tissue anthocyanin content. LcMYB1 transcripts were only detected in anthocyanin-accumulating tissues and were positively correlated with anthocyanin accumulation in the pericarps of 12 genotypes. ABA and sunlight exposure promoted, whereas CPPU and bagging inhibited the expression of LcMYB1 and anthocyanin accumulation in the pericarp. Cis-elements associated with light responsiveness and abscisic acid responsiveness were identified in the promoter region of LcMYB1. Among the 6 structural genes tested, only LcUFGT was highly correlated with LcMYB1. These results suggest that LcMYB1 controls anthocyanin biosynthesis in litchi and LcUFGT might be the structural gene that is targeted and regulated by LcMYB1. Furthermore, the overexpression of LcMYB1 induced anthocyanin accumulation in all tissues in tobacco, confirming the function of LcMYB1 in the regulation of anthocyanin biosynthesis. The upregulation of NtAn1b in response to LcMYB1 overexpression seems to be essential for anthocyanin accumulation in the leaf and pedicel. In the reproductive tissues of transgenic tobacco, however, increased anthocyanin accumulation is independent of tobacco's endogenous MYB and bHLH transcriptional factors, but associated with the upregulation of specific structural genes.

LcMYB1 is a key determinant of differential anthocyanin accumulation among genotypes, tissues, developmental phases and ABA and light stimuli in Litchi chinensis.

Science.gov (United States)

Lai, Biao; Li, Xiao-Jing; Hu, Bing; Qin, Yong-Hua; Huang, Xu-Ming; Wang, Hui-Cong; Hu, Gui-Bing

2014-01-01

The red coloration of litchi fruit depends on the accumulation of anthocyanins. The anthocyanins level in litchi fruit varies widely among cultivars, developmental stages and environmental stimuli. Previous studies on various plant species demonstrate that anthocyanin biosynthesis is controlled at the transcriptional level. Here, we describe a litchi R2R3-MYB transcription factor gene, LcMYB1, which demonstrates a similar sequence as other known anthocyanin regulators. The transcription levels of the LcMYB1 and anthocyanin biosynthetic genes were investigated in samples with different anthocyanin levels. The expression of LcMYB1 was strongly associated with tissue anthocyanin content. LcMYB1 transcripts were only detected in anthocyanin-accumulating tissues and were positively correlated with anthocyanin accumulation in the pericarps of 12 genotypes. ABA and sunlight exposure promoted, whereas CPPU and bagging inhibited the expression of LcMYB1 and anthocyanin accumulation in the pericarp. Cis-elements associated with light responsiveness and abscisic acid responsiveness were identified in the promoter region of LcMYB1. Among the 6 structural genes tested, only LcUFGT was highly correlated with LcMYB1. These results suggest that LcMYB1 controls anthocyanin biosynthesis in litchi and LcUFGT might be the structural gene that is targeted and regulated by LcMYB1. Furthermore, the overexpression of LcMYB1 induced anthocyanin accumulation in all tissues in tobacco, confirming the function of LcMYB1 in the regulation of anthocyanin biosynthesis. The upregulation of NtAn1b in response to LcMYB1 overexpression seems to be essential for anthocyanin accumulation in the leaf and pedicel. In the reproductive tissues of transgenic tobacco, however, increased anthocyanin accumulation is independent of tobacco's endogenous MYB and bHLH transcriptional factors, but associated with the upregulation of specific structural genes.

Small scale affinity purification and high sensitivity reversed phase nanoLC-MS N-glycan characterization of mAbs and fusion proteins.

Science.gov (United States)

Higel, Fabian; Seidl, Andreas; Demelbauer, Uwe; Sörgel, Fritz; Frieß, Wolfgang

2014-01-01

N-glycosylation is a complex post-translational modification with potential effects on the efficacy and safety of therapeutic proteins and known influence on the effector function of biopharmaceutical monoclonal antibodies (mAbs). Comprehensive characterization of N-glycosylation is therefore important in biopharmaceutical development. In early development, e.g. during pool or clone selection, however, only minute protein amounts of multiple samples are available for analytics. High sensitivity and high throughput methods are thus needed. An approach based on 96-well plate sample preparation and nanoLC-MS of 2- anthranilic acid or 2-aminobenzoic acid (AA) labeled N-glycans for the characterization of biopharmaceuticals in early development is reported here. With this approach, 192 samples can be processed simultaneously from complex matrices (e.g., cell culture supernatant) to purified 2-AA glycans, which are then analyzed by reversed phase nanoLC-MS. Attomolar sensitivity has been achieved by use of nanoelectrospray ionization, resulting in detailed glycan maps of mAbs and fusion proteins that are exemplarily shown in this work. Reproducibility, robustness and linearity of the approach are demonstrated, making use in a routine manner during pool or clone selection possible. Other potential fields of application, such as glycan biomarker discovery from serum samples, are also presented.

Screening for anabolic steroids in urine of forensic cases using fully automated solid phase extraction and LC-MS-MS.

Science.gov (United States)

Andersen, David W; Linnet, Kristian

2014-01-01

A screening method for 18 frequently measured exogenous anabolic steroids and the testosterone/epitestosterone (T/E) ratio in forensic cases has been developed and validated. The method involves a fully automated sample preparation including enzyme treatment, addition of internal standards and solid phase extraction followed by analysis by liquid chromatography-tandem mass spectrometry (LC-MS-MS) using electrospray ionization with adduct formation for two compounds. Urine samples from 580 forensic cases were analyzed to determine the T/E ratio and occurrence of exogenous anabolic steroids. Extraction recoveries ranged from 77 to 95%, matrix effects from 48 to 78%, overall process efficiencies from 40 to 54% and the lower limit of identification ranged from 2 to 40 ng/mL. In the 580 urine samples analyzed from routine forensic cases, 17 (2.9%) were found positive for one or more anabolic steroids. Only seven different steroids including testosterone were found in the material, suggesting that only a small number of common steroids are likely to occur in a forensic context. The steroids were often in high concentrations (>100 ng/mL), and a combination of steroids and/or other drugs of abuse were seen in the majority of cases. The method presented serves as a fast and automated screening procedure, proving the suitability of LC-MS-MS for analyzing anabolic steroids. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

A new method for information retrieval in two-dimensional grating-based X-ray phase contrast imaging

International Nuclear Information System (INIS)

Wang Zhi-Li; Gao Kun; Chen Jian; Ge Xin; Tian Yang-Chao; Wu Zi-Yu; Zhu Pei-Ping

2012-01-01

Grating-based X-ray phase contrast imaging has been demonstrated to be an extremely powerful phase-sensitive imaging technique. By using two-dimensional (2D) gratings, the observable contrast is extended to two refraction directions. Recently, we have developed a novel reverse-projection (RP) method, which is capable of retrieving the object information efficiently with one-dimensional (1D) grating-based phase contrast imaging. In this contribution, we present its extension to the 2D grating-based X-ray phase contrast imaging, named the two-dimensional reverse-projection (2D-RP) method, for information retrieval. The method takes into account the nonlinear contributions of two refraction directions and allows the retrieval of the absorption, the horizontal and the vertical refraction images. The obtained information can be used for the reconstruction of the three-dimensional phase gradient field, and for an improved phase map retrieval and reconstruction. Numerical experiments are carried out, and the results confirm the validity of the 2D-RP method

Intra-specific genetic relationship analyses of Elaeagnus angustifolia based on RP-HPLC biochemical markers

Institute of Scientific and Technical Information of China (English)

无

2006-01-01

Elaeagnus angustifolia Linn. has various ecological, medicinal and economical uses. An approach was established using RP-HPLC (reversed-phase high-performance liquid chromatography) to classify and analyse the intra-specific genetic relationships of seventeen populations of E. angustifolia, collected from the Xinjiang areas of China. Chromatograms of alcohol-soluble proteins produced by seventeen populations ofE. angustifolia, were compared. Each chromatogram of alcohol-soluble proteins came from a single seed of one wild plant only. The results showed that when using a Waters Delta Pak. C18, 5 μm particle size reversed phase column (150 mm×3.9 mm), a linear gradient of 25%～60% solvent B with flow rate of 1 ml/min and run time of 67 min, the chromatography yielded optimum separation ofE. angustifolia alcohol-soluble proteins. Representative peaks in each population were chosen according to peak area and occurrence in every seed. The converted data on the elution peaks of each population were different and could be used to represent those populations. GSC (genetic similarity coefficients) of 41% to 62% showed a medium degree of genetic diversity among the populations in these eco-areas. Cluster analysis showed that the seventeen populations ofE. angustifolia could be divided into six clusters at the GSC=0.535 level and indicated the general and unique biochemical markers of these clusters. We suggest that E. angustifolia distribution in these eco-areas could be classified into six variable species. RP-HPLC was shown to be a rapid, repeatable and reliable method for E. angustifolia classification and identification and for analysis of genetic diversity.

Reversed-phase liquid chromatographic profile of free amino acids in strawberry-tree (Arbutus unedo L.) honey.

Science.gov (United States)

Spano, Nadia; Piras, Irene; Ciulu, Marco; Floris, Ignazio; Panzanelli, Angelo; Pilo, Maria I; Piu, Paola C; Sanna, Gavino

2009-01-01

The typical profile of the free amino acids contained in strawberry-tree (Arbutus unedo L.) honey is reported for the first time. An optimized reversed-phase liquid chromatographic (RP-LC) method with phenyl isothiocyanate precolumn derivatization was used. Fourteen free amino acids were identified and quantified in 16 analytical samples. Proline (65.63%) was found to be the most abundant free amino acid, followed by glutamic acid (6.49%), arginine (5.21%), alanine (5.17%), and phenylalanine (4.97%). The total free amino acid content of strawberry-tree honey (average value, 436 mg/kg) was found to be low in comparison to amounts cited in the literature concerning unifloral honeys. The analytical method was optimized and fully validated in terms of detection and quantitation limits, precision (by testing repeatability and reproducibility), linearity, and bias (by means of recovery tests). The acceptability of the validation protocol results was verified using Horwitz's mathematical model and AOAC guidelines.

Nucleosynthesis of proton-rich nuclei. Experimental results on the rp-process

International Nuclear Information System (INIS)

Galaviz, D; Amthor, A M; Bazin, D; Becerril, A D; Brown, B A; Cole, A; Cook, J M; Elliot, T; Estrade, A; Gade, A; Glasmacher, T; Lorusso, G; Matos, M; Montes, F; Mueller, W; Chen, A A; Fueloep, Z S; Heger, A; Howard, M E; Kessler, R

2010-01-01

We report in this study the nuclear properties of proton-rich isotopes located along the rp-process path. The experiments have recently been performed at the National Superconducting Cyclotron Laboratory (NSCL) at Michigan State University. The level properties above the proton separation energy of the nuclei 30 S, 36 K and 37 Ca were measured with precision of < 10 keV. This will allow a reduction in the determination of the astrophysical (p,γ) reaction rate under rp-process conditions.

Quantitation and Identification of Intact Major Milk Proteins for High-Throughput LC-ESI-Q-TOF MS Analyses.

Directory of Open Access Journals (Sweden)

Delphine Vincent

Full Text Available Cow's milk is an important source of proteins in human nutrition. On average, cow's milk contains 3.5% protein. The most abundant proteins in bovine milk are caseins and some of the whey proteins, namely beta-lactoglobulin, alpha-lactalbumin, and serum albumin. A number of allelic variants and post-translationally modified forms of these proteins have been identified. Their occurrence varies with breed, individuality, stage of lactation, and health and nutritional status of the animal. It is therefore essential to have reliable methods of detection and quantitation of these proteins. Traditionally, major milk proteins are quantified using liquid chromatography (LC and ultra violet detection method. However, as these protein variants co-elute to some degree, another dimension of separation is beneficial to accurately measure their amounts. Mass spectrometry (MS offers such a tool. In this study, we tested several RP-HPLC and MS parameters to optimise the analysis of intact bovine proteins from milk. From our tests, we developed an optimum method that includes a 20-28-40% phase B gradient with 0.02% TFA in both mobile phases, at 0.2 mL/min flow rate, using 75°C for the C8 column temperature, scanning every 3 sec over a 600-3000 m/z window. The optimisations were performed using external standards commercially purchased for which ionisation efficiency, linearity of calibration, LOD, LOQ, sensitivity, selectivity, precision, reproducibility, and mass accuracy were demonstrated. From the MS analysis, we can use extracted ion chromatograms (EICs of specific ion series of known proteins and integrate peaks at defined retention time (RT window for quantitation purposes. This optimum quantitative method was successfully applied to two bulk milk samples from different breeds, Holstein-Friesian and Jersey, to assess differences in protein variant levels.

Development and Validation of a LC/MS/MS Method for the Determination of Duloxetine in Human Plasma and Its Application to Pharmacokinetic Study

Directory of Open Access Journals (Sweden)

D. Chandrapal Reddy

2012-01-01

Full Text Available A selective, high sensitive and high throughput liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS method has been developed and validated for the chromatographic separation and quantitation of duloxetine in human EDTA plasma using fluoxetine (IS as an internal standard. Analyte and IS were extracted from human plasma by liquid-liquid extraction using MTBE-n Hexane (80:20.The eluted samples were chromatographed on X-terra RP8 (50 mmx4.6 mm, 5 μm particle size column by using mixture of 30 mM ammonium formate (pH-5.0±0.05 and acetonitrile as an isocratic mobile phase at a flow rate of 0.40 mL/min and analyzed by mass spectrometer in the multiple reaction monitoring (MRM using the respective m/z 298.08→154.0 for duloxetine and 310.02→148.07 for IS. The linearity of the response/ concentration curve was established in human plasma over the concentration range 0.100-100.017 ng/mL. The lower detection limit (LOD,S/N>3 was 0.04 ng/mL and the lower limit of quantization (LOQ,S/N>10 was 0.100 ng/mL. This LC-MS/MS method was validated with Intra-batch and Inter-batch precision of 5.21-7.02. The Intra-batch and Inter-batch accuracy was 97.14-103.50 respectively. Recovery of duloxetine in human plasma is 80.31% and ISTD recovery is 81.09%. The main pharmacokinetic parameters were Tmax (hr = (7.25±1.581, Cmax (ng/mL (44.594±18.599, AUC0→t, = (984.702±526.502 and AUC0→∞, (1027.147±572.790 respectively.

Development and validation of an rp-hplc method for simultaneous determination of Ramipril and Amlodipine in tablets

Directory of Open Access Journals (Sweden)

Shi-Ying Dai

2013-12-01

Full Text Available An rp-hplc method for the simultaneous determination of Ramipril (RP and Amlodipine (AL in tablets was developed and validated by Chinese Pharmacopoeia 2010. The linearity of the proposed method was investigated in the range of 0.01–0.25 mg/mL (r2=0.9998 for RP and 0.014–0.36 mg/mL (r2=0.9997 for AL. The limits of detection (LOD were 0.06 μg/mL and 0.02 μg/mL for RP and AL, and the limits of quantitation (LOQ were 0.2 μg/mL and 0.07 μg/mL, respectively. Some major impurities and degradation products did not disturb the detection of RP and AL and the assay can thus be considered stability-indicating. Keywords: Ramipril, Amlodipine, RP-HPLC, Stability-indicating

Differential benefits of rock phosphate (RP) by tomato ...

African Journals Online (AJOL)

plant as affected by nitrogen froms and soil types. ... Moneymaker in minirhizotrons at Hohenheim to assess root-induced chemical changes in the ... from rhizosphere pH treatment with C-horizon of Luvisol and there was no RP benefit to plant.

Autophagy in hypothalamic AgRP neurons regulates food intake and energy balance

OpenAIRE

Kaushik, Susmita; Rodriguez-Navarro, Jose Antonio; Arias, Esperanza; Kiffin, Roberta; Sahu, Srabani; Schwartz, Gary J.; Cuervo, Ana Maria; Singh, Rajat

2011-01-01

Macroautophagy is a lysosomal degradative pathway that maintains cellular homeostasis by turning over cellular components. Here, we demonstrate a role for autophagy in hypothalamic agouti-related peptide (AgRP) neurons in the regulation of food intake and energy balance. We show that starvation-induced hypothalamic autophagy mobilizes neuron-intrinsic lipids to generate endogenous free fatty acids, which in turn regulate AgRP levels. The functional consequences of inhibiting autophagy are the...

Neutron Radiography at the RP-10 reactor

International Nuclear Information System (INIS)

Hinostrosa, H.; Ravello, Y.; Cornejo, N.; Mendoza, M.; Montoya, M.

1992-01-01

The facility of neutron radiography at the RP-10 peruvian research reactor is described. The factor of collimation L/D is 149; the Cadmium ratio for the gold in the inspection's area is 4.5, and the thermal neutrons flux on the sample is 3,14 x 10 6 n/cm 2 s (author). 5 refs. 5 fig

An excitatory paraventricular nucleus to AgRP neuron circuit that drives hunger.

Science.gov (United States)

Krashes, Michael J; Shah, Bhavik P; Madara, Joseph C; Olson, David P; Strochlic, David E; Garfield, Alastair S; Vong, Linh; Pei, Hongjuan; Watabe-Uchida, Mitsuko; Uchida, Naoshige; Liberles, Stephen D; Lowell, Bradford B

2014-03-13

Hunger is a hard-wired motivational state essential for survival. Agouti-related peptide (AgRP)-expressing neurons in the arcuate nucleus (ARC) at the base of the hypothalamus are crucial to the control of hunger. They are activated by caloric deficiency and, when naturally or artificially stimulated, they potently induce intense hunger and subsequent food intake. Consistent with their obligatory role in regulating appetite, genetic ablation or chemogenetic inhibition of AgRP neurons decreases feeding. Excitatory input to AgRP neurons is important in caloric-deficiency-induced activation, and is notable for its remarkable degree of caloric-state-dependent synaptic plasticity. Despite the important role of excitatory input, its source(s) has been unknown. Here, through the use of Cre-recombinase-enabled, cell-specific neuron mapping techniques in mice, we have discovered strong excitatory drive that, unexpectedly, emanates from the hypothalamic paraventricular nucleus, specifically from subsets of neurons expressing thyrotropin-releasing hormone (TRH) and pituitary adenylate cyclase-activating polypeptide (PACAP, also known as ADCYAP1). Chemogenetic stimulation of these afferent neurons in sated mice markedly activates AgRP neurons and induces intense feeding. Conversely, acute inhibition in mice with caloric-deficiency-induced hunger decreases feeding. Discovery of these afferent neurons capable of triggering hunger advances understanding of how this intense motivational state is regulated.

History of Sulphur Content Effects on the Thermal Stability of RP-1 under Heated Conditions

National Research Council Canada - National Science Library

Irvine, Solveig

2004-01-01

.... Since the main fuel used in modern liquid hydrocarbon systems is RP-1, there is concern that Standard Grade RP-1 may not be a suitable propellant for future-generation rocket engines due to concern...

Perbandingan Passive LC Filter Dan Passve Single Tuned Filter Untuk Mereduksi Harmonisa Variable Speed Drive Dengan Beban Motor Induksi Tiga Fasa

OpenAIRE

Mustamam

2016-01-01

Most of the tools that used as speed control of three phase induction motors rotation is 1 phase Variable Speed Drive (VSD), where VSD is a harmonic generator. Therefore the harmonics that arise in the VSD can be reduced by using passive LC filters fasif namely passive single tuned filter that meet the standards IEC61000-3-2 Class A.The results obtained through passive LC filter could reduce the total distortion of harmonic current (THDi) from 102.9% to 23.78% while passive sin...

Lacticin LC14, a new bacteriocin produced by Lactococcus lactis BMG6.14: isolation, purification and partial characterization.

Science.gov (United States)

Lasta, Samar; Ouzari, Hadda; Andreotti, Nicolas; Fajloun, Ziad; Mansuelle, Pascal; Boudabous, Abdellatif; Sampieri, Francois; Sabatier, Jean Marc

2012-08-01

A new bacteriocin, lacticin LC14, produced by Lactococcus lactis BMG6.14, was isolated and characterized. It was purified to homogeneity from overnight broth culture by ammonium sulfate precipitation, Sep-Pak chromatography, and two steps of reversed-phase HPLC. Lacticin LC14 showed bactericidal-type antimicrobial activity against several lactic acid bacteria and pathogenic strains including Listeria monocytogenes. It was inactivated by proteinase K and pronase E, but was resistant to papain, lysozyme, lipase and catalase. Lacticin LC14 was heat resistant, stable over a wide range of pH (2-10) and after treatment by solvents and detergents. Its N-terminal end was found unreactive towards Edman sequencing. Based on MALDI-TOF mass spectrometry, its molecular mass was 3333.7 Da. LC14 amino acid composition revealed a high proportion of hydrophobic residues, but no modified ones. LC14 may be able to challenge other well known other bacteriocins in probiotic and therapeutic applications.

Novel USH2A compound heterozygous mutations cause RP/USH2 in a Chinese family.

Science.gov (United States)

Liu, Xiaowen; Tang, Zhaohui; Li, Chang; Yang, Kangjuan; Gan, Guanqi; Zhang, Zibo; Liu, Jingyu; Jiang, Fagang; Wang, Qing; Liu, Mugen

2010-03-17

To identify the disease-causing gene in a four-generation Chinese family affected with retinitis pigmentosa (RP). Linkage analysis was performed with a panel of microsatellite markers flanking the candidate genetic loci of RP. These loci included 38 known RP genes. The complete coding region and exon-intron boundaries of Usher syndrome 2A (USH2A) were sequenced with the proband DNA to screen the disease-causing gene mutation. Restriction fragment length polymorphism (RFLP) analysis and direct DNA sequence analysis were done to demonstrate co-segregation of the USH2A mutations with the family disease. One hundred normal controls were used without the mutations. The disease-causing gene in this Chinese family was linked to the USH2A locus on chromosome 1q41. Direct DNA sequence analysis of USH2A identified two novel mutations in the patients: one missense mutation p.G1734R in exon 26 and a splice site mutation, IVS32+1G>A, which was found in the donor site of intron 32 of USH2A. Neither the p.G1734R nor the IVS32+1G>A mutation was found in the unaffected family members or the 100 normal controls. One patient with a homozygous mutation displayed only RP symptoms until now, while three patients with compound heterozygous mutations in the family of study showed both RP and hearing impairment. This study identified two novel mutations: p.G1734R and IVS32+1G>A of USH2A in a four-generation Chinese RP family. In this study, the heterozygous mutation and the homozygous mutation in USH2A may cause Usher syndrome Type II or RP, respectively. These two mutations expand the mutant spectrum of USH2A.

Modification of radiation therapy by RP-170, hypoxic cell sensitizer and OK-432, biological response modifier (BRM)

International Nuclear Information System (INIS)

Murayama, Chieko; Mori, Tomoyuki; Tamauchi, Hidekazu

1995-01-01

Studies have shown that the combination therapy with RP-170, irradiation and OK-432 could produce significantly higher antitumor effect. The multiple local administration of OK-432 showed superior sensitizing effect in tumor growth delay, tumor control and survival rate. DMF from TCD 50 assay was 2.09 which might be equivalent to that from irradiation with over several hundred mg/kg of RP-170. Additionally combined with 50 mg/kg RP-170, nearly available dose in clinical use, the combined treatment with RP-170, irradiation and OK-432 produced DMF of 1.4, which could be gotten by over 100 mg/kg of RP-170. The success of this combination therapy on murine tumors might also suggest a possibility for a great benefit in clinical radiotherapy. More detailed investigations, histological and immunological research, are under progress. 14 refs., 2 tabs., 3 figs

Validated RP-HPLC/DAD Method for the Quantification of Insect Repellent Ethyl 2-Aminobenzoate in Membrane-Moderated Matrix Type Monolithic Polymeric Device.

Science.gov (United States)

Islam, Johirul; Zaman, Kamaruz; Chakrabarti, Srijita; Sharma Bora, Nilutpal; Mandal, Santa; Pratim Pathak, Manash; Srinivas Raju, Pakalapati; Chattopadhyay, Pronobesh

2017-07-01

A simple, accurate and sensitive reversed-phase high-performance liquid chromatographic (RP-HPLC) method has been developed for the estimation of ethyl 2-aminobenzoate (EAB) in a matrix type monolithic polymeric device and validated as per the International Conference on Harmonization guidelines. The analysis was performed isocratically on a ZORBAX Eclipse plus C18 analytical column (250 × 4.4 mm, 5 μm) and a diode array detector (DAD) using acetonitrile and water (75:25 v/v) as the mobile phase by keeping the flow-rate constant at 1.0 mL/min. Determination of EAB was not interfered in the presence of excipients. Inter- and intra-day relative standard deviations were not higher than 2%. Mean recovery was between 98.7 and 101.3%. Calibration curve was linear in the concentration range of 0.5-10 µg/mL. Limits of detection and quantification were 0.19 and 0.60 µg/mL, respectively. Thus, the present report put forward a novel method for the estimation of EAB, an emerging insect repellent, by using RP-HPLC technique. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

SFC-MS/MS as an orthogonal technique for improved screening of polar analytes in anti-doping control.

Science.gov (United States)

Parr, Maria Kristina; Wuest, Bernhard; Naegele, Edgar; Joseph, Jan F; Wenzel, Maxi; Schmidt, Alexander H; Stanic, Mijo; de la Torre, Xavier; Botrè, Francesco

2016-09-01

HPLC is considered the method of choice for the separation of various classes of drugs. However, some analytes are still challenging as HPLC shows limited resolution capabilities for highly polar analytes as they interact insufficiently on conventional reversed-phase (RP) columns. Especially in combination with mass spectrometric detection, limitations apply for alterations of stationary phases. Some highly polar sympathomimetic drugs and their metabolites showed almost no retention on different RP columns. Their retention remains poor even on phenylhexyl phases that show different selectivity due to π-π interactions. Supercritical fluid chromatography (SFC) as an orthogonal separation technique to HPLC may help to overcome these issues. Selected polar drugs and metabolites were analyzed utilizing SFC separation. All compounds showed sharp peaks and good retention even for the very polar analytes, such as sulfoconjugates. Retention times and elution orders in SFC are different to both RP and HILIC separations as a result of the orthogonality. Short cycle times could be realized. As temperature and pressure strongly influence the polarity of supercritical fluids, precise regulation of temperature and backpressure is required for the stability of the retention times. As CO2 is the main constituent of the mobile phase in SFC, solvent consumption and solvent waste are considerably reduced. Graphical Abstract SFC-MS/MS vs. LC-MS/MS.

Development and Validation of a Stability-Indicating RP-HPLC ...

African Journals Online (AJOL)

Development and Validation of a Stability-Indicating RP-HPLC Method for ... of Paracetamol, Tramadol HCl and Domperidone in a Combined Dosage Form. ... testing, as well as for quality control of the combined drugs in pharmaceutical ...

Oncolytic Herpes Virus rRp450 Shows Efficacy in Orthotopic Xenograft Group 3/4 Medulloblastomas and Atypical Teratoid/Rhabdoid Tumors

Directory of Open Access Journals (Sweden)

Adam W. Studebaker

2017-09-01

Full Text Available Pediatric brain tumors including medulloblastoma and atypical teratoid/rhabdoid tumor are associated with significant mortality and treatment-associated morbidity. While medulloblastoma tumors within molecular subgroups 3 and 4 have a propensity to metastasize, atypical teratoid/rhabdoid tumors frequently afflict a very young patient population. Adjuvant treatment options for children suffering with these tumors are not only sub-optimal but also associated with many neurocognitive obstacles. A potentially novel treatment approach is oncolytic virotherapy, a developing therapeutic platform currently in early-phase clinical trials for pediatric brain tumors and recently US Food and Drug Administration (FDA-approved to treat melanoma in adults. We evaluated the therapeutic potential of the clinically available oncolytic herpes simplex vector rRp450 in cell lines derived from medulloblastoma and atypical teratoid/rhabdoid tumor. Cells of both tumor types were supportive of virus replication and virus-mediated cytotoxicity. Orthotopic xenograft models of medulloblastoma and atypical teratoid/rhabdoid tumors displayed significantly prolonged survival following a single, stereotactic intratumoral injection of rRp450. Furthermore, addition of the chemotherapeutic prodrug cyclophosphamide (CPA enhanced rRp450’s in vivo efficacy. In conclusion, oncolytic herpes viruses with the ability to bioactivate the prodrug CPA within the tumor microenvironment warrant further investigation as a potential therapy for pediatric brain tumors.

Structure-retention and mobile phase-retention relationships for reversed-phase high-performance liquid chromatography of several hydroxythioxanthone derivatives in binary acetonitrile-water mixtures

International Nuclear Information System (INIS)

Amiri, Ali Asghar; Hemmateenejad, Bahram; Safavi, Afsaneh; Sharghi, Hashem; Beni, Ali Reza Salimi; Shamsipur, Mojtaba

2007-01-01

The reversed-phase high-performance liquid chromatographic (RP-HPLC) behavior of some newly synthesized hydroxythioxanthone derivatives using binary acetonitrile-water mixtures as mobile phase has been examined. First, the variation in the retention time of each molecule as a function of mobile phase properties was studied by Kamlet-Taft solvatochromic equations. Then, the influences of molecular structure of the hydroxythioxanthone derivatives on their retention time in various mobile phase mixtures were investigated by quantitative structure-property relationship (QSPR) analysis. Finally, a unified model containing both the molecular structure parameters and mobile phase properties was developed to describe the chromatographic behavior of the systems studied. Among the solvent properties, polarity/polarizability parameter (π * ) and hydrogen-bond basicity (β), and among the solute properties, the most positive local charge (MPC), the sum of positive charges on hydrogen atoms contributing in hydrogen bonding (SPCH) and lipophilicity index (log P) were identified as controlling factors in the RP-HPLC behavior of hydroxythioxanthone derivatives in actonitrile-water binary solvents

RP-2 Thermal Stability and Heat Transfer Investigation for Hydrocarbon Boost Engines

Science.gov (United States)

VanNoord, J. L.; Stiegemeier, B. R.

2010-01-01

A series of electrically heated tube tests were performed at the NASA Glenn Research Center s Heated Tube Facility to investigate the use of RP-2 as a fuel for next generation regeneratively cooled hydrocarbon boost engines. The effect that test duration, operating condition and test piece material have on the overall thermal stability and materials compatibility characteristics of RP-2 were evaluated using copper and 304 stainless steel test sections. The copper tests were run at 1000 psia, heat flux up to 6.0 Btu/in.2-sec, and wall temperatures up to 1180 F. Preliminary results, using measured wall temperature as an indirect indicator of the carbon deposition process, show that in copper test pieces above approximately 850 F, RP-2 begins to undergo thermal decomposition resulting in local carbon deposits. Wall temperature traces show significant local temperature increases followed by near instantaneous drops which have been attributed to the carbon deposition/shedding process in previous investigations. Data reduction is currently underway for the stainless steel test sections and carbon deposition measurements will be performed in the future for all test sections used in this investigation. In conjunction with the existing thermal stability database, these findings give insight into the feasibility of cooling a long life, high performance, high-pressure liquid rocket combustor and nozzle with RP-2.

Precipitate microstructure evolution in exposed IN738LC superalloy

Energy Technology Data Exchange (ETDEWEB)

Strunz, Pavel, E-mail: strunz@ujf.cas.cz [Nuclear Physics Institute ASCR, CZ-25068 Řež near Prague (Czech Republic); Petrenec, Martin [Institute of Physics of Materials of the AS CR, Brno (Czech Republic); Gasser, Urs [Laboratory for Neutron Scattering, PSI, CH-5232 Villigen (Switzerland); Tobiáš, Jiří; Polák, Jaroslav [Institute of Physics of Materials of the AS CR, Brno (Czech Republic); Šaroun, Jan [Nuclear Physics Institute ASCR, CZ-25068 Řež near Prague (Czech Republic)

2014-03-15

Highlights: • Evolution of γ′-phase morphology in IN738LC Ni-base superalloy was examined by SANS. • In situ tests at high temperatures revealed trimodal precipitate distribution. • Formation, dissolution and (slow) kinetics of small γ′ precipitates was determined. • Equilibrium volume fraction of γ′ phase is significantly higher than 45%. • The small γ′ precipitates arise regardless the application of the mechanical load. -- Abstract: Nickel base superalloy IN738LC has been studied after low-cycle fatigue by Small Angle Neutron Scattering (SANS). Samples subjected to high-temperature low-cycle fatigue were annealed at various temperatures to change the size and the distribution of precipitates. Ex and in situ SANS and TEM studies were performed. It was found that additional precipitates are formed either during slow cooling from high temperatures or after reheating above 570 °C. Their size and distribution were evaluated. The precipitates arise regardless the application of the mechanical load. Nevertheless, these small precipitates influence low-cycle fatigue resistance. From the SANS data, it can be also deduced that the equilibrium volume fraction of γ′-precipitates at temperatures from room temperature to 825 °C is significantly higher than 45%. The kinetics of formation of small and medium-size γ′ precipitates at 700 and 800 °C was determined as well.

Determination of the appetite suppressant P57 in Hoodia gordonii plant extracts and dietary supplements by liquid chromatography/electrospray ionization mass spectrometry (LC-MSD-TOF) and LC-UV methods.

Science.gov (United States)

Avula, Bharathi; Wang, Yan-Hong; Pawar, Rahul S; Shukla, Yatin J; Schaneberg, Brian; Khan, Ikhlas A

2006-01-01

Hoodia gordonii is traditionally used in South Africa for its appetite suppressant properties. P57AS3 (P57), an oxypregnane steroidal glycoside, is the only reported active constituent from this plant as an appetite suppressant. Effective quality control of these extracts or products requires rapid methods to determine P57 content. New methods of liquid chromatography/mass spectrometry (LC/MS) and LC-UV for analysis of P57 from H. gordonii have been developed. The quantitative determination of P57 was achieved with a Phenomenex Gemini (Torrance, CA) reversed-phase column using gradient mobile phase of water and acetonitrile, both containing 0.1% acetic acid. The method was validated for linearity, repeatability, and limits of detection and quantification. Good results were obtained in terms of repeatability (relative standard deviation <5.0%) and recovery (98.5-103.5%). The developed methods were applied to the determination of P57 for H. gordonii plant samples, one related genus (Opuntia ficus-indica), and dietary supplements that claim to contain H. gordonii.

Surface modeling and chemical solution deposition of SrO(SrTiO3)n Ruddlesden-Popper phases

International Nuclear Information System (INIS)

Zschornak, M.; Gemming, S.; Gutmann, E.; Weissbach, T.; Stoecker, H.; Leisegang, T.; Riedl, T.; Traenkner, M.; Gemming, T.; Meyer, D.C.

2010-01-01

Strontium titanate (STO) is a preferred substrate material for functional oxide growth, whose surface properties can be adjusted through the presence of Ruddlesden-Popper (RP) phases. Here, density functional theory (DFT) is used to model the (1 0 0) and (0 0 1) surfaces of SrO(SrTiO 3 ) n RP phases. Relaxed surface structures, electronic properties and stability relations have been determined. In contrast to pure STO, the near-surface SrO-OSr stacking fault can be employed to control surface roughness by adjusting SrO and TiO 2 surface rumpling, to stabilize SrO termination in an SrO-rich surrounding or to increase the band gap in the case of TiO 2 termination. RP thin films have been epitaxially grown on (0 0 1) STO substrates by chemical solution deposition. In agreement with DFT results, the fraction of particular RP phases n = 1-3 changes with varying heating rate and molar ratio Sr:Ti. This is discussed in terms of bulk formation energy.

Quantification of amino acids and peptides in an ionic liquid based aqueous two-phase system by LC-MS analysis.

Science.gov (United States)

Oppermann, Sebastian; Oppermann, Christina; Böhm, Miriam; Kühl, Toni; Imhof, Diana; Kragl, Udo

2018-04-25

Aqueous two-phase systems (ATPS) occur by the mixture of two polymers or a polymer and an inorganic salt in water. It was shown that not only polymers but also ionic liquids in combination with inorganic cosmotrophic salts are able to build ATPS. Suitable for the formation of ionic liquid-based ATPS systems are hydrophilic water miscible ionic liquids. To understand the driving force for amino acid and peptide distribution in IL-ATPS at different pH values, the ionic liquid Ammoeng 110™ and K 2 HPO 4 have been chosen as a test system. To quantify the concentration of amino acids and peptides in the different phases, liquid chromatography and mass spectrometry (LC-MS) technologies were used. Therefore the peptides and amino acids have been processed with EZ:faast™-Kit from Phenomenex for an easy and reliable quantification method even in complex sample matrices. Partitioning is a surface-dependent phenomenon, investigations were focused on surface-related amino acid respectively peptide properties such as charge and hydrophobicity. Only a very low dependence between the amino acids or peptides hydrophobicity and the partition coefficient was found. Nevertheless, the presented results show that electrostatic respectively ionic interactions between the ionic liquid and the amino acids or peptides have a strong impact on their partitioning behavior.

[Determination of aristolochic acid A in Guanxinsuhe preparations by RP-HPLC].

Science.gov (United States)

Li, Lin; Gao, Hui-Min; Wang, Zhi-Min; Wang, Wei-Hao

2006-01-01

To establish a determination method of aristolochic acid A in Guanxisuhe preparations by RP-HPLC. The instrument used was Hewlett-Packard 1100 HPLC with a Alltech C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase was methanol-water-acetic acid (68: 32:1) and the flow rate was 1.0 mL x min(-1). The UV detection wavelength was 390 nm and the column temperature was at 35 degrees C. The extracted solvent for the preparations was methanol solution contained 10% formic acid. The calibration curve was linear (r = 0.999 9) within the range of 0.119-1.89 microg for aristolochic acid A. The average recovery 99.0%, RSD 0.63%. The method with good linear relationship was convenient, quick, accurate, and suitable for the quality control of the aristolochic acid A in Guanxinsuhe and other traditional Chinese medicines containing aristolochic acid A.

Mass measurement on the rp-process waiting point {sup 72}Kr

Energy Technology Data Exchange (ETDEWEB)

Rodriguez, D. [Gesellschaft fuer Schwerionenforschung mbH, Darmstadt (Germany); Kolhinen, V.S. [Jyvaeskylae Univ. (Finland); Audi, G. [CSNSM-IN2P3-Centre National de la Recherche Scientifique (CNRS), 91 - Orsay (FR)] [and others

2004-06-01

The mass of one of the three major waiting points in the astrophysical rp-process {sup 72}Kr was measured for the first time with the Penning trap mass spectrometer ISOLTRAP. The measurement yielded a relative mass uncertainty of {delta}m/m=1.2 x 10{sup -7} ({delta}m=8 keV). Other Kr isotopes, also needed for astrophysical calculations, were measured with more than one order of magnitude improved accuracy. We use the ISOLTRAP masses of{sup 72-74}Kr to reanalyze the role of the {sup 72}Kr waiting point in the rp-process during X-ray bursts. (orig.)

A validated specific stability-indicating RP-HPLC assay method for Ambrisentan and its related substances.

Science.gov (United States)

Narayana, M B V; Chandrasekhar, K B; Rao, B M

2014-09-01

A validated specific stability-indicating reverse-phase liquid chromatographic method was developed for the quantitative determination of Ambrisentan as well as its related substances in bulk samples, pharmaceutical dosage forms in the presence of degradation products and its related impurities. Forced degradation studies were performed on bulk samples of Ambrisentan as per the ICH-prescribed stress conditions using acid, base, oxidative, thermal stress and photolytic degradation to show the stability-indicating power of the LC method. Significant degradation in acidic, basic stress conditions was observed and no degradation was observed in other stress conditions. The chromatographic method was optimized using the samples generated from the forced degradation studies and the impurity-spiked solution. Good resolution between the peaks corresponds to Ambrisentan-related impurities and degradation products from the analyte were achieved on a SunFire C18 column using a mobile phase consisting of a mixture of potassium dihydrogen orthophosphate at a pH adjusted to 2.5 with ortho-phosphoric acid in water and a mixture of acetonitrile:methanol using a simple linear gradient. The detection was carried out at 225 nm. The limit of detection and the limit of quantification for the Ambrisentan and its related impurities were established. The stressed test solutions were assayed against the qualified working standard of Ambrisentan and the mass balance in each case was between 98.9 and 100.3%, indicating that the developed LC method was stability indicating. Validation of the developed LC method was carried out as per the ICH requirements. The developed method was found to be suitable to check the quality of bulk samples of Ambrisentan at the time of batch release and also during its storage (long-term and accelerated stability). © The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Loss of function mutations in RP1 are responsible for retinitis pigmentosa in consanguineous familial cases

Science.gov (United States)

Kabir, Firoz; Ullah, Inayat; Ali, Shahbaz; Gottsch, Alexander D.H.; Naeem, Muhammad Asif; Assir, Muhammad Zaman; Khan, Shaheen N.; Akram, Javed; Riazuddin, Sheikh; Ayyagari, Radha; Hejtmancik, J. Fielding

2016-01-01

Purpose This study was undertaken to identify causal mutations responsible for autosomal recessive retinitis pigmentosa (arRP) in consanguineous families. Methods Large consanguineous families were ascertained from the Punjab province of Pakistan. An ophthalmic examination consisting of a fundus evaluation and electroretinography (ERG) was completed, and small aliquots of blood were collected from all participating individuals. Genomic DNA was extracted from white blood cells, and a genome-wide linkage or a locus-specific exclusion analysis was completed with polymorphic short tandem repeats (STRs). Two-point logarithm of odds (LOD) scores were calculated, and all coding exons and exon–intron boundaries of RP1 were sequenced to identify the causal mutation. Results The ophthalmic examination showed that affected individuals in all families manifest cardinal symptoms of RP. Genome-wide scans localized the disease phenotype to chromosome 8q, a region harboring RP1, a gene previously implicated in the pathogenesis of RP. Sanger sequencing identified a homozygous single base deletion in exon 4: c.3697delT (p.S1233Pfs22*), a single base substitution in intron 3: c.787+1G>A (p.I263Nfs8*), a 2 bp duplication in exon 2: c.551_552dupTA (p.Q185Yfs4*) and an 11,117 bp deletion that removes all three coding exons of RP1. These variations segregated with the disease phenotype within the respective families and were not present in ethnically matched control samples. Conclusions These results strongly suggest that these mutations in RP1 are responsible for the retinal phenotype in affected individuals of all four consanguineous families. PMID:27307693

A small potassium current in AgRP/NPY neurons regulates feeding behavior and enery metabolism

Science.gov (United States)

Neurons that co-express agouti-related peptide (AgRP) and neuropeptide Y (NPY) are indispensable for normal feeding behavior. Firing activities of AgRP/NPY neurons are dynamically regulated by energy status and coordinate appropriate feeding behavior to meet nutritional demands. However, intrinsic m...

Evaluation of operational incidents in the research reactor RP-10 according to scale INES; Evaluacion de incidentes operacionales en el reactor de investigacion RP-10 segun escala INES

Energy Technology Data Exchange (ETDEWEB)

Arrieta, Rolando W.B.; Vela Mora, Mariano, E-mail: rarrieta@ipen.gob.pe, E-mail: mvela@ipen.gob.pe [Instituto Peruano de energia Nuclear, Lima (Peru). Dept. de Operacion de Reactores

2013-07-01

This report presents the evaluation of the events in 2011 in the RP-10 Nuclear Reactor Nuclear Center Huarangal from the point of view of safety. To classify these events produced is used Scale International Nuclear and Radiological Event Scale (INES) to facilitate a common understanding between the technical community, the media and the general public. From the results we can say that in 2011 all related to security events that occurred in the RP -10 are classified as 'below scale' or no safety significance. (author)

Improved LC-MS/MS method for the quantification of hepcidin-25 in clinical samples.

Science.gov (United States)

Abbas, Ioana M; Hoffmann, Holger; Montes-Bayón, María; Weller, Michael G

2018-06-01

Mass spectrometry-based methods play a crucial role in the quantification of the main iron metabolism regulator hepcidin by singling out the bioactive 25-residue peptide from the other naturally occurring N-truncated isoforms (hepcidin-20, -22, -24), which seem to be inactive in iron homeostasis. However, several difficulties arise in the MS analysis of hepcidin due to the "sticky" character of the peptide and the lack of suitable standards. Here, we propose the use of amino- and fluoro-silanized autosampler vials to reduce hepcidin interaction to laboratory glassware surfaces after testing several types of vials for the preparation of stock solutions and serum samples for isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS). Furthermore, we have investigated two sample preparation strategies and two chromatographic separation conditions with the aim of developing a LC-MS/MS method for the sensitive and reliable quantification of hepcidin-25 in serum samples. A chromatographic separation based on usual acidic mobile phases was compared with a novel approach involving the separation of hepcidin-25 with solvents at high pH containing 0.1% of ammonia. Both methods were applied to clinical samples in an intra-laboratory comparison of two LC-MS/MS methods using the same hepcidin-25 calibrators with good correlation of the results. Finally, we recommend a LC-MS/MS-based quantification method with a dynamic range of 0.5-40 μg/L for the assessment of hepcidin-25 in human serum that uses TFA-based mobile phases and silanized glass vials. Graphical abstract Structure of hepcidin-25 (Protein Data Bank, PDB ID 2KEF).

TLR4 accessory molecule RP105 (CD180 regulates monocyte-driven arteriogenesis in a murine hind limb ischemia model.

Directory of Open Access Journals (Sweden)

Antonius J N M Bastiaansen

Full Text Available AIMS: We investigated the role of the TLR4-accessory molecule RP105 (CD180 in post-ischemic neovascularization, i.e. arteriogenesis and angiogenesis. TLR4-mediated activation of pro-inflammatory Ly6Chi monocytes is crucial for effective neovascularization. Immunohistochemical analyses revealed that RP105+ monocytes are present in the perivascular space of remodeling collateral arterioles. As RP105 inhibits TLR4 signaling, we hypothesized that RP105 deficiency would lead to an unrestrained TLR4-mediated inflammatory response and hence to enhanced blood flow recovery after ischemia. METHODS AND RESULTS: RP105-/- and wild type (WT mice were subjected to hind limb ischemia and blood flow recovery was followed by Laser Doppler Perfusion Imaging. Surprisingly, we found that blood flow recovery was severely impaired in RP105-/- mice. Immunohistochemistry showed that arteriogenesis was reduced in these mice compared to the WT. However, both in vivo and ex vivo analyses showed that circulatory pro-arteriogenic Ly6Chi monocytes were more readily activated in RP105-/- mice. FACS analyses showed that Ly6Chi monocytes became activated and migrated to the affected muscle tissues in WT mice following induction of hind limb ischemia. Although Ly6Chi monocytes were readily activated in RP105-/- mice, migration into the ischemic tissues was hampered and instead, Ly6Chi monocytes accumulated in their storage compartments, bone marrow and spleen, in RP105-/- mice. CONCLUSIONS: RP105 deficiency results in an unrestrained inflammatory response and monocyte over-activation, most likely due to the lack of TLR4 regulation. Inappropriate, premature systemic activation of pro-inflammatory Ly6Chi monocytes results in reduced infiltration of Ly6Chi monocytes in ischemic tissues and in impaired blood flow recovery.

Lepeophtheirus salmonis: characterization of prostaglandin E(2) in secretory products of the salmon louse by RP-HPLC and mass spectrometry.

Science.gov (United States)

Fast, M D; Ross, N W; Craft, C A; Locke, S J; MacKinnon, S L; Johnson, S C

2004-01-01

Lepeophtheirus salmonis is an ectoparasitic copepod that causes serious disease outbreaks in both wild and farmed salmonids. As the relationship between L. salmonis and its hosts is not well understood, the current investigation was undertaken to investigate whether any immunomodulatory compounds could be identified from secretions of L. salmonis. By incubating live L. salmonis adults with the neurotransmitter dopamine in seawater, we were able to obtain secretions from the parasite. These were analyzed by RP-HPLC column, as well as LC-MS. L. salmonis secretions contained a compound with the same retention time and mass of PGE(2). The identity of this compound as PGE(2) was confirmed by MS-in source dissociation. The concentrations of PGE(2) in L. salmonis secretions ranged from 0.2 to 12.3 ng/individual and varied with incubation temperature and time kept off the host. Prostaglandin E(2) is a potent vasodilator and thought to aid in parasite evasion from host immune responses. This is the first reported evidence of prostaglandin production in parasitic copepod secretions and its implications for the host-parasite relationship are discussed.

Determination of anabolic steroids in human urine by automated in-tube solid-phase microextraction coupled with liquid chromatography-mass spectrometry.

Science.gov (United States)

Saito, Keita; Yagi, Katsuharu; Ishizaki, Atsushi; Kataoka, Hiroyuki

2010-09-05

A simple, rapid and sensitive method was developed for determining the presence of seven anabolic steroids (boldenone, nandrolone, testosterone, methyltestosterone, epiandrosterone, androsterone, and atnozolol) in human urine. Glucuronide-conjugates of these compounds were hydrolyzed with beta-glucuronidase. The anabolic steroids were analyzed by on-line in-tube solid-phase microextraction (SPME) coupled with liquid chromatography-mass spectrometry (LC-MS). The steroids were separated within 14 min by high performance liquid chromatography using a Chromolith RP-18e column and 5 mM ammonium formate/methanol (35/65, v/v) as a mobile phase at a flow rate of 1.0 mL/min. Electrospray ionization conditions in the positive ion mode were optimized for the MS detection of these compounds. The optimum in-tube SPME conditions were 20 draw/eject cycles with a sample size of 40 microL using a Supel-Q PLOT capillary column for the extraction. The extracted compounds could be desorbed readily from the capillary column by flow of the mobile phase, and no carryover was observed. Using the in-tube SPME LC-MS with SIM mode detection, good linearity of the calibration curve (r>0.995) was obtained in the concentration range of 0.5-20 ng/mL, except for stanozolol. The detection limits (S/N=3) of anabolic steroids were in the range 9-182 pg/mL and the proposed method showed 20-33-fold higher sensitivity than the direct injection method. The within-day and between-day precisions were below 4.0% and 7.3% (n=5), respectively. This method was applied successfully to the analysis of urine samples without the interference peaks. The recovery rates of anabolic steroids spiked into urine samples were above 85%. This method is useful to analyze the urinary levels of these compounds in anti-doping tests. 2010 Elsevier B.V. All rights reserved.

RpA, an extracellular protease similar to the metalloprotease of serralysin family, is required for pathogenicity of Ralstonia pickettii.

Science.gov (United States)

Chen, C-M; Liu, J-J; Chou, C-W; Lai, C-H; Wu, L-T

2015-10-01

To investigate the biochemical and functional properties of an extracellular protease, RpA, in Ralstonia pickettii WP1 isolated from water supply systems. An extracellular protease was identified and characterized from R. pickettii WP1. A mutant strain WP1M2 was created from strain WP1 by mini-Tn5 transposition. The culture filtrates from WP1M2 had a lower cytotoxic effect than the parental WP1 on several mammalian cell lines. Cloning and sequence analysis revealed the Tn5 transposon inserted at a protease gene (rpA) which is 81% homologous to prtA and aprX genes of Pseudomonas fluorescens. The rpA gene encodes a 482-residue protein showing sequence similarity to metalloproteases of the serralysin family. The RpA protein was expressed in Escherichia coli using a pET expression vector and purified as a 55 kDa molecular weight protein. Furthermore, the protease activity of RpA was inhibited by protease inhibitor and heat treatment. The in vitro cytotoxic activity of R. pickettii culture filtrates was attributed to RpA protease. An extracellular protease, RpA, was identified from R. pickettii WP1 isolated from water supply system. The RpA metalloproteases is required for the pathogenicity of R. pickettii to mammalian cell lines. © 2015 The Society for Applied Microbiology.

Somato-dendritic localization and signaling by leptin receptors in hypothalamic POMC and AgRP neurons.

Directory of Open Access Journals (Sweden)

Sangdeuk Ha

Full Text Available Leptin acts via neuronal leptin receptors to control energy balance. Hypothalamic pro-opiomelanocortin (POMC and agouti-related peptide (AgRP/Neuropeptide Y (NPY/GABA neurons produce anorexigenic and orexigenic neuropeptides and neurotransmitters, and express the long signaling form of the leptin receptor (LepRb. Despite progress in the understanding of LepRb signaling and function, the sub-cellular localization of LepRb in target neurons has not been determined, primarily due to lack of sensitive anti-LepRb antibodies. Here we applied light microscopy (LM, confocal-laser scanning microscopy (CLSM, and electron microscopy (EM to investigate LepRb localization and signaling in mice expressing a HA-tagged LepRb selectively in POMC or AgRP/NPY/GABA neurons. We report that LepRb receptors exhibit a somato-dendritic expression pattern. We further show that LepRb activates STAT3 phosphorylation in neuronal fibers within several hypothalamic and hindbrain nuclei of wild-type mice and rats, and specifically in dendrites of arcuate POMC and AgRP/NPY/GABA neurons of Leprb (+/+ mice and in Leprb (db/db mice expressing HA-LepRb in a neuron specific manner. We did not find evidence of LepRb localization or STAT3-signaling in axon-fibers or nerve-terminals of POMC and AgRP/NPY/GABA neurons. Three-dimensional serial EM-reconstruction of dendritic segments from POMC and AgRP/NPY/GABA neurons indicates a high density of shaft synapses. In addition, we found that the leptin activates STAT3 signaling in proximity to synapses on POMC and AgRP/NPY/GABA dendritic shafts. Taken together, these data suggest that the signaling-form of the leptin receptor exhibits a somato-dendritic expression pattern in POMC and AgRP/NPY/GABA neurons. Dendritic LepRb signaling may therefore play an important role in leptin's central effects on energy balance, possibly through modulation of synaptic activity via post-synaptic mechanisms.

LC-NMR, NMR, and LC-MS identification and LC-DAD quantification of flavonoids and ellagic acid derivatives in Drosera peltata.

Science.gov (United States)

Braunberger, Christina; Zehl, Martin; Conrad, Jürgen; Fischer, Sonja; Adhami, Hamid-Reza; Beifuss, Uwe; Krenn, Liselotte

2013-08-01

The herb of Drosera peltata, commonly named the shield sundew, is used as an antitussive in phytotherapy, although the plants' composition has not been determined in detail so far. Hence, in this study, we present a validated, sensitive, reliable, and cheap narrow-bore LC-DAD method for the simultaneous quantification of flavonoids and ellagic acid derivatives in this herbal drug. In addition, the structures of 13 compounds have been elucidated by LC-MS, LC-NMR, and offline NMR experiments after isolation: herbacetin-3-O-glucoside (1), gossypitrin (2), ellagic acid (3), quercetin-7-O-glucoside (4), isoquercitrin (5), kaempferol-3-O-(6″-O-galloyl)-glucoside (6), herbacetin-7-O-glucoside (7), astragalin (8), gossypetin (9), herbacetin (10), quercetin (11), 3,3'-di-O-methyl ellagic acid (12), and kaempferol (13). Compounds 1, 2, 4, 5, 6, 7, and 10 have been identified in D. peltata for the first time, and compounds 1, 4, 6, 7, and 10 have not been detected in any Drosera species before. Copyright © 2013 Elsevier B.V. All rights reserved.

Axure RP 6 Prototyping Essentials

CERN Document Server

Schwartz, Ezra

2012-01-01

Axure RP 6 Prototyping Essentials is a detailed, practical primer on the leading rapid prototyping tool. Short on jargon and high on concepts, real-life scenarios and step-by-step guidance through hands-on examples, this book will show you how to integrate Axure into your UX workflow. This book is written for UX practitioners, business analysts, product managers, and anyone else who is involved in UX projects. The book assumes that you have no or very little familiarity with Axure. It will help you if you are evaluating the tool for an upcoming project or are required to quickly get up to spee

Three novel and the common Arg677Ter RP1 protein truncating mutations causing autosomal dominant retinitis pigmentosa in a Spanish population

Directory of Open Access Journals (Sweden)

Antiñolo Guillermo

2006-04-01

Full Text Available Abstract Background Retinitis pigmentosa (RP, a clinically and genetically heterogeneous group of retinal degeneration disorders affecting the photoreceptor cells, is one of the leading causes of genetic blindness. Mutations in the photoreceptor-specific gene RP1 account for 3–10% of cases of autosomal dominant RP (adRP. Most of these mutations are clustered in a 500 bp region of exon 4 of RP1. Methods Denaturing gradient gel electrophoresis (DGGE analysis and direct genomic sequencing were used to evaluate the 5' coding region of exon 4 of the RP1 gene for mutations in 150 unrelated index adRP patients. Ophthalmic and electrophysiological examination of RP patients and relatives according to pre-existing protocols were carried out. Results Three novel disease-causing mutations in RP1 were detected: Q686X, K705fsX712 and K722fsX737, predicting truncated proteins. One novel missense mutation, Thr752Met, was detected in one family but the mutation does not co-segregate in the family, thereby excluding this amino acid variation in the protein as a cause of the disease. We found the Arg677Ter mutation, previously reported in other populations, in two independent families, confirming that this mutation is also present in a Spanish population. Conclusion Most of the mutations reported in the RP1 gene associated with adRP are expected to encode mutant truncated proteins that are approximately one third or half of the size of wild type protein. Patients with mutations in RP1 showed mild RP with variability in phenotype severity. We also observed several cases of non-penetrant mutations.

Protective role of AgRP neuron's PDK1 against salt-induced hypertension.

Science.gov (United States)

Zhang, Boyang; Nakata, Masanori; Lu, Ming; Nakae, Jun; Okada, Takashi; Ogawa, Wataru; Yada, Toshihiko

2018-06-12

In the hypothalamic arcuate nucleus (ARC), orexigenic agouti-related peptide (AgRP) neurons regulate feeding behavior and energy homeostasis. The 3-phosphoinositide-dependent protein kinase-1 (PDK1) in AgRP neurons serves as a major signaling molecule for leptin and insulin, the hormones regulating feeding behavior, energy homeostasis and circulation. However, it is unclear whether PDK1 in AGRP neurons is also involved in regulation of blood pressure. This study explored it by generating and analyzing AgRP neuron-specific PDK1 knockout (Agrp-Pdk1 flox/flox ) mice and effect of high salt diet on blood pressure in KO and WT mice was analyzed. Under high salt diet feeding, systolic blood pressure (SBP) of Agrp-Pdk1 flox/flox mice was significantly elevated compared to Agrp-Cre mice. When the high salt diet was switched to control low salt diet, SBP of Agrp-Pdk1 flox/flox mice returned to the basal level observed in Agrp-Cre mice within 1 week. In Agrp-Pdk1 flox/flox mice, urinary noradrenalin excretion and NUCB2 mRNA expression in hypothalamic paraventricular nucleus (PVN) were markedly upregulated. Moreover, silencing of NUCB2 in the PVN counteracted the rises in urinary noradrenalin excretions and SBP. These results demonstrate a novel role of PDK1 in AgRP neurons to counteract the high salt diet-induced hypertension by preventing hyperactivation of PVN nesfatin-1 neurons. Copyright © 2018 Elsevier Inc. All rights reserved.

Development of a definition for Rapid Progression (RP) of renal function in HIV-positive persons

DEFF Research Database (Denmark)

Kamara, David A; Nielsen, Lene Ryom; Ross, Michael

2014-01-01

No consensus exists on how to define abnormally rapid deterioration in renal function (Rapid Progression, RP). We developed an operational definition of RP in HIV-positive persons with baseline estimated glomerular filtration rate (eGFR) >90ml/min/1.73m2 (using Cockcroft Gault) in the Data...

Development and validation of a simple, rapid and sensitive LC-MS/MS method for the measurement of urinary neurotransmitters and their metabolites.

Science.gov (United States)

Yan, Jingya; Kuzhiumparambil, Unnikrishnan; Bandodkar, Sushil; Solowij, Nadia; Fu, Shanlin

2017-12-01

Neurotransmitters play crucial roles in physiological functions and their imbalances have demonstrated association in the pathology of several diseases. The measurement of neurotransmitters possesses a great potential as a significant clinical tool. This study presents the development and validation of an LC-MS/MS method for simultaneous quantification of multi-class neurotransmitters associated with dopamine, tryptophan and glutamate-γ-aminobutyric acid pathways. A total of ten neurotransmitters and their metabolites (dopamine, epinephrine, metanephrine, tryptophan, serotonin, kynurenic acid, kynurenine, anthranilic acid, GABA, glutamic acid) were determined based on a simple and rapid 'dilute and shoot' method using minimal urine volume. The chromatographic separation was achieved using a Poroshell 120 Bonus-RP LC Column in combination with a gradient elution within an 8.5-min time frame. The method exhibited good sensitivity as the limits of quantification ranged between 0.025 and 0.075 μg/mL with acceptable matrix effects ( 0.98). The accuracy and precision for all analytes were within tolerances, at neurotransmitter concentrations in urine of healthy donors. Furthermore, the undertaken stability experiments indicated that acidified urine specimens allowed the analytes to be stable for prolonged durations in comparison to those untreated. The study also reveals the performance of the method is unaffected by the absence of expensive deuterated reference standards under the experimental conditions employed which further simplifies the analytical procedures and provides a significant cost saving for running the assay. Graphical abstract The quantification of multi-class neurotransitters associated with the dopamine, tryptophan and GABA-glutamate pathways using a simple 'dilute and shoot' LC-MS/MS method.

Development and Validation of a RP-HPLC Method for the ...

African Journals Online (AJOL)

Development and Validation of a RP-HPLC Method for the Simultaneous Determination of Rifampicin and a Flavonoid Glycoside - A Novel ... range, accuracy, precision, limit of detection, limit of quantification, robustness and specificity.

Formation of Escherichia coli Hfr strains by integrative suppression with the P group plasmid RP1.

OpenAIRE

Martin, R R; Thorlton, C L; Unger, L

1981-01-01

Hfr strains of Escherichia coli were obtained by integrative suppression of a dnaA(Ts) mutation by the Inc P-1 plasmid RP1 without prior creation of an unnatural homology between the plasmid and the E. coli chromosome. Unmodified RP1 mobilized the polarized transfer of the chromosome in a counterclock-wise direction from a distinct origin between 81 min (pyrE) and 82 min (dnaA) with pyrE as a leading marker. Inheritance of RP1-Hfr chromosomal and antibiotic resistance genes was due to recombi...

A new high-throughput LC-MS method for the analysis of complex fructan mixtures

DEFF Research Database (Denmark)

Verspreet, Joran; Hansen, Anders Holmgaard; Dornez, Emmie

2014-01-01

In this paper, a new liquid chromatography-mass spectrometry (LC-MS) method for the analysis of complex fructan mixtures is presented. In this method, columns with a trifunctional C18 alkyl stationary phase (T3) were used and their performance compared with that of a porous graphitized carbon (PGC...

Identification and quantification of flavonoids and ellagic acid derivatives in therapeutically important Drosera species by LC-DAD, LC-NMR, NMR, and LC-MS.

Science.gov (United States)

Zehl, Martin; Braunberger, Christina; Conrad, Jürgen; Crnogorac, Marija; Krasteva, Stanimira; Vogler, Bernhard; Beifuss, Uwe; Krenn, Liselotte

2011-06-01

Droserae herba is a drug commonly used for treatment of convulsive or whooping cough since the seventeenth century. Because of the contribution of flavonoids and ellagic acid derivatives to the therapeutic activity of Droserae herba, an LC-DAD method has been developed for quantification of these analytes in four Drosera species used in medicine (Drosera anglica, D. intermedia, D. madagascariensis, and D. rotundifolia). During elaboration of the method 13 compounds, including three substances not previously described for Drosera species, were detected and unambiguously identified by means of extensive LC-MS and LC-NMR experiments and by off-line heteronuclear 2D NMR after targeted isolation. The most prominent component of D. rotundifolia and D. anglica, 2″-O-galloylhyperoside, with myricetin-3-O-β-glucopyranoside and kaempferol-3-O-(2″-O-galloyl)-β-galactopyranoside, were identified for the very first time in this genus. The LC-DAD method for quantification was thoroughly validated, and enables, for the first time, separation and precise analysis of these analytes in Droserae herba. Simple sample preparation and use of a narrow-bore column guarantee low cost and simplicity of the suggested system, which is excellently suited to quality control of the drug or herbal medicinal products containing this drug.

Metabolism of the new psychoactive substances N,N-diallyltryptamine (DALT) and 5-methoxy-DALT and their detectability in urine by GC-MS, LC-MSn, and LC-HR-MS-MS.

Science.gov (United States)

Michely, Julian A; Helfer, Andreas G; Brandt, Simon D; Meyer, Markus R; Maurer, Hans H

2015-10-01

N,N-Diallyltryptamine (DALT) and 5-methoxy-DALT (5-MeO-DALT) are synthetic tryptamine derivatives commonly referred to as so-called new psychoactive substances (NPS). They have psychoactive effects that may be similar to those of other tryptamine derivatives. The objectives of this work were to study the metabolic fate and detectability, in urine, of DALT and 5-MeO-DALT. For metabolism studies, rat urine obtained after high-dose administration was prepared by precipitation and analyzed by liquid chromatography-high-resolution mass spectrometry (LC-HR-MS-MS). On the basis of the metabolites identified, several aromatic and aliphatic hydroxylations, N-dealkylation, N-oxidation, and combinations thereof are proposed as the main metabolic pathways for both compounds. O-Demethylation of 5-MeO-DALT was also observed, in addition to extensive glucuronidation or sulfation of both compounds after phase I transformation. The cytochrome P450 (CYP) isoenzymes predominantly involved in DALT metabolism were CYP2C19, CYP2D6, and CYP3A4; those mainly involved in 5-MeO-DALT metabolism were CYP1A2, CYP2C19, CYP2D6, and CYP3A4. For detectability studies, rat urine was screened by GC-MS, LC-MS(n), and LC-HR-MS-MS after administration of low doses. LC-MS(n) and LC-HR-MS-MS were deemed suitable for monitoring consumption of both compounds. The most abundant targets were a ring hydroxy metabolite of DALT, the N,O-bis-dealkyl metabolite of 5-MeO-DALT, and their glucuronides. GC-MS enabled screening of DALT by use of its main metabolites only.

Simultaneous RP-HPLC determination of sparfloxacin and dexamethasone in pharmaceutical formulations

Directory of Open Access Journals (Sweden)

Syed Naeem Razzaq

2013-06-01

Full Text Available The present study describes the development and subsequent validation of simple and accurate stability indicating RP-HPLC method for the determination of sparfloxacin and dexamethasone in pharmaceutical formulations in the presence of their stress-induced degradation products. Both the drugs and their stress-induced degradation products were separated within 10 minutes using C8 column and mixture of methanol and 0.02 M phosphate buffer pH 3.0 (60:40 v/v, respectively as mobile phase at 270 nm using diode array detector. Regression analysis showed linearity in the range of 15-105 µg/mL for sparfloxacin and 5-35 µg/mL for dexamethasone. All the analytes were adequately resolved with acceptable tailing. Peak purity of the two drugs was also greater than 0.9999, showing no co-elution peaks. The developed method was applied for simultaneous determination of sparfloxacin and dexamethasone in pharmaceutical formulations for stability studies.

Quantitative analysis of rutin, quercetin, naringenin, and gallic acid by validated RP- and NP-HPTLC methods for quality control of anti-HBV active extract of Guiera senegalensis.

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Alam, Perwez; Parvez, Mohammad K; Arbab, Ahmed H; Al-Dosari, Mohammed S

2017-12-01

Guiera senegalensis J.F. Gmel (Combretaceae) is a folk medicinal plant used in various metabolic and infectious diseases. In addition to its antiviral activities against herpes and fowlpox, the anti-HBV efficacy is very recently reported. To develop and validate simple, sensitive RP-/NP-HPTLC methods for quantitative determination of biomarkers rutin, quercetin, naringenin, and gallic acid in the anti-HBV active G. senegalensis leaves ethanol-extract. RP-HPTLC (rutin & quercetin; phase- acetonitrile:water, 4:6) and NP-HPTLC (naringenin & gallic acid; phase- toluene:ethyl acetate:formic acid, 6:4:0.8) were performed on glass-backed silica gel plates 60F 254 -RP18 and 60F 254 , respectively. The methods were validated according to the ICH guidelines. Well-separated and compact spots (R f ) of rutin (0.52 ± 0.006), quercetin (0.23 ± 0.005), naringenin (0.56 ± 0.009) and gallic acid (0.28 ± 0.006) were detected. The regression equations (Y) were 12.434x + 443.49, 10.08x + 216.85, 11.253x + 973.52 and 11.082x + 446.41 whereas the coefficient correlations (r 2 ) were 0.997 ± 0.0004, 0.9982 ± 0.0001, 0.9974 ± 0.0004 and 0.9981 ± 0.0001, respectively. The linearity ranges (ng/spot) were 200-1400 (RP-HPTLC) and 100-1200 (NP-HPTLC). The LOD/LOQ (ng/band) were 33.03/100.1 (rutin), 9.67/29.31 (quercetin), 35.574/107.8 (naringenin), and 12.32/37.35 (gallic acid). Gallic acid (7.01 μg/mg) was the most abundant biomarker compared to rutin (2.42 μg/mg), quercetin (1.53 μg/mg) and naringenin (0.14 μg/mg) in the extract. The validated NP-/RP-HPTLC methods were simple, accurate, and sensitive for separating and quantifying antiviral biomarkers in G. senegalensis, and endorsed its anti-HBV activity. The developed methods could be further employed in the standardization and quality-control of herbal formulations.

Unified pH values of liquid chromatography mobile phases.

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Suu, Agnes; Jalukse, Lauri; Liigand, Jaanus; Kruve, Anneli; Himmel, Daniel; Krossing, Ingo; Rosés, Martí; Leito, Ivo

2015-03-03

This work introduces a conceptually new approach of measuring pH of mixed-solvent liquid chromatography (LC) mobile phases. Mobile phase pH is very important in LC, but its correct measurement is not straightforward, and all commonly used approaches have deficiencies. The new approach is based on the recently introduced unified pH (pH(abs)) scale, which enables direct comparison of acidities of solutions made in different solvents based on chemical potential of the proton in the solutions. This work represents the first experimental realization of the pH(abs) concept using differential potentiometric measurement for comparison of the chemical potentials of the proton in different solutions (connected by a salt bridge), together with earlier published reference points for obtaining the pH(abs) values (referenced to the gas phase) or pH(abs)(H₂O) values (referenced to the aqueous solution). The liquid junction potentials were estimated in the framework of Izutsu's three-component method. pH(abs) values for a number of common LC and LC-MS mobile phases have been determined. The pH(abs) scale enables for the first time direct comparison of acidities of any LC mobile phases, with different organic additives, different buffer components, etc. A possible experimental protocol of putting this new approach into chromatographic practice has been envisaged and its applicability tested. It has been demonstrated that the ionization behavior of bases (cationic acids) in the mobile phases can be better predicted by using the pH(abs)(H₂O) values and aqueous pKa values than by using the alternative means of expressing mobile phase acidity. Description of the ionization behavior of acids on the basis of pH(abs)(H₂O) values is possible if the change of their pKa values with solvent composition change is taken into account.

Analysis of U-47700, a Novel Synthetic Opioid, in Human Urine by LC-MS-MS and LC-QToF.

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Fleming, Steven W; Cooley, Justin C; Johnson, Leonard; Frazee, C Clinton; Domanski, Kristina; Kleinschmidt, Kurt; Garg, Uttam

2017-04-01

The illicit drug market has rapidly evolved from synthetic cannabinoids to cathinone derivatives and now a new emerging threat of synthetic opioids. These compounds were mostly developed by pharmaceutical companies during drug discovery. The new psychoactive substances are not routinely covered in drug screening and may go undetected. Recently fentanyl analogous, AH-7921, MT-45 and now U-47700 have been encountered in clinical and forensic casework. U-47700 is gaining popularity on drug user forms as a legal alternative to heroin. It is a µ-receptor agonist that is part of the trans-1-2-diamine opioid analgesic drug class developed by The Upjohn Company in an attempt to develop a non-addicting analgesic. A LC-MS-MS method was developed and validated to detect and quantify U-47700. Additional analysis was conducted with an LC-QToF to identify the presence of the parent drug and metabolites. A total of four cases have been evaluated by the LC-MS-MS methodology which has an analytical range of 1-1,250 ng/mL and limit of detection of 1 ng/mL. The concentration of U-47700 in urine specimens ranged from below the limit of quantification to 224 ng/mL. The ToF analysis detected the presence of suspected phase I demethylated metabolites that may assist future analysis of this compound. The prevalence of designer opioids in casework highlights the importance of analysis for new psychoactive substances. Traditional opiates/opioids were not detected in the presented cases, but the available case histories revealed an opioid toxidrome. These findings suggest that U-47700 drug may cause significant morbidity and mortality within the United States as an emerging drug threat. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Rapid extraction combined with LC-tandem mass spectrometry (CREM-LC/MS/MS) for the determination of ciguatoxins in ciguateric fish flesh.

Science.gov (United States)

Lewis, Richard J; Yang, Aijun; Jones, Alun

2009-07-01

Ciguatera is a significant food borne disease caused by potent polyether toxins known as ciguatoxins, which accumulate in the flesh of ciguateric fish at risk levels above 0.1 ppb. The management of ciguatera has been hindered by the lack of analytical methods to detect and quantify clinically relevant levels of ciguatoxin in easily prepared crude extracts of fish. Here we report a ciguatoxin rapid extraction method (CREM) that allows the rapid preparation of fish flesh extracts for the detection and quantification of ciguatoxin by gradient reversed-phase liquid chromatography-tandem mass spectrometry (LC/MS/MS). CREM-LC/MS/MS delivers a linear response to P-CTX-1 spiked into fish prior to extraction. A similar response was obtained for P-CTX-1 spiked after extraction, indicating >95% extraction efficiency was achieved overall and 85% at the limit of quantification (0.1 ppb). Using this approach, levels >or=0.1 ppb P-CTX-1 could be detected and quantified from an extract of 2g fish flesh, making it suitable as a confirmatory assay for suspect ciguateric carnivorous fish in the Pacific Ocean. The approach is designed to simplify the extraction and analysis of multiple samples per day.

Phosphorylation of mTOR and S6RP predicts the efficacy of everolimus in patients with metastatic renal cell carcinoma

International Nuclear Information System (INIS)

Li, Siming; Kong, Yan; Si, Lu; Chi, Zhihong; Cui, Chuanliang; Sheng, Xinan; Guo, Jun

2014-01-01

The incidence of renal cell cancer (RCC) has been increasing for the past decade, and the 5-year survival for patients with metastatic RCC (mRCC) is rather low. Everolimus (RAD001), a new inhibitor for mammalian target of rapamycin (mTOR), is generally well tolerated, and demonstrates clinical benefit to patients with anti-VEGF-refractory mRCC. However, factors for selection of patients who may benefit from everolimus remain largely unknown. Here we aimed to explore potential molecular indicators for mRCC patients who may benefit from everolimus treatment. Paraffin-embedded tumor tissue specimens derived from 18 mRCC patients before everolimus treatment, who participated the phase 1b trial of everolimus in VEGF receptor (VEGFR)-tyrosine kinase inhibitor (TKI)-refractory Chinese patients with mRCC (clinicaltrials.gov, NCT01152801), were examined for the expression levels of phosphorylated AKT, mTOR, eukaryotic initiation factor 4E (eIF4E) binding protein-1 (4EBP1) and 40S ribosomal protein S6 (S6RP) by immunohistochemistry. Clinical benefit rate (complete response [CR], partial response [PR], plus stable disease [SD] ≥ 6 months) and progression-free survival time (PFS) were correlated with expression levels of these mTOR-associated molecules. In these 18 patients, there were 1 PR, 15 SDs (including 9 SDs ≥ 6 months), and 2 progressive diseases (PD). The clinical benefit rate (CBR) was 55.6% (10/18), and the median PFS time was 8.4 months. Patients with positive expression of phospho-mTOR showed a better CBR (71.4% versus 0%, P = 0.023) and PFS time (11.3 versus 3.7 months, P = 0.001) than those patients with negative expression. The median PFS of patients with positive phospho-S6RP expression was longer (11.3 versus 3.7 months, P = 0.002) than that of patients negative for phospho-S6RP expression. However, expression levels of phospho-4EBP1 and phospho-AKT were unassociated to efficacy of everolimus treatment with respect to CBR and PFS. Co-expression of

A novel electrophototrophic bacterium Rhodopseudomonas palustris strain RP2, exhibits hydrocarbonoclastic potential in anaerobic environments

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Krishnaveni Venkidusamy

2016-07-01

Full Text Available An electrophototrophic, hydrocarbonoclastic bacterium Rhodopseudomonas palustris stain RP2 was isolated from the anodic biofilms of hydrocarbon fed microbial electrochemical remediation systems (MERS. Salient properties of the strain RP2 were direct electrode respiration, dissimilatory metal oxide reduction, spore formation, anaerobic nitrate reduction, free living diazotrophy and the ability to degrade n-alkane components of petroleum hydrocarbons in anoxic, photic environments. In acetate fed microbial electrochemical cells, a maximum current density of 305±10 mA/m2 (1000Ω was generated (power density 131.65±10 mW/m2 by strain RP2 with a coulombic efficiency of 46.7 ± 1.3%. Cyclic voltammetry studies showed that anaerobically grown cells of strain RP2 is electrochemically active and likely to transfer electrons extracellularly to solid electron acceptors through membrane bound compounds, however, aerobically grown cells lacked the electrochemical activity. The ability of strain RP2 to produce current (maximum current density 21±3 mA/m2; power density 720±7 µW/m2, 1000Ω using petroleum hydrocarbon (PH as a sole energy source was also examined using an initial concentration of 800 mg l-1 of diesel range hydrocarbons (C9- C36 with a concomitant removal of 47.4 ± 2.7% hydrocarbons in MERS. Here, we also report the first study that shows an initial evidence for the existence of a hydrocarbonoclastic behavior in the strain RP2 when grown in different electron accepting and illuminated conditions (anaerobic and MERS degradation. Such observations reveal the importance of photoorganotrophic growth in the utilization of hydrocarbons from contaminated environments. Identification of such novel petrochemical hydrocarbon degrading electricigens, not only expands the knowledge on the range of bacteria known for the hydrocarbon bioremediation but also shows a biotechnological potential that goes well beyond its applications to MERS.

The Effect of Hydrogen Addition on the Combustion Characteristics of RP-3 Kerosene/Air Premixed Flames

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Wen Zeng

2017-07-01

Full Text Available Experimental studies have been performed to investigate the effects of hydrogen addition on the combustion characteristics of Chinese No.3 jet fuel (RP-3 kerosene/air premixed flames. Experiments were carried out in a constant volume chamber and the influences of the initial temperatures of 390 and 420 K, initial pressures of 0.1 and 0.3 MPa, equivalence ratios of 0.6–1.6 and hydrogen additions of 0.0–0.5 on the laminar burning velocities, and Markstein numbers of Hydrogen (H2/RP-3/air mixtures were investigated. The results show that the flame front surfaces of RP-3/air mixtures remain smooth throughout the entire flame propagation process at a temperature of 390 K, pressure of 0.3 MPa, equivalence ratio of 1.3 and without hydrogen addition, but when the hydrogen addition increases from 0.0 to 0.5 under the same conditions, flaws and protuberances occur at the flame surfaces. It was also found that with the increase of the equivalence ratio from 0.9 to 1.5, the laminar burning velocities of the mixtures increase at first and then decrease, and the highest laminar burning velocity was measured at an equivalence ratio of 1.2. Meanwhile, with the increase of hydrogen addition, laminar burning velocities of H2/RP-3/air mixtures increase. However, the Markstein numbers of H2/RP-3/air mixtures decrease with the increase of hydrogen addition, which means that the flames of H2/RP-3/air mixtures become unstable with the increase of hydrogen addition.

PAP-1, the mutated gene underlying the RP9 form of dominant retinitis pigmentosa, is a splicing factor

International Nuclear Information System (INIS)

Maita, Hiroshi; Kitaura, Hirotake; Keen, T. Jeffrey; Inglehearn, Chris F.; Ariga, Hiroyoshi; Iguchi-Ariga, Sanae M.M.

2004-01-01

PAP-1 is an in vitro phosphorylation target of the Pim-1 oncogene. Although PAP-1 binds to Pim-1, it is not a substrate for phosphorylation by Pim-1 in vivo. PAP-1 has recently been implicated as the defective gene in RP9, one type of autosomal dominant retinitis pigmentosa (adRP). However, RP9 is a rare disease and only two missense mutations have been described, so the report of a link between PAP-1 and RP9 was tentative. The precise cellular role of PAP-1 was also unknown at that time. We now report that PAP-1 localizes in nuclear speckles containing the splicing factor SC35 and interacts directly with another splicing factor, U2AF35. Furthermore, we used in vitro and in vivo splicing assays to show that PAP-1 has an activity, which alters the pattern of pre-mRNA splicing and that this activity is dependent on the phosphorylation state of PAP-1. We used the same splicing assay to examine the activities of two mutant forms of PAP-1 found in RP9 patients. The results showed that while one of the mutations, H137L, had no effect on splicing activity compared with that of wild-type PAP-1, the other, D170G, resulted in both a defect in splicing activity and a decreased proportion of phosphorylated PAP-1. The D170G mutation may therefore cause RP by altering splicing of retinal genes through a decrease in PAP-1 phosphorylation. These results demonstrate that PAP-1 has a role in pre-mRNA splicing and, given that three other splicing factors have been implicated in adRP, this finding provides compelling further evidence that PAP-1 is indeed the RP9 gene

Validation of LC-TOF-MS screening for drugs, metabolites, and collateral compounds in forensic toxicology specimens.

Science.gov (United States)

Guale, Fessessework; Shahreza, Shahriar; Walterscheid, Jeffrey P; Chen, Hsin-Hung; Arndt, Crystal; Kelly, Anna T; Mozayani, Ashraf

2013-01-01

Liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS) analysis provides an expansive technique for identifying many known and unknown analytes. This study developed a screening method that utilizes automated solid-phase extraction to purify a wide array of analytes involving stimulants, benzodiazepines, opiates, muscle relaxants, hypnotics, antihistamines, antidepressants and newer synthetic "Spice/K2" cannabinoids and cathinone "bath salt" designer drugs. The extract was applied to LC-TOF-MS analysis, implementing a 13 min chromatography gradient with mobile phases of ammonium formate and methanol using positive mode electrospray. Several common drugs and metabolites can share the same mass and chemical formula among unrelated compounds, but they are structurally different. In this method, the LC-TOF-MS was able to resolve many isobaric compounds by accurate mass correlation within 15 ppm mass units and a narrow retention time interval of less than 10 s of separation. Drug recovery yields varied among spiked compounds, but resulted in overall robust area counts to deliver an average match score of 86 when compared to the retention time and mass of authentic standards. In summary, this method represents a rapid, enhanced screen for blood and urine specimens in postmortem, driving under the influence, and drug facilitated sexual assault forensic toxicology casework.

Top-down approach for the direct characterization of low molecular weight heparins using LC-FT-MS.

Science.gov (United States)

Li, Lingyun; Zhang, Fuming; Zaia, Joseph; Linhardt, Robert J

2012-10-16

Low molecular heparins (LMWHs) are structurally complex, heterogeneous, polydisperse, and highly negatively charged mixtures of polysaccharides. The direct characterization of LMWH is a major challenge for currently available analytical technologies. Electrospray ionization (ESI) liquid chromatography-mass spectrometry (LC-MS) is a powerful tool for the characterization complex biological samples in the fields of proteomics, metabolomics, and glycomics. LC-MS has been applied to the analysis of heparin oligosaccharides, separated by size exclusion, reversed phase ion-pairing chromatography, and chip-based amide hydrophilic interaction chromatography (HILIC). However, there have been limited applications of ESI-LC-MS for the direct characterization of intact LMWHs (top-down analysis) due to their structural complexity, low ionization efficiency, and sulfate loss. Here we present a simple and reliable HILIC-Fourier transform (FT)-ESI-MS platform to characterize and compare two currently marketed LMWH products using the top-down approach requiring no special sample preparation steps. This HILIC system relies on cross-linked diol rather than amide chemistry, affording highly resolved chromatographic separations using a relatively high percentage of acetonitrile in the mobile phase, resulting in stable and high efficiency ionization. Bioinformatics software (GlycReSoft 1.0) was used to automatically assign structures within 5-ppm mass accuracy.

Rugged Large Volume Injection for Sensitive Capillary LC-MS Environmental Monitoring

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Hanne Roberg-Larsen

2017-08-01

Full Text Available A rugged and high throughput capillary column (cLC LC-MS switching platform using large volume injection and on-line automatic filtration and filter back-flush (AFFL solid phase extraction (SPE for analysis of environmental water samples with minimal sample preparation is presented. Although narrow columns and on-line sample preparation are used in the platform, high ruggedness is achieved e.g., injection of 100 non-filtrated water samples did not result in a pressure rise/clogging of the SPE/capillary columns (inner diameter 300 μm. In addition, satisfactory retention time stability and chromatographic resolution were also features of the system. The potential of the platform for environmental water samples was demonstrated with various pharmaceutical products, which had detection limits (LOD in the 0.05–12.5 ng/L range. Between-day and within-day repeatability of selected analytes were <20% RSD.

The retinitis pigmentosa-mutated RP2 protein exhibits exonuclease activity and translocates to the nucleus in response to DNA damage

International Nuclear Information System (INIS)

Yoon, Jung-Hoon; Qiu Junzhuan; Cai Sheng; Chen Yuan; Cheetham, Michael E.; Shen Binghui; Pfeifer, Gerd P.

2006-01-01

Retinitis pigmentosa (RP) is a genetically heterogeneous disease characterized by degeneration of the retina. Mutations in the RP2 gene are linked to the second most frequent form of X-linked retinitis pigmentosa. RP2 is a plasma membrane-associated protein of unknown function. The N-terminal domain of RP2 shares amino acid sequence similarity to the tubulin-specific chaperone protein co-factor C. The C-terminus consists of a domain with similarity to nucleoside diphosphate kinases (NDKs). Human NDK1, in addition to its role in providing nucleoside triphosphates, has recently been described as a 3' to 5' exonuclease. Here, we show that RP2 is a DNA-binding protein that exhibits exonuclease activity, with a preference for single-stranded or nicked DNA substrates that occur as intermediates of base excision repair pathways. Furthermore, we show that RP2 undergoes re-localization into the nucleus upon treatment of cells with DNA damaging agents inducing oxidative stress, most notably solar simulated light and UVA radiation. The data suggest that RP2 may have previously unrecognized roles as a DNA damage response factor and 3' to 5' exonuclease

Analysis of nicotine and cotinine in hair by on-line in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry as biomarkers of exposure to tobacco smoke.

Science.gov (United States)

Inukai, Takehito; Kaji, Sanae; Kataoka, Hiroyuki

2018-07-15

Smoking not only increases the risk of lung cancer but is strongly related to the onset of cardiovascular disease. Particularly, passive smoking due to sidestream smoke is a critical public health problem. To assess active and passive exposure to tobacco smoke, we developed a simple and sensitive method, consisting of on-line in-tube solid phase microextraction (SPME) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS), to determine nicotine and its metabolite cotinine in hair samples. These compounds were separated within 5 min using a Polar-RP80A column and detected in the positive ion mode by multiple reaction monitoring. The optimum in-tube SPME conditions were 25 draw/eject cycles of 40 μL of sample at a flow rate of 200 μL/min using a Carboxen 1006 capillary column as an extraction device. The extracted compounds in the stationary phase on the inner wall of the capillary could be dissolved easily into the mobile phase and transferred to an LC column. Using the in-tube SPME LC-MS/MS method, the calibration curves were linear in the 5-1000 pg/mL ranges for nicotine and cotinine, and the detection limits (signal to noise ratio of 3) were 0.45 and 0.13 pg/mL, respectively. The intra-day and inter-day precisions were below 3.4% and 6.0% (n = 5), respectively. This method was utilized successfully to analyze pg/mg levels of nicotine and cotinine in 1 mg of hairs without interference peaks, and good recoveries were obtained. The concentration of cotinine in hair was two orders of magnitude lower than that of nicotine, but a good positive correlation was found between the concentrations of these compounds. This method can automate the extraction, concentration and analysis of samples, and is useful for the assessment of long-term exposure to tobacco smoke. Copyright © 2018 Elsevier B.V. All rights reserved.

Comparative analysis of native and permethylated human milk oligosaccharides by liquid chromatography coupled to high resolution mass spectrometry.

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Oursel, Stéphanie; Cholet, Sophie; Junot, Christophe; Fenaille, François

2017-12-15

Human milk oligosaccharides (HMOs) represent the third most abundant components of milk after lactose and lipids. HMOs are indigestible by the suckling infant but can act as prebiotics and have significant biological functions regarding the organism defense against pathogens (such as bacteria or viruses) by preventing interactions with their receptors. Although constituted of only five distinct monosaccharide building blocks, HMOs are highly structurally diverse compounds with many co-existing structural isomers. Here we report the development and comparison of two distinct glycomic platforms based on liquid chromatography coupled to high resolution mass spectrometry (LC-MS) for analyzing HMOs. We have implemented and thoroughly compared the LC-MS of permethylated and native HMOs on reversed phase (RP) and porous graphitic carbon (PGC) columns for their ability to resolve the natural heterogeneity of milk oligosaccharides at the highest sensitivity. Our data essentially underlines the usefulness of analyzing HMOs as permethylated derivatives especially for getting more precise structural information at high sensitivity. For instance, permethylation annihilates gas-phase fucose migration during MS/MS experiments, thus facilitating spectra interpretation and giving access to relevant information regarding oligosaccharide branching and isomer distinction. At the opposite, LC-MS profiling of native HMOs (using PGC) in milk performed best in terms of detected species, while also being much faster in terms of sample preparation. Although less efficient than PGC chromatography, RPLC proved successful for separating pairs of permethylated isomeric HMOs. A key advantage of RP over PGC liquid chromatography is that retention times can be correlated to molecular weights, which can greatly facilitate further HMO identification using retention time prediction. Altogether these data lead us to think that LC-MS analysis of native HMOs (using PGC) can be used as first

A Study of Method Development, Validation, and Forced Degradation for Simultaneous Quantification of Paracetamol and Ibuprofen in Pharmaceutical Dosage Form by RP-HPLC Method

OpenAIRE

Jahan, Md. Sarowar; Islam, Md. Jahirul; Begum, Rehana; Kayesh, Ruhul; Rahman, Asma

2014-01-01

A rapid and stability-indicating reversed phase high-performance liquid chromatography (RP-HPLC) method was developed for simultaneous quantification of paracetamol and ibuprofen in their combined dosage form especially to get some more advantages over other methods already developed for this combination. The method was validated according to United States Pharmacopeia (USP) guideline with respect to accuracy, precision, specificity, linearity, solution stability, robustness, sensitivity, and...

Determination of two capsaicinoids in analgesic transdermal patches using RP-HPLC and UV spectroscopy

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F. Kobarfard

2017-11-01

Full Text Available Background and objectives: At the present time, a considerable frontier in the administration of therapeutic medications is transdermal drug delivery. Methods: In this study, a rapid, precise, sensitive and selective reversed-phasehigh performance liquid chromatography (RP-HPLC method has been evaluated, developed and validated to separate and quantitate capsaicin and dihydrocapsaicin (main active agents in analgesic dermal patches produced in Iran. Results: After isolation from laminated adhesive patches, capsaicinoids were analyzed on Lichrospher C18 analytical columns with reversed phase, using a mobile phase composition of methanol and distilled water (70:30 v/v and without any buffer (pH=6.5. The flow rate was 1 mL/min and the UV detector was operating at 281 nm. The assay was found to be linear over the range of 0.1-1.0 mg/mL. All validation parameters were within the acceptable range. Conclusion: It seems that the developed method was fairly sensitive and reliable in measuring capsaicinoids in commercially available analgesic transdermal patches in Iran.

A simple RP-HPLC method for related substances of zoledronic acid in pharmaceutical products

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L. Maheswara Reddy

2017-02-01

Full Text Available A novel, selective and sensitive reverse phase-high performance liquid chromatography (RP-HPLC method has been developed for the validated estimation of imidazol-1-yl-acetic acid in zoledronic acid formulations. The separation was achieved on a 5 μ C18 column (250 × 4.6 mm using a mobile phase that consists of the buffer (4.5 g of di-potassium hydrogen phosphate anhydrous and 2.0 g of tetra butyl ammonium hydrogen sulphate (TBAHS in 1000 mL of water and methanol in the ratio of 900:100 v/v. The flow rate was maintained at 1.0 mL min−1. The detection of the constituents was done at 215 nm using a UV detector. The retention times of imidazol-1-yl-acetic acid and zoledronic acid were 7.2 and 10.2 min respectively. Recovery studies were satisfactory and the correlation coefficient, 0.999 indicates linearity of the method within the limits. The developed method can be applicable for regular qualitative analysis.

β-Decay half-lives and nuclear structure of exotic proton-rich waiting point nuclei under rp-process conditions

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Nabi, Jameel-Un; Böyükata, Mahmut

2016-03-01

We investigate even-even nuclei in the A ∼ 70 mass region within the framework of the proton-neutron quasi-particle random phase approximation (pn-QRPA) and the interacting boson model-1 (IBM-1). Our work includes calculation of the energy spectra and the potential energy surfaces V (β , γ) of Zn, Ge, Se, Kr and Sr nuclei with the same proton and neutron number, N = Z. The parametrization of the IBM-1 Hamiltonian was performed for the calculation of the energy levels in the ground state bands. Geometric shape of the nuclei was predicted by plotting the potential energy surfaces V (β , γ) obtained from the IBM-1 Hamiltonian in the classical limit. The pn-QRPA model was later used to compute half-lives of the neutron-deficient nuclei which were found to be in very good agreement with the measured ones. The pn-QRPA model was also used to calculate the Gamow-Teller strength distributions and was found to be in decent agreement with the measured data. We further calculate the electron capture and positron decay rates for these N = Z waiting point (WP) nuclei in the stellar environment employing the pn-QRPA model. For the rp-process conditions, our total weak rates are within a factor two compared with the Skyrme HF +BCS +QRPA calculation. All calculated electron capture rates are comparable to the competing positron decay rates under rp-process conditions. Our study confirms the finding that electron capture rates form an integral part of the weak rates under rp-process conditions and should not be neglected in the nuclear network calculations.

75 FR 65366 - Recovery Policy RP9524.2, Landslides and Slope Stability Related to Public Facilities

Science.gov (United States)

2010-10-22

...] Recovery Policy RP9524.2, Landslides and Slope Stability Related to Public Facilities AGENCY: Federal... the final Recovery Policy RP9524.2, Landslides and Slope Stability Related to Public Facilities, which... facilities threatened by landslides or slope failures; as well as the eligibility of permanent repairs to...

75 FR 29569 - Recovery Policy RP9526.1, Hazard Mitigation Funding Under Section 406 (Stafford Act)

Science.gov (United States)

2010-05-26

...] Recovery Policy RP9526.1, Hazard Mitigation Funding Under Section 406 (Stafford Act) AGENCY: Federal... the final Recovery Policy RP9526.1, Hazard Mitigation Funding Under Section 406 (Stafford Act), which... mitigation discretionary funding available under Section 406 of the Robert T. Stafford Disaster Relief and...

On-line coupled LC-LC-GC for irradiation detection in complex lipid matrices

International Nuclear Information System (INIS)

Schulzki, G.; Spiegelberg, A.; Helle, N.; Boegl, K.W.; Schreiber, G.A.

1993-01-01

Since sample preparation with HPLC coupled on-line to the GC has been performed for only a few weeks in our laboratory, the results presented give a first look at what can be done by means of this technique. Even difficult samples as the described fish species, where an unequivocal identification regarding an irradiation treatment seemed to become a hopeless enterprise, could be managed. Because of the greater variety of fatty acids in fish ''new'' radiation-induced hydrocarbons were available. According to the theory of Nawar in addition to 16:2 and 17:2 hydrocarbons we have looked for in irradiated meat, further alkadienes appeared in irradiated fish, which were 14:2, 18:2 and 20:2. Analysis of the alkadiene-fraction, transferred to the GC after a two step LC clean up, resulted in an unequivocal identification of all fish samples as well as the fruits and sponge cake. For fruits and sponge cake the detection limit seems to be clearly below 0.5 kGy. It can further be lowered by increasing the amount of lipid whereas the upper limit for a certain LC column has to be determined. In contrast to these samples only qualitative results were obtained for fish. In the case of sponge cake for the first time irradiation of a component of a heat treated food was detected. Further investigations regarding reproducibility, dose dependence and detection limit have to be done. On-line coupled (LC-)LC-GC was proved to be a highly efficient method for analysis of complex samples. In contrast to off-line Florisil column chromatography only a small part of the initial lipid material is needed because the complete hydrocarbon fraction is transferred on-line to the GC. This offers the possibility to analyze even foods with a low fat content like various seafoods. Classification of the hydrocarbon fraction by a two step LC may facilitate the identification of the radiolytic products also if no mass spectrometric detection system is available. (orig./vhe)

The Relevance of AgRP Neuron-Derived GABA Inputs to POMC Neurons Differs for Spontaneous and Evoked Release

OpenAIRE

Rau, Andrew R.; Hentges, Shane T.

2017-01-01

Hypothalamic agouti-related peptide (AgRP) neurons potently stimulate food intake, whereas proopiomelanocortin (POMC) neurons inhibit feeding. Whether AgRP neurons exert their orexigenic actions, at least in part, by inhibiting anorexigenic POMC neurons remains unclear. Here, the connectivity between GABA-releasing AgRP neurons and POMC neurons was examined in brain slices from male and female mice. GABA-mediated spontaneous IPSCs (sIPSCs) in POMC neurons were unaffected by disturbing GABA re...

Development and validation of a stability-indicating RP-HPLC method for determination of atomoxetine hydrochloride in tablets.

Science.gov (United States)

Patel, Sejal K; Patel, Natvarlal J

2010-01-01

This paper describes the development of a stability-indicating RP-HPLC method for the determination of atomoxetine hydrochloride (ATX) in the presence of its degradation products generated from forced decomposition studies. The drug substance was subjected to stress conditions of acid, base, oxidation, wet heat, dry heat, and photodegradation. In stability tests, the drug was susceptible to acid, base, oxidation, and dry and wet heat degradation. It was found to be stable under the photolytic conditions tested. The drug was successfully separated from the degradation products formed under stress conditions on a Phenomenex C18 column (250 x 4.6 mm id, 5 microm particle size) by using acetonitrile-methanol-0.032 M ammonium acetate (55 + 05 + 40, v/v/v) as the mobile phase at 1.0 mL/min and 40 degrees C. Photodiode array detection at 275 nm was used for quantitation after RP-HPLC over the concentration range of 0.5-5 microg/mL with a mean recovery of 100.8 +/- 0.4% for ATX. Statistical analysis demonstrated that the method is repeatable, specific, and accurate for the estimation of ATX. Because the method effectively separates the drug from its degradation products, it can be used as a stability-indicating method.

Phase effects in masking by harmonic complexes: speech recognition.

Science.gov (United States)

Deroche, Mickael L D; Culling, John F; Chatterjee, Monita

2013-12-01

Harmonic complexes that generate highly modulated temporal envelopes on the basilar membrane (BM) mask a tone less effectively than complexes that generate relatively flat temporal envelopes, because the non-linear active gain of the BM selectively amplifies a low-level tone in the dips of a modulated masker envelope. The present study examines a similar effect in speech recognition. Speech reception thresholds (SRTs) were measured for a voice masked by harmonic complexes with partials in sine phase (SP) or in random phase (RP). The masker's fundamental frequency (F0) was 50, 100 or 200 Hz. SRTs were considerably lower for SP than for RP maskers at 50-Hz F0, but the two converged at 100-Hz F0, while at 200-Hz F0, SRTs were a little higher for SP than RP maskers. The results were similar whether the target voice was male or female and whether the masker's spectral profile was flat or speech-shaped. Although listening in the masker dips has been shown to play a large role for artificial stimuli such as Schroeder-phase complexes at high levels, it contributes weakly to speech recognition in the presence of harmonic maskers with different crest factors at more moderate sound levels (65 dB SPL). Copyright © 2013 Elsevier B.V. All rights reserved.

A Validated RP-HPLC Method for the Determination of Atazanavir in Pharmaceutical Dosage Form

Directory of Open Access Journals (Sweden)

K. Srinivasu

2011-01-01

Full Text Available A validated RP HPLC method for the estimation of atazanavir in capsule dosage form on YMC ODS 150 × 4.6 mm, 5 μ column using mobile phase composition of ammonium dihydrogen phosphate buffer (pH 2.5 with acetonitrile (55:45 v/v. Flow rate was maintained at 1.5 mL/min with 288 nm UV detection. The retention time obtained for atazanavir was at 4.7 min. The detector response was linear in the concentration range of 30 - 600 μg/mL. This method has been validated and shown to be specific, sensitive, precise, linear, accurate, rugged, robust and fast. Hence, this method can be applied for routine quality control of atazanavir in capsule dosage forms as well as in bulk drug.

One-pot preparation of a mixed-mode organic-silica hybrid monolithic capillary column and its application in determination of endogenous gibberellins in plant tissues.

Science.gov (United States)

Zhang, Zheng; Hao, Yan-Hong; Ding, Jun; Xu, Sheng-Nan; Yuan, Bi-Feng; Feng, Yu-Qi

2015-10-16

A newly improved one-pot method, based on "thiol-ene" click chemistry and sol-gel approach in microemulsion system, was developed for the preparation of C8/PO(OH)2-silica hybrid monolithic capillary column. The prepared monolith possesses large specific surface area, narrow mesopore size distribution and high column efficiency. The monolithic column was demonstrated to have cation exchange/reversed-phase (CX/RP) mixed-mode retention for analytes on nano-liquid chromatography (nano-LC). On the basis of the developed nano-LC system with MS detector coupled to pipette tip solid phase extraction (PT-SPE) and derivatization process, we then realized simultaneous determination of 10 gibberellins (GAs) with low limits of detection (LODs, 0.003-0.025 ng/mL). Furthermore, 6 endogenous GAs in only 5mg rice leaves (fresh weight) were successfully detected and quantified. The developed PT-SPE-nano-LC-MS strategy may offer promising applications in the determination of low abundant bioactive molecules from complex matrix. Copyright © 2015 Elsevier B.V. All rights reserved.

AMPK is essential for energy homeostasis regulation and glucose sensing by POMC and AgRP neurons.

Science.gov (United States)

Claret, Marc; Smith, Mark A; Batterham, Rachel L; Selman, Colin; Choudhury, Agharul I; Fryer, Lee G D; Clements, Melanie; Al-Qassab, Hind; Heffron, Helen; Xu, Allison W; Speakman, John R; Barsh, Gregory S; Viollet, Benoit; Vaulont, Sophie; Ashford, Michael L J; Carling, David; Withers, Dominic J

2007-08-01

Hypothalamic AMP-activated protein kinase (AMPK) has been suggested to act as a key sensing mechanism, responding to hormones and nutrients in the regulation of energy homeostasis. However, the precise neuronal populations and cellular mechanisms involved are unclear. The effects of long-term manipulation of hypothalamic AMPK on energy balance are also unknown. To directly address such issues, we generated POMC alpha 2KO and AgRP alpha 2KO mice lacking AMPK alpha2 in proopiomelanocortin- (POMC-) and agouti-related protein-expressing (AgRP-expressing) neurons, key regulators of energy homeostasis. POMC alpha 2KO mice developed obesity due to reduced energy expenditure and dysregulated food intake but remained sensitive to leptin. In contrast, AgRP alpha 2KO mice developed an age-dependent lean phenotype with increased sensitivity to a melanocortin agonist. Electrophysiological studies in AMPK alpha2-deficient POMC or AgRP neurons revealed normal leptin or insulin action but absent responses to alterations in extracellular glucose levels, showing that glucose-sensing signaling mechanisms in these neurons are distinct from those pathways utilized by leptin or insulin. Taken together with the divergent phenotypes of POMC alpha 2KO and AgRP alpha 2KO mice, our findings suggest that while AMPK plays a key role in hypothalamic function, it does not act as a general sensor and integrator of energy homeostasis in the mediobasal hypothalamus.

Transduced PEP-1-ribosomal protein S3 (rpS3) ameliorates 12-O-tetradecanoylphorbol-13-acetate-induced inflammation in mice

International Nuclear Information System (INIS)

Ahn, Eun Hee; Kim, Dae Won; Kang, Hye Won; Shin, Min Jae; Won, Moo Ho; Kim, Joon; Kim, Dong Joon; Kwon, Oh-Shin; Kang, Tae-Cheon; Han, Kyu Hyung; Park, Jinseu; Eum, Won Sik; Choi, Soo Young

2010-01-01

This study investigated the preventive effect of ribosomal protein S3 (rpS3) on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema in mice. A cell permeable expression vector PEP-1-rpS3 was constructed. Topical application of the vector markedly inhibited TPA-induced expression levels of cyclooxygenase-2 (COX-2) and pro-inflammatory cytokines. Application of PEP-1-rpS3 also resulted in a significant reduction in the activation of nuclear factor-kappa B (NF-kB) and mitogen-activated protein kinase (MAPK) in TPA-treated ears. These results indicate that PEP-1-rpS3 inhibits inflammatory response cytokines and enzymes by blocking NF-kB and MAPK, prompting the suggestion that PEP-1-rpS3 can be used as a therapeutic agent against skin inflammation.

Separation of thorium (IV) from lanthanide concentrate (LC) and water leach purification (WLP) residue

International Nuclear Information System (INIS)

AL-Areqi, Wadeeah M.; Majid, Amran Ab.; Sarmani, Sukiman

2014-01-01

Thorium (IV) content in industrial residue produced from rare earth elements production industry is one of the challenges to Malaysian environment. Separation of thorium from the lanthanide concentrate (LC) and Water Leach Purification (WLP) residue from rare earth elements production plant is described. Both materials have been tested by sulphuric acid and alkaline digestions. Th concentrations in LC and WLP were determined to be 1289.7 ± 129 and 1952.9±17.6 ppm respectively. The results of separation show that the recovery of Th separation from rare earth in LC after concentrated sulphuric acid dissolution and reduction of acidity to precipitate Th was found 1.76-1.20% whereas Th recovery from WLP was less than 4% after concentrated acids and alkali digestion processes. Inductively Coupled Plasma-Mass Spectroscopy (ICP-MS) was used to determine Th concentrations in aqueous phase during separation stages. This study indicated that thorium maybe exists in refractory and insoluble form which is difficult to separate by these processes and stays in WLP residue as naturally occurring radioactive material (NORM)

Investigating the Retention Mechanisms of Liquid Chromatography Using Solid-Phase Extraction Cartridges

Science.gov (United States)

O'Donnell, Mary E.; Musial, Beata A.; Bretz, Stacey Lowery; Danielson, Neil D.; Ca, Diep

2009-01-01

Liquid chromatography (LC) experiments for the undergraduate analytical laboratory course often illustrate the application of reversed-phase LC to solve a separation problem, but rarely compare LC retention mechanisms. In addition, a high-performance liquid chromatography instrument may be beyond what some small colleges can purchase. Solid-phase…

Accounting for inertia in modal choices: some new evidence using a RP/SP dataset

DEFF Research Database (Denmark)

Cherchi, Elisabetta; Manca, Francesco

2011-01-01

effect is stable along the SP experiments. Inertia has been studied more extensively with panel datasets, but few investigations have used RP/SP datasets. In this paper we extend previous work in several ways. We test and compare several ways of measuring inertia, including measures that have been...... proposed for both short and long RP panel datasets. We also explore new measures of inertia to test for the effect of “learning” (in the sense of acquiring experience or getting more familiar with) along the SP experiment and we disentangle this effect from the pure inertia effect. A mixed logit model...... is used that allows us to account for both systematic and random taste variations in the inertia effect and for correlations among RP and SP observations. Finally we explore the relation between the utility specification (especially in the SP dataset) and the role of inertia in explaining current choices....

In-situ phase transition from microemulsion to liquid crystal with the potential of prolonged parenteral drug delivery.

Science.gov (United States)

Ren, Xiazhong; Svirskis, Darren; Alany, Raid G; Zargar-Shoshtari, Sara; Wu, Zimei

2012-07-15

This study is the first to investigate and demonstrate the potential of microemulsions (MEs) for sustained release parenteral drug delivery, due to phase transition behavior in aqueous environments. Phase diagrams were constructed with Miglyol 812N oil and a blend of (co)surfactants Solutol HS 15 and Span 80 with ethanol. Liquid crystal (LC) and coarse emulsion (CE) regions were found adjacent to the ME region in the water-rich corner of the phase diagram. Two formulations were selected, a LC-forming ME and a CE-forming ME and each were investigated with respect to their rheology, particle size, drug release profiles and particularly, the phase transition behavior. The spreadability in an aqueous environment was determined and release profiles from MEs were generated with gamma-scintigraphy. The CE-forming ME dispersed readily in an aqueous environment, whereas the LC-forming ME remained in a contracted region possibly due to the transition of ME to LC at the water/ME interface. Gamma-scintigraphy showed that the LC-forming ME had minimal spreadability and a slow release of (99m)Tc in the first-order manner, suggesting phase conversion at the interface. In conclusion, owing to the potential of phase transition, LC-forming MEs could be used as extravascular injectable drug delivery vehicles for prolonged drug release. Copyright © 2012 Elsevier B.V. All rights reserved.

Low Thermal Conductivity of RE-Doped SrO(SrTiO3)1 Ruddlesden Popper Phase Bulk Materials Prepared by Molten Salt Method

Science.gov (United States)

Putri, Yulia Eka; Said, Suhana Mohd; Refinel, Refinel; Ohtaki, Michitaka; Syukri, Syukri

2018-04-01

The SrO(SrTiO3)1 (Sr2TiO4) Ruddlesden Popper (RP) phase is a natural superlattice comprising of alternately stacking perovskite-type SrTiO3 layers and rock salt SrO layers along the crystallographic c direction. This paper discusses the properties of the Sr2TiO4 and (La, Sm)-doped Sr2TiO4 RP phase synthesized via molten salt method, within the context of thermoelectric applications. A good thermoelectric material requires high electrical conductivity, high Seebeck coefficient and low thermal conductivity. All three conditions have the potential to be fulfilled by the Sr2TiO4 RP phase, in particular, the superlattice structure allows a higher degree of phonon scattering hence resulting in lowered thermal conductivity. In this work, the Sr2TiO4 RP phase is doped with Sm and La respectively, which allows injection of charge carriers, modification of its electronic structure for improvement of the Seebeck coefficient, and most significantly, reduction of thermal conductivity. The particles with submicron size allows excessive phonon scattering along the boundaries, thus reduces the thermal conductivity by fourfold. In particular, the Sm-doped sample exhibited even lower lattice thermal conductivity, which is believed to be due to the mismatch in the ionic radius of Sr and Sm. This finding is useful as a strategy to reduce thermal conductivity of Sr2TiO4 RP phase materials as thermoelectric candidates, by employing dopants of differing ionic radius.

Photogalvanic self-pulsing of electro-wetting of LC droplet on LN-crystal

International Nuclear Information System (INIS)

Kukhtarev, N; Kukhtareva, T; Geng, J; Zhang, XZ; Xu, JJ

2017-01-01

Photogalvanic current photoinduced by CW laser illumination of Fe-doped LN reveals quasi-periodic pulsing due to microplasmas discharges. This transformation of constant external influence into periodic response is interesting as example of self-excited systems, resembling pulsations of biological systems (including heart beats). Microplasmas is also interesting for potential applications in various fields, such as surface treatment, sterilization, water splitting, light sources, micro jets. Self-pulsing regime of microplasmas discharges in microhollow cathode discharges was observed and modeled in plasmas physics. In this paper, we analyze self-pulsing of photogalvanic current visualized by dynamic electrowetting in a simple LC-cell, formed by a nematic LC droplet placed on Fe-doped LN crystal. We have developed model of self-pulsing regime based on nonlinear discharge resistance. Experimental phase-space plot may be used to find parameters of the nonlinear discharge resistance. In the dynamic modeling contributions from both photogalvanic and pyroelectric effect are included. The analysis of dynamic regime reveals that a current pulse (in microsecond range) have asymmetric shape with extremely sharp rise with longer decay time. These self-pulsations are visualized by reflection of laser light from a LC droplet placed on LN-crystal. Reflected interference patterned was modulated by the dynamic electro-wetting effect. (paper)

Nuclear binding around the RP-process waiting points $^{68}$Se and $^{72}$Kr

CERN Multimedia

2002-01-01

Encouraged by the success of mass determinations of nuclei close to the Z=N line performed at ISOLTRAP during the year 2000 and of the recent decay spectroscopy studies on neutron-deficient Kr isotopes (IS351 collaboration), we aim to measure masses and proton separation energies of the bottleneck nuclei defining the flow of the astrophysical rp-process beyond A$\\sim$70. In detail, the program includes mass measurements of the rp-process waiting point nuclei $^{68}$Se and $^{72}$Kr and determination of proton separation energies of the proton-unbound $^{69}$Br and $^{73}$Rb via $\\beta$-decays of $^{69}$Kr and $^{73}$Sr, respectively. The aim of the project is to complete the experimental database for astrophysical network calculations and for the liquid-drop type of mass models typically used in the modelling of the astrophysical rp process in the region. The first beamtime is scheduled for the August 2001 and the aim is to measure the absolute mass of the waiting-point nucleus $^{72}$Kr.

Weak interaction rates for Kr and Sr waiting-point nuclei under rp-process conditions

International Nuclear Information System (INIS)

Sarriguren, P.

2009-01-01

Weak interaction rates are studied in neutron deficient Kr and Sr waiting-point isotopes in ranges of densities and temperatures relevant for the rp process. The nuclear structure is described within a microscopic model (deformed QRPA) that reproduces not only the half-lives but also the Gamow-Teller strength distributions recently measured. The various sensitivities of the decay rates to both density and temperature are discussed. Continuum electron capture is shown to contribute significantly to the weak rates at rp-process conditions.

Chromatographic lipophilicity determination using large volume injections of the solvents non-miscible with the mobile phase.

Science.gov (United States)

Sârbu, Costel; Naşcu-Briciu, Rodica Domnica; Casoni, Dorina; Kot-Wasik, Agata; Wasik, Andrzej; Namieśnik, Jacek

2012-11-30

A new perspective in the lipophilicity evaluation through RP-HPLC is permitted by analysis of the retention factor (k) obtained by injecting large volumes of test samples prepared in solvents immiscible with mobile phase. The experiment is carried out on representative groups of compounds with increased toxicity (mycotoxins and alkaloids) and amines with important biological activity (naturally occurring monoamine compounds and related drugs), which are covering a large interval of lipophilicity. The stock solution of each compound was prepared in hexane and the used mobile phases were mixtures of methanol or acetonitrile and water, in suited volume ratio. The injected volume was between 10 and 100 μL, while the used stationary phases were RP-18 and RP-8. On both reverse stationary phases the retention factors were linearly decreasing while the injection volume was increasing. In all cases, the linear models were highly statistically significant. On the basis of the obtained results new lipophilicity indices were purposed and discussed. The developed lipophilicity indices and the computationally expressed ones are correlated at a high level of statistical significance. Copyright © 2012 Elsevier B.V. All rights reserved.

A solid-phase technique for preparation of no-carrier-added technetium-99m radiopharmaceuticals: application to the streptavidin/biotin system

International Nuclear Information System (INIS)

Dunn-Dufault, Robert; Pollak, Alfred; Fitzgerald, Jane; Thornback, John R.; Ballinger, James R.

2000-01-01

A high effective specific activity (HESA) formulation of a biotin-containing 99m Tc ligand [RP488: dimethyl-Gly-Ser-Cys(Acm)-Lys(Biotin)-Gly] conveniently prepared from solid phase was compared to a typical low effective specific activity (LESA) solution formulation to demonstrate improved targeting to streptavidin in an in vitro assay and in an in vivo rat model. RP488 was coupled to a maleimide-functionalized polyethylene glycol resin via a thiol ether linkage and labeled with 99m Tc-gluconate at room temperature, followed by elution of the HESA 99m Tc-RP488 in saline (minimum specific activity ∼ 1000 TBq/mmol by amino acid analysis). Both HESA and LESA 99m Tc-RP488 labeled at > 90% purity. In vitro, HESA 99m Tc-RP488 incubated with streptavidin-agarose was bound quantitatively, but there was competition from addition of increasing amounts of cold RP488. In rats, radiotracer uptake was evident at the site of implantation of streptavidin-agarose beads for the HESA dose, less uptake of low effective specific activity (LESA) material, and no appreciable uptake in the control rats of the LESA or HESA dose. The target-to-background ratio for HESA 99m Tc-RP488 was 5.4 times that of the control. The solid-phase technology offers a convenient way to prepare high specific activity receptor-targeting 99m Tc radiopharmaceuticals

Rare itemsets mining algorithm based on RP-Tree and spark framework

Science.gov (United States)

Liu, Sainan; Pan, Haoan

2018-05-01

For the issues of the rare itemsets mining in big data, this paper proposed a rare itemsets mining algorithm based on RP-Tree and Spark framework. Firstly, it arranged the data vertically according to the transaction identifier, in order to solve the defects of scan the entire data set, the vertical datasets are divided into frequent vertical datasets and rare vertical datasets. Then, it adopted the RP-Tree algorithm to construct the frequent pattern tree that contains rare items and generate rare 1-itemsets. After that, it calculated the support of the itemsets by scanning the two vertical data sets, finally, it used the iterative process to generate rare itemsets. The experimental show that the algorithm can effectively excavate rare itemsets and have great superiority in execution time.

On-the-job training as new element in RP training: experiences from the ENETRAP pilot module in Karlsruhe

International Nuclear Information System (INIS)

Moebius, Siegurd; Bickel, Angela; Schmitt-Hannig, Annemarie; Coeck, Michele

2008-01-01

A suitable qualification for responsible personnel in Radiation Protection (RP) must be in general a combination of theoretical knowledge, and the ability (competency) to practice RP. While the theoretical knowledge is acquired by suitable education and by attending training courses, competency and skills can only be obtained by appropriate on-the-job training (OJT) followed by a period of work experience. From the feedback of questionnaires from 30 EU countries within the framework of the EU supported ENETRAP project it can be concluded that OJT provides better chances for future job opportunities and increases international flexibility. As result we recommend covering OJT together with education and training in the Basic Safety Standards and their guidelines for implementation. OJT should be specified by its content (syllabus, learning objectives), availability of necessary facilities and infrastructures as precondition for OJT, assessment of the competence of the participant, format of certificate, recognition of OJT, and responsibilities of host organisation and trainees. OJT should remain a key element in the remodelled 'European RP Training'. Based on these recommendations a two weeks training module on 'Occupational RP: Specificities of Waste Management and Decommissioning' designed for radiation protection professionals has been compiled at the Karlsruhe Training Centre FTU. While the first week of the pilot course focuses on the theoretical knowledge ranging from waste classification to decontamination techniques and transport of radioactive materials, the second week is addressed to practical training as OJT from clearance of radioactive waste, operative RP in the Decontamination Department to RP work in a Research Reactor under decommissioning. Special emphasis is devoted to RP aspects and active involvement of the participants in workshops and case studies. The results of the pilot run with participants from 8 different European countries are reported

Development and Validation of RP-HPLC Method for the Determination of Adefovir Dipivoxil in Bulk and in Pharmaceutical Formulation

Directory of Open Access Journals (Sweden)

Zaheer Ahmed

2009-01-01

Full Text Available A rapid and sensitive RP-HPLC method with UV detection (262 nm for routine analysis of adefovir dipivoxil in bulk and in pharmaceutical formulation was developed. Chromatography was performed with mobile phase containing a mixture of acetonitrile and phosphate buffer (50:50, v/v with flow rate 1.0 mL min-l. In the range of 5.0-100 µg/mL, the linearity of adefovir dipivoxil shows a correlation co-efficient of 0.9999. The proposed method was validated by determining sensitivity accuracy, precision, robustness stability, specificity, selectivity and system suitability parameters.

Development, Validation and Application of RP-HPLC Method: Simultaneous Determination of Antihistamine and Preservatives with Paracetamol in Liquid Formulations and Human Serum

OpenAIRE

Hasan, Najmul; Chaiharn, Mathurot; Toor, Umair Ali; Mirani, Zulfiqar Ali; Sajjad, Ghulam; Sher, Nawab; Aziz, Mubashir; Siddiqui, Farhan Ahmed

2016-01-01

In this article we describe development and validation of stability indicating, accurate, specific, precise and simple Ion-pairing RP-HPLC method for simultaneous determination of paracetamol and cetirizine HCl along with preservatives i.e. propylparaben, and methylparaben in pharmaceutical dosage forms of oral solution and in serum. Acetonitrile: Buffer: Sulfuric Acid (45:55:0.3 v/v/v) was the mobile phase at flow rate 1.0 mL min-1 using a Hibar? Lichrosorb? C18 column and monitored at wavel...

Analysis of perfluorinated phosponic acids and perfluorooctane sulfonic acid in water, sludge and sediment by LC-MS/MS

NARCIS (Netherlands)

Esperza, X.; Moyano, E.; de Boer, J.; Galceran, M.T.; van Leeuwen, S.P.J.

2011-01-01

Residues of perfluorinated phosphonic acids (PFPAs) and perfluorooctane sulfonic acid (PFOS) were investigated in various Dutch surface waters, sludge and sediments. For this purpose, a liquid chromatographic (LC) method was optimized by testing several columns with different mobile phases.

Determination of 8 Synthetic Food Dyes by Solid Phase Extraction ...

African Journals Online (AJOL)

Keywords: Synthetic colors, Food, Fruit flavored drinks, Solid phase extraction, RP-HPLC. Tropical Journal of ..... food dyes by thin-layer chromatography-fast atom bombardment ... food dyes in soft drinks containing natural pigments by.

Evaluation of operational incidents in the research reactor RP-10 according to scale INES

International Nuclear Information System (INIS)

Arrieta, Rolando W.B.; Vela Mora, Mariano

2013-01-01

This report presents the evaluation of the events in 2011 in the RP-10 Nuclear Reactor Nuclear Center Huarangal from the point of view of safety. To classify these events produced is used Scale International Nuclear and Radiological Event Scale (INES) to facilitate a common understanding between the technical community, the media and the general public. From the results we can say that in 2011 all related to security events that occurred in the RP -10 are classified as 'below scale' or no safety significance. (author)

AgRP Neurons Can Increase Food Intake during Conditions of Appetite Suppression and Inhibit Anorexigenic Parabrachial Neurons.

Science.gov (United States)

Essner, Rachel A; Smith, Alison G; Jamnik, Adam A; Ryba, Anna R; Trutner, Zoe D; Carter, Matthew E

2017-09-06

To maintain energy homeostasis, orexigenic (appetite-inducing) and anorexigenic (appetite suppressing) brain systems functionally interact to regulate food intake. Within the hypothalamus, neurons that express agouti-related protein (AgRP) sense orexigenic factors and orchestrate an increase in food-seeking behavior. In contrast, calcitonin gene-related peptide (CGRP)-expressing neurons in the parabrachial nucleus (PBN) suppress feeding. PBN CGRP neurons become active in response to anorexigenic hormones released following a meal, including amylin, secreted by the pancreas, and cholecystokinin (CCK), secreted by the small intestine. Additionally, exogenous compounds, such as lithium chloride (LiCl), a salt that creates gastric discomfort, and lipopolysaccharide (LPS), a bacterial cell wall component that induces inflammation, exert appetite-suppressing effects and activate PBN CGRP neurons. The effects of increasing the homeostatic drive to eat on feeding behavior during appetite suppressing conditions are unknown. Here, we show in mice that food deprivation or optogenetic activation of AgRP neurons induces feeding to overcome the appetite suppressing effects of amylin, CCK, and LiCl, but not LPS. AgRP neuron photostimulation can also increase feeding during chemogenetic-mediated stimulation of PBN CGRP neurons. AgRP neuron stimulation reduces Fos expression in PBN CGRP neurons across all conditions. Finally, stimulation of projections from AgRP neurons to the PBN increases feeding following administration of amylin, CCK, and LiCl, but not LPS. These results demonstrate that AgRP neurons are sufficient to increase feeding during noninflammatory-based appetite suppression and to decrease activity in anorexigenic PBN CGRP neurons, thereby increasing food intake during homeostatic need. SIGNIFICANCE STATEMENT The motivation to eat depends on the relative balance of activity in distinct brain regions that induce or suppress appetite. An abnormal amount of activity in

Solution structure of LC4 transmembrane segment of CCR5.

Directory of Open Access Journals (Sweden)

Kazuhide Miyamoto

Full Text Available CC-chemokine receptor 5 (CCR5 is a specific co-receptor allowing the entry of human immunodeficiency virus type 1 (HIV-1. The LC4 region in CCR5 is required for HIV-1 entry into the cells. In this study, the solution structure of LC4 in SDS micelles was elucidated by using standard 1H two-dimensional NMR spectroscopy, circular dichroism, and fluorescence quenching. The LC4 structure adopts two helical structures, whereas the C-terminal part remains unstructured. The positions in which LC4 binds to the HIV-1 inhibitory peptide LC5 were determined by docking calculations in addition to NMR data. The poses showed the importance of the hydrophobic interface of the assembled structures. The solution structure of LC4 elucidated in the present work provides a structural basis for further studies on the HIV-1 inhibitory function of the LC4 region.

Solution structure of LC4 transmembrane segment of CCR5.

Science.gov (United States)

Miyamoto, Kazuhide; Togiya, Kayo

2011-01-01

CC-chemokine receptor 5 (CCR5) is a specific co-receptor allowing the entry of human immunodeficiency virus type 1 (HIV-1). The LC4 region in CCR5 is required for HIV-1 entry into the cells. In this study, the solution structure of LC4 in SDS micelles was elucidated by using standard 1H two-dimensional NMR spectroscopy, circular dichroism, and fluorescence quenching. The LC4 structure adopts two helical structures, whereas the C-terminal part remains unstructured. The positions in which LC4 binds to the HIV-1 inhibitory peptide LC5 were determined by docking calculations in addition to NMR data. The poses showed the importance of the hydrophobic interface of the assembled structures. The solution structure of LC4 elucidated in the present work provides a structural basis for further studies on the HIV-1 inhibitory function of the LC4 region.

Silver ion chromatography for peak resolution enhancement: Application to the preparative separation of two sesquiterpenes using online heart-cutting LC-LC technique.

Science.gov (United States)

Yang, Yang; Zhang, Yongmin; Wei, Chong; Li, Jing; Sun, Wenji

2018-09-01

Silver ion chromatography, utilizing columns packed with silver ions bonded to silica gel, has proved to be an invaluable technique for the analysis of some positional isomers. In this work, silver ion chromatography by combination with online heart-cutting LC-LC technique for the preparative separation of two sesquiterpenes positional isomers from a natural product was investigated. On the basis of the evaluation that silver ion content impacts on the separation, the laboratory-made silver ion columns, utilizing silica gel impregnated with 15% silver nitrate as column packing materials, were used for peak resolution improvement of these two isomers and the preparative separation of them in heart-cutting LC-LC. The relationship among the maximal sample load, flow rate and peak resolution in the silver ion column were optimized, and the performance of the silver ion column was compared with conventional C 18 column and silica gel column. Based on the developed chromatographic conditions, online heart-cutting LC-LC chromatographic separation system in combination with a silica gel column and a silver ion column that was applied to preparative separation of these two isomers from a traditional Chinese medicine, Inula racemosa Hook.f., was established. The results showed that the online heart-cutting LC-LC technique by combination of a silica gel column and a silver ion column for the preparative separation of these two positional isomers from this natural plant was superior to the preparative separation performed on a single-column system with C 18 column or silica gel column. Copyright © 2018 Elsevier B.V. All rights reserved.

Analytical method development and validation of simultaneous estimation of rabeprazole, pantoprazole, and itopride by reverse-phase high-performance liquid chromatography

Directory of Open Access Journals (Sweden)

Senthamil Selvan Perumal

2014-12-01

Full Text Available A simple, selective, rapid, and precise reverse-phase high-performance liquid chromatography (RP-HPLC method for the simultaneous estimation of rabeprazole (RP, pantoprazole (PP, and itopride (IP has been developed. The compounds were well separated on a Phenomenex C18 (Luna column (250 mm × 4.6 mm, dp = 5 μm with C18 guard column (4 mm × 3 mm × 5 μm with a mobile phase consisting of buffer containing 10 mM potassium dihydrogen orthophosphate (adjusted to pH 6.8: acetonitrile (70:30 v/v at a flow rate of 1.0 mL/min and ultraviolet detection at 288 nm. The retention time of RP, PP, and IP were 5.35, 7.92, and 11.16 minutes, respectively. Validation of the proposed method was carried out according to International Conference on Harmonisation (ICH guidelines. Linearity range was obtained for RP, PP, and IP over the concentration range of 2.5–25, 1–30, and 3–35 μg/mL and the r2 values were 0.994, 0.978, and 0.991, respectively. The calculated limit of detection (LOD values were 1, 0.3, and 1 μg/mL and limit of quantitation (LOQ values were 2.5, 1, and 3 μg/mL for RP, PP, and IP correspondingly. Thus, the current study showed that the developed reverse-phase liquid chromatography method is sensitive and selective for the estimation of RP, PP, and IP in combined dosage form.

Analytical method development and validation of simultaneous estimation of rabeprazole, pantoprazole, and itopride by reverse-phase high-performance liquid chromatography.

Science.gov (United States)

Perumal, Senthamil Selvan; Ekambaram, Sanmuga Priya; Raja, Samundeswari

2014-12-01

A simple, selective, rapid, and precise reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of rabeprazole (RP), pantoprazole (PP), and itopride (IP) has been developed. The compounds were well separated on a Phenomenex C 18 (Luna) column (250 mm × 4.6 mm, dp = 5 μm) with C 18 guard column (4 mm × 3 mm × 5 μm) with a mobile phase consisting of buffer containing 10 mM potassium dihydrogen orthophosphate (adjusted to pH 6.8): acetonitrile (70:30 v/v) at a flow rate of 1.0 mL/min and ultraviolet detection at 288 nm. The retention time of RP, PP, and IP were 5.35, 7.92, and 11.16 minutes, respectively. Validation of the proposed method was carried out according to International Conference on Harmonisation (ICH) guidelines. Linearity range was obtained for RP, PP, and IP over the concentration range of 2.5-25, 1-30, and 3-35 μg/mL and the r 2 values were 0.994, 0.978, and 0.991, respectively. The calculated limit of detection (LOD) values were 1, 0.3, and 1 μg/mL and limit of quantitation (LOQ) values were 2.5, 1, and 3 μg/mL for RP, PP, and IP correspondingly. Thus, the current study showed that the developed reverse-phase liquid chromatography method is sensitive and selective for the estimation of RP, PP, and IP in combined dosage form. Copyright © 2014. Published by Elsevier B.V.

Solid phase extraction method for determination of mitragynine in ...

African Journals Online (AJOL)

phase high performance liquid chromatography (RP-HPLC) to determine mitragynine (MG) in rat and human urine, and to investigate the influence of caffeine (CF) on urinary excretion of MG in rats. Methods: A two-dimensional wash-elute ...

Characterization of rhamnolipids by liquid chromatography/mass spectrometry after solid-phase extraction.

Science.gov (United States)

Behrens, Beate; Engelen, Jeannine; Tiso, Till; Blank, Lars Mathias; Hayen, Heiko

2016-04-01

Rhamnolipids are surface-active agents with a broad application potential that are produced in complex mixtures by bacteria of the genus Pseudomonas. Analysis from fermentation broth is often characterized by laborious sample preparation and requires hyphenated analytical techniques like liquid chromatography coupled to mass spectrometry (LC-MS) to obtain detailed information about sample composition. In this study, an analytical procedure based on chromatographic method development and characterization of rhamnolipid sample material by LC-MS as well as a comparison of two sample preparation methods, i.e., liquid-liquid extraction and solid-phase extraction, is presented. Efficient separation was achieved under reversed-phase conditions using a mixed propylphenyl and octadecylsilyl-modified silica gel stationary phase. LC-MS/MS analysis of a supernatant from Pseudomonas putida strain KT2440 pVLT33_rhlABC grown on glucose as sole carbon source and purified by solid-phase extraction revealed a total of 20 congeners of di-rhamnolipids, mono-rhamnolipids, and their biosynthetic precursors 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs) with different carbon chain lengths from C8 to C14, including three rhamnolipids with uncommon C9 and C11 fatty acid residues. LC-MS and the orcinol assay were used to evaluate the developed solid-phase extraction method in comparison with the established liquid-liquid extraction. Solid-phase extraction exhibited higher yields and reproducibility as well as lower experimental effort.

Gradient HPLC of antibiotics in urine, ground water, chicken muscle, hospital wastewater, and pharmaceutical samples using C-18 and RP-amide columns.

Science.gov (United States)

Kumar, Ashwini; Kumar Malik, Ashok; Kumar Tewary, Dhananjay; Singh, Baldev

2008-02-01

A simple and highly sensitive high pressure liquid chromatographic (HPLC-UV) method has been developed for the determination of ofloxacin, lomefloxacin, cinoxacin, and nalidixic acid, in mobile phase citrate buffer (0.001 M) of pH 4.5 prepared in water (X), methanol (Y), and ACN (Z) using gradient at a flow rate of 1.0 mL/min by direct UV absorbance detection at lambda = 280 nm. Separation of analytes was studied on the C-18 and RP-amide columns and best results were observed on the RP-amide column with LODs (3.3 x S/m) 0.89, 0.55, 0.67, and 1.41 ng/mL for ofloxacin, lomefloxacin, cinoxacin, and nalidixic acid, respectively, and better RSD than the C-18 column. The recovery of Fluoroquinolones (FQs) in urine, ground water, hospital wastewater, and chicken muscle using this method is more than 90%. The method was successfully applied to the analysis of ofloxacin, lomefloxacin, cinoxacin, and nalidixic acid in urine, ground water, pharmaceutical dosage forms, hospital wastewater, and chicken muscle.

Improved soil characterization for pipe piles in sand in API RP-2A

International Nuclear Information System (INIS)

Hossain, M.K.; Briaud, J.L.

1993-01-01

In the offshore, most foundations are steel pipe piles and most of them are designed using the API RP 2A guidelines. For axial capacity of piles in sand the current guidelines in many cases show definite discrepancies when compared against actual load capacities of piles. An updated data base analysis shows that there are three major weaknesses in the current guidelines with respect to soil characterization: (a) the consideration of the lateral earth pressure coefficient, K, as a constant (1.0 or 0.8); (b) the consideration of the unit point bearing resistance, q, as a linear function of depth; and (c) the absence of an unambiguous soil parameter determination process based on reliable in-situ test results. In this paper, specific modifications to the current API RP 2A guidelines are proposed on the basis of a data base analysis to account for the discrepancies arising from (a), (b), and (c) above. These modifications will reduce the discrepancies in the current API RP 2A method and increase the accuracy of the prediction of axial capacity of pipe piles in sand. Furthermore this will make the method fundamentally more consistent with soil behavior in deep foundations

Transcription of four Rhopalosiphum padi (L.) heat shock protein genes and their responses to heat stress and insecticide exposure.

Science.gov (United States)

Li, Yuting; Zhao, Qi; Duan, Xinle; Song, Chunman; Chen, Maohua

2017-03-01

The bird cherry-oat aphid, Rhopalosiphum padi (L.), a worldwide destructive pest, is more heat tolerant than other wheat aphids, and it has developed resistance to different insecticides. Heat shock proteins (HSPs) play an important role in coping with environmental stresses. To investigate Hsp transcriptional responses to heat and insecticide stress, four full-length Hsp genes from R. padi (RpHsp60, RpHsc70, RpHsp70-1, and RpHsp70-2) were cloned. Four RpHsps were expressed during all R. padi developmental stages, but at varying levels. The mRNA levels of RpHsps were increased under thermal stress and reached maximal induction at a lower temperature (36°C) in the alate morph than in the apterous morph (37°C or 38°C). RpHsp expressions under heat stress suggest that RpHsp70-1 and RpHsp70-2 are inducible in both apterous and alate morphs, RpHsc70 is only heat-inducible in apterous morph, and RpHsp60 exhibits poor sensitivity to heat stress. The pretreatment at 37°C significantly increase both the survival rate and the RpHsps expression level of R. padi at subsequent lethal temperature. Under exposure to two sublethal concentrations (LC 10 and LC 30 ) of beta-cypermethrin, both RpHsp70-1 and RpHsp70-2 expressions were induced and reached a maximum 24h after exposure. In contrast, expression of RpHsp60 was not induced by either sublethal concentration of beta-cypermethrin. Moreover, the responses of RpHsp70-1 and RpHsp70-2 to heat shock were more sensitive than those to beta-cypermethrin. These results suggest that induction of RpHsp expression is related to thermal tolerance, and that RpHsp70-1 and RpHsp70-2 are the primary genes involved in the response to both heat and pesticide stress. Copyright © 2016 Elsevier Inc. All rights reserved.

Numerical Investigation on Supercritical Heat Transfer of RP3 Kerosene Flowing inside a Cooling Channel of Scramjet

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Ning Wang

2014-06-01

Full Text Available Supercritical convective heat transfer characteristics of hydrocarbon fuel play a fundamental role in the active cooling technology of scramjet. In this paper, a 2D-axisymmetric numerical study of supercritical heat transfer of RP3 flowing inside the cooling channels of scramjet has been conducted. The main thermophysical properties of RP3, including density, specific heat, and thermal conductivity, are obtained from experimental data, while viscosity is evaluated from a commercial code with a ten-species surrogate. Effects of heat flux, mass flow rate, and inlet temperature on supercritical heat transfer processes have been investigated. Results indicate that when the wall temperature rises above the pseudocritical temperature of RP3, heat transfer coefficient decreases as a result of drastic decrease of the specific heat. The conventional heat transfer correlations, that is, Gnielinski formula, are no longer proper for the supercritical heat transfer of RP3. The modified Jackson and Hall formula, which was proposed for supercritical CO2 and water, gives good prediction except when the wall temperature is near or higher than the pseudocritical temperature.

Phase Analysis of the Cellulose Triacetate-Nitromethane System

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Anna B. Shipovskaya

2012-01-01

Full Text Available A comprehensive study was made on the cellulose triacetate-nitromethane system to explore its phase separation within ranges 2–25 wt.% and −5÷+80°C by means of polarization light and electron microscopy, the turbidity spectrum method, differential thermal and X-ray analyses, and rheological techniques. The physical state of the polymer was identified within the phase coexistence boundaries on the phase diagram which included three types of phase separation (amorphous (with a UCST at Tcr=57∘C and ccr=7.3 wt.%, crystal, and liquid crystal. The boundaries of the regions determining the coexistence of the liquid crystal (LC and the partly crystal phase were found to be inside the region of amorphous liquid-liquid phase separation. For cellulose ester-solvent systems, this state diagram is the first experimental evidence for the possibility of coexistence of several phases with amorphous, LC, and crystal polymer ordering.

Regulation of the autophagy protein LC3 by phosphorylation

Science.gov (United States)

Cherra, Salvatore J.; Kulich, Scott M.; Uechi, Guy; Balasubramani, Manimalha; Mountzouris, John; Day, Billy W.

2010-01-01

Macroautophagy is a major catabolic pathway that impacts cell survival, differentiation, tumorigenesis, and neurodegeneration. Although bulk degradation sustains carbon sources during starvation, autophagy contributes to shrinkage of differentiated neuronal processes. Identification of autophagy-related genes has spurred rapid advances in understanding the recruitment of microtubule-associated protein 1 light chain 3 (LC3) in autophagy induction, although braking mechanisms remain less understood. Using mass spectrometry, we identified a direct protein kinase A (PKA) phosphorylation site on LC3 that regulates its participation in autophagy. Both metabolic (rapamycin) and pathological (MPP+) inducers of autophagy caused dephosphorylation of endogenous LC3. The pseudophosphorylated LC3 mutant showed reduced recruitment to autophagosomes, whereas the nonphosphorylatable mutant exhibited enhanced puncta formation. Finally, autophagy-dependent neurite shortening induced by expression of a Parkinson disease–associated G2019S mutation in leucine-rich repeat kinase 2 was inhibited by dibutyryl–cyclic adenosine monophosphate, cytoplasmic expression of the PKA catalytic subunit, or the LC3 phosphorylation mimic. These data demonstrate a role for phosphorylation in regulating LC3 activity. PMID:20713600

Photo polymerization-induced vertical phase separation and homeotropic alignment in liquid crystal and polymer mixtures

International Nuclear Information System (INIS)

Kang, Hyo; Joo, Sangwoo; Kang, Daeseung

2012-01-01

We presented a novel method for the homeotropic alignment of LC by using the irradiation of UV light on the LC/NOA65 mixture cell, in which the photo-initiated-polymerization-induced phase separation lowers the surface energy. When the amount of polymer content is sufficiently small, the gravel and network patterns were formed at the substrates via the vertical phase separation. We found that surface roughness plays an important role in the formation of the homeotropic alignment of LC. We also observed the alignment transition of the cells by varying the mixing ratio of LC/NOA65 or the UV radiation time. Furthermore, the present proposed method has great potential for application in display devices. For decades, studies on the alignment of liquid crystal (LC) molecules have been of significant interest due to their immediate applications for display devices and the intriguing physiochemical properties they exhibit at the surface of mixtures. Usually, homeotropic (or vertical) alignment, in which the long axes of the LC molecules are oriented in a direction perpendicular to the surface, is achieved by using surfactants such as lecithin, silanes or polyimide. Recently homeotropic alignment of liquid crystal molecules was achieved by irradiating photosensitive polymers, by doping nanoparticles into LC, by utilizing nano/micro patterns, or by incorporating self-assembled monolayers (SAMs). However, a clear understanding about the alignment mechanism is still elusive. In this paper, we report a novel method for homeotropic alignment of LC by utilizing the phase separation of LC/polymer mixtures

Preparation of polyhedral oligomeric silsesquioxane based hybrid monoliths by ring-opening polymerization for capillary LC and CEC.

Science.gov (United States)

Lin, Hui; Zhang, Zhenbin; Dong, Jing; Liu, Zhongshan; Ou, Junjie; Zou, Hanfa

2013-09-01

A new organic-inorganic hybrid monolith was prepared by the ring-opening polymerization of octaglycidyldimethylsilyl polyhedral oligomeric silsesquioxane (POSS) with 1,4-butanediamine (BDA) using 1-propanol, 1,4-butanediol, and PEG 10,000 as a porogenic system. Benefiting from the moderate phase separation process, the resulting poly(POSS-co-BDA) hybrid monolith possessed a uniform microstructure and exhibited excellent performance in chromatographic applications. Neutral, acidic, and basic compounds were successfully separated on the hybrid monolith in capillary LC (cLC), and high column efficiencies were achieved in all of the separations. In addition, as the amino groups could generate a strong EOF, the hybrid monolith was also applied in CEC for the separation of neutral and polar compounds, and a satisfactory performance was obtained. These results demonstrate that the poly(POSS-co-BDA) hybrid monolith is a good separation media in chromatographic separations of various types of compounds by both cLC and CEC. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Entangled Coherent States Generation in two Superconducting LC Circuits

International Nuclear Information System (INIS)

Chen Meiyu; Zhang Weimin

2008-01-01

We proposed a novel pure electronic (solid state) device consisting of two superconducting LC circuits coupled to a superconducting flux qubit. The entangled coherent states of the two LC modes is generated through the measurement of the flux qubit states. The interaction of the flux qubit and two LC circuits is controlled by the external microwave control lines. The geometrical structure of the LC circuits is adjustable and makes a strong coupling between them achievable. This entangled coherent state generator can be realized by using the conventional microelectronic fabrication techniques which increases the feasibility of the experiment.

Multi-residue analysis method for analysis of pharmaceuticals using liquid chromatography-time of flight/mass spectrometry (LC-TOF/MS) in water sample

Science.gov (United States)

Al-Qaim, Fouad Fadhil; Abdullah, Md Pauzi; Othman, Mohamed Rozali

2013-11-01

In this work, a developed method using solid - phase extraction (SPE) followed by liquid chromatography - time of flight mass spectrometry (LC-ESI-TOF/MS) was developed and validated for quantification and confirmation of eleven pharmaceuticals with different therapeutic classes in water samples, Malaysia. These compounds are caffeine (CAF), prazosin (PRZ), enalapril (ENL), carbamazepine (CBZ), nifedipine (NFD), levonorgestrel (LNG), simvastatin (SMV), hydrochlorothiazide (HYD), gliclazide (GLIC), diclofenac-Na (DIC-Na) and mefenamic acid (MEF). LC was performed on a Dionex Ultimate 3000/LC 09115047 (USA) system. Chromatography was performed on a Thermo Scientific C18 (250 mm × 2.1 mm, i.d.: 5μm) column. Several parameters were optimised such as; mobile phase, gradient elution, collision energy and solvent elution for extraction of compounds from water. The recoveries obtained ranged from 30-148 % in river water. Five pharmaceutical compounds were detected in the surface water samples: caffeine, prazosin, enalpril, diclofenac-Na and mefenamic acid. The developed method is precise and accepted recoveries were got. In addition, this method is suitable to identify and quantify trace concentrations of pharmaceuticals in surface water.

Simultaneous Estimation of Gemcitabine Hydrochloride and Capecitabine Hydrochloride in Combined Tablet Dosage Form by RP-HPLC Method

Directory of Open Access Journals (Sweden)

V. Rajesh

2011-01-01

Full Text Available A new reverse phase high performance liquid chromatography (RP-HPLC method has been developed for the simultaneous estimation of gemcitabine hydrochloride and capecitabine hydrochloride in combined tablet dosage form. An inertsil ODS-3 C-18 column having dimensions of 250×4.6 mm and particle size of 5 µm, with mobile phase containing a mixture of acetonitrile : water : triethyelamine in the ratio of (70 : 28 : 2v/v was used. The pH of mobile phase was adjusted to 4.0 with ortho-phosphoric acid. The flow rate was 1 mL/min and the column effluents were monitored at 260 nm. The retention time for gemcitabine hydrochloride and capecitabine hydrochloride was found to be 2.76 and 2.3 min respectively. The proposed method was validated in terms of linearity, accuracy, precision, limit of detection, limit of quantitation and robustness. The method was found to be linear in the range of 10-50 µg/mL and 4-24 µg/mL for gemcitabine hydrochloride and capecitabine hydrochloride, with regression coefficient r = 0.999 and r = 0.999, respectively.

75 FR 24740 - Recovery Fact Sheet RP9580.102, Permanent Relocation

Science.gov (United States)

2010-05-05

... DEPARTMENT OF HOMELAND SECURITY Federal Emergency Management Agency [Docket ID FEMA-2010-0028] Recovery Fact Sheet RP9580.102, Permanent Relocation AGENCY: Federal Emergency Management Agency, DHS... Management Agency (FEMA). DATES: This fact sheet is effective April 14, 2010. ADDRESSES: This final fact...

RP-HPLC×HILIC chromatography for quantifying ertapenem sodium with a look at green chemistry.

Science.gov (United States)

Pedroso, Tahisa M; Medeiros, Ana C D; Salgado, Herida R N

2016-11-01

Ertapenem sodium is a polar and ionizable compound; therefore, it has little retention on traditional C18 columns in reverse-phase high-performance liquid chromatography, even using a highly-aqueous mobile phase that can result in dewetting in the stationary phase. Thus, the most coherent process for ERTM is to develop a method for Hydrophilic Interaction Chromatography. However, for the traditional methods in HILIC, the use of a highly organic mobile phase is necessary; usually an amount exceeding 80% acetonitrile is necessary. On the other hand, the RP-HPLC mode is considered for the analysis technique, which is more often used for quantification of substances, and new columns are often introduced to analyze different groups of compounds. Two new analytical methods have been developed for routine analysis. The proposed chromatographic method was adequate and advantageous by presenting simplicity, linearity, precision, accuracy, robustness, detection limits, and satisfactory quantification. Analytical methods are constantly undergoing changes and improvements. Researchers worldwide are rapidly adopting Green Chemistry. The development of new pharmaceutical methods based in Green chemistry has been encouraged by universities and the pharmaceutical industry. Issues related to green chemistry are in evidence and they have been featured in international journals of high impact. The methods described here have economic advantages and they feature an eco-friendly focus, which is discussed in this work. This work was developed with an environmental conscience, always looking to minimize the possible generated organic waste. Therefore, discussion on this aspect is included. Copyright © 2016 Elsevier B.V. All rights reserved.

Characterization and adjustment of the neutron radiography facility of the RP-10 nuclear reactor; Caracterizacion y puesta a punto de la facilidad de neutrografia del reactor nuclear RP-10

Energy Technology Data Exchange (ETDEWEB)

Ravello R, Y R

2001-07-01

The main aim of this work was to characterize and adjust the neutron radiography facility of the RP-10 nuclear reactor, and therefore be able to offer with this technique services to the industry and research centers in general. This technique will be complemented with others such as x-rays and gamma radiography. First, the shielding capacity of the facility was analyzed, proving that it complies with the radiological safety requirements established by the radiological safety code. Then gamma filtration tests were conducted in order to implement the direct method for image formation, optical density curves were built according to the thickness of the gamma filter, the type of film and the type of irradiation. Also, the indirect method for image formation was implemented for two types of converters: indium and dysprosium. Growth curves for optical density were also made according to contact time between converter-film, for different types of films. The resolution of the facility was also analyzed using two methods: Klasens (1946) and Harms (1986). Harms method came closer to the resolution of the human eye when compared to the Klasens method. Finally, the application fields of neutron radiography are presented, including those conducted at the neutron radiography facility of the RP-10 nuclear reactor. With this work, the RP-10 neutron radiography facility is ready to offer inspection and research services.

Stability Indicating RP-HPLC Method for Determination of Valsartan in Pure and Pharmaceutical Formulation

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S. K. Patro

2010-01-01

Full Text Available A simple, rapid and accurate and stability indicating RP-HPLC method was developed for the determination of valsartan in pure and tablet forms. The method showed a linear response for concentrations in the range of 50-175 µg/mL using 0.01 M NH4H2PO4 (pH 3.5 buffer: methanol [50:50] as the mobile phase with detection at 210 nm and a flow rate of 1 mL/min and retention time 11.041 min. The method was statistically validated for accuracy, precision, linearity, ruggedness, robustness, forced degradation, solution stability and selectivity. Quantitative and recovery studies of the dosage form were also carried out and analyzed; the % RSD from recovery studies was found to be less than 1. Due to simplicity, rapidity and accuracy of the method, we believe that the method will be useful for routine quality control analysis.

Analysis of glutathione in supernatants and lysates of a human proximal tubular cell line from perfusion culture upon intoxication with cadmium chloride by HPLC and LC-ESI-MS

NARCIS (Netherlands)

Hahn, Hans; Huck, Christian W; Rainer, Matthias; Najam-ul-Haq, Muhammad; Bakry, Rania; Abberger, Thomas; Jennings, Paul; Pfaller, Walter; Bonn, Günther K

A simple and highly effective reversed-phase (RP) high-performance liquid chromatography (HPLC) method is described for analysing glutathione (GSH) and glutathione disulfide (GSSG) in out-flowing supernatants and lysates of perfusion cell cultures of human kidney cells (HK-2 cells) continuously

Matrix removal in state of the art sample preparation methods for serum by charged aerosol detection and metabolomics-based LC-MS.

Science.gov (United States)

Schimek, Denise; Francesconi, Kevin A; Mautner, Anton; Libiseller, Gunnar; Raml, Reingard; Magnes, Christoph

2016-04-07

Investigations into sample preparation procedures usually focus on analyte recovery with no information provided about the fate of other components of the sample (matrix). For many analyses, however, and particularly those using liquid chromatography-mass spectrometry (LC-MS), quantitative measurements are greatly influenced by sample matrix. Using the example of the drug amitriptyline and three of its metabolites in serum, we performed a comprehensive investigation of nine commonly used sample clean-up procedures in terms of their suitability for preparing serum samples. We were monitoring the undesired matrix compounds using a combination of charged aerosol detection (CAD), LC-CAD, and a metabolomics-based LC-MS/MS approach. In this way, we compared analyte recovery of protein precipitation-, liquid-liquid-, solid-phase- and hybrid solid-phase extraction methods. Although all methods provided acceptable recoveries, the highest recovery was obtained by protein precipitation with acetonitrile/formic acid (amitriptyline 113%, nortriptyline 92%, 10-hydroxyamitriptyline 89%, and amitriptyline N-oxide 96%). The quantification of matrix removal by LC-CAD showed that the solid phase extraction method (SPE) provided the lowest remaining matrix load (48-123 μg mL(-1)), which is a 10-40 fold better matrix clean-up than the precipitation- or hybrid solid phase extraction methods. The metabolomics profiles of eleven compound classes, comprising 70 matrix compounds showed the trends of compound class removal for each sample preparation strategy. The collective data set of analyte recovery, matrix removal and matrix compound profile was used to assess the effectiveness of each sample preparation method. The best performance in matrix clean-up and practical handling of small sample volumes was showed by the SPE techniques, particularly HLB SPE. CAD proved to be an effective tool for revealing the considerable differences between the sample preparation methods. This detector can

Clinical study with a new radiopharmaceutical (RP-517) for the in vivo labeling of neutrophils with 99mTc

International Nuclear Information System (INIS)

Parysow, O.; Mele, A.; Sarmiento, A.; Zubillaga, M.; Caro, R.; Calmanovici, G.; Nicolini, J.; Lazewatsky, J.; Camin, L.

2002-01-01

RP-517 is a 99m Tc labeled antagonist to the LTB 4 receptor which, with a single i.v. injection, allows the in vivo labeling of neutrophils for the detection and localization of infectious and/or inflammatory foci. Aim: To determine: 1) the safety and tolerance of the radiopharmaceutical in humans; 2) its pharmacokinetics and dosimetry; 3) its preliminary efficacy for the detection and localization of infectious foci. Materials and Methods: 20 subjects (14M, 6F) received 592 ± 111 MBq (16 ± 3 mCi) of RP-517 intravenously. All subjects were evaluated (medical history, physical exam, vital signs, ECG, blood and urine analysis) before and at several times up to 24 hours post-injection of RP-517. Blood samples were taken to determine pharmacokinetics and blood clearance. Urine and feces were collected to measure excretion. For dosimetry determination, whole body images, one transmission pre-injection and sequential emission images post-injection, were obtained with a dual head gamma camera (ADAC) in 2 patients and 14 volunteers. Sequential focal images were obtained in 4 patients. The presence of infection was determined by culture and/or follow-up. Results: There were no adverse clinical events or clinically significant changes in any of the laboratory analysis parameters, ECG or vital signs. RP-517 clears the blood mainly through the hepatobiliary system. The critical organ is the gall bladder wall, followed by the intestinal wall. A suggested maximum injected dose of about 740MBq of RP-517 would deliver ≤ 50 mGy to the critical organ. There were 6 patients with suspicion of infection: in 2 of them the infection was proven; a third subject had 2 suspected infectious foci, one was proven, the other one was equivocal; at the time of imaging, there was no infections process in the other 3 subjects. RP-517 images showed a clear and persistent uptake in the 3 proven infectious foci, was negative in the equivocal one and in the three patients without infection

Comparison of a polymeric pseudostationary phase in EKC with ODS stationary phase in RP-HPLC.

Science.gov (United States)

Ni, Xinjiong; Zhang, Min; Xing, Xiaoping; Cao, Yuhua; Cao, Guangqun

2018-01-01

Poly(stearyl methacrylate-co-methacrylic acid) (P(SMA-co-MAA)) was induced as pseudostationary phase (PSP) in electrokinetic chromatography (EKC). The n-octadecyl groups in SMA were the same as that in octadecylsilane (ODS) C18 column. Thus, the present work focused on the comparison of selectivity between polymeric PSP and ODS stationary phase (SP), and the effect of organic modifiers on the selectivity of polymeric PSP and ODS SP. 1-butanol could directly interacted with PSP as a Class I modifier, and improved both of the methylene selectivity and polar group selectivity. When the analysis times were similar, the polymeric PSP exhibited better methylene selectivity and polar group selectivity. Although the hydrophobic groups were similar, the substituted benzenes elution order was different between polymeric PSP and ODS SP. Linear solvation energy relationships (LSER) model analysis found that polymeric PSP and ODS SP exhibited two same key factors in selectivity: hydrophobic interaction and hydrogen bonding acidity. But polymeric PSP exhibited relatively strong n- and π-electrons interaction to the analytes. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Stability-indicating RP-HPLC method for simultaneous determination of gatifloxacin and flurbiprofen in binary combination

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Islam Ullah Khan

2014-04-01

Full Text Available A stability-indicating RP-HPLC method is presented for determination of gatifloxacin and flurbiprofen in binary combination. Gatifloxacin, flurbiprofen and their degradation products were detected at 254 nm using a BDS Hypersil C8 (250 X 4.6 mm, 5 µm column and mixture of 20 mM phosphate buffer (pH 3.0 and methanol 30:70 v/v as mobile phase. Response was linear over the range of 15-105 mg mL-1 for gatifloxacin (r² > 0.998 and of 1.5-10.5 mg mL-1 for flurbiprofen (r² > 0.999. The developed method efficiently separated the analytical peaks from degradation products (peak purity index > 0.9999. The method developed can be applied successfully for determination of gatifloxacin and flurbiprofen in human serum, urine, pharmaceutical formulations, and their stability studies.

LC-MS/MS Peptide Mapping with Automated Data Processing for Routine Profiling of N-Glycans in Immunoglobulins

Science.gov (United States)

Shah, Bhavana; Jiang, Xinzhao Grace; Chen, Louise; Zhang, Zhongqi

2014-06-01

Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. These methods require time-consuming sample preparations and do not provide site-specific glycosylation information. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). Experimental conditions were optimized to achieve accurate profiling of glycoforms. Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans.

An improved method for fast and selective separation of carotenoids by LC-MS.

Science.gov (United States)

Abate-Pella, Daniel; Freund, Dana M; Slovin, Janet P; Hegeman, Adrian D; Cohen, Jerry D

2017-11-01

Carotenoids are a large class of compounds that are biosynthesized by condensation of isoprene units in plants, fungi, bacteria, and some animals. They are characteristically highly conjugated through double bonds, which lead to many isomers as well susceptibility to oxidation and other chemical modifications. Carotenoids are important because of their potent antioxidant activity and are the pigments responsible for color in a wide variety of foods. Human consumption is correlated to many health benefits including prevention of cancer, cardiovascular disease, and age-related disease. Extreme hydrophobicity, poor stability, and low concentration in biological samples make these compounds difficult to analyze and difficult to develop analytical methods for aimed towards identification and quantification. Examples in the literature frequently report the use of exotic stationary phases, solvents, and additives, such as ethyl acetate, dichloromethane, and methyl tert-butyl ether that are incompatible with liquid chromatography mass spectrometry (LC-MS). In order to address these issues, we implemented the use of LC-MS friendly conditions using a low-hydrophobicity cyano-propyl column (Agilent Zorbax SB-CN). We successfully differentiated between isomeric carotenoids by optimizing two gradient methods and using a mixture of 11 standards and LC-MS in positive ionization mode. Three complex biological samples from strawberry leaf, chicken feed supplement, and the photosynthetic bacterium Chloroflexus aurantiacus were analyzed and several carotenoids were resolved in these diverse backgrounds. Our results show this methodology is a significant improvement over other alternatives for analyzing carotenoids because of its ease of use, rapid analysis time, high selectivity, and, most importantly, its compatibility with typical LC-MS conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

Solution Structure of LC4 Transmembrane Segment of CCR5

OpenAIRE

Miyamoto, Kazuhide; Togiya, Kayo

2011-01-01

CC-chemokine receptor 5 (CCR5) is a specific co-receptor allowing the entry of human immunodeficiency virus type 1 (HIV-1). The LC4 region in CCR5 is required for HIV-1 entry into the cells. In this study, the solution structure of LC4 in SDS micelles was elucidated by using standard 1H two-dimensional NMR spectroscopy, circular dichroism, and fluorescdence quenching. The LC4 structure adopts two helical structures, whereas the C-terminal part remains unstructured. The positions in which LC4 ...

Experimental asymmetric phase-covariant quantum cloning of polarization qubits

Czech Academy of Sciences Publication Activity Database

Soubusta, Jan; Bartůšková, L.; Černoch, Antonín; Dušek, M.; Fiurášek, J.

2008-01-01

Roč. 78, č. 5 (2008), 052323/1-052323/7 ISSN 1050-2947 R&D Projects: GA MŠk(CZ) 1M06002 Grant - others:GAMŠk(CZ) LC06007 Program:LC Institutional research plan: CEZ:AV0Z10100522 Keywords : phase-covariant cloning * quantum information processing Subject RIV: BH - Optics, Masers, Lasers Impact factor: 2.908, year: 2008

Measurement of trace levels of antibiotics in river water using on-line enrichment and triple-quadrupole LC-MS/MS.

Science.gov (United States)

Dinh, Quoc Tuc; Alliot, Fabrice; Moreau-Guigon, Elodie; Eurin, Joëlle; Chevreuil, Marc; Labadie, Pierre

2011-09-15

This study presents the development of an automated on-line solid phase extraction (SPE)-liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of 23 antibiotics in environmental water samples. After optimisation of LC-MS/MS conditions, SPE parameters such as sorbent type, sample pH or sample volume were optimised. Antibiotic recoveries ranged from 64% to 98% and compared favourably with those achieved using off-line SPE. Limits of detection were in the range 0.5-13.7 ng L(-1). This on-line SPE-LC-MS/MS procedure was applied to the analysis of water samples taken in three rivers within the Seine River basin, near Paris (France). The obtained results revealed the occurrence of 12 antibiotics, including tylosin, erythromycin, tetracycline, amoxicillin, trimethoprim, sulfamethoxazole, oxolinic acid, flumequine, norfloxacin, ciprofloxacin, ofloxacin, and vancomycin (2-1435 ng L(-1)). Copyright © 2011 Elsevier B.V. All rights reserved.

Twenty years of Radiology in RP-10 nuclear reactor protection; Veinte anos de proteccion radiologica en el reactor nuclear RP-10

Energy Technology Data Exchange (ETDEWEB)

Zapata, Alejandro L.; Ramos, Fernando T.; Arrieta, Rolando W.B.; Vela Mora, Mariano, E-mail: lzapata@ipen.gob.pe, E-mail: framos@ipen.gob.pe, E-mail: rarrieta@ipen.gob.pe, E-mail: mvela@ipen.gob.pe [Instituto Peruano de Energia Nuclear (IPEN), Lima (Peru)

2013-07-01

In this report we present the results about radiation controls during 1990 - 2010, carried out in the Nuclear Reactor RP-10 of the Nuclear Center of Huarangal. These controls and radiological evaluation are of much utility for the radio personnel protection of this one and other reactors, since it allows to compares these variables with respect to the time. From the results obtained in monitoring and radiation controls, we conclude that in no case it has been reached the limits allowed by the Peruvian Regulating Authority. (author)

ANALYSIS OF BASIC PSYCHOTROPIC DRUGS IN BIOLOGICAL FLUIDS AND TISSUES BY REVERSED-PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY.

Science.gov (United States)

Petruczynik, Anna; Waksmundzka-Hajnos, Monika

2017-03-01

The review of the RP HPLC analysis of basic psychotropic drugs is presented. It contains sample preparation methods with centrifugation, protein precipitation, liquid-liquid extraction (LLE), dispersive liquid-liquid microextraction (DLLME), solid-phase extraction (SPE), solid-phase microextraction (SPME), microwave-assisted extraction (MAE) and RP-HPLC analysis. Chromatographic behavior of basic drugs in aqueous media - eluents used in reversed phase systems is discussed. Methods of blocking of residue surface silanols' interaction are mentioned. Analytical methods used for the analysis are divided into parts according with the above methods: the use of low-pH eluents, the use of high-pH eluents, the use of silanol blockers, special stationary phases for basic analytes. Literature connected with the sample preparation methods and analytical systems for the drug analysis are cited in details and presented also in Table 1.

Phase 1 study of the investigational Aurora A kinase inhibitor alisertib (MLN8237) in East Asian cancer patients: pharmacokinetics and recommended phase 2 dose.

Science.gov (United States)

Venkatakrishnan, Karthik; Kim, Tae Min; Lin, Chia-Chi; Thye, Lim Soon; Chng, Wee Joo; Ma, Brigette; Chen, Ming Huang; Zhou, Xiaofei; Liu, Hua; Kelly, Virginia; Kim, Won Seog

2015-08-01

This phase 1 study assessed the pharmacokinetics (PK), maximum tolerated dose (MTD)/recommended phase 2 dose (RP2D), safety, and preliminary efficacy of the investigational Aurora A kinase inhibitor, alisertib, in East Asian patients with advanced solid tumors or lymphomas. Patients received alisertib twice-daily (BID) for 7 days in 21-day cycles. Doses were escalated (3 + 3) from 30 mg BID based on cycle 1 dose-limiting toxicities (DLTs) until the MTD, followed by expansion for PK/safety characterization. Thirty-six patients (61 % Chinese, 36 % Korean, 3 % Malay) received alisertib (30 mg BID, n = 30; 40 mg BID, n = 6; median, 2.5 cycles). Alisertib exposures increased approximately dose proportionally, and mean half-life was 16 h. Geometric mean apparent oral clearance (2.65 L/h) was 40 % lower than previous estimates in Western patients, resulting in approximately 70 % higher mean dose-normalized, steady-state exposures (735 nM*h/mg) in East Asian patients. Two patients experienced DLTs at 40 mg BID (grade 3 stomatitis; grade 4 neutropenia); the MTD/RP2D was 30 mg BID. Common toxicities (grade ≥3 at RP2D) were neutropenia (50 %), diarrhea (13 %), and stomatitis (10 %). One patient with extranodal T-/NK-cell lymphoma (nasal type) achieved a partial response and 18 (51 %) had stable disease. The MTD/RP2D of alisertib in East Asian patients (30 mg BID) was lower than in Western patients (50 mg BID), consistent with higher systemic exposures in the East Asian population. Alisertib was generally well tolerated and showed signs of antitumor activity in East Asian cancer patients.

Octanol/water partitioning simulation by RP-HPLC for structurally diverse acidic drugs: comparison of three columns in the presence and absence of n-octanol as the mobile phase additive.

Science.gov (United States)

Giaginis, Costas; Theocharis, Stamatios; Tsantili-Kakoulidou, Anna

2013-12-01

The advantageous effect of n-octanol as a mobile phase additive for lipophilicity assessment of structurally diverse acidic drugs both in the neutral and ionized form was explored. Two RP C18 columns, ABZ+ and Aquasil, were used for the determination of logkw indices, and the results were compared with those previously reported on a base-deactivated silica column. At pH 2.5, the use of n-octanol-saturated buffer as the mobile phase aqueous component led to high-quality 1:1 correlation between logkw and logP for the ABZ+ column, while inferior statistics were obtained for Aquasil. At physiological pH, the correlations were significantly improved if strongly ionized acidic drugs were treated separately from weakly ionized ones. In the latter case, 1:1 correlations between logD7.4 and logkw(oct) indices were obtained in the presence of 0.25% n-octanol. Concerning strongly ionized compounds, adequate correlations were established under the same conditions; however, slopes were significantly lower than unity, while large negative intercepts were obtained. According to the absolute difference (diff = logD7.4 – logkw) pattern, base-deactivated silica showed a better performance than ABZ+, however, the latter seems more efficient for the lipophilicity assessment of highly lipophilic acidic compounds. Aquasil may be the column of choice if logD7.4<3 with the limitation, however, that very hydrophilic compounds cannot be measured.

LC-MS/MS analysis of uncommon paracetamol metabolites derived through in vitro polymerization and nitration reactions in liquid nitrogen.

Science.gov (United States)

Trettin, Arne; Jordan, Jens; Tsikas, Dimitrios

2014-09-01

Paracetamol (acetaminophen, APAP) is a commonly used analgesic drug. Known paracetamol metabolites include the glucuronide, sulfate and mercapturate. N-Acetyl-benzoquinonimine (NAPQI) is considered the toxic intermediate metabolite of paracetamol. In vitro and in vivo studies indicate that paracetamol is also metabolized to additional poorly characterized metabolites. For example, metabolomic studies in urine samples of APAP-treated mice revealed metabolites such as APAP-sulfate-APAP and APAP-S-S-APAP in addition to the classical phase II metabolites. Here, we report on the development and application of LC-MS and LC-MS/MS approaches to study reactions of unlabelled and (2)H-labelled APAP with unlabelled and (15)N-labelled nitrite in aqueous phosphate buffers (pH 7.4) upon their immersion into liquid nitrogen (-196°C). In mechanistic studies, these reactions were also studied in aqueous buffer prepared in (18)O-labelled water. LC-MS and LC-MS/MS analyses were performed on a reverse-phase material (C18) using gradient elution (2mM ammonium acetate/acetonitrile), in positive and negative electrospray mode. We identified a series of APAP metabolites including di-, tri- and tetra-APAP, mono- and di-nitro-APAP and nitric ester of di-APAP. Our study indicates that nitrite induces oxidation, i.e., polymerization and nitration of APAP, when buffered APAP/nitrite solutions are immersed into liquid nitrogen. These reactions are specific for nitrite with respect to nitrate and do not proceed via intermediate formation of NAPQI. Potassium ions and physiological saline but not thiols inhibit nitrite- and shock-freeze-induced reactions of paracetamol. The underlying mechanism likely involves in situ formation of NO2 radicals from nitrite secondary to profound pH reduction (down to pH 1) and disproportionation. Polymeric paracetamol species can be analyzed as pentafluorobenzyl derivatives by LC-MS but not by GC-MS. Copyright © 2013 Elsevier B.V. All rights reserved.

Determination of linsidomine in human plasma by tandem LC-MS with ESI.

Science.gov (United States)

Sutherland, F C; de Jager, A D; Swart, K J; Hundt, H K; Scanes, T; Hundt, A F

2000-04-01

A sensitive method for the determination of linsidomine in plasma was developed, using high-performance liquid chromatographic (HPLC) separation with tandem mass spectrometric detection. Linsidomine was derivatised with propyl chloroformate and extracted with tert-butyl methyl ether/1,2-dichloroethane (55:45, v/v), back-extracted into HCl (0.01 M) followed by alkalinisation and back-extraction into ether; the final ether extract evaporated, reconstituted in mobile phase and then separated on a Phenomenex Luna C18 (2) 5 micron 2.1 x 150 mm column with a mobile phase consisting of methanol water formic acid (98/100%) (400:600:0.05, v/v/v) at a flow-rate of 0.4 ml min(-1). Detection was achieved by a Finnigan MAT mass spectrometer (LCQ) at unit resolution in the selected reaction monitoring (SRM) mode monitoring the transition of the protonated molecular ion m/z 257.0 to the product ion m/z 86.0. The mean recovery for linsidomine was 51% with a lower limit of quantification of 0.70 ng/ml using 1 ml plasma for extraction. This LC-MS/MS method for the determination of linsidomine in human plasma allows for better specificity and a higher sample throughput than the traditional LC-UV methods. It also demonstrates the profound effect that the composition of acidic modifiers and matrix constituents can have on the electrospray ionisation (ESI) of the analyte.

MOLECULARLY IMPRINTED SOLID PHASE EXTRACTION FOR TRACE ANALYSIS OF DIAZINON IN DRINKING WATER

Directory of Open Access Journals (Sweden)

M. Rahiminejad ، S. J. Shahtaheri ، M. R. Ganjali ، A. Rahimi Forushani ، F. Golbabaei

2009-04-01

Full Text Available Amongst organophosphate pesticides, the one most widely used and common environmental contaminant is diazinon; thus methods for its trace analysis in environmental samples must be developed. Use of diazinon imprinted polymers such as sorbents in solid phase extraction, is a prominent and novel application area of molecular imprinted polymers. For diazinon extraction, high performance liquid chromatography analysis was demonstrated in this study. During optimization of the molecular imprinted solid phase extraction procedure for efficient solid phase extraction of diazinon, Plackett-Burman design was conducted. Eight experimental factors with critical influence on molecular imprinted solid phase extraction performance were selected, and 12 different experimental runs based on Plackett-Burman design were carried out. The applicability of diazinon imprinted polymers as the sorbent in solid phase extraction, presented obtained good recoveries of diazinon from LC-grade water. An increase in pH caused an increase in the recovery on molecular imprinted solid phase extraction. From these results, the optimal molecular imprinted solid phase extraction procedure was as follows: solid phase extraction packing with 100 mg diazinon imprinted polymers; conditioning with 5 mL of methanol and 6 mL of LC-grade water; sample loading containing diazinon (pH=10; washing with 1 mL of LC-grade water, 1 mL LC- grade water containing 30% acetonitrile and 0.5 mL of acetonitrile, respectively; eluting with 1 mL of methanol containing 2% acetic acid. The percentage recoveries obtained by the optimized molecular imprinted solid phase extraction were more than 90% with drinking water spiked at different trace levels of diazinon. Generally speaking, the molecular imprinted solid phase extraction procedure and subsequent high performance liquid chromatography analysis can be a relatively fast and proper approach for qualitative and quantitative analysis of diazinon in

On-line comprehensive two-dimensional normal-phase liquid chromatography × reversed-phase liquid chromatography for preparative isolation of Peucedanum praeruptorum.

Science.gov (United States)

Wang, Xin-Yuan; Li, Jia-Fu; Jian, Ya-Mei; Wu, Zhen; Fang, Mei-Juan; Qiu, Ying-Kun

2015-03-27

A new on-line comprehensive preparative two-dimensional normal-phase liquid chromatography × reversed-phase liquid chromatography (2D NPLC × RPLC) system was developed for the separation of complicated natural products. It was based on the use of a silica gel packed medium-pressure column as the first dimension and an ODS preparative HPLC column as the second dimension. The two dimensions were connected with normal-phase (NP) and reversed-phase (RP) enrichment units, involving a newly developed airflow assisted adsorption (AAA) technique. The instrument operation and the performance of this NPLC × RPLC separation method were illustrated by gram-scale isolation of ethanol extract from the roots of Peucedanum praeruptorum. In total, 19 compounds with high purity were obtained via automated multi-step preparative separation in a short period of time using this system, and their structures were comprehensively characterized by ESI-MS, (1)H NMR, and (13)C NMR. Including two new compounds, five isomers in two groups with identical HPLC and TLC retention values were also obtained and identified by 1D NMR and 2D NMR. This is the first report of an NPLC × RPLC system successfully applied in an on-line preparative process. This system not only solved the interfacing problem of mobile-phase immiscibility caused by NP and RP separation, it also exhibited apparent advantages in separation efficiency and sample treatment capacity compared with conventional methods. Copyright © 2015 Elsevier B.V. All rights reserved.

Simultaneous analysis of hepatotoxic pyrrolizidine alkaloids and N-oxides in comfrey root by LC-ion trap mass spectrometry.

Science.gov (United States)

Wuilloud, Jorgelina C A; Gratze, Samuel R; Gamble, Bryan M; Wolnik, Karen A

2004-02-01

The purpose of the current study was to develop a LC-MS(n) method for the analysis of pyrrolizidine alkaloids (PAs) in comfrey. Published data presents an extensive list of PAs and their N-oxides present in comfrey. However, standards are not commercially available for any of the PAs typically present in comfrey. Those PAs that are not stereoisomers were readily resolved on a C(18) column using a water-acetonitrile gradient as the mobile phase. The use of a selective technique, LC-MS/MS, allowed us to identify groups of PAs and their N-oxides, as well as identify the number of PAs present in each group, including those that were not completely resolved chromatographically.

RP105 Protects Against Apoptosis in Ischemia/Reperfusion-Induced Myocardial Damage in Rats by Suppressing TLR4-Mediated Signaling Pathways

Directory of Open Access Journals (Sweden)

Jun Yang

2015-07-01

Full Text Available Background: Myocardial apoptosis is heavily implicated in the myocardial damage caused by ischemia-reperfusion (I/R. Toll-like receptor 4 (TLR4 is a potent inducer of these apoptotic cascades. In contrast, the radioprotective 105 kDa protein (RP105 is a specific negative regulator of TLR4 signaling pathways. However, the precise mechanisms by which RP105 inhibits myocardium apoptosis via TLR4-associated pathways during I/R is not fully understood. Methods: We utilized a rat model of myocardial ischemic reperfusion injury (MIRI. Animals were pre-treated with Ad-EGFP adenovirus, Ad-EGFP-RP105 adenovirus, saline, or nothing (sham. After three days, rats underwent a 30min left anterior descending coronary artery occlusion and a 4h reperfusion. Mycardial tissue was assessed by immunohistochemistry, TUNEL-staining, Western blot, quantitative RT-PCR, and a morphometric assay. Results: RP105 overexpression resulted in a reduction in infarct size, fewer TUNEL-positive cardiomyocytes, and a reduction in mitochondrial-associated apoptosis cascade activity. Further, RP105 overexpression repressed I/R-induced myocardial injury by attenuating myocardial apoptosis. This was mediated by inhibiting TLR4 activation and the phosphorylation of P38MAPK and the downstream transcription factor AP-1. Conclusion: RP105 overexpression leads to the de-activation of TLR4, P38MAPK, and AP-1 signaling pathways, and subsequently represses apoptotic cascades and ensuing damage of myocardial ischemic reperfusion. These findings may become the basis of a novel therapeutic approach for reducing of cardiac damage caused by MIRI.

Analysis of membrane proteome by data-dependent LC-MS/MS combined with data-independent LC-MSE technique

Directory of Open Access Journals (Sweden)

Joseph Kwon

2010-03-01

Full Text Available Proteomics work resembles the search for a needle in a haystack. The identification of protein biomarker requires the removal of the false protein data from the whole protein mixture. For high quality proteomic data, even a strict filtration step using the false discovery rate (FDR is insufficient for obtaining perfect protein information from the biological samples. In this study, the cyanobacterial whole membrane fraction was applied to the data-dependent analysis (DDA mode of LC-MS/MS, which was used along with the data-independent LC-MSE technique in order to evaluate the membrane proteomic data. Furthermore, the identified MSE-information (MSE-i data based on the peptide mass and the retention time were validated by the other database search, i.e., the probability-based MASCOT and de novo search engine PEAKS. In this present study, 208 cyanobacterial proteins with FDR of 5% were identified using the data-independent nano-UPLC/MSE acquisition with the Protein Lynx Global Server (PLGS, and 56 of these proteins were the predicted membrane proteins. When a total of 208 MSE-i proteomic data were applied to the DDA mode of LC-MS/MS, the number of identified membrane proteins was 26 and 33 from MASCOT and PEAKS with a FDR of 5%, respectively. The number of totally overlapped membrane proteins was 25. Therefore, the data-independent LC-MSE identified more proteins with a high confidence.

Application of Ionic Liquids in High Performance Reversed-Phase Chromatography

Directory of Open Access Journals (Sweden)

Wentao Bi

2009-06-01

Full Text Available Ionic liquids, considered “green” chemicals, are widely used in many areas of analytical chemistry due to their unique properties. Recently, ionic liquids have been used as a kind of novel additive in separation and combined with silica to synthesize new stationary phase as separation media. This review will focus on the properties and mechanisms of ionic liquids and their potential applications as mobile phase modifier and surface-bonded stationary phase in reversed-phase high performance liquid chromatography (RP-HPLC. Ionic liquids demonstrate advantages and potential in chromatographic field.

Estimation of Rabeprazole Sodium and Itopride Hydrochloride in Tablet Dosage Form Using Reverse Phase High Performance Liquid Chromatography

Directory of Open Access Journals (Sweden)

Shaik Harum Rasheed

2011-01-01

Full Text Available A reversed phase high performance liquid chromatography (RP-HPLC method was developed, validated and used for the quantitative determination of rabeprazole sodium (RP and itopride hydrochloride (IH, from its tablet dosage form. Chromatographic separation was performed on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm, with a mobile phase comprising of a mixture of 50 mM ammonium acetate buffer and methanol (20:80v/v, pH 4.5 adjusted with acetic acid, at a flow rate of 1.3 mL/min with detection at 286 nm. Separation was completed in less than 10 min. As per International Conference on Harmonization (ICH guidelines the method was validated for linearity, accuracy, precision, limit of quantitation and limit of detection. Linearity of RP was found to be in the range of 37.5-375 μg/mL and IH was found to be in the range of 5-50 μg/mL. The correlation coefficients were 0.9997 and 0.9995 for RB and IH respectively. The accuracy of the developed method was found to be 98.6-100.7 for RP and 99.42 -100.81 for IH. The experiment shows the developed method is free from interference of excipients. It indicates the developed RP-HPLC method is simple, linear, precise and accurate and it can be conveniently adopted for the routine quality control analysis of the tablet dosage form.

Putative digenic inheritance of heterozygous RP1L1 and C2orf71 null mutations in syndromic retinal dystrophy

DEFF Research Database (Denmark)

Liu, Yangfan P.; Bosch, Daniëlle G.M.; Siemiatkowska, Anna M.

2017-01-01

Background: Retinitis pigmentosa (RP) is the most common cause of inherited retinal degeneration and can occur in non-syndromic and syndromic forms. Syndromic RP is accompanied by other symptoms such as intellectual disability, hearing loss, or congenital abnormalities. Both forms are known to ex...

Stability indicating RP-LC-PDA method for the quantitative analysis of saxagliptin in pharmaceutical dosage form

Directory of Open Access Journals (Sweden)

Laís Engroff Scheeren

2015-06-01

Full Text Available Saxagliptin is a potent and selective inhibitor of the enzyme dipeptidyl peptidase 4. It is effective in the treatment of type 2 diabetes mellitus because it stimulates the pancreas to produce insulin. In the present study, a liquid chromatography method was developed and validated to quantify the drug in tablets. This method was based on the isocratic elution of saxagliptin, using a mobile phase consisting of 0.1% phosphoric acid at pH 3.0 - methanol (70: 30, v/v at a flow rate of 1 mL.min-1 with UV detection at 225 nm. The chromatographic separation was achieved in 8 minutes on a Waters XBridge C18 column (250 mm x 4.6 mm, 5µm maintained at ambient temperature. The proposed method proved to be specific and robust for the quality control of saxagliptin in pharmaceutical dosage forms, showing good linearity in the range of 15.0 - 100.0 µg.mL-1 (r>0.999, precision (RSD

[Determination of content and entrapment efficiency of 20 (S)-protopanaxadiol in pharmacosomes by RP-HPLC method].

Science.gov (United States)

Han, Meihua; Chen, Jing; Chen, Shilin; Wang, Xiangtao

2009-05-01

To establish a RP-HPLC method for content and entrapment efficiency of 20 (S)-protopanaxadiol in pharmacosomes. The separation was performed with a COSMOSIL 5 C18-MS-II column (4.6 mm x 250 mm, 5 mmicrom) using methanol-water (95:5) as the mobile phase and detected at 203 nm. The flow rate was 1.0 mL x min(-1) and 50 microL sample solution was injected for each time. The calibration curve was linear within the range 0.1-0.5 mg x mL(-1) (r = 0. 9999) , the intra-day RSD and inter-day RSD were less than 2% and the average recovery was between 101.44%-103.11% (n = 3). The method is simple, accurate, sensitive and applicable for determination of content and entrapment efficiency of 20 (S)-protopanaxadiol pharmacosomes.

Efficient preparation of incensole and incensole acetate, and quantification of these bioactive diterpenes in Boswellia papyrifera by a RP-DAD-HPLC method.

Science.gov (United States)

Paul, Michael; Jauch, Johann

2012-03-01

Incensole and incensole acetate, found in incense, are encouraging potent bioactive diterpenic cembrenoids, inhibiting Nuclear Factor-kappaB activation. Furthermore, incensole acetate elicits psycho-activity in mice by activating the TRPV3 channels in the brain. Starting from crude extracts of the incense species Boswellia papyrifera Hochst., a convenient procedure for the efficient large-scale synthesis of incensole and its acetate is presented. Additionally, a reversed-phase, diode-array-detection, high-performance liquid chromatography (RP-DAD-HPLC) method for the quantification of incensole and incensole acetate is reported, indicating that these two compounds are typical biomarkers for B. papyrifera.

Azadirachtin-A and tetrahydroazadirachtin-A concentrates: preparation, LC-MS characterization and insect antifeedant/IGR activity against Helicoverpa armigera (Hübner).

Science.gov (United States)

Sharma, Vandana; Walia, Suresh; Dhingra, Swaran; Kumar, Jitendra; Parmar, Balraj S

2006-10-01

A 60% azadirachtin-A concentrate has been obtained through repeated precipitation with hexane from a methanolic solution of a 20% concentrate. Azadirachtin-A (90%) has been obtained by medium-pressure liquid chromatography of the 60% concentrate with an RP-18 column and a methanol + water (1 + 1 by volume) solvent system. Catalytic hydrogenation of the 60 and 90% azadirachtin concentrates yielded the corresponding tetrahydroazadirachtin concentrates. Dihydroazadirachtin and tetrahydroazadirachtin formed during the first 5 h of hydrogenation were identified by LC-ESI-MS on the basis of their unique mass fragmentation pattern. The efficacy of tetrahydroazadirachtin concentrates in inhibiting the feeding and growth of Helicoverpa armigera (Hübner) larvae has been compared with that of azadirachtin concentrates. They were in general more active and deterred larvae from feeding at all concentrations. Tetrahydroazadirachtin-A (90%) and azadirachtin-A (90%) with respective IC(50) values of 280 and 390 mg L(-1) were effective as insect growth regulators, while tetrahydroazadirachtin-A (90%) displayed higher antifeedant activity (AI(50) 14 mg L(-1)) against the test insect.

Nanomaterials as stationary phases and supports in liquid chromatography.

Science.gov (United States)

Beeram, Sandya R; Rodriguez, Elliott; Doddavenkatanna, Suresh; Li, Zhao; Pekarek, Allegra; Peev, Darin; Goerl, Kathryn; Trovato, Gianfranco; Hofmann, Tino; Hage, David S

2017-10-01

The development of various nanomaterials over the last few decades has led to many applications for these materials in liquid chromatography (LC). This review will look at the types of nanomaterials that have been incorporated into LC systems and the applications that have been explored for such systems. A number of carbon-based nanomaterials and inorganic nanomaterials have been considered for use in LC, ranging from carbon nanotubes, fullerenes and nanodiamonds to metal nanoparticles and nanostructures based on silica, alumina, zirconia and titanium dioxide. Many ways have been described for incorporating these nanomaterials into LC systems. These methods have included covalent immobilization, adsorption, entrapment, and the synthesis or direct development of nanomaterials as part of a chromatographic support. Nanomaterials have been used in many types of LC. These applications have included the reversed-phase, normal-phase, ion-exchange, and affinity modes of LC, as well as related methods such as chiral separations, ion-pair chromatography and hydrophilic interaction liquid chromatography. Both small and large analytes (e.g., dyes, drugs, amino acids, peptides and proteins) have been used to evaluate possible applications for these nanomaterial-based methods. The use of nanomaterials in columns, capillaries and planar chromatography has been considered as part of these efforts. Potential advantages of nanomaterials in these applications have included their good chemical and physical stabilities, the variety of interactions many nanomaterials can have with analytes, and their unique retention properties in some separation formats. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Development of a definition for Rapid Progression (RP) of renal function in HIV-positive persons : the D:A:D study

NARCIS (Netherlands)

Kamara, David A; Ryom, Lene; Ross, Michael; Kirk, Ole; Reiss, Peter; Morlat, Philippe; Moranne, Olivier; Fux, Christoph A; Mocroft, Amanda; Sabin, Caroline; Lundgren, Jens D; Smith, Colette J; Schölvinck, Elisabeth H.

2014-01-01

BACKGROUND: No consensus exists on how to define abnormally rapid deterioration in renal function (Rapid Progression, RP). We developed an operational definition of RP in HIV-positive persons with baseline estimated glomerular filtration rate (eGFR) >90 ml/min/1.73 m2 (using Cockcroft Gault) in the

Development of a definition for Rapid Progression (RP) of renal function in HIV-positive persons: the D:A:D study

NARCIS (Netherlands)

Kamara, David A.; Ryom, Lene; Ross, Michael; Kirk, Ole; Reiss, Peter; Morlat, Philippe; Moranne, Olivier; Fux, Christoph A.; Mocroft, Amanda; Sabin, Caroline; Lundgren, Jens D.; Smith, Colette J.; Powderly, B.; Shortman, N.; Moecklinghoff, C.; Reilly, G.; Franquet, X.; Ryom, L.; Sabin, C. A.; Kamara, D.; Smith, C.; Phillips, A.; Mocroft, A.; Tverland, J.; Mansfeld, M.; Nielsen, J.; Raben, D.; Lundgren, J. D.; Brandt, R. Salbøl; Rickenbach, M.; Fanti, I.; Krum, E.; Hillebregt, M.; Geffard, S.; Sundström, A.; Prins, J. M.; Kuijpers, T. W.; Scherpbier, H. J.; van der Meer, J. T. M.; Wit, F. W. M. N.; Godfried, M. H.; Nellen, F. J. B.; Lange, J. M. A.; Geerlings, S. E.; van Vugt, M.; Pajkrt, D.; Grijsen, M. L.; Wiersinga, W. J.; Goorhuis, A.; Hovius, J. W. R.

2014-01-01

No consensus exists on how to define abnormally rapid deterioration in renal function (Rapid Progression, RP). We developed an operational definition of RP in HIV-positive persons with baseline estimated glomerular filtration rate (eGFR) >90 ml/min/1.73 m2 (using Cockcroft Gault) in the Data

Development of a definition for Rapid Progression (RP) of renal function in HIV-positive persons: the D:A:D study

NARCIS (Netherlands)

Kamara, D.A.; Ryom, L.; Ross, M.; Kirk, O.; Reiss, P.; Morlat, P.; Moranne, O.; Fux, C.A.; Mocroft, A.; Sabin, C.; Lundgren, J.D.; Smith, C.J.; Koopmans †, P.P.; Keuter, M.; Ven, A.J.A.M. van der; Hofstede, H.J.M. ter; Dofferhoff, A.S.M.; Warris, A.; Crevel, R. van; et al.,

2014-01-01

BACKGROUND: No consensus exists on how to define abnormally rapid deterioration in renal function (Rapid Progression, RP). We developed an operational definition of RP in HIV-positive persons with baseline estimated glomerular filtration rate (eGFR) >90 ml/min/1.73 m2 (using Cockcroft Gault) in the

The role of GluN2A and GluN2B NMDA receptor subunits in AgRP and POMC neurons on body weight and glucose homeostasis.

Science.gov (United States)

Üner, Aykut; Gonçalves, Gabriel H M; Li, Wenjing; Porceban, Matheus; Caron, Nicole; Schönke, Milena; Delpire, Eric; Sakimura, Kenji; Bjørbæk, Christian

2015-10-01

Hypothalamic agouti-related peptide (AgRP) and pro-opiomelanocortin (POMC) expressing neurons play critical roles in control of energy balance. Glutamatergic input via n-methyl-d-aspartate receptors (NMDARs) is pivotal for regulation of neuronal activity and is required in AgRP neurons for normal body weight homeostasis. NMDARs typically consist of the obligatory GluN1 subunit and different GluN2 subunits, the latter exerting crucial differential effects on channel activity and neuronal function. Currently, the role of specific GluN2 subunits in AgRP and POMC neurons on whole body energy and glucose balance is unknown. We used the cre-lox system to genetically delete GluN2A or GluN2B only from AgRP or POMC neurons in mice. Mice were then subjected to metabolic analyses and assessment of AgRP and POMC neuronal function through morphological studies. We show that loss of GluN2B from AgRP neurons reduces body weight, fat mass, and food intake, whereas GluN2B in POMC neurons is not required for normal energy balance control. GluN2A subunits in either AgRP or POMC neurons are not required for regulation of body weight. Deletion of GluN2B reduces the number of AgRP neurons and decreases their dendritic length. In addition, loss of GluN2B in AgRP neurons of the morbidly obese and severely diabetic leptin-deficient Lep (ob/ob) mice does not affect body weight and food intake but, remarkably, leads to full correction of hyperglycemia. Lep (ob/ob) mice lacking GluN2B in AgRP neurons are also more sensitive to leptin's anti-obesity actions. GluN2B-containing NMDA receptors in AgRP neurons play a critical role in central control of body weight homeostasis and blood glucose balance via mechanisms that likely involve regulation of AgRP neuronal survival and structure, and modulation of hypothalamic leptin action.

Development of RP-HPLC, Stability Indicating Method for Degradation Products of Linagliptin in Presence of Metformin HCl by Applying 2 Level Factorial Design; and Identification of Impurity-VII, VIII and IX and Synthesis of Impurity-VII

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Sushant B. Jadhav

2017-06-01

Full Text Available The novel reverse phase-high performance liquid chromatography (RP-HPLC, stability indicating method was developed for determination of linagliptin (LGP and its related substances in linagliptin and metformin HCl (MET HCl tablets by implementing design of experiment to understand the critical method parameters and their relation with critical method attributes; to ensure robustness of the method. The separation of nine specified impurities was achieved with a Zorbax SB-Aq 250 × 4.6 mm, 5 µm column, using gradient elution and a detector wavelength of 225 nm, and validated in accordance with International Conference on Harmonization (ICH guidelines and found to be accurate, precise, reproducible, robust, and specific. The drug was found to be degrading extensively in heat, humidity, basic, and oxidation conditions and was forming degradation products during stability studies. After slight modification in the buffer and the column, the same method was used for liquid chromatography–mass spectrometry (LC-MS and ultra-performance liquid chromatography -time-of-flight/mass spectrometry UPLC-TOF/MS analysis, to identify m/z and fragmentation of maximum unspecified degradation products i.e., Impurity-VII (7, Impurity-VIII (8, and Impurity-IX (9 formed during stability studies. Based on the results, a degradation pathway for the drug has been proposed and synthesis of Impurity-VII (7 is also discussed to ensure an in-depth understanding of LGP and its related degradation products and optimum performance during the lifetime of the product.

Phase synchronization of instrumental music signals

Science.gov (United States)

Mukherjee, Sayan; Palit, Sanjay Kumar; Banerjee, Santo; Ariffin, M. R. K.; Bhattacharya, D. K.

2014-06-01

Signal analysis is one of the finest scientific techniques in communication theory. Some quantitative and qualitative measures describe the pattern of a music signal, vary from one to another. Same musical recital, when played by different instrumentalists, generates different types of music patterns. The reason behind various patterns is the psycho-acoustic measures - Dynamics, Timber, Tonality and Rhythm, varies in each time. However, the psycho-acoustic study of the music signals does not reveal any idea about the similarity between the signals. For such cases, study of synchronization of long-term nonlinear dynamics may provide effective results. In this context, phase synchronization (PS) is one of the measures to show synchronization between two non-identical signals. In fact, it is very critical to investigate any other kind of synchronization for experimental condition, because those are completely non identical signals. Also, there exists equivalence between the phases and the distances of the diagonal line in Recurrence plot (RP) of the signals, which is quantifiable by the recurrence quantification measure τ-recurrence rate. This paper considers two nonlinear music signals based on same raga played by two eminent sitar instrumentalists as two non-identical sources. The psycho-acoustic study shows how the Dynamics, Timber, Tonality and Rhythm vary for the two music signals. Then, long term analysis in the form of phase space reconstruction is performed, which reveals the chaotic phase spaces for both the signals. From the RP of both the phase spaces, τ-recurrence rate is calculated. Finally by the correlation of normalized tau-recurrence rate of their 3D phase spaces and the PS of the two music signals has been established. The numerical results well support the analysis.

Gradient Scouting in Reversed-Phase HPLC Revisited

Science.gov (United States)

Alcazar, A.; Jurado, J. M.; Gonzalez, A. G.

2011-01-01

Gradient scouting is the best way to decide the most suitable elution mode in reversed-phase high-performance liquid chromatography (RP-HPLC). A simple rule for this decision involves the evaluation of the ratio [delta]t/t[subscript G] (where [delta]t is the difference in the retention time between the last and the first peak and t[subscript G] is…

Anti-LC1 autoantibodies in patients with chronic hepatitis C virus infection.

Science.gov (United States)

Béland, Kathie; Lapierre, Pascal; Marceau, Gabriel; Alvarez, Fernando

2004-03-01

Various autoantibodies have been reported in patients chronically infected by hepatitis C virus. 2% to 10% of theses patients have anti-liver-kidney microsome type 1 (anti-LKM1) autoantibodies. In type 2 autoimmune hepatitis, anti-LKM1 autoantibodies are frequently associated with anti-liver-cytosol type 1 (anti-LC1) autoantibodies. To determine the prevalence of anti-LC1 autoantibodies in a hepatitis C-positive population and characterize their reactivity. 146 patients suffering from liver diseases, of which 99 were chronically infected by hepatitis C virus, were tested by Western blotting and immunoprecipitation to detect and characterize anti-LC1 autoantibodies. 12% of this hepatitis C population had anti-LC1 autoantibodies. LC1 positivity by Western blotting was 30% of LC1+ sera. Epitopes were found throughout the protein but linear epitopes were situated in the 395-541 amino acid region of formiminotransferase cyclodeaminase. Three putative conformational epitopes were identified by phage display. Anti-LC1 autoantibodies are as prevalent as anti-LKM1 autoantibodies in patients infected with hepatitis C virus and their production is not dependent of anti-LKM1 autoantibodies formation. Autoantibody reactivity against the anti-LC1 antigen is different in hepatitis C than in type 2 autoimmune hepatitis. Anti-LC1 autoantibodies can now be regarded as a serological marker of autoimmunity in chronic hepatitis C infection.

Greening Reversed-Phase Liquid Chromatography Methods Using Alternative Solvents for Pharmaceutical Analysis

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Moussa Yabré

2018-05-01

Full Text Available The greening of analytical methods has gained increasing interest in the field of pharmaceutical analysis to reduce environmental impacts and improve the health safety of analysts. Reversed-phase high-performance liquid chromatography (RP-HPLC is the most widely used analytical technique involved in pharmaceutical drug development and manufacturing, such as the quality control of bulk drugs and pharmaceutical formulations, as well as the analysis of drugs in biological samples. However, RP-HPLC methods commonly use large amounts of organic solvents and generate high quantities of waste to be disposed, leading to some issues in terms of ecological impact and operator safety. In this context, greening HPLC methods is becoming highly desirable. One strategy to reduce the impact of hazardous solvents is to replace classically used organic solvents (i.e., acetonitrile and methanol with greener ones. So far, ethanol has been the most often used alternative organic solvent. Others strategies have followed, such as the use of totally aqueous mobile phases, micellar liquid chromatography, and ionic liquids. These approaches have been well developed, as they do not require equipment investments and are rather economical. This review describes and critically discusses the recent advances in greening RP-HPLC methods dedicated to pharmaceutical analysis based on the use of alternative solvents.

Rp58 and p27kip1 coordinate cell cycle exit and neuronal migration within the embryonic mouse cerebral cortex.

Science.gov (United States)

Clément, Olivier; Hemming, Isabel Anne; Gladwyn-Ng, Ivan Enghian; Qu, Zhengdong; Li, Shan Shan; Piper, Michael; Heng, Julian Ik-Tsen

2017-05-15

During the development of the mammalian cerebral cortex, newborn postmitotic projection neurons are born from local neural stem cells and must undergo radial migration so as to position themselves appropriately to form functional neural circuits. The zinc finger transcriptional repressor Rp58 (also known as Znf238 or Zbtb18) is critical for coordinating corticogenesis, but its underlying molecular mechanism remains to be better characterised. Here, we demonstrate that the co-expression of Rp58 and the cyclin dependent kinase inhibitor (CDKI) p27 kip1 is important for E14.5-born cortical neurons to coordinate cell cycle exit and initiate their radial migration. Notably, we find that the impaired radial positioning of Rp58-deficient cortical neurons within the embryonic (E17.5) mouse cortex, as well as their multipolar to bipolar transition from the intermediate zone to the cortical plate can be restored by forced expression of p27 kip1 in concert with suppression of Rnd2, a downstream target gene of Rp58. Furthermore, the restorative effects of p27 kip1 and Rnd2 abrogation are reminiscent of suppressing RhoA signalling in Rp58-deficient cells. Our findings demonstrate functional interplay between a transcriptional regulator and a CDKI to mediate neuroprogenitor cell cycle exit, as well as to promote radial migration through a molecular mechanism consistent with suppression of RhoA signalling.

Individual Phosphatidylcholine Species Analysis by RP-HPLC-ELSD for Determination of Polyenylphosphatidylcholine in Lecithins.

Science.gov (United States)

Lee, Wei-Ju; Weng, Shun-Hsiang; Su, Nan-Wei

2015-04-22

Polyenylphosphatidylcholine (PPC), a subgroup of the bioactive agents in phosphatidylcholine (PC), has been indicated to possess liver-protective effects. This study aimed to investigate a promising and feasible method to determine PC molecular species with a reverse phase (RP) high-performance liquid chromatograph (HPLC) equipped with an evaporative light scattering detector (ELSD). Chromatography was achieved using a C30 column and an isocratic mobile phase consisting of acetonitrile/methanol/triethylamine (40/58/2, v/v/v) at a flow rate of 1 mL/min, and ELSD detection was performed using 80 °C for the drift tube and an air flow rate of 1.8 L/min. To identify individual peaks on the chromatogram, MALDI-TOF-MS was employed for initial detection, and then the results were used to investigate the relationship between the retention time and fatty acyl chains of each PC molecule. A linear correlation was observed between the retention time and theoretical carbon number (TCN) of individual PC species. The compositions of PC molecular species in soybean and sunflower lecithins were similar to each other, and the major PC molecular species were 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (LLPC), 1-oleoyl-2-linoleoyl-sn-glycero-3-phosphocholine (OLPC), and 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (PLPC). The contents of LLPC in soybean PC and sunflower PC were 40.6% and 64.3%, respectively.

A validated method using RP-HPLC for quantification of reserpine in the Brazilian tree Rauvolfia sellowii Müll. Arg. (Apocynaceae

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Leopoldo C. Baratto

2012-01-01

Full Text Available This study describes a simple, fast and reproducible method using RP-HPLC-UV, in a gradient system, for quantification of reserpine in Rauvolfia sellowii stem bark. The analysis were carried out on a C18 column; mobile phase was water and acetonitrile, and separations were carried out in 10 min, flow rate of 1.0 mL min-1, 25 ºC and 268 nm. The validation data showed that the method was specific, accurate, precise and robust. Results were linear over a range of 0.625-40.0 μg mL-1, and the mean recovery was 95.1%. The amount of reserpine found in the dried stem bark was 0.01% (m/m.

ACADEMIC TRAINING (R.P. Walker)

CERN Multimedia

Françoise Benz

2002-01-01

15, 16, 17 May LECTURE SERIES from 11.00 to 12.00 hrs - Council room, bldg. 503 on 15 May, Auditorium, bldg. 500 on 16 and 17 May Introduction to free electron lasers by R.P. Walker / Rutherford Laboratory, UK The Free-electron laser (FEL) is a source of coherent electromagnetic radiation based on a relativistic electron beam. First operated 25 years ago, the FEL has now reached a stage of maturity for operation in the infra-red region of the spectrum and several facilities provide intense FEL radiation beams for research covering a wide range of disciplines. Several projects both underway and proposed aim at pushing the minimum wavelength from its present limit around 100 nm progressively down to the 1 Angstrom region where the X-ray FEL would open up many new and exciting research possibilities. Other developments aim at increasing power levels to the 10's of kW level. In this series of lectures we give an introduction to the basic principles of FELs and their different modes of operation, and summarise the...

Multiclass determination and confirmation of antibiotic residues in honey using LC-MS/MS.

Science.gov (United States)

Lopez, Mayda I; Pettis, Jeffery S; Smith, I Barton; Chu, Pak-Sin

2008-03-12

A multiclass method has been developed for the determination and confirmation in honey of tetracyclines (chlortetracycline, doxycycline, oxytetracycline, and tetracycline), fluoroquinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, and sarafloxacin), macrolides (tylosin), lincosamides (lincomycin), aminoglycosides (streptomycin), sulfonamides (sulfathiazole), phenicols (chloramphenicol), and fumagillin residues using liquid chromatography tandem mass spectrometry (LC-MS/MS). Erythromycin (a macrolide) and monensin (an ionophore) can be detected and confirmed but not quantitated. Honey samples (approximately 2 g) are dissolved in 10 mL of water and centrifuged. An aliquot of the supernatant is used to determine streptomycin. The remaining supernatant is filtered through a fine-mesh nylon fabric and cleaned up by solid phase extraction. After solvent evaporation and sample reconstitution, 15 antibiotics are assayed by LC-MS/MS using electrospray ionization (ESI) in positive ion mode. Afterward, chloramphenicol is assayed using ESI in negative ion mode. The method has been validated at the low part per billion levels for most of the drugs with accuracies between 65 and 104% and coefficients of variation less than 17%. The evaluation of matrix effects caused by honey of different floral origin is presented.

Stability indicating RP-HPLC method for simultaneous determination of piroxicam and ofloxacin in binary combination.

Science.gov (United States)

John, Peter; Azeem, Waqar; Ashfaq, Muhammad; Khan, Islam Ullah; Razzaq, Syed Naeem

2015-09-01

A simple and precise RP-HPLC method was developed for simultaneous determination of piroxicam and ofloxacin in pharmaceutical formulations and human serum. Optimum separations of piroxicam, ofloxacin and stress-induced degradation products were achieved by use of Hypersil BDS C8 column (250 x 4.6mm, 5 μm). The mobile phase was a mixture of acetonitrile: 0.012M K2HPO4: 0.008M sodium citrate (both buffers mixed and pH adjusted to 2.8) (50:25:25 v/v/v) delivered at flow rate of 1.5 mL min⁻¹ using DAD at 254 nm. Response was linear function of concentration over the ranges of 70-130 mg mL⁻¹ for piroxicam and ofloxacin (r² ≥ 0.999). The method efficiently separated the analytical peaks from degradation products with acceptable tailing and resolution. The developed method was successfully used for concurrent analysis of piroxicam and ofloxacin in pharmaceutical formulations, human serum and in vitro drug interaction studies.

Flexible Bistable Smectic-A Liquid Crystal Device Using Photolithography and Photoinduced Phase Separation

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Yang Lu

2012-01-01

Full Text Available A flexible bistable smectic-A liquid crystal (SmA LC device using pixel-isolated mode was demonstrated, in which SmA LC molecules were isolated in pixels by vertical polymer wall and horizontal polymer layer. The above microstructure was achieved by using ultraviolet (UV photolithography and photoinduced phase separation. The polymer wall was fabricated by photolithography, and then the SmA LC was encapsulated in pixels between polymer wall through UV-induced phase separation, in which the polymer wall acts as supporting structure from mechanical pressure and maintains the cell gap from bending, and the polymer layer acts as adhesive for tight attachment of two substrates. The results demonstrated that all the intrinsic bistable properties of the SmA LC are preserved, and good electrooptical characteristics such as high contrast ratio and excellent stability of the bistable states were characterized. This kind of SmA bistable flexible display has high potential to be used as electronic paper, smart switchable reflective windows, and so forth.

Determination of Insecticidal Effect (LC50 and LC90) of Organic Fatty Acids Mixture (C8910+Silicone) Against Aedes aegypti and Aedes albopictus (Diptera: Culicidae).

Science.gov (United States)

Dunford, James C; Falconer, Aneika; Leite, Laura N; Wirtz, Robert A; Brogdon, William G

2016-05-01

Emerging and re-emerging vector-borne diseases such as chikungunya and dengue and associated Aedes vectors are expanding their historical ranges; thus, there is a need for the development of novel insecticides for use in vector control programs. The mosquito toxicity of a novel insecticide and repellent consisting of medium-chain carbon fatty acids (C8910) was examined. Determination of LC 50 and LC 90 was made against colony-reared Aedes aegypti (L.) and Aedes albopictus (Skuse) using probit analysis on mortality data generated by Centers for Disease Control and Prevention bottle bioassays. Six different concentrations of C8910 + silicone oil yielded an LC 50 of 160.3 µg a.i/bottle (147.6-182.7) and LC 90 of 282.8 (233.2-394.2) in Ae. aegypti; five concentrations yielded an LC 50 of 125.4 (116.1-137.6) and LC 90 of 192.5 (165.0-278.9) in Ae. albopictus. Further development of C8910 and similar compounds could provide vector control specialists novel insecticides for controlling insect disease vectors. Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.

Transient phases during crystallization of solution-processed organic thin films

Science.gov (United States)

Wan, Jing; Li, Yang; Ulbrandt, Jeffery; Smilgies, Detlef-M.; Hollin, Jonathan; Whalley, Adam; Headrick, Randall

We report an in-situ study of 2,7-dioctyl[1]benzothieno[3,2-b][1]benzothiophene (C8-BTBT) organic semiconductor thin film deposition from solution via hollow pen writing, which exhibits multiple transient phases during crystallization. Under high writing speed (25 mm/s) the films have an isotropic morphology, although the mobilities range up to 3.0 cm2/V.s. To understand the crystallization in this highly non-equilibrium regime, we employ in-situ microbeam grazing incidence wide-angle X-ray scattering combined with optical video microscopy at different deposition temperatures. A sequence of crystallization was observed in which a layered liquid-crystalline (LC) phase of C8-BTBT precedes inter-layer ordering. For films deposited above 80ºC, a transition from LC phase to a transient crystalline state that we denote as Cr1 occurs after a temperature-dependent incubation time, which is consistent with classical nucleation theory. After an additional ~ 0.5s, Cr1 transforms to the final stable structure Cr2. Based on these results, we demonstrate a method to produce large crystalline grain size and high carrier mobility during high-speed processing by controlling the nucleation rate during the transformation from the LC phase. Nsf DMR-1307017, NSF DMR-1332208.

ICPD-a new peak detection algorithm for LC/MS.

Science.gov (United States)

Zhang, Jianqiu; Haskins, William

2010-12-01

The identification and quantification of proteins using label-free Liquid Chromatography/Mass Spectrometry (LC/MS) play crucial roles in biological and biomedical research. Increasing evidence has shown that biomarkers are often low abundance proteins. However, LC/MS systems are subject to considerable noise and sample variability, whose statistical characteristics are still elusive, making computational identification of low abundance proteins extremely challenging. As a result, the inability of identifying low abundance proteins in a proteomic study is the main bottleneck in protein biomarker discovery. In this paper, we propose a new peak detection method called Information Combining Peak Detection (ICPD ) for high resolution LC/MS. In LC/MS, peptides elute during a certain time period and as a result, peptide isotope patterns are registered in multiple MS scans. The key feature of the new algorithm is that the observed isotope patterns registered in multiple scans are combined together for estimating the likelihood of the peptide existence. An isotope pattern matching score based on the likelihood probability is provided and utilized for peak detection. The performance of the new algorithm is evaluated based on protein standards with 48 known proteins. The evaluation shows better peak detection accuracy for low abundance proteins than other LC/MS peak detection methods.

Simultaneous and accurate determination of water- and fat-soluble vitamins in multivitamin tablets by using an RP-HPLC method

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Semahat Kucukkolbasi

2013-01-01

Full Text Available In the present study, a reversed-phase high-performance liquid chromatographic (RP-HPLC procedure was developed and validated for the simultaneous determination of seven water-soluble vitamins (thiamine, riboflavin, niacin, cyanocobalamin, ascorbic acid, folic acid, and p-aminobenzoic acid and four fat-soluble vitamins (retinol acetate, cholecalciferol, α-tocopherol, and phytonadione in multivitamin tablets. The linearity of the method was excellent (R² > 0.999 over the concentration range of 10 - 500 ng mL-1. The statistical evaluation of the method was carried out by performing the intra- and inter-day precision. The accuracy of the method was tested by measuring the average recovery; values ranged between 87.4% and 98.5% and were acceptable quantitative results that corresponded with the label claims.

78 FR 65690 - Trees and Plantings Associated With Eligible Facilities, RP9524.5

Science.gov (United States)

2013-11-01

...] Trees and Plantings Associated With Eligible Facilities, RP9524.5 AGENCY: Federal Emergency Management... policy Trees and Plantings Associated with Eligible Facilities. The Federal Emergency Management Agency... trees, shrubs, and other plantings, including limited eligibility for replacement of grass and sod...

Molecular characterization and bio-functional property determination using SDS-PAGE and RP-HPLC of protein fractions from two Nigella species.

Science.gov (United States)

Alu'datt, Muhammad H; Rababah, Taha; Alhamad, Mohammad N; Alodat, Moh'd; Al-Mahasneh, Majdi A; Gammoh, Sana; Ereifej, Khalil; Almajwal, Ali; Kubow, Stan

2017-09-01

This study aimed to investigate the molecular and bio-functional properties of protein fractions from Nigella damascena and Nigella arvensis, including the albumin, globulin, glutein-1, glutein-2 and prolamin fractions. Protein subunits were not observed in globulin and prolamin fractions. No peaks appeared in RP-HPLC chromatograms of globulin for either species. Two predominant peaks were observed in the RP-HPLC profiles of all protein fractions. Proteins separated by RP-HPLC have potential inhibitory and antioxidant activities in all fractions. Optimum ACE-inhibitory and antioxidant activities of proteins separated by RP-HPLC were observed in glutein-2 and albumin, respectively, for both species. For pepsin and combined pepsin-trypsin hydrolyses, the highest degree of hydrolysis (DH) was obtained in glutein-2 fraction of Nigella arvensis. Highest ACE-inhibitory activity of hydrolyzed protein fractions was found at 4h via pepsin hydrolysis in globulin fraction of Nigella damascena. Highest antioxidant activities of hydrolyzed protein fractions were found in glutelin-2 for both species. Copyright © 2017 Elsevier Ltd. All rights reserved.

Characterization and adjustment of the neutron radiography facility of the RP-10 nuclear reactor

International Nuclear Information System (INIS)

Ravello R, Y.R.

2001-01-01

The main aim of this work was to characterize and adjust the neutron radiography facility of the RP-10 nuclear reactor, and therefore be able to offer with this technique services to the industry and research centers in general. This technique will be complemented with others such as x-rays and gamma radiography. First, the shielding capacity of the facility was analyzed, proving that it complies with the radiological safety requirements established by the radiological safety code. Then gamma filtration tests were conducted in order to implement the direct method for image formation, optical density curves were built according to the thickness of the gamma filter, the type of film and the type of irradiation. Also, the indirect method for image formation was implemented for two types of converters: indium and dysprosium. Growth curves for optical density were also made according to contact time between converter-film, for different types of films. The resolution of the facility was also analyzed using two methods: Klasens (1946) and Harms (1986). Harms method came closer to the resolution of the human eye when compared to the Klasens method. Finally, the application fields of neutron radiography are presented, including those conducted at the neutron radiography facility of the RP-10 nuclear reactor. With this work, the RP-10 neutron radiography facility is ready to offer inspection and research services

File list: Unc.PSC.20.AllAg.EpiLC [Chip-atlas[Archive

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Full Text Available Unc.PSC.20.AllAg.EpiLC mm9 Unclassified Pluripotent stem cell EpiLC SRX1074910,SRX1...074907 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.20.AllAg.EpiLC.bed ...

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Full Text Available Unc.PSC.50.AllAg.EpiLC mm9 Unclassified Pluripotent stem cell EpiLC SRX1074910,SRX1...074907 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.50.AllAg.EpiLC.bed ...

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File list: Unc.PSC.05.AllAg.EpiLC [Chip-atlas[Archive

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Full Text Available Unc.PSC.05.AllAg.EpiLC mm9 Unclassified Pluripotent stem cell EpiLC SRX1074910,SRX1...074907 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.PSC.05.AllAg.EpiLC.bed ...

Simultaneous determination of hyoscine N-butyl bromide and paracetamol in their binary mixture by RP-HPLC method

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Nouruddin W. Ali

2017-05-01

Full Text Available RP-HPLC chromatographic method was developed for the determination of hyoscine N-butyl bromide (HBB and Paracetamol (PAR. In this chromatographic method, HBB and PAR were separated using C18 (25 cm × 4.6 mm i.d. 5 μm particle size column as a stationary phase and water: methanol (50:50, V/V pH adjusted to 3.9 with CF3COOH acid as a mobile phase, maintaining the flow rate at 1.0 mL min−1 with UV detection at 210 nm. The proposed method was successfully applied for the determination of HBB and PAR in pure form over a concentration range of 2.0–50.0 μg mL−1 for HBB with mean percentage recovery of 100.10 ± 0.475 and over a concentration range of 5.0–200.0 μg mL−1 for PAR with mean percentage recovery of 99.87 ± 0.942 and in their pharmaceutical formulations (Buscopan plus® tablets, Buscamol® tablets and Buscopan plus® suppositories.

Isolation and quantitation of metallothionein isoforms using reversed-phase high-performance liquid chromatography

International Nuclear Information System (INIS)

Richards, M.P.; Darcey, S.E.; Steele, N.C.

1986-01-01

Reversed-phase HPLC (RP-HPLC) was used to isolate and quantify metallothionein (MT) isoforms from a variety of animal species and tissues. Separations were performed on C 18 radially compressed cartridge columns, eluted with a 2-step linear gradient of acetonitrile in 10 mM sodium phosphate, pH 7.0. Isoforms were detected by UV absorbance (214 nm) and by on-line interfacing with an atomic absorption spectrophotometer (HPLC-AA) to determine bound Zn, Cd and Cu. Rabbit liver and horse kidney MT's exhibited 7 distinct peaks on RP-HPLC, 2 of which were predominant (MT1 and 2). Pig liver and kidney MT2 yielded 2 subspecies on RP-HPLC, while MT1 yielded a single peak. Avian liver MT was unique from mammalian MT's in that MT2 was about tenfold more abundant than MT1. RP-HPLC and HPLC-AA were used to isolate and quantitate MT isoforms and their Zn content directly from cytosol. Quantitation was achieved by peak area integration and extrapolation from a standard curve of purified avian liver MT2. Both RP-HPLC and HPLC-AA had a lower detection limit of 1 + g of peptide and .1 μg of Zn. Recoveries (92-98%) were determined with labeled ( 35 S) MT and MT of known Zn content. Cytoplasmic MT-Zn in avian embryo hepatocytes cultured with added Zn was quantitated using HPLC-AA. In conclusion, both RP-HPLC and HPLC-AA are rapid and powerful separation techniques for the isolation, quantitation and characterization of the isoproteins comprising the MT gene family

Lipopolysaccharide (LPS) and tumor necrosis factor alpha (TNFα) blunt the response of Neuropeptide Y/Agouti-related peptide (NPY/AgRP) glucose inhibited (GI) neurons to decreased glucose.

Science.gov (United States)

Hao, Lihong; Sheng, Zhenyu; Potian, Joseph; Deak, Adam; Rohowsky-Kochan, Christine; Routh, Vanessa H

2016-10-01

A population of Neuropeptide Y (NPY) neurons which co-express Agouti-related peptide (AgRP) in the arcuate nucleus of the hypothalamus (ARC) are inhibited at physiological levels of brain glucose and activated when glucose levels decline (e.g. glucose-inhibited or GI neurons). Fasting enhances the activation of NPY/AgRP-GI neurons by low glucose. In the present study we tested the hypothesis that lipopolysaccharide (LPS) inhibits the enhanced activation of NPY/AgRP-GI neurons by low glucose following a fast. Mice which express green fluorescent protein (GFP) on their NPY promoter were used to identify NPY/AgRP neurons. Fasting for 24h and LPS injection decreased blood glucose levels. As we have found previously, fasting increased c-fos expression in NPY/AgRP neurons and increased the activation of NPY/AgRP-GI neurons by decreased glucose. As we predicted, LPS blunted these effects of fasting at the 24h time point. Moreover, the inflammatory cytokine tumor necrosis factor alpha (TNFα) blocked the activation of NPY/AgRP-GI neurons by decreased glucose. These data suggest that LPS and TNFα may alter glucose and energy homeostasis, in part, due to changes in the glucose sensitivity of NPY/AgRP neurons. Interestingly, our findings also suggest that NPY/AgRP-GI neurons use a distinct mechanism to sense changes in extracellular glucose as compared to our previous studies of GI neurons in the adjacent ventromedial hypothalamic nucleus. Copyright © 2016 Elsevier B.V. All rights reserved.

LC-lens array with light field algorithm for 3D biomedical applications

Science.gov (United States)

Huang, Yi-Pai; Hsieh, Po-Yuan; Hassanfiroozi, Amir; Martinez, Manuel; Javidi, Bahram; Chu, Chao-Yu; Hsuan, Yun; Chu, Wen-Chun

2016-03-01

In this paper, liquid crystal lens (LC-lens) array was utilized in 3D bio-medical applications including 3D endoscope and light field microscope. Comparing with conventional plastic lens array, which was usually placed in 3D endoscope or light field microscope system to record image disparity, our LC-lens array has higher flexibility of electrically changing its focal length. By using LC-lens array, the working distance and image quality of 3D endoscope and microscope could be enhanced. Furthermore, the 2D/3D switching ability could be achieved if we turn off/on the electrical power on LClens array. In 3D endoscope case, a hexagonal micro LC-lens array with 350um diameter was placed at the front end of a 1mm diameter endoscope. With applying electric field on LC-lens array, the 3D specimen would be recorded as from seven micro-cameras with different disparity. We could calculate 3D construction of specimen with those micro images. In the other hand, if we turn off the electric field on LC-lens array, the conventional high resolution 2D endoscope image would be recorded. In light field microscope case, the LC-lens array was placed in front of the CMOS sensor. The main purpose of LC-lens array is to extend the refocusing distance of light field microscope, which is usually very narrow in focused light field microscope system, by montaging many light field images sequentially focusing on different depth. With adjusting focal length of LC-lens array from 2.4mm to 2.9mm, the refocusing distance was extended from 1mm to 11.3mm. Moreover, we could use a LC wedge to electrically shift the optics axis and increase the resolution of light field.

Development of Chiral LC-MS Methods for small Molecules and Their Applications in the Analysis of Enantiomeric Composition and Pharmacokinetic Studies

Energy Technology Data Exchange (ETDEWEB)

Desai, Meera Jay [Iowa State Univ., Ames, IA (United States)

2004-01-01

The purpose of this research was to develop sensitive LC-MS methods for enantiomeric separation and detection, and then apply these methods for determination of enantiomeric composition and for the study of pharmacokinetic and pharmacodynamic properties of a chiral nutraceutical. Our first study, evaluated the use of reverse phase and polar organic mode for chiral LC-API/MS method development. Reverse phase methods containing high water were found to decrease ionization efficiency in electrospray, while polar organic methods offered good compatibility and low limits of detection with ESI. The use of lower flow rates dramatically increased the sensitivity by an order of magnitude. Additionally, for rapid chiral screening, the coupled Chirobiotic column afforded great applicability for LC-MS method development. Our second study, continued with chiral LC-MS method development in this case for the normal phase mode. Ethoxynonafluorobutane, a fluorocarbon with low flammability and no flashpoint, was used as a substitute solvent for hexane/heptane mobile phases for LC-APCI/MS. Comparable chromatographic resolutions and selectivities were found using ENFB substituted mobile phase systems, although, peak efficiencies were significantly diminished. Limits of detection were either comparable or better for ENFB-MS over heptane-PDA detection. The miscibility of ENFB with a variety of commonly used organic modifiers provided for flexibility in method development. For APCI, lower flow rates did not increase sensitivity as significantly as was previously found for ESI-MS detection. The chiral analysis of native amino acids was evaluated using both APCI and ESI sources. For free amino acids and small peptides, APCI was found to have better sensitivities over ESI at high flow rates. For larger peptides, however, sensitivity was greatly improved with the use of electrospray. Additionally, sensitivity was enhanced with the use of non-volatile additives, This optimized method was then

利用SPSS软件计算杀虫剂的LC50%Calculatiang the LC50 of insecticides with software SPSS

Institute of Scientific and Technical Information of China (English)

贾春生

2006-01-01

利用实例和SPSS 130软件上的Probit过程,介绍如何进行LC50计算,并对主要输出结果进行了解释.并通过与其它软件比较,表明应用SPSS软件进行LC50等计算简便、快速、准确.

Rapid Quantitative Analysis of Naringenin in the Fruit Bodies of Inonotus vaninii by Two-phase Acid Hydrolysis Followed by Reversed Phase-high Performance Liquid Chromatography-ultra Violet.

Science.gov (United States)

Guohua, Xia; Pan, Ruirong; Bao, Rui; Ge, Yanru; Zhou, Cunshan; Shen, Yuping

2017-01-01

Sanghuang is one of mystical traditional Chinese medicines recorded earliest 2000 years ago, that included various fungi of Inonotus genus and was well-known for antitumor effect in modern medicine. Inonotus vaninii is grown in natural forest of Northeastern China merely and used as Sanghuang commercially, but it has no quality control specification until now. This study was to establish a rapid method of two-phase acid hydrolysis followed by reversed phase-high performance liquid chromatography-ultra violet (RP-HPLC-UV) to quantify naringenin in the fruit body of I. vaninii . Sample solution was prepared by pretreatment of raw material in two-phase acid hydrolysis and the hydrolysis technology was optimized. After reconstitution, analysis was performed using RP-HPLC-UV. The method validation was investigated and the naringenin content of sample and comparison were determined. The naringenin was obtained by two-phase acid hydrolysis method, namely, 10.0 g of raw material was hydrolyzed in 200 mL of 1% sulfuric acid aqueous solution (v/v) and 400 mL of chloroform in oil bath at 110°C for 2 h. Good linearity ( r = 0.9992) was achieved between concentration of analyte and peak area. The relative standard deviation (RSD) of precision was 2.47% and the RSD of naringenin contents for repeatability was 3.13%. The accuracy was supported with recoveries at 96.37%, 97.30%, and 99.31%. The sample solution prepared using the proposed method contained higher content of naringenin than conventional method and was stable for 8 h. Due to the high efficiency of sample preparation and high reliability of the HPLC method, it is feasible to use this method for routine analysis of naringenin in the fungus. A convenient two-phase acid hydrolysis was employed to produce naringenin from raw material, and then an efficient and reliable reversed phase-high performance liquid chromatography-ultra violet method was established to monitor naringenin in the fruit bodies of Inonotus vaninii

A LC-MS-MS method for determination of low doxazosin concentrations in plasma after oral administration to dogs.

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Erceg, Marijana; Cindric, Mario; Pozaic Frketic, Lidija; Vertzoni, Maria; Cetina-Cizmek, Biserka; Reppas, Christos

2010-02-01

A rapid and sensitive reversed phase liquid chromatography- tandem mass spectrometry (LC-MS-MS) method is developed for the determination of doxazosin in canine plasma. The samples are prepared by precipitation of proteins using a mixture of methanol and acetonitrile, followed by freezing and evaporation of the organic solvent. The remaining dry residue is redissolved in mobile phase and analyzed by LC-MS-MS with positive electrospray ionization using the selected reactions monitoring mode. An XTerra MS C(18) column, a mobile phase composed of acetonitrile and 2mM ammonium acetate with gradient elution, and a flow rate of 400 microL/min are employed. The elution times for prazosin (internal standard) and doxazosin are approximately 8 and 10 min, respectively. Calibration curves are linear in the 1-20 ng/mL concentration range. Limits of detection and quantification are 0.4 ng/mL and 1.2 ng/mL, respectively. Recovery is higher than 94%. Intra- and inter-day relative standard deviations are below 7% and 8%, respectively. The method is applied for the determination of doxazosin plasma levels following a single administration of doxazosin base and doxazosin mesylate tablets (2 mg dose) to dogs in the fed state. The results indicate possible superiority of the mesylate salt on the plasma input rates of doxazosin.

Chiral chromatography studies of chemical behavior of cinacalcet on polysaccharide chiral reversed-phase HPLC stationary phases.

Science.gov (United States)

Dousa, Michal; Brichác, Jirí

2012-01-01

A rapid HPLC method for the analytical resolution of cinacalcet enantiomers was developed. Four chiral columns (two amylose and two cellulose type) were evaluated in RP systems. Excellent enantioseparation with a resolution of more than 6 was achieved on Chiralpak AY (amylose 5-chloro-2-methylphenylcarbamate chiral stationary phase) using 10 mM triethylamine (pH 8.0)-acetonitrile (40 + 60, v/v) mobile phase. Validation of the HPLC method, including linearity, LOD, LOQ, precision, accuracy, and selectivity, was performed according to the International Conference on Harmonization guidelines. The method was successfully applied for the determination of (S)-cinacalcet in enantiopure active pharmaceutical ingredient (R)-cinacalcet.

RP-HPLC Method Development and Validation for the Determination and Stability Indicative Studies of Montelukast in Bulk and its Pharmaceutical Formulations

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A. Patnaik

2012-01-01

Full Text Available A simple, precise, accurate, economical and reproducible HPLC method for estimation of montelukast in tablet dosage form has been developed. Quantitative HPLC was performed with Shimadzu LC2010c HT with Winchrom Software with UV-Visible detector (SPD-IOA, PUMP (LC-IOAT and (LC-IOATvp. Phenomenex C8, 5 μm, 25 cm × 4.6 mm i.d. column was used in the study. The mobile phase of ACN: Acetate buffer= 6.5:3.5 of pH 3 were used in this study. The conditions optimized were: flow rate (1 mL/minute, wavelength (222 nm and run time was 20 min. Retention time was found to be 3.08 min. The linearity was found to be in the concentration range of 10-100 μg/mL. The developed method was evaluated in the assay of commercially available tablet moni containing 10 mg of montelukast. The amount of drug in tablet was found to be 10.34 mg/tab for the brand. Results of analysis were validated statistically and by recovery studies. The recovery studies 99.67% was indicative of the accuracy of proposed method. The precision was calculated as repeatability, inter and intraday variation (%RSD for the drug. By using the method, stability of the drug has been studied.

RESTING PLASMA AgRP LEVELS RESPONSE TO EXERCISE-CONJUGATED DIET AND ONLY DIET IN OVERWEIGHT AND OBESE SEDENTARY FEMALES

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Mahmoud Delphan

2012-01-01

Full Text Available Agouti-related peptide (AgRP is an orexigenic neuropeptide produced mainly in the arcuate nucleus (ARC of the hypothalamus, which leads to positive energy balance. The purpose of the present study was to investigate the resting plasma AgRP levels’ response to exercise-conjugated diet and only diet in overweight and obese sedentary female college students. Thirty subjects were randomly divided into two experimental groups and one control group. Experimental group 1 experienced 12 days of researchers’ proposed diet (D, experimental group 2 experienced 12 days of researchers’ proposed diet (every day with 12-day running training (the intensity was 60 to 70�0heart rate maximum for 50 minutes every other day (AED and the control group remained sedentary with a normal diet in this period. Blood samples were collected 24 hours before and after the enforced protocol. Body fat percentage (BF�20of subjects was measured using the 3-point method. Plasma AgRP levels were measured using the enzyme-linked immunosorbent assay (ELISA method. The data were analysed by Leven, Kolmogorov-Smirnov, one-way analysis of variance (ANOVA and Tukey post-hoc test. At the end of the protocols, the resting plasma AgRP in the D group increased significantly (p0.05. Body weight and BMI were significantly reduced for both experimental groups (P<0.001. Body fat percentage in the AED group was decreased (p<0.001 and no change was observed in the D group. The present results indicate that the AED is better than the D protocol because it leads to stability of AgRP, which may prevent lipogenesis and ultimately reduce body fat percentage.

Identification of a novel p.R1443W mutation in RP1 gene associated with retinitis pigmentosa sine pigmento

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Li Ma

2013-08-01

Full Text Available AIM: To screen mutations in the retinitis pigmentosa 1 (RP1 gene and the rhodopsin (RHO gene in Chinese patients with retinitis pigmentosa sine pigmento (RPSP and describe the genotype-phenotype relationship of the mutations.METHODS:Twenty affected, unrelated Chinese individuals with RPSP (4 autosomal dominant RPSP, 12 autosomal recessive RPSP and 4 unknown inheritance pattern were recruited between 2009 and 2012. The clinical features were determined by complete ophthalmologic examinations. Polymerase chain reaction (PCR and direct DNA sequencing were used to screen the entire coding region and splice junctions of the RP1 gene and the RHO gene. The cosegregation analysis and population frequency studies were performed for patients with identified mutations.RESULTS: Five variants in the RP1 gene and one in the RHO gene were detected in 20 probands. Four missense changes (rs444772, rs446227, rs414352, rs441800 and one non-coding variant (rs56340615 were common SNPs and none of them showed a significant relationship with RPSP. A missense mutation p.R1443W was identified in the RP1 gene in three affected individuals from a family with autosomal dominant RPSP and was found to cosegregate with the phenotype in this family, suggestive of pathogenic. In addition, population frequency analysis showed the p.R1443W mutation was absent in 300 healthy controls.CONCLUSION: The identification of p.R1443W mutation cosegregating in a family with autosomal dominant RPSP highlights an atypical phenotype of the RP1 gene mutation, while RHO gene is not associated with the pathogenesis of RPSP in this study. To our knowledge, this is the fist mutation identified to associate with RPSP.

Identification of a novel p.R1443W mutation in RP1 gene associated with retinitis pigmentosa sine pigmento.

Science.gov (United States)

Ma, Li; Sheng, Xun-Lun; Li, Hui-Ping; Zhang, Fang-Xia; Liu, Ya-Ni; Rong, Wei-Ning; Zhang, Jian-Ling

2013-01-01

To screen mutations in the retinitis pigmentosa 1 (RP1) gene and the rhodopsin (RHO) gene in Chinese patients with retinitis pigmentosa sine pigmento (RPSP) and describe the genotype-phenotype relationship of the mutations. Twenty affected, unrelated Chinese individuals with RPSP (4 autosomal dominant RPSP, 12 autosomal recessive RPSP and 4 unknown inheritance pattern) were recruited between 2009 and 2012. The clinical features were determined by complete ophthalmologic examinations. Polymerase chain reaction (PCR) and direct DNA sequencing were used to screen the entire coding region and splice junctions of the RP1 gene and the RHO gene. The cosegregation analysis and population frequency studies were performed for patients with identified mutations. Five variants in the RP1 gene and one in the RHO gene were detected in 20 probands. Four missense changes (rs444772, rs446227, rs414352, rs441800) and one non-coding variant (rs56340615) were common SNPs and none of them showed a significant relationship with RPSP. A missense mutation p.R1443W was identified in the RP1 gene in three affected individuals from a family with autosomal dominant RPSP and was found to cosegregate with the phenotype in this family, suggestive of pathogenic. In addition, population frequency analysis showed the p.R1443W mutation was absent in 300 healthy controls. The identification of p.R1443W mutation cosegregating in a family with autosomal dominant RPSP highlights an atypical phenotype of the RP1 gene mutation, while RHO gene is not associated with the pathogenesis of RPSP in this study. To our knowledge, this is the fist mutation identified to associate with RPSP.

Education and training in radiological protection: the activities of the isRP

International Nuclear Information System (INIS)

Coeck, M.

2005-01-01

The International School for Radiological Protection, a task force within SCK-CEN, co-ordinates and organises training programs on all aspects of radiological protection. IsRP courses are directed as well to the private sector as to the political and academic world and the general public. International meetings, publications and recommendations with regard to safety culture increasingly stress the importance of education and training in the field of radiological protection. In addition, the need to standardise and harmonise the recognition of skills and practices on a national and European level is emerging. In this sense, the objectives of isRP are threefold : (1) to continue the organisation of open courses and training-on-demand of which the programmes are made up according to the background level and training requirements of the participants, and this for the Belgian medical and industrial sector; (2) to come to a closer cooperation with national universities and high schools and relevant international institutions; (3) to contribute to a better harmonisation of training practice and of skills recognition on a national and European level

NPY/AgRP neurons are not essential for feeding responses to glucoprivation.

Science.gov (United States)

Luquet, Serge; Phillips, Colin T; Palmiter, Richard D

2007-02-01

Animals respond to hypoglycemia by eating and by stimulating gluconeogenesis. These responses to glucose deprivation are initiated by glucose-sensing neurons in the brain, but the neural circuits that control feeding behavior are not well established. Neurons in the arcuate region of the hypothalamus that express neuropeptide Y (NPY) and agouti-related protein (AgRP) have been implicated in mediating the feeding response to glucoprivation. We devised a method to selectively ablate these neurons in neonatal mice and then tested adult mice for their feeding responses to fasting, mild hypoglycemia, 2-deoxy-d-glucose and a ghrelin receptor agonist. Whereas the feeding response to the ghrelin receptor agonist was completely abrogated, the feeding response to glucoprivation was normal. The feeding response after a fast was attenuated when standard chow was available but normal with more palatable solid or liquid diet. We conclude that NPY/AgRP neurons are not necessary for generating or mediating the orexigenic response to glucose deficiency, but they are essential for the feeding response to ghrelin and refeeding on standard chow after a fast.

Treatment of Brodie's Syndrome using parasymphyseal distraction through virtual surgical planning and RP assisted customized surgical osteotomy guide-A mock surgery report

Science.gov (United States)

Dahake, Sandeep; Kuthe, Abhaykumar; Mawale, Mahesh

2017-10-01

This paper aims to describe virtual surgical planning (VSP), computer aided design (CAD) and rapid prototyping (RP) systems for the preoperative planning of accurate treatment of the Brodie's Syndrome. 3D models of the patient's maxilla and mandible were separately generated based on computed tomography (CT) image data and fabricated using RP. During the customized surgical osteotmy guide (CSOG) design process, the correct position was identified and the geometry of the CSOG was generated based on affected mandible of the patient and fabricated by a RP technique. Surgical approach such as preoperative planning and simulation of surgical procedures was performed using advanced software. The VSP and RP assisted CSOG was used to avoid the damage of the adjacent teeth and neighboring healthy tissues. Finally the mock surgery was performed on the biomodel (i.e. diseased RP model) of mandible with reference to the normal maxilla using osteotomy bur with the help of CSOG. Using this CSOG the exact osteotomy of the mandible and the accurate placement of the distractor were obtained. It ultimately improved the accuracy of the surgery in context of the osteotomy and distraction. The time required in cutting the mandible and placement of the distractor was found comparatively less than the regular free hand surgery.

A Thermal Degradation (Thermolysis) Study of Rotenone Extracted from Derris elliptica Roots Using Reverse-Phase High Performance Liquid Chromatography (RP-HPLC)

International Nuclear Information System (INIS)

Saiful Irwan Zubairi; Mohamad Roji Sarmidi; Ramlan Abdul Aziz

2015-01-01

Bio-pesticides are becoming increasingly important as pest management tools in various cropping systems in the tropics essentially to remedy problems associated with the indiscriminate use of hard and non-environmental friendly inorganic pesticide. In these past few decades, many bio-pesticidal products, both microbial-based (bacteria, fungi, microsprodia, entomopathogenic nematodes and viruses) and plant-based botanicals (rotenone and azadiracthin) have been studied for their use against insect pests in the tropics. In this study, the effects of the concentration process with respect to the yield of rotenone (mg) and its concentration (mg/mL) are presented extensively. The raw plants were collected from Kota Johor Lama, Johor and sorted to obtain the roots and stems. Only the roots and stems were utilized as raw materials of the extraction process. The rotenone from roots and stems was extracted using the normal soaking extraction (NSE) at 28 to 30 degree Celsius with 95 % (v/v) of acetone as a solvent and the solvent-to-solid ratio of 10 mL/ g. The extraction was carried out for 24 h. Next, the liquid crude extract was concentrated using the rotary evaporator at 50 degree Celsius and 80 mbar of vacuum pressure to remove approximately 90 % of solvent. The fractions of the liquid crude extract were collected (15 min/ mL/ fraction), diluted (1/100 with acetone) and cleaned up (to remove any fine debris) prior to determination of rotenone content (mg) and concentration (mg/mL) by using the reverse-phase high performance liquid chromatography (RP-HPLC). Finally, the results showed that there was a significant effect of thermal degradation or dissipation of rotenone content at higher operating temperature (greater than 40 degree Celsius) with a rapid rotenone reduction for the first 15 min of exposure. The possibilities for better exploitation and identification of the effective operating parameters based on the above mentioned results will be perhaps discussed in the

New Inhibitors of the DENV-NS5 RdRp from Carpolepis laurifolia as Potential Antiviral Drugs for Dengue Treatment

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Paul Coulerie

2014-05-01

Full Text Available Since a few decades the dengue virus became a major public health concern and no treatment is available yet. In order to propose potential antidengue compounds for chemotherapy we focused on DENV RNA polymerase (DENV-NS5 RdRp which is specific and essential for the virus replication. Carpolepis laurifolia belongs to the Myrtaceae and is used as febrifuge in traditional kanak medicine. Leaf extract of this plant has been identified as a hit against the DENV-NS5 RdRp. Here we present a bioguided fractionation of the leaf extract of C. laurifolia which is also the first phytochemical evaluation of this plant. Five flavonoids, namely quercetin (1, 6-methyl-7-methoxyapigenin (2, avicularin (3, quercitrin (4 and hyperoside (5, together with betulinic acid (6, were isolated from the leaf extract of C. laurifolia. All isolated compounds were tested individually against the DENV-NS5 RdRp and compared with four other commercial flavonoids: isoquercitrin (7, spiraeoside (8, quercetin-3,4’-di-O-glucoside (9 and rutine (10. Compounds 3, 4, 6, 8 and 10 displayed IC 50 ranging from 1.7 to 2.1 µM, and were the most active against the DENV-NS5 RdRp.

Sensitive determination of nitrophenol isomers by reverse-phase high-performance liquid chromatography in conjunction with liquid-liquid extraction

Science.gov (United States)

A method for the highly sensitive determination of 2-, 3- and 4- nitrophenols was developed using reverse-phase high-performance liquid chromatography (RP-HPLC) with a UV photodiode array detector. Using a reverse-phase column and 40% aqueous acetonitrile as an eluent (i.e. isocratic elution), the i...

High-precision mass measurements for the rp-process at JYFLTRAP

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Canete Laetitia

2017-01-01

Full Text Available The double Penning trap JYFLTRAP at the University of Jyväskylä has been successfully used to achieve high-precision mass measurements of nuclei involved in the rapid proton-capture (rp process. A precise mass measurement of 31Cl is essential to estimate the waiting point condition of 30S in the rp-process occurring in type I x-ray bursts (XRBs. The mass-excess of 31C1 measured at JYFLTRAP, -7034.7(3.4 keV, is 15 more precise than the value given in the Atomic Mass Evaluation 2012. The proton separation energy Sp determined from the new mass-excess value confirmed that 30S is a waiting point, with a lower-temperature limit of 0.44 GK. The mass of 52Co effects both 51Fe(p,γ52Co and 52Co(p,γ53Ni reactions. The mass-excess value measured, - 34 331.6(6.6 keV is 30 times more precise than the value given in AME2012. The Q values for the 51Fe(p,γ52Co and 52Co(p,γ53Ni reactions are now known with a high precision, 1418(11 keV and 2588(26 keV respectively. The results show that 52Co is more proton bound and 53Ni less proton bound than what was expected from the extrapolated value.

[Analysis of phenylethanoid glycosides of Herba cistanchis by RP-HPLC].

Science.gov (United States)

Tu, P F; Wang, B; Deyama, T; Zhang, Z G; Lou, Z C

1997-04-01

The Chinese drug "Rou Cong-rong" (Herba Cistanchis) is one of the commonly used drugs in Chinese traditional medicine. It is used to reinforce the vital function of kidney, especially that of the sexual organs and induce laxation, for the treatment of impotence, premature ejaculation in men, infertility, morbid leukorrhea, profuse metrorrhagia in women, and chronic constipation in the aged. This paper deals with the qualitative and quantitative analysis of phenylethanoid glycosides of four species and one variety of Genus Cistanche and 23 lots of commercial crude drugs of Herba Cistanchis by RP-HPLC. The results were as follows: the chemical constituents of Cistanche deserticola Ma, C. salsa (C. A. Mey) G. Beck, C. salsa var. albiflora P. F. Tu et Z. C. Lou and C. tubulosa were similar while those of C. sinensis were different from the others; the contents of echinacoside and acteoside of C. salsa, which were 2.13% and 1.51%, were the highest of the genus Cistanche. An ODS column (Alltima C18, 5 microns, 250 x 4.6 mm) was employed. Linear gradient elution of acetonitrile--1.5% acetic acid was used as mobile phase, and concentration of acetontrile was from 8% to 20% (0-60 min) in the qualitative analysis, and from 11.5 to 20% (0-35 min) in the quantitative analysis. The flow rate was 1.2 ml.min-1. The detection wavelength was set at 335 nm.

Identification of a nuclear localization signal in the retinitis pigmentosa-mutated RP26 protein, ceramide kinase-like protein

International Nuclear Information System (INIS)

Inagaki, Yuichi; Mitsutake, Susumu; Igarashi, Yasuyuki

2006-01-01

Retinitis pigmentosa (RP) is a genetically heterogeneous disease characterized by degeneration of the retina. A mutation in a new ceramide kinase (CERK) homologous gene, named CERK-like protein (CERKL), was found to cause autosomal recessive retinitis pigmentosa (RP26). Here, we show a point mutation of one of two putative nuclear localization signal (NLS) sequences inhibited the nuclear localization of the protein. Furthermore, the tetra-GFP-tagged NLS, which cannot passively enter the nucleus, was observed not only in the nucleus but also in the nucleolus. Our results provide First evidence of the active nuclear import of CERKL and suggest that the identified NLS might be responsible for nucleolar retention of the protein. As recent studies have shown other RP-related proteins are localized in the nucleus or the nucleolus, our identification of NLS in CERKL suggests that CERKL likely plays important roles for retinal functions in the nucleus and the nucleolus

Keefektifan Model Pembelajaran LC 5E Dan TSTS Berbantuan LKPD Terhadap Hasil Belajar

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Ririn Asmawati

2014-06-01

Full Text Available AbstrakTujuan penelitian ini adalah untuk mengetahui apakah hasil belajar pada pembelajaran Learning Cycle 5E (LC 5E dan kooperatif Two Stay Two Stray (TSTS berbantuan LKPD dapat mencapai KKM yang ditetapkan sekolah; untuk mengetahui apakah rata-rata hasil belajar pada pembelajaran LC 5E dan TSTS berbantuan LKPD lebih tinggi daripada pem-belajaran ekspositori, serta apakah rata-rata hasil belajar pada pembelajaran LC 5E berban-tuan LKPD lebih baik daripada pembelajaran TSTS berbantuan LKPD. Data penelitian diperoleh dengan metode tes dan observasi. Analisis data hasil belajar meliputi uji nor-malitas, uji homogenitas, uji perbedaan rata-rata, uji scheffe, dan uji proporsi. Hasil pene-litian menunjukkan bahwa hasil belajar pada pembelajaran LC 5E berbantuan LKPD telah mencapai KKM, sedangkan hasil belajar pada pembelajaran TSTS berbantuan LKPD dan pembelajaran ekspositori belum mencapai KKM. Selain itu, rata-rata hasil belajar pada pembelajaran LC 5E dan TSTS berbantuan LKPD lebih tinggi daripada pembelajaran eks-positori, sedangkan rata-rata hasil belajar pada pembelajaran LC 5E berbantuan LKPD le-bih baik daripada pembelajaran TSTS berbantuan LKPD. Kata kunci:       hasil belajar; LC 5E; TSTS  AbstractThe purposes of this research are to find out whether the learning outcomes in the Learning Cycle 5E (LC 5E and the cooperative Two Stay Two Stray (TSTS assisted LKPD may attain the KKM settled by the school; to find out whether the average of the learning outcome in the LC 5E and TSTS assisted LKPD is higher than the expository learning, and whether the average of the learning outcome in the LC 5E assisted LKPD is better than TSTS assisted LKPD. The data of the research were obtained by testing and observation methods. The data analysis of learning outcomes included normality test, homogeneity test, mean difference test, Scheffe test, and proportions test. The research outcome indicated that the learning outcome in the implementation of

The Structural Basis for Lipid and Endotoxin Binding in RP105-MD-1, and Consequences for Regulation of Host Lipopolysaccharide Sensitivity.

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Ortiz-Suarez, Maite L; Bond, Peter J

2016-01-05

MD-1 is a member of the MD-2-related lipid-recognition (ML) family, and associates with RP105, a cell-surface protein that resembles Toll-like receptor 4 (TLR4). The RP105⋅MD-1 complex has been proposed to play a role in fine-tuning the innate immune response to endotoxin such as bacterial lipopolysaccharide (LPS) via TLR4⋅MD-2, but controversy surrounds its mechanism. We have used atomically detailed simulations to reveal the structural basis for ligand binding and consequent functional dynamics of MD-1 and the RP105 complex. We rationalize reports of endogenous phospholipid binding, by showing that they prevent collapse of the malleable MD-1 fold, before refining crystallographic models and uncovering likely binding modes for LPS analogs. Subsequent binding affinity calculations reveal that endotoxin specificity arises from the entropic cost of expanding the MD-1 cavity to accommodate bulky lipid tails, and support the role of MD-1 as a "sink" that sequesters endotoxin from TLR4 and stabilizes RP105/TLR4 interactions. Copyright © 2016 Elsevier Ltd. All rights reserved.

High-Temperature Nucleosynthesis Processes on the Proton-Rich Side of Stability: the Alpha-Rich Freezeout and the rp^2-Process

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Meyer, Bradley S.

2001-10-01

Nucleosynthesis on the proton-rich side of stability has at least two intriguing aspects. First, the most abundant of the stable iron-group isotopes, such as ^48Ti, ^52Cr, and ^56,57Fe, are synthesized as proton-rich, radioactive parents in alpha-rich freezeouts from equilibrium. The production of these radioactive progenitors depends in large measure on reactions on the proton-rich side of stability. The second intriguing aspect is that explosive nucleosynthesis in a hydrogen-rich environment (namely, the rp-process) may be associated with exotic astrophysical settings, such as x-ray bursts, and may be responsible for production of some of the light p-process nuclei (for example, ^92,94Mo and ^96,98Ru). We have developed web-based tools to help nuclear physicists determine which nuclear reactions on the proton-rich side of stability govern the nucleosynthesis in these processes. For the alpha-rich freezeout, one may determine the effect of any one of 2,140 reactions on the yield of any isotope in the nuclear reaction network with the web calculator. As a relevant example, I will discuss the governing role of ^57Ni (n,p)^57Co in the synthesis of the important astronomical observable ^57Co. As for explosive, proton-rich burning, I will discuss the synthesis of p-process nuclei in the repetitive rp-process (the rp^2-process). movies/rp.html>Movies of the rp^2-process illustrate its important features and give some indications of the important nuclear reactions.

Effect of Mobile Phase Additives on the Resolution of Four Bioactive Compounds by RP-HPLC

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Shengnan Li

2010-05-01

Full Text Available The use of mobile phase additives enhances the separation and resolution of the bioactive compounds on the C18 column. Chlorogenic acid, caffeic acid, rutin, and scoparone from Herba Artemisiae Scopariae were investigated as the target compounds. Acetic acid, triethylamine, inorganic salts, and several ionic liquids were added as mobile phase additives into methanol/water (40:60, v/v. The result revealed that a mobile phase with 0.01 mol/L of ionic liquid [BMIM][BF4] enabled the optimum separation of the four target compounds.

Characterisation of the ester-substituted products of the reaction of p-t-butyl calix[4]arene and ethyl bromoacetate using LC-UV-MS and LC-DAD.

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McMahon, Gillian; Wall, Rachel; Nolan, Kieran; Diamond, Dermot

2002-07-19

A series of derivatisation reactions between p-t-butyl calix[4]arene and ethyl bromoacetate were carried out in order to prepare 1,3 diester substituted calix[4]arene. Mass spectral data, obtained from direct injection of samples, indicated that the reactions were rich in the desired product. Since the ultra violet (UV) spectra of the desired product and possible impurities are very similar, liquid chromatography (LC) chromatographic data seemed to corroborate these results. However, when on-line LC-UV-MS was carried out and each LC peak subjected to MS analysis as it eluted, a very different picture emerged. It was found that many of these reactions actually contained high levels of the monoester product which, having less affinity for sodium in the MS, is therefore seriously underestimated in any direct injection assay. LC-diode array detection (DAD) methods were also used to help successfully identify and characterise the compounds being formed in these complex reactions. The overall results obtained in this paper allowed the optimal reaction conditions to be determined for this reaction. LC-MS analysis of the chromatographic peaks also identified the presence of two isomers of the diester substituted calix[4]arene (1,3 and 1,2 diesters). The combination of LC and UV/MS detection is required for accurate analysis of the products of such reactions.

Polar and nonpolar organic polymer-based monolithic columns for capillary electrochromatography and high-performance liquid chromatography.

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Rathnasekara, Renuka; Khadka, Shantipriya; Jonnada, Murthy; El Rassi, Ziad

2017-01-01

This review article is a continuation of the previous reviews on the area of monolithic columns covering the progress made in the field over the last couple of years from the beginning of the second half of 2014 until the end of the first half of 2016. It summarizes and evaluates the evolvement of both polar and nonpolar organic monolithic columns and their use in hydrophilic interaction LC and CEC and reversed-phase chromatography and RP-CEC. The review article discusses the results reported in a total of 62 references. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Simultaneous Determination of Florfenicol and Diclazuril in Compound Powder by RP-HPLC-UV Method

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Leilei Guo

2014-01-01

Full Text Available A RP-HPLC-UV method was developed and validated for simultaneous determination of florfenicol and diclazuril in compound powder. The separation involved using a SinoChoom ODS-BP C18 (5 μm, 4.6 mm × 250 mm analytical column. The mobile phase was a mixture of acetonitrile-0.2% phosphoric acid (pH was adjusted to 3.0 with triethylamine. The ratio of acetonitrile and 0.2% phosphoric acid in the mobile phase was 60 : 40 (v/v from 0 minutes to 6 minutes and 70 : 30 (v/v from 6.1 minutes to 15 minutes. The flow rate was 1 mL/min. The temperature of the analytical column was maintained at 30°C. The detection was monitored at 225 nm and 277 nm for florfenicol and diclazuril, respectively. The excipients in the compound powder did not interfere with the drug peaks. The calibration curves of florfenicol and diclazuril were fairly linear over the concentration ranges between 50.0–500.0 μg/mL (r=0.9995 and 10.0–100.0 μg/mL (r=0.9992, respectively. The RSD of both the intraday and interday variations was below 2.1% for florfenicol and diclazuril. The method was successfully validated according to International Conference on Harmonisation and proved to be suitable for the simultaneous determination of florfenicol and diclazuril in compound powder.

Trans Ova Genetics, L.C.

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The EPA is providing notice of a proposed Administrative Penalty Assessment against Trans Ova Genetics, L.C., a business located at 2938 380th Street Sioux Center, IA 51250, for alleged violations at the Trans Ova Genetics, L.C.’s facility located in 12425

LC-MS (/MS) in clinical toxicology screening methods.

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Viette, Véronique; Hochstrasser, Denis; Fathi, Marc

2012-01-01

Toxicological screening is the analysis of biological samples to detect and identify unknown compounds. The high selectivity and sensitivity of liquid chromatography (LC) coupled to mass spectrometry (MS) or tandem mass spectrometry (MS/MS) technology provide an attractive alternative to the current methods (LC-UV, GC/MS, etc.). For these reasons, an increasing number of applications are being published. This paper is a brief overview of LC-MS(/MS) screening methods developed for clinical toxicology in recent years. Various sample treatments, chromatographic separations and detection by mass spectrometry can be combined to obtain screening methods adapted to the constraints and needs of clinical toxicology laboratories. Currently the techniques are in the hands of specialists, mainly in academic institutions. However, the evolution in technology should allow application of these techniques as a tool in toxicology laboratories, thus allowing a more widespread exploitation of their potential.

RP-HPLC-DAD method for the determination of phenylepherine, paracetamol, caffeine and chlorpheniramine in bulk and marketed formulation

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A.P. Dewani

2014-11-01

Full Text Available A simple, specific and accurate isocratic RP-HPLC-DAD method was developed for the simultaneous determination of phenylephrine, paracetamol, caffeine and chlorpheniramine in bulk and tablet dosage form. The four contents are present in variable concentrations and have variable chromatographic behavior making the process of analysis very difficult. For present studies a reversed-phase C-18 column (150 mm × 4.5 mm i.d., particle size 5 μm with mobile phase consisting of acetonitrile, methanol and 10 Mm phosphate buffer 16:22:62 (v/v (pH of buffer 2.5 ± 0.02, adjusted with ortho phosphoric acid was used. The flow rate was 1.0 ml/min and eluents were monitored at 280 nm. The mean retention times of phenylephrine, paracetamol, caffeine and chlorpheniramine were found to be 1.8, 3.1, 5.2 and 10.9 min, respectively. The method was validated in terms of linearity, range, specificity, accuracy, precision and robustness. The proposed method was successfully applied to the estimation of phenylephrine, paracetamol, caffeine and chlorpheniramine in combined tablet dosage form.

Development and validation of a fast RP-HPLC method for determination of methotrexate entrapment efficiency in polymeric nanocapsules.

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Sartori, Tatiane; Seigi Murakami, Fabio; Pinheiro Cruz, Ariane; Machado de Campos, Angela

2008-07-01

A rapid and effective isocratic chromatographic procedure is successfully developed to determinate methotrexate (MTX) entrapment efficiency (EE) in polymeric nanocapsules using reversed-phase high-performance liquid chromatography. The method employed a RP-C(18) Shimadzu Shim-pack CLC-ODS (150 mm x 4.6 mm, 5 microm) column with mobile phase constituted by a mixture of water-acetonitrile-tetrahydrofuran (65:30:5 v/v/v; pH 3.0) at a flow rate of 0.8 mL/min. The eluate is monitored with a UV detector set at 313 nm. The parameters used in the validation process are: linearity, specificity, precision, accuracy, and limit of quantitation (LOQ). The linearity is evaluated by a calibration curve in the concentration range of 10-50 microg/mL and presented a correlation coefficient of 0.9998. The polymers (PLA or PLA-PEG), oil, and surfactants used in the nanocapsule formulation did not interfere with analysis and the recovery was quantitative. The intra and inter-day assay relative standard deviation were less than 0.72%. Results are satisfactory, and the method proved to be adequate for the determination of methotrexate in nanocapsules formulations.

Simultaneous Determination of Cyclosporine A, Tacrolimus, Sirolimus, and Everolimus in Whole-Blood Samples by LC-MS/MS

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Mustafa Karapirli

2012-01-01

Full Text Available Objectives. Cyclosporine A (CyA, tacrolimus (TRL, sirolimus (SIR, and everolimus (RAD are immunosuppressive drugs frequently used in organ transplantation. Our aim was to confirm a robust sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS method for determination of CyA, TRL, SIR, and RAD in whole-blood samples. Materials and Methods. We used an integrated online solid-phase extraction-LC-MS/MS system and atmospheric pressure ionization tandem mass spectrometry (API-MS/MS in the multiple reaction monitoring (MRM detection mode. CyA, TRL, SIR, and RAD were simultaneously analyzed in whole blood treated with precipitation reagent taken from transplant patients. Results. System performance parameters were suitable for using this method as a high-throughput technique in clinical practice. The high concentration of one analyte in the sample did not affect the concentration of other analytes. Total analytical time was 2.5 min, and retention times of all analytes were shorter than 2 minutes. Conclusion. This LC-MS/MS method can be preferable for therapeutic drug monitoring of these immunosuppressive drugs (CyA, TRL, SRL, and RAD in whole blood. Sample preparation was too short and simple in this method, and it permits robust, rapid, sensitive, selective, and simultaneous determination of these drugs.

Comparison of ultra-violet and inductively coupled plasma-atomic emission spectrometry for the on-line quantification of selenium species after their separation by reversed-phase liquid chromatography

International Nuclear Information System (INIS)

Tsopelas, Fotios N.; Ochsenkuehn-Petropoulou, Maria Th.; Mergias, Ioannis G.; Tsakanika, Lambrini V.

2005-01-01

An analytical approach for selenium speciation using liquid chromatography (LC) coupled with ultra-violet (UV) and inductively coupled plasma-atomic emission spectrometry (ICP-AES) was developed. The separation of the investigated selenium species, selenites, selenates, selenomethionine, selenocystine, selenocystamine and dimethyldiselenide was accomplished in less than 6 min on a BIO Wide Pore RP-18 column using sodium salt of n-octanesulfonic acid as ion-pairing modifier. The on-line detection of the separated selenium species was performed using UV spectrometry at the optimum wavelength of 192 nm, obtained by the UV spectra of the investigated individual selenium species. ICP-AES was also used as element specific on-line detector, after its coupling with the chromatographic system. The UV and ICP-AES detectors were compared for their suitability, including sensitivity and detection limits, for the on-line quantification of the six selenium species. The developed LC-UV as well as LC-ICP-AES techniques were successfully applied to a selenized yeast candidate reference material, after its enzymatic extraction with protease XIV. It was found that the described LC-UV technique is suitable for the determination of selenomethionine, the main selenium compound in the yeast, with an accuracy of 5%, although the UV detector is not element specific and it is rarely used for selenium speciation. This finding can prove valuable for routine laboratories to perform selenium speciation in such materials

Corrosión de componentes de aluminio en el reactor RP-10

OpenAIRE

Morales Larrea, Soledad; Tenorio de la Cruz, Favio

1983-01-01

Se analiza la velocidad de corrosión, por pérdida de peso de placas de aluminio a diversos valores de pH y temperatura en solución acuosa. El estudio se hace simulando las condiciones de trabajo (pH y temperatura) a las que pueden verse sometidas las vainas de aluminio de los elementos combustibles del reactor RP-10.

Twenty years of Radiology in RP-10 nuclear reactor protection

International Nuclear Information System (INIS)

Zapata, Alejandro L.; Ramos, Fernando T.; Arrieta, Rolando W.B.; Vela Mora, Mariano

2013-01-01

In this report we present the results about radiation controls during 1990 - 2010, carried out in the Nuclear Reactor RP-10 of the Nuclear Center of Huarangal. These controls and radiological evaluation are of much utility for the radio personnel protection of this one and other reactors, since it allows to compares these variables with respect to the time. From the results obtained in monitoring and radiation controls, we conclude that in no case it has been reached the limits allowed by the Peruvian Regulating Authority. (author)

Central control of circadian phase in arousal-promoting neurons.

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Carrie E Mahoney

Full Text Available Cells of the dorsomedial/lateral hypothalamus (DMH/LH that produce hypocretin (HCRT promote arousal in part by activation of cells of the locus coeruleus (LC which express tyrosine hydroxylase (TH. The suprachiasmatic nucleus (SCN drives endogenous daily rhythms, including those of sleep and wakefulness. These circadian oscillations are generated by a transcriptional-translational feedback loop in which the Period (Per genes constitute critical components. This cell-autonomous molecular clock operates not only within the SCN but also in neurons of other brain regions. However, the phenotype of such neurons and the nature of the phase controlling signal from the pacemaker are largely unknown. We used dual fluorescent in situ hybridization to assess clock function in vasopressin, HCRT and TH cells of the SCN, DMH/LH and LC, respectively, of male Syrian hamsters. In the first experiment, we found that Per1 expression in HCRT and TH oscillated in animals held in constant darkness with a peak phase that lagged that in AVP cells of the SCN by several hours. In the second experiment, hamsters induced to split their locomotor rhythms by exposure to constant light had asymmetric Per1 expression within cells of the middle SCN at 6 h before activity onset (AO and in HCRT cells 9 h before and at AO. We did not observe evidence of lateralization of Per1 expression in the LC. We conclude that the SCN communicates circadian phase to HCRT cells via lateralized neural projections, and suggests that Per1 expression in the LC may be regulated by signals of a global or bilateral nature.

A compact, all solid-state LC high voltage generator.

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Fan, Xuliang; Liu, Jinliang

2013-06-01

LC generator is widely applied in the field of high voltage generation technology. A compact and all solid-state LC high voltage generator based on saturable pulse transformer is proposed in this paper. First, working principle of the generator is presented. Theoretical analysis and circuit simulation are used to verify the design of the generator. Experimental studies of the proposed LC generator with two-stage main energy storage capacitors are carried out. And the results show that the proposed LC generator operates as expected. When the isolation inductance is 27 μH, the output voltage is 1.9 times larger than the charging voltage on single capacitor. The multiplication of voltages is achieved. On the condition that the primary energy storage capacitor is charged to 857 V, the output voltage of the generator can reach to 59.5 kV. The step-up ratio is nearly 69. When self breakdown gas gap switch is used as main switch, the rise time of the voltage pulse on load resistor is 8.7 ns. It means that the series-wound inductance in the discharging circuit is very small in this system. This generator can be employed in two different applications.

Dopamine serotonin stabilizer RP5063: A randomized, double-blind, placebo-controlled multicenter trial of safety and efficacy in exacerbation of schizophrenia or schizoaffective disorder.

Science.gov (United States)

Cantillon, Marc; Prakash, Arul; Alexander, Ajay; Ings, Robert; Sweitzer, Dennis; Bhat, Laxminarayan

2017-11-01

The study objectives were to evaluate the efficacy, safety, tolerability, and pharmacokinetics of RP5063 versus placebo. The study was conducted in adults with acute exacerbation of schizophrenia or schizoaffective disorder. This 28-day, multicenter, placebo-controlled, double-blind study randomized 234 subjects to RP5063 15, 30, or 50mg; aripiprazole; or placebo (3:3:3:1:2) once daily. The aripiprazole arm was included solely to show assay sensitivity and was not powered to show efficacy. The primary endpoint was change from baseline to Day 28/EOT (End-of-Treatment) in Positive and Negative Syndrome Scale (PANSS) total score; secondary endpoints included PANSS subscales, improvement ≥1 point on the Clinical Global Impressions-Severity (CGI-S), depression and cognition scales. The primary analysis of PANSS Total showed improvement by a mean (SE) of -20.23 (2.65), -15.42 (2.04), and -19.21 (2.39) in the RP5063 15, 30, and 50mg arms, versus -11.41 (3.45) in the placebo arm. The difference between treatment and placebo reached statistical significance for the 15mg (p=0.021) and 50mg (p=0.016) arms. Improvement with RP5063 was also seen for multiple secondary efficacy outcomes. Discontinuation for any reason was much lower for RP5063 (14%, 25%, 12%) versus placebo (26%) and aripiprazole (35%). The most common treatment-emergent adverse events (TEAE) in the RP5063 groups were insomnia and agitation. There were no significant changes in body weight, electrocardiogram, or incidence of orthostatic hypotension; there was a decrease in blood glucose, lipid profiles, and prolactin levels. In conclusion, the novel dopamine serotonin stabilizer, RP5063 is an efficacious and well-tolerated treatment for acute exacerbation of schizophrenia or schizoaffective disorder. Copyright © 2017. Published by Elsevier B.V.

Discrimination of Four Marine Biofilm-Forming Bacteria by LC-MS Metabolomics and Influence of Culture Parameters.

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Favre, Laurie; Ortalo-Magné, Annick; Greff, Stéphane; Pérez, Thierry; Thomas, Olivier P; Martin, Jean-Charles; Culioli, Gérald

2017-05-05

Most marine bacteria can form biofilms, and they are the main components of biofilms observed on marine surfaces. Biofilms constitute a widespread life strategy, as growing in such structures offers many important biological benefits. The molecular compounds expressed in biofilms and, more generally, the metabolomes of marine bacteria remain poorly studied. In this context, a nontargeted LC-MS metabolomics approach of marine biofilm-forming bacterial strains was developed. Four marine bacteria, Persicivirga (Nonlabens) mediterranea TC4 and TC7, Pseudoalteromonas lipolytica TC8, and Shewanella sp. TC11, were used as model organisms. The main objective was to search for some strain-specific bacterial metabolites and to determine how culture parameters (culture medium, growth phase, and mode of culture) may affect the cellular metabolism of each strain and thus the global interstrain metabolic discrimination. LC-MS profiling and statistical partial least-squares discriminant analyses showed that the four strains could be differentiated at the species level whatever the medium, the growth phase, or the mode of culture (planktonic vs biofilm). A MS/MS molecular network was subsequently built and allowed the identification of putative bacterial biomarkers. TC8 was discriminated by a series of ornithine lipids, while the P. mediterranea strains produced hydroxylated ornithine and glycine lipids. Among the P. mediterranea strains, TC7 extracts were distinguished by the occurrence of diamine derivatives, such as putrescine amides.

Determination of ximelagatran, melagatran and two intermediary metabolites in plasma by mixed-mode solid phase extraction and LC-MS/MS.

Science.gov (United States)

Dunér, Kristina; Bäckström, Jonas; Magnell, Niklas; Svennberg, Henrik; Ahnoff, Martin; Logren, Ulrika

2007-06-01

An analytical method was developed for the determination of ximelagatran, an oral direct thrombin inhibitor, its active metabolite melagatran, and the two intermediate metabolites, OH-melagatran and ethyl-melagatran in human plasma. Extraction of plasma was carried out on a mixed mode bonded sorbent material (C8/SO(3)(-)). All four analytes, including their isotope-labelled internal standards, were eluted at high ionic strength with a mixture of 50% methanol and 50% buffer (0.25 M ammonium acetate and 0.05 M formic acid, pH 5.3) with an extraction recovery above 80%. The extracts were demonstrated to be clean in terms of a low concentration of albumin and lysoPC. The sample extraction was fully automated and performed in 96-well plates using a Tecan Genesis pipetting robot. Analysis of the extracts were performed with liquid chromatography followed by positive electrospray ionization mass spectrometry. The low organic content and the low pH of the extracts allowed for, after dilution 1:3 with buffer, direct injection onto the LC-column. The four analytes were separated on a C18 analytical LC-column using gradient elution with the acetonitrile concentration varying from 10 to 30% (v/v) and the ammonium acetate and acetic acid concentration kept constant at 10 and 5 mmol/L, respectively, at a flow rate of 0.75 mL/min. Linearity was achieved over the calibrated range 0.010-4.0 micromol/L with accuracy and relative standard deviation in the range 96.9-101.2% and 6.6-17.1%, respectively at LLOQ, and in the range 94.7-102.6% and 2.7-6.8%, respectively at concentrations above 3 x LLOQ. The method replaces a manual method, and displays the advantages of having a fully automated sample clean-up, no evaporation/reconstitution step, high recovery, and complete LC-separation of all four analytes.

Improving lactose digestion and symptoms of lactose intolerance with a novel galacto-oligosaccharide (RP-G28): a randomized, double-blind clinical trial.

Science.gov (United States)

Savaiano, Dennis A; Ritter, Andrew J; Klaenhammer, Todd R; James, Gareth M; Longcore, Amy T; Chandler, Justin R; Walker, W Allan; Foyt, Howard L

2013-12-13

Lactose intolerance (LI) is a common medical problem with limited treatment options. The primary symptoms are abdominal pain, diarrhea, bloating, flatulence, and cramping. Limiting dairy foods to reduce symptoms contributes to low calcium intake and the risk for chronic disease. Adaptation of the colon bacteria to effectively metabolize lactose is a novel and potentially useful approach to improve lactose digestion and tolerance. RP-G28 is novel galacto-oligosaccharide (GOS) being investigated to improve lactose digestion and the symptoms of lactose intolerance in affected patients. A randomized, double-blind, parallel group, placebo-controlled study was conducted at 2 sites in the United States. RP-G28 or placebo was administered to 85 patients with LI for 35 days. Post-treatment, subjects reintroduced dairy into their daily diets and were followed for 30 additional days to evaluate lactose digestion as measured by hydrogen production and symptom improvements via a patient-reported symptom assessment instrument. Lactose digestion and symptoms of LI trended toward improvement on RP-G28 at the end of treatment and 30 days post-treatment. A reduction in abdominal pain was also demonstrated in the study results. Fifty percent of RP-G28 subjects with abdominal pain at baseline reported no abdominal pain at the end of treatment and 30 days post treatment (p = 0.0190). RP-G28 subjects were also six times more likely to claim lactose tolerance post-treatment once dairy foods had been re-introduced into their diets (p = 0.0389). Efficacy trends and favorable safety/tolerability findings suggest that RP-G28 appears to be a potentially useful approach for improving lactose digestion and LI symptoms. The concurrent reduction in abdominal pain and improved overall tolerance could be a meaningful benefit to lactose intolerant individuals.

Improving lactose digestion and symptoms of lactose intolerance with a novel galacto-oligosaccharide (RP-G28): a randomized, double-blind clinical trial

Science.gov (United States)

2013-01-01

Background Lactose intolerance (LI) is a common medical problem with limited treatment options. The primary symptoms are abdominal pain, diarrhea, bloating, flatulence, and cramping. Limiting dairy foods to reduce symptoms contributes to low calcium intake and the risk for chronic disease. Adaptation of the colon bacteria to effectively metabolize lactose is a novel and potentially useful approach to improve lactose digestion and tolerance. RP-G28 is novel galacto-oligosaccharide (GOS) being investigated to improve lactose digestion and the symptoms of lactose intolerance in affected patients. Methods A randomized, double-blind, parallel group, placebo-controlled study was conducted at 2 sites in the United States. RP-G28 or placebo was administered to 85 patients with LI for 35 days. Post-treatment, subjects reintroduced dairy into their daily diets and were followed for 30 additional days to evaluate lactose digestion as measured by hydrogen production and symptom improvements via a patient-reported symptom assessment instrument. Results Lactose digestion and symptoms of LI trended toward improvement on RP-G28 at the end of treatment and 30 days post-treatment. A reduction in abdominal pain was also demonstrated in the study results. Fifty percent of RP-G28 subjects with abdominal pain at baseline reported no abdominal pain at the end of treatment and 30 days post treatment (p = 0.0190). RP-G28 subjects were also six times more likely to claim lactose tolerance post-treatment once dairy foods had been re-introduced into their diets (p = 0.0389). Conclusions Efficacy trends and favorable safety/tolerability findings suggest that RP-G28 appears to be a potentially useful approach for improving lactose digestion and LI symptoms. The concurrent reduction in abdominal pain and improved overall tolerance could be a meaningful benefit to lactose intolerant individuals. Study registration ClinicalTrials.gov NCT01113619. PMID:24330605

Two phase transitions in Nuclear Physics

International Nuclear Information System (INIS)

Bes, D.R.

1985-01-01

The status of the art of the problem associated with two phase transitions in the nuclear matter, viz.: the disappearance of the nuclear superfluiditiy with the raising of the rotation velocity and the appearance of an octupolar deformation in the actinide zone, is presented. (L.C.) [pt

Analysis of benzodiazepines and their metabolites using DBS cards and LC-MS/MS.

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Lee, Heesang; Park, Yujin; Jo, Jiyeong; In, Sangwhan; Park, Yonghoon; Kim, Eunmi; Pyo, Jaesung; Choe, Sanggil

2015-10-01

Dried Blood Spot (DBS) has been used a blood extraction method for inherited metabolic disorder screening since 1960s. With introduction of LC-MS/MS, not only DBS could be used to analysis drugs in small blood volume, but in various fields, such as toxicology, drug therapeutic monitoring, drug diagnostic screening, and illicit drugs. In toxicology field, many drugs (e.g. benzodiazepines, acetaminophen, small molecule drugs) have been tested with DBS. Compared with earlier blood extraction methods (SPE and LLE), DBS has lots of advantages; lower blood volume (less than 50μL), shorter analysis time caused by a more concise analysis procedure and lower cost. We optimized the DBS procedure and LC-MS/MS conditions for 18 benzodiazepines, seven benzodiazepine metabolites, and one z-drug (zolpidem) analysis in blood. 30μL of whole blood was spotted on FTA DMPK card C and dried for 2h in a desiccator. A 6-mm disk was punched and vortexed for 1min in a centrifuge tube with 300μL methanol/acetonitrile mixture (1:1, v/v). After evaporation, redissolved in 100μL mobile phase of LC-MS/MS and 5μL was injected. In the analysis for 26 target compounds in blood, all of the method validation parameters - LLOD, LLOQ, accuracy (intra- and inter-assay), and precision (intra- and inter-assay) - were satisfied with method validation criteria, within 15%. The results of matrix effect, recovery, and process efficiency were good. We developed a fast and reliable sample preparation method using DBS for 26 benzodiazepines, benzodiazepine metabolites, and z-drug (zolpidem). Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Development of Isocratic RP-HPLC Method for Separation and Quantification of L-Citrulline and L-Arginine in Watermelons

Directory of Open Access Journals (Sweden)

Rasdin Ridwan

2018-01-01

Full Text Available Watermelons (Citrullus lanatus are known to have sufficient amino acid content. In this study, watermelons grown and consumed in Malaysia were investigated for their amino acid content, L-citrulline and L-arginine, by the isocratic RP-HPLC method. Flesh and rind watermelons were juiced, and freeze-dried samples were used for separation and quantification of L-citrulline and L-arginine. Three different mobile phases, 0.7% H3P04, 0.1% H3P04, and 0.7% H3P04 : ACN (90 : 10, were tested on two different columns using Zorbax Eclipse XDB-C18 and Gemini C18 with a flow rate of 0.5 mL/min and a detection wavelength at 195 nm. Efficient separation with reproducible resolution of L-citrulline and L-arginine was achieved using 0.1% H3P04 on the Gemini C18 column. The method was validated and good linearity of L-citrulline and L-arginine was obtained with R2 = 0.9956, y=0.1664x+2.4142 and R2=0.9912, y=0.4100x+3.4850, respectively. L-citrulline content showed the highest concentration in red watermelon of flesh and rind juice extract (43.81 mg/g and 45.02 mg/g, whereas L-arginine concentration was lower than L-citrulline, ranging from 3.39 to 11.14 mg/g. The isocratic RP-HPLC method with 0.1% H3P04 on the Gemini C18 column proved to be efficient for separation and quantification of L-citrulline and L-arginine in watermelons.

In tube-solid phase microextraction-nano liquid chromatography: Application to the determination of intact and degraded polar triazines in waters and recovered struvite.

Science.gov (United States)

Serra-Mora, P; Jornet-Martinez, N; Moliner-Martinez, Y; Campíns-Falcó, P

2017-09-01

In-tube solid-phase microextraction (IT-SPME) coupled to miniaturized liquid chromatography (LC) techniques are attractive mainly due to the column efficiency improvement, sensitivity enhancement and reduction of solvent consumption. In addition, the nanomaterials based sorbents can play a key role in the improvement of the extraction efficiency taking into account their interesting physical and chemical properties. Thus, in this work the performance of IT-SPME coupled to nano LC (NanoLC) has been compared with the performance of IT-SPME coupled to capillary LC (CapLC) with similar configurations for the determination of polar triazines including their degradation products. In both cases, a DAD detector was used. Different extractive phases such as TRB-5, TRB-5/c-SWNTs, TRB-5/c-MWNTs capillary columns have been tested. The dimensions of the capillary columns were 0.32mm id×40cm length and 0.1 or 0.075mm i.d.×15cm length for the couplings with CapLC and NanoLC, respectively. The processed volume was 4mL for CapLC and 0.5mL for NanoLC. The elution was carried out with ACN:H 2 O (30:70, v/v). IT-SPME-NanoLC has shown a higher performance than IT-SPME-CapLC for the target analytes demonstrating the enhancement of the extraction efficiency with the former configuration. A new phase TEOS-MTEOS-SiO 2 NPs has been also proposed for IT-SPME-NanoLC, which improves the retention of polar compounds. Compared with previously published works, improved LODs were achieved (0.025-0.5μgL -1 ). The practical application of the proposed procedure has been demonstrated for the analysis of water samples and recovered struvite samples from wastewater treatment plants. Therefore, the proposed procedure can be an alternative method for regulatory purposes. Copyright © 2017 Elsevier B.V. All rights reserved.

New theory for competing interactions and microstructures in partially-ordered (liquid-crystalline) phases

International Nuclear Information System (INIS)

Dowell, F.

1987-01-01

A summary of results from a unique statistical-physics theory to predict and explain competing interactions and resulting microstructures in some partially-ordered [in this case, liquid-crystalline (LC)] phases is presented. The static aspects of both partial orientational and partial positional ordering of the molecules into various microstructures in these phases (including the incommensurate smectic-Ad phase) can be understood in terms of various competing interactions (both entropic and energetic) involved in the packing together of the different molecular sub-units at given pressures and temperatures. These microstructures are predicted and explained (using no ad hoc or arbitrarily adjustable parameter) as a function of molecule chemical structure [including lengths and shapes (from bond lengths and angles), intramolecular rotations, site-site polarizabilities and pair potentials, dipole moments, etc]. Theoretical results are presented for the nematic, re-entrant nematic, smectic-Ad, and smectic-Al LC phases and the isotropic phase

RP process studies with radioactive beams at ATLAS

Energy Technology Data Exchange (ETDEWEB)

Rehm, K E [Argonne National Lab., Physics Div., Argonne, IL (United States)

1998-06-01

Reactions of interest to nuclear astrophysics have been studied with radioactive beams at the ATLAS accelerator. Using a modified ISOL technique, beams of {sup 18}F(T{sub 1/2}=110 min) and {sup 56}Ni(T{sub 1/2}=6.1 d) have been produced. The reactions {sup 18}F(p,{alpha}){sup 15}O, {sup 18}F(p,{gamma}){sup 19}Ne, and {sup 56}Ni(d,p){sup 57}Ni have been investigated. The results indicate that the {sup 18}F(p,{gamma}) route is a small contributor to the breakout from the hot CNO cycle into the rp process, while the {sup 56}Ni(p,{gamma}){sup 57}Cu rate is about ten times larger than previously assumed. (orig.)

Analysis of Theobromine and Related Compounds by Reversed Phase High-Performance Liquid Chromatography with Ultraviolet Detection: An Update (1992–2011

Directory of Open Access Journals (Sweden)

Sandra Aparecida de Assis

2011-01-01

Full Text Available Theobromine and its related compounds, such as caffeine and theophylline, are secondary metabolites that belong to the alkaloids and have economic and cultural importance. These alkaloids have demonstrated stimulatory effects on the central nervous, gastrointestinal, cardiovascular, renal and respiratory systems, resulting in 'energy arousal', increased motivation to work, increased alertness and increased cognitive function. Several analytical methods have been used to analyse these compounds, but reversed phase high-performance liquid chromatography (RP-HPLC is the most commonly applied because of its efficiency, sensitivity, specificity and speed. This review describes the analyses of theobromine-related compounds by RP-HPLC with ultraviolet detection (UV in four sources: food, beverages, biological fluids and plants. Many RP-HPLC methods have been developed and optimized for the detection and quantification of these natural compounds. Elution under isocratic conditions is the most frequent method, with a water, methanol and acetonitrile mixture modified with acetic, phosphoric or formic acid as the mobile phase. For xanthine analysis, the use of reversed phase high-performance liquid chromatography with an ultraviolet/diode array detector (UV/DAD is particularly suitable as derivation is not required; it allows the analysis of absorbance at all wavelengths, it is simple and rapid.

File list: InP.PSC.50.AllAg.EpiLC [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available InP.PSC.50.AllAg.EpiLC mm9 Input control Pluripotent stem cell EpiLC SRX823839,SRX5...5,SRX823838,SRX823831,SRX823826,SRX590324 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.PSC.50.AllAg.EpiLC.bed ...

Passive Reactor Compensated Cascaded H-Bridge Multilevel LC-StatCom

DEFF Research Database (Denmark)

Farivar, Glen; Townsend, Christopher David; Hredzak, Branislav

2017-01-01

The cascaded H-bridge (CHB) low-capacitance StatCom (LC-StatCom) has a limited operating area in the inductive region compared to a conventional StatCom's V–I characteristic. This limitation for operation in the inductive region is considered to be the biggest disadvantage of CHB LC-StatCom. In t...

Neutral Higgs Sector of the MSSM without $R_{p}$

CERN Document Server

Davidson, Sacha; Rius, N

2000-01-01

We analyse the neutral scalar sector of the MSSM without R-parity. Our analysis is performed for a one-generation model in terms of ``basis-independent'' parameters, and includes one-loop corrections due to large yukawa couplings. We concentrate on the consequences of large $R_p$ violating masses in the soft sector, which mix the Higgses with the sleptons, because these are only constrained by their one-loop contributions to neutrino masses. We focus on the effect of $R_p$-violation on the Higgs mass and branching ratios. We find that the experimental lower bound on the lightest CP-even Higgs in this model can be lower than in the MSSM.

Development and validation of an RP-HPLC method for quantification of trans-resveratrol in the plant extracts

Directory of Open Access Journals (Sweden)

Cvetković Žika S.

2015-01-01

Full Text Available New, simple, cost effective, accurate and reproducible RP-HPLC method was developed and validated for the quantification of trans-resveratrol in the extracts of grape exocarp and seeds. The method has proved to be simpler and faster than available methods. Methanol was used as a mobile phase with a flow rate of 1.0 cm3 min-1, while the quantification was effected at 306 nm. The separation was performed at 35°C using a C18 column. The results showed that the peak area response was linear in the concentration range of 1-40 μg cm-3. The values of LOD and LOQ were found to be 0.125 and 0.413 μg cm-3, respectively. The antioxidant activity of the extracts was determined using DPPH assay. The ability of DPPH radicals inhibition decreases in the following order: the extract of grape exocarp > trans-resveratrol standard > the extract of grape seeds. [Projekat Ministarstva nauke Republike Srbije, br. TRp-34012

A validated stability-indicating RP-HPLC method for levofloxacin in the presence of degradation products, its process related impurities and identification of oxidative degradant.

Science.gov (United States)

Lalitha Devi, M; Chandrasekhar, K B

2009-12-05

The objective of current study was to develop a validated specific stability indicating reversed-phase liquid chromatographic method for the quantitative determination of levofloxacin as well as its related substances determination in bulk samples, pharmaceutical dosage forms in the presence of degradation products and its process related impurities. Forced degradation studies were performed on bulk sample of levofloxacin as per ICH prescribed stress conditions using acid, base, oxidative, water hydrolysis, thermal stress and photolytic degradation to show the stability indicating power of the method. Significant degradation was observed during oxidative stress and the degradation product formed was identified by LCMS/MS, slight degradation in acidic stress and no degradation was observed in other stress conditions. The chromatographic method was optimized using the samples generated from forced degradation studies and the impurity spiked solution. Good resolution between the peaks corresponds to process related impurities and degradation products from the analyte were achieved on ACE C18 column using the mobile phase consists a mixture of 0.5% (v/v) triethyl amine in sodium dihydrogen orthophosphate dihydrate (25 mM; pH 6.0) and methanol using a simple linear gradient. The detection was carried out at 294 nm. The limit of detection and the limit of quantitation for the levofloxacin and its process related impurities were established. The stressed test solutions were assayed against the qualified working standard of levofloxacin and the mass balance in each case was in between 99.4 and 99.8% indicating that the developed LC method was stability indicating. Validation of the developed LC method was carried out as per ICH requirements. The developed LC method was found to be suitable to check the quality of bulk samples of levofloxacin at the time of batch release and also during its stability studies (long term and accelerated stability).

Identification of phase I and II metabolites of the new designer drug α-pyrrolidinohexiophenone (α-PHP) in human urine by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS).

Science.gov (United States)

Paul, Michael; Bleicher, Sergej; Guber, Susanne; Ippisch, Josef; Polettini, Aldo; Schultis, Wolfgang

2015-11-01

Pyrrolidinophenones represent one emerging class of newly encountered drugs of abuse, also known as 'new psychoactive substances', with stimulating psychoactive effects. In this work, we report on the detection of the new designer drug α-pyrrolidinohexiophenone (α-PHP) and its phase I and II metabolites in a human urine sample of a drug abuser. Determination and structural elucidation of these metabolites have been achieved by liquid chromatography electrospray ionisation quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS). By tentative identification, the exact and approximate structures of 19 phase I metabolites and nine phase II glucuronides were elucidated. Major metabolic pathways revealed the reduction of the ß-keto moieties to their corresponding alcohols, didesalkylation of the pyrrolidine ring, hydroxylation and oxidation of the aliphatic side chain leading to n-hydroxy, aldehyde and carboxylate metabolites, and oxidation of the pyrrolidine ring to its lactam followed by ring cleavage and additional hydroxylation, reduction and oxidation steps and combinations thereof. The most abundant phase II metabolites were glucuronidated ß-keto-reduced alcohols. Besides the great number of metabolites detected in this sample, α-PHP is still one of the most abundant ions together with its ß-keto-reduced alcoholic dihydro metabolite. Monitoring of these metabolites in clinical and forensic toxicology may unambiguously prove the abuse of the new designer drug α-PHP. Copyright © 2015 John Wiley & Sons, Ltd.

New 2LC-Y DC-DC converter topologies for high-voltage/low-current renewable applications

DEFF Research Database (Denmark)

Bhaskar, Mahajan Sagar; Padmanaban, Sanjeevikumar; Maroti, Pandav Kiran

2017-01-01

(2LC-LVD, 2LC-2LVD, 2LC-2LCVD and 2LC-2LCmVD) converters are presented in this treatise which offer an effective solution for renewable applications requiring a very high voltage conversion ratio such as a Photovoltaic Multilevel Inverter (PV-MLI) system, hybrid electrical drives and automotive...

[3H]52770 RP, a platelet-activating factor receptor antagonist, and tritiated platelet-activating factor label a common specific binding site in human polymorphonuclear leukocytes

International Nuclear Information System (INIS)

Marquis, O.; Robaut, C.; Cavero, I.

1988-01-01

In human polymorphonuclear leukocytes (PMNs), the tritiated platelet activating factor ([ 3 H]PAF) labels in a saturable manner a single class of binding sites with a Kd of 3.5 +/- 0.5 nM (n = 7) and a maximum binding capacity (Bmax) of 206 +/- 13 fmol/2.5 X 10(6) PMNs (n = 7). 52770 RP, a nonphospholipid antagonist of PAF receptors, fully and competitively displaced the [ 3 H]PAF from its binding sites with a Ki of 7.0 +/- 0.7 nM (n = 4). The high potency and the low solubility in cellular membranes of this compound led us to prepare [ 3 H]52770 RP. This ligand was characterized by a binding which was rapid, reversible, confined to a single site, saturable, specific and stereoselective. Its Kd and Bmax were 4.2 +/- 0.3 nM and 181 +/- 11 fmol/2.5 X 10(6) PMNs, respectively. The stereoselectivity of the binding was suggested by the 600- and 1050-fold higher potency of the d-enantiomer with respect to l-52770 RP in displacing [ 3 H]52770 RP or [ 3 H]PAF, respectively. Several PAF analogs (e.g., lyso-PAF, 2-O-methyl-lyso-PAF), which are poorly active as PAF receptor agonists in functional tests, were weak displacers of [ 3 H]PAF and [ 3 H]52770 RP. Furthermore, for a series of 14 known PAF receptor agonists or antagonists belonging to different chemical families, there was an excellent correlation (r = 0.98) between their ability to displace [ 3 H]PAF and [ 3 H]52770 RP. Thus, [ 3 H]52770 RP and [ 3 H]PAF appear to interact with the same binding site on human PMNs which is proposed to be the PAF receptor mediating functional responses

new topology for single-phase, three-level, spwm vsi with lc filter

African Journals Online (AJOL)

level PWM inverter. However, this is not the case with single-phase PWM inverters. In these days, the popular single-phase inverters adopt the full-bridge type using approximate sinusoidal modulation technique. The output voltage in them has two values: zero and pos- itive supply dc voltage levels in the positive half cycle.

Can (Or Should) RP Serve as the Model for the Teaching of English ...

African Journals Online (AJOL)

This paper presents the case for and against the continued adoption of RP, but argues that the spread of English to many parts of the world should change our perception of what constitutes Standard English – that the speech of a tiny minority in Britain can no longer be considered the norm by which all others must be ...

Könyvtükörben : Paul-Eerik Rummo / Károly Árpás

Index Scriptorium Estoniae

Árpás, Károly

2003-01-01

Luuletõlked tekstis: "hidegtől kék..." ("külmast siniste..."); "Az én dalom te vagy..."; Liivre gondolva mindig 1-2 (Ikka Liivist mõteldes); Epiloog: tempora mutantur (tõlkinud Károly Árpás)

Determination of phenolic derivatives of antipyrine in plasma with solid-phase extraction and high-performance liquid chromatography-atmospheric-pressure chemical ionization mass spectrometry

NARCIS (Netherlands)

Coolen, S.A.J.; Ligor, T.; Lieshout, van H.P.M.; Huf, F.A.

1999-01-01

This manuscript describes a method to determine antipyrine and its phenolic derivatives in plasma by reversed-phase high-performance liquid chromatography–mass spectrometry (RP-HPLC–MS). The sample pretreatment consisted of a C18 solid-phase extraction (SPE), to remove the salts and proteins. The

Characterization of foot-and-mouth disease virus's viral peptides with LC-ESI-MS

International Nuclear Information System (INIS)

Pzdemir, Z.O.; Bulut, E.K.; Mustafeva, Z.; Karahan, M.

2010-01-01

Peptides and proteins play a central role in numerous biological and physiological processes in living organisms. Viral capsid peptides are part of the viruses' outer shell of genetic materials. Viruses are recognized by immune system via capsid peptides. Depending on this property of capsid peptides, prototypes synthetic peptide-based vaccine can be developed. In this work, we synthesized three different viral peptide sequences of foot-and-mouth disease virus with microwave enhanced solid phase synthesis method. These peptides were characterized by using liquid chromatography electro spray interface mass spectrometry (LC-ESI-MS) with electro spray ionization. We briefly describe the essential facts for peptide characterization. (author)

Sensitometric effects of varying the intensifying screens used with Agfa Dentus ST8G and RP6 panoramic radiographic films.

Science.gov (United States)

Wakoh, M; Farman, A G; Scarfe, W C; Kitagawa, H; Kuroyanagi, K

1997-07-01

To compare the sensitometric effects and information yield of varying the intensifying screens used with both Dentus ST8G and RP6 Agfa Gevaert, Dormagen, Germany panoramic radiographic films. Four screen-film combinations were employed for each of the two film types. The screens used were blue fluorescing PX-III (Kasei Optonix, Tokyo, Japan) and Special (Siemens AG, Bensheim, Germany), as well as green fluorescing Lanex Regular (Eastman Kodak, Rochester, NY, USA) and Trimax T16 (3M, Mineapolis, Minnesota, USA). The density response for each screen-film combination was evaluated using the characteristic curves generated. Information yield, as determined by the radiographic detection of defects in an aluminium test object, was evaluated by nine observers. The characteristic curves for ST8G were different when green and blue fluorescing screens were used; however, those for RP6 varied little irrespective of the choice of intensifying screens. Observers were able to perceive defects at significantly lower radiation exposures for ST8G combined with green fluorescing screens compared with blue emitting screens. RP6 with all screen combinations provided similar image detail perceptibility at comparable exposures with ST8G with green-fluorescing screens. RP6 is suitable for use with either the spectrally matched blue emitting screens or green-emitting screens. ST8G radiographic film should always be matched to rare earth screens.

LC Quadrature Generation in Integrated Circuits

DEFF Research Database (Denmark)

Christensen, Kåre Tais

2001-01-01

Today quadrature signals for IQ demodulation are provided through RC polyphase networks, quadrature oscillators or double frequency VCOs. This paper presents a new method for generating quadrature signals in integrated circuits using only inductors and capacitors. This LC quadrature generation...

Evaluation of Carbon Nanotubes Functionalized Polydimethylsiloxane Based Coatings for In-Tube Solid Phase Microextraction Coupled to Capillary Liquid Chromatography

OpenAIRE

Neus Jornet-Martínez; Pascual Serra-Mora; Yolanda Moliner-Martínez; Rosa Herráez-Hernández; Pilar Campíns-Falcó

2015-01-01

In the present work, the performance of carbon nanotubes (c-CNTs) functionalized polydimethylsiloxane (PDMS) based coatings as extractive phases for in-tube solid phase microextraction (IT-SPME) coupled to Capillary LC (CapLC) has been evaluated. Carboxylic-single walled carbon nanotubes (c-SWNTs) and carboxylic-multi walled carbon nanotubes (c-MWNTs) have been immobilized on the activated surface of PDMS capillary columns. The effect of different percentages of diphenyl groups in the PDMS ex...

Theoretical aspects of gradient reversed-phase high performance liquid chromatography of styrene-butylacrylate block copolymers

NARCIS (Netherlands)

Kolarova, L.; Jandera, P.; Vonk, E.C.; Claessens, H.A.

2004-01-01

Butylacrylate – styrene co-polymers prepared by atom transfer radical polymeratization were separated on an octadecyl silica column by gradient elution with tetrahydrofuran in water, up to the molar masses 10,000. In reversed-phase high performance liquid chromatography (RP-HPLC), the retention of

RP-LC and TLC Densitometric Determination of Paracetamol and Pamabrom in Presence of Hazardous Impurity of Paracetamol and Application to Pharmaceuticals

Directory of Open Access Journals (Sweden)

Ola Mohamed El-Houssini

2013-01-01

Full Text Available Two simple, accurate and reproducible methods were developed and validated for the simultaneous determination of paracetamol (PARA and pamabrom (PAMB in pure form and in tablets. The first method was based on reserved-phase high-performance liquid chromatography, on a Thermo Hypersil ODS column using methanol:0.01 M sodium hexane sulfonate:formic acid (67.5:212.5:1 v/v/v as the mobile phase. The flow rate was 2 mL/min and the column temperature was adjusted to 35 °C. Quantification was achieved with UV detection at 277 nm over concentration range of 100-600 and 4-24 μg/mL, with mean percentage recoveries were found to be 99.90 ± 0.586 and 99.26 ± 0.901 for PARA and PAMB, respectively. The second method was based on thin-layer chromatography separation of PARA and PAMB followed by densitometric measurement of the spots at 254 nm and 277 nm for PARA and PAMB respectively. Separation was carried out on aluminum sheet of silica gel 60F 254 using dichloromethane:methanol:glacial acetic acid (7.5:1:0.5 v/v/v as the mobile phase over concentration range of 1-10 and 0.32-3.20 μg per spot, with mean percentage recovery of 100.52 ± 1.332 and 99.71 ± 1.478 for PARA and PAMB, respectively. The methods retained their accuracy in presence of up to 50% of P-aminophenol and could be successfully applied in tablets.

LC/MS Guided Isolation of Alkaloids from Lotus Leaves by pH-Zone-Refining Counter-Current Chromatography

Directory of Open Access Journals (Sweden)

Rui-Lin Hu

2011-03-01

Full Text Available The traditional methods used in natural product separation primarily target the major components and the minor components may thus be lost during the separation procedure. Consequently, it’s necessary to develop efficient methods for the preparative separation and purification of relatively minor bioactive components. In this paper, a LC/MS method was applied to guide the separation of crude extract of lotus (Nelumbo nucifera Gaertn. leaves whereby a minor component was identified in the LC/MS analysis. Afterwards, an optimized pH-zone-refining CCC method was performed to isolate this product, identified as N-demethylarmepavine. The separation procedure was carried out with a biphasic solvent system composed of hexane-ethyl acetate-methyl alcohol-water (1:6:1:6, v/v with triethylamine (10 mM added to the upper organic phase as a retainer and hydrochloric acid (5 mM to the aqueous mobile phase eluent. Two structurally similar compounds – nuciferine and roemerine – were also obtained from the crude lotus leaves extract. In total 500 mg of crude extract furnished 7.4 mg of N-demethylarmepavine, 45.3 mg of nuciferine and 26.6 mg of roemerine with purities of 90%, 92% and 96%, respectively. Their structures were further identified by HPLC/ESI-MSn, FTICR/MS and the comparison with reference compounds.

Liquid chromatography-tandem mass spectrometry detection of the quaternary ammonium compound mebezonium as an active ingredient in t61.

Science.gov (United States)

Kirschbaum, Katrin M; Grellner, Wolfgang; Rochholz, Gertrud; Musshoff, Frank; Madea, Burkhard

2011-03-01

Quaternary ammonium compounds pose an analytical challenge. Mebezonium, a muscle-relaxing agent contained in veterinary euthanasia solution T61, was analyzed in body fluids, organs, and injection sites of a veterinarian by liquid chromatography-tandem mass spectrometry (LC-MS-MS) method. Additionally, embutramide and tetracaine, which are two other active ingredients contained in T61, methadone, xylazine, and analgesics were detected by LC-MS-MS and high-performance liquid chromatography-ultraviolet detection methods. For detection of mebezonium a solid-phase extraction (SPE) combined with ionpairing reagent heptafluorobutyric acid was developed. Separation was achieved on Phenomenex Synergi Hydro RP C(18) column combined with ammonium formate buffer and acetonitrile (pH 3.5). To enrich other drugs, liquid-liquid extraction procedures were used. Most of these drugs were separated on a Restek Allure PFP Propyl column using the mentioned mobile phase. Mebezonium and embutramide were detected in femoral vein serum in concentrations of 10.9 and 2.0 mg/L, respectively. The concentration of xylazine and methadone in serum was 2.0 and 0.4 mg/L, respectively. The LC-MS-MS method with SPE combined with an ion-pairing reagent allowed the quantitation of mebezonium. Methadone was detected in toxic concentrations and was, in combination with xylazine and T61, considered to be the cause of death.

High-throughput screening and confirmation of 22 banned veterinary drugs in feedstuffs using LC-MS/MS and high-resolution Orbitrap mass spectrometry.

Science.gov (United States)

Wang, Xufeng; Liu, Yanghong; Su, Yijuan; Yang, Jianwen; Bian, Kui; Wang, Zongnan; He, Li-Min

2014-01-15

A new analytical strategy based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with accurate mass high-resolution Orbitrap mass spectrometry (HR-Orbitrap MS) was performed for high-throughput screening, confirmation, and quantification of 22 banned or unauthorized veterinary drugs in feedstuffs according to Bulletin 235 of the Ministry of Agriculture, China. Feed samples were extracted with acidified acetonitrile, followed by cleanup using solid-phase extraction cartridge. The extracts were first screened by LC-MS/MS in a single selected reaction monitoring mode. The suspected positive samples were subjected to a specific pretreatment for confirmation and quantification of analyte of interest with LC-MS/MS and HR-Orbitrap MS. Mean recoveries for all target analytes (except for carbofuran and chlordimeform, which were about 35 and 45%, respectively) ranged from 52.2 to 90.4%, and the relative standard deviations were screening of real samples obtained from local feed markets and confirmation of the suspected target analytes. It provides a high-throughput, sensitive, and reliable screening, identification, and quantification of banned veterinary drugs in routine monitoring programs of feedstuffs.

Development and Validation of a Simultaneous RP-HPLCUV/DAD Method for Determination of Polyphenols in Gels Containing S. terebinthifolius Raddi (Anacardiaceae)

Science.gov (United States)

Carvalho, Melina G.; Aragão, Cícero F. S; Raffin, Fernanda N.; de L. Moura, Túlio F. A.

2017-01-01

Topical gels containing extracts of Schinus terebinthifolius have been used to treat bacterial vaginosis. It has been reported that this species has antimicrobial, anti-inflammatory and anti-ulcerogenic properties, which can be attributed to the presence of phenolic compounds. In this work, a sensitive and selective reversed-phase HPLC-UV/DAD method for the simultaneous assay of six polyphenols that could be present in S. terebinthifolius was developed. The method was shown to be accurate and precise. Peak purity and similarity index both exceeded 0.99. Calibration curves were linear over the concentration range studied, with correlation coefficients between 0.9931 and 0.9974. This method was used to determine the polyphenol content of a hydroalcoholic extract and pharmacy-compounded vaginal gel. Although the method is useful to assess the 6 phenolic compounds, some compounds could not be detected in the products. SUMMARY A sensitive, selective, accurate and precise reversed-phase HPLC-UV/DAD method for the simultaneous assay of six polyphenols in S. terebinthifolius Raddi Abbreviations used: RP-HPLC-UV/DAD: Reverse Phase High Performance Liquid Chromatograph with Ultraviolet and Diode Array Detector, HPLC: High Performance Liquid Chromatograph, HPLC-UV: High Performance Liquid Chromatograph with Ultraviolet Detector, ANVISA: Brazilian National Health Surveillance Agency, LOD: Limit of detection, LOQ: Limit of quantitation PMID:28539726

Stability indicating RP-HPLC method for simultaneous determination of pantoprazole sodium and itopride hydrochloride in bulk and capsule

Directory of Open Access Journals (Sweden)

Krishna R. Gupta

2011-03-01

Full Text Available A stability indicating reversed-phase HPLC method has been developed and subsequently validated for simultaneous estimation of pantoprazole present as pantoprazole sodium sesquihydrate (PSS, and itopride hydrochloride from their combination product. The proposed RP-HPLC method utilizes a Phenomenex® C18, 5 µm, 250 mm X 4.6 mm i.d. column, mobile phase consisting of phosphate buffer and acetonitrile in the proportion of 55:45 (v/v with apparent pH adjusted to 5.0, and UV detection at 289.0 nm using a UV detector. PAN, ITH and their combination drug product were exposed to thermal, photolytic, hydrolytic and oxidative stress conditions, and the stressed samples were analyzed by the proposed method. The described method was linear over a range of 4-20 µg/mL for PAN and 15-75 µg/mL for ITH. The mean recoveries were 100.02 and 99.88 for PAN and ITH, respectively. Chromatographic peak purity data of PAN and ITH indicated no co-eluting peaks with the main peaks of drugs which demonstrated the specificity of assay method for their estimation in presence of degradation products. The proposed method can be useful in the quality control of combination drug products.

Mutual couling reduction using a lumped LC circuit

DEFF Research Database (Denmark)

Thaysen, Jesper; Jakobsen, Kaj Bjarne

2004-01-01

A technique to reduce the mutual coupling between two Planar Inverted F Antennas (PIFA) is presented in this paper. By the use of a parallel LC circuit it is possible to reduce the mutual coupling between two antennas. This results in a 16 % improvement in the radiation efficiency.......A technique to reduce the mutual coupling between two Planar Inverted F Antennas (PIFA) is presented in this paper. By the use of a parallel LC circuit it is possible to reduce the mutual coupling between two antennas. This results in a 16 % improvement in the radiation efficiency....

Interaction of HSP20 with a viral RdRp changes its sub-cellular localization and distribution pattern in plants.

Science.gov (United States)

Li, Jing; Xiang, Cong-Ying; Yang, Jian; Chen, Jian-Ping; Zhang, Heng-Mu

2015-09-11

Small heat shock proteins (sHSPs) perform a fundamental role in protecting cells against a wide array of stresses but their biological function during viral infection remains unknown. Rice stripe virus (RSV) causes a severe disease of rice in Eastern Asia. OsHSP20 and its homologue (NbHSP20) were used as baits in yeast two-hybrid (YTH) assays to screen an RSV cDNA library and were found to interact with the viral RNA-dependent RNA polymerase (RdRp) of RSV. Interactions were confirmed by pull-down and BiFC assays. Further analysis showed that the N-terminus (residues 1-296) of the RdRp was crucial for the interaction between the HSP20s and viral RdRp and responsible for the alteration of the sub-cellular localization and distribution pattern of HSP20s in protoplasts of rice and epidermal cells of Nicotiana benthamiana. This is the first report that a plant virus or a viral protein alters the expression pattern or sub-cellular distribution of sHSPs.

A Study of Method Development, Validation, and Forced Degradation for Simultaneous Quantification of Paracetamol and Ibuprofen in Pharmaceutical Dosage Form by RP-HPLC Method

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Md. Sarowar Jahan

2014-01-01

Full Text Available A rapid and stability-indicating reversed phase high-performance liquid chromatography (RP-HPLC method was developed for simultaneous quantification of paracetamol and ibuprofen in their combined dosage form especially to get some more advantages over other methods already developed for this combination. The method was validated according to United States Pharmacopeia (USP guideline with respect to accuracy, precision, specificity, linearity, solution stability, robustness, sensitivity, and system suitability. Forced degradation study was validated according to International Conference on Harmonisation (ICH. For this, an isocratic condition of mobile phase comprising phosphate buffer (pH 6.8 and acetonitrile in a ratio of 65:35, v/v at a flow rate of 0.7 mL/minute over RP C18 (octadecylsilane (ODS, 150 × 4.6 mm, 5 μm, Phenomenex Inc. column at ambient temperature was maintained. The method showed excellent linear response with correlation coefficient ( R 2 values of 0.999 and 1.0 for paracetamol and ibuprofen respectively, which were within the limit of correlation coefficient ( R 2 > 0.995. The percent recoveries for two drugs were found within the acceptance limit of (97.0-103.0%. Intra-and inter-day precision studies of the new method were less than the maximum allowable limit percentage of relative standard deviation (%RSD ≤ 2.0. Forced degradation of the drug product was carried out as per the ICH guidelines with a view to establishing the stability-indicating property of this method and providing useful information about the degradation pathways, degradation products, and how the quality of a drug substance and drug product changes with time under the influence of various stressing conditions. The degradation of ibuprofen was within the limit (5-20%, according to the guideline of ICH, while paracetamol showed <20% degradation in oxidation and basic condition.

A Study of Method Development, Validation, and Forced Degradation for Simultaneous Quantification of Paracetamol and Ibuprofen in Pharmaceutical Dosage Form by RP-HPLC Method.

Science.gov (United States)

Jahan, Md Sarowar; Islam, Md Jahirul; Begum, Rehana; Kayesh, Ruhul; Rahman, Asma

2014-01-01

A rapid and stability-indicating reversed phase high-performance liquid chromatography (RP-HPLC) method was developed for simultaneous quantification of paracetamol and ibuprofen in their combined dosage form especially to get some more advantages over other methods already developed for this combination. The method was validated according to United States Pharmacopeia (USP) guideline with respect to accuracy, precision, specificity, linearity, solution stability, robustness, sensitivity, and system suitability. Forced degradation study was validated according to International Conference on Harmonisation (ICH). For this, an isocratic condition of mobile phase comprising phosphate buffer (pH 6.8) and acetonitrile in a ratio of 65:35, v/v at a flow rate of 0.7 mL/minute over RP C18 (octadecylsilane (ODS), 150 × 4.6 mm, 5 μm, Phenomenex Inc.) column at ambient temperature was maintained. The method showed excellent linear response with correlation coefficient (R (2)) values of 0.999 and 1.0 for paracetamol and ibuprofen respectively, which were within the limit of correlation coefficient (R (2) > 0.995). The percent recoveries for two drugs were found within the acceptance limit of (97.0-103.0%). Intra-and inter-day precision studies of the new method were less than the maximum allowable limit percentage of relative standard deviation (%RSD) ≤ 2.0. Forced degradation of the drug product was carried out as per the ICH guidelines with a view to establishing the stability-indicating property of this method and providing useful information about the degradation pathways, degradation products, and how the quality of a drug substance and drug product changes with time under the influence of various stressing conditions. The degradation of ibuprofen was within the limit (5-20%, according to the guideline of ICH), while paracetamol showed degradation in oxidation and basic condition.

Medical implants by using RP and investment casting technologies

Directory of Open Access Journals (Sweden)

Milan Horacek

2011-02-01

Full Text Available The paper deals with the production technology of knee joint replacement by using rapid prototyping technology. The aim of the work is to outline the manufacturing technology intended for prototype production with the use of rapid prototyping and investment casting technology for use in orthopaedics and the surgery of knee joint replacement. The research results should make an effective contribution in the attempts to minimize the invasive surgical procedure, shorten the production of knee joint replacement as well as reduce the cost. At present, the research is focused on the preparation of STL data from CT (Computed Tomography and verification of the production technology of prototypes made using available RP technology and its evaluation.

Identification of regioisomers and enantiomers of triacylglycerols in different yeasts using reversed- and chiral-phase LC–MS

Czech Academy of Sciences Publication Activity Database

Řezanka, Tomáš; Kolouchová, I.; Čejková, A.; Cajthaml, Tomáš; Sigler, Karel

2013-01-01

Roč. 36, č. 20 (2013), s. 3310-3320 ISSN 1615-9306 R&D Projects: GA ČR(CZ) GAP503/11/0215 Institutional support: RVO:61388971 Keywords : Atmospheric pressure chemical ionization mass spectrometry * Chiral LC * Reversed phase LC Subject RIV: EE - Microbiology, Virology Impact factor: 2.594, year: 2013

Development and validation of reverse phase high performance liquid chromatography for citral analysis from essential oils.

Science.gov (United States)

Gaonkar, Roopa; Yallappa, S; Dhananjaya, B L; Hegde, Gurumurthy

2016-11-15

Citral is a widely used monoterpene aldehyde in aromatherapy, food and pesticide industries. A new validated reverse phase high performance liquid chromatography (RP - HPLC) procedure for the detection and quantification of cis-trans isomers of citral was developed. The RP-HPLC analysis was carried out using Enable C - 18G column (250×4.6mm, 5μ), with acetonitrile and water (70: 30) mobile phase in isocratic mode at 1mL/min flow. A photodiode array (PDA) detector was set at 233nm for the detection of citral. The method showed linearity, selectivity and accuracy for citral in the range of 3-100μg/mL. In order to compare the new RP-HPLC method with the available methods, one of the commercially available essential oil from Cymbopogon flexuosus was analyzed using new RP-HPLC method and the same was analyzed using GC-MS for the comparison of the method for the detection of citral. The GC-MS analysis was done using mass selective detector (MSD) showed citral content to be of 72.76%; wherein the new method showed to contain that same at 74.98%. To prove the application of the new method, essential oils were extracted from lemongrass, lemon leaves and mosambi peels by steam distillation. The citral content present in the essential and also in the condensate was analyzed. The method was found to be suitable for the analysis of citral in essential oils and water based citral formulations with a very good resolution of its components geranial and neral. Copyright © 2016 Elsevier B.V. All rights reserved.

Role of AMPK in regulation of LC3 lipidation as a marker of autophagy in skeletal muscle

DEFF Research Database (Denmark)

Fritzen, Andreas Mæchel; Frøsig, Christian; Jeppesen, Jacob Fuglsbjerg

2016-01-01

During induction of the autophagosomal degradation process, LC3-I is lipidated to LC3-II and associates to the cargo isolation membrane allowing for autophagosome formation. Lipidation of LC3 results in an increased LC3-II/LC3-I ratio, and this ratio is an often used marker for autophagy in vario...

Exploring Practical Responses of M3LC for Learning Literacy

Science.gov (United States)

Nasrullah; Baharman

2018-01-01

This study aims to explore the responses of participants toward Mathematics-Language Literacy Learning Courseware (M3LC) for learning literacy. There are five practical aspects concerned by involving 30 participants in the focus group discussion. In the beginning, participants were given some response sheet and introduced to M3LC by watching learning video of M3LC. At the end, they were asked to concern about response sheet and give comments related what they saw during the introduction session. The results show that the responses of users’ agree and strongly agree are still higher than those of users’ disagree or strongly disagree, with below 30% of responses are in the fair category. It means that the participants tend to give a positive opinion that M3LC is a useful courseware since it is qualified to satisfy 5 practical aspects, including knowledge use, knowledge construction, evaluation practice, social programming, and valuing to support literacy learning. In future, the implementation of using this courseware can be enhanced to further recognition of literacy level so that students can be well-prepared before starting learning activities in the classroom.

On the supression of electron phase-bunching in gyroresonant interactions in the magnetosphere

International Nuclear Information System (INIS)

Serra, F.M.

1985-01-01

The gyroresonant interaction between a whistler mode monochromatic wave and energetic electrons may cause a spatial and temporal coherence of resonant electrons. It is shown using a simple test particle model that a perturbing second wave can destroy the coherence by inhibiting phase bunching of the first-wave resonant electron. (R.P.)

[Quantative analysis of eupatilin and jaceosidin in folium of Artemisia argyi from different areas in China by RP-HPLC based on ancient medicine books].

Science.gov (United States)

Huang, Xian-Zhang; Kang, Li-Ping; Gao, Li; Zhang, Yuan; Guo, Lan-Ping; Huang, Lu-Qi

2017-09-01

In order to evaluate the quality of Artemisia argyi from Qichun, Ningbo, Anguo and Nanyang, the contents of eupatilin and jaceosidin were determined by RP-HPLC. The determination was performed on Agilent Eclipse XDB-C₁₈ (4.6 mm×250 mm, 5 μm) with mobile phase consisted of acetonitrile-0.2% phosphoric acid(35∶65) at the flow rate 1.0 mL•min ⁻¹. The detection wavelength was 350 nm and the column temperature was 25 ℃. The results showed that the amount of eupatilin and jaceosidin had a clear linear relationship in the range of 0.003-0.126 g•L ⁻¹ (r=0.999 9) and 0.005-0.200 g•L ⁻¹ (r=0.999 9), and the average recovery rates for them were 99.14% (n=6, RSD 1.2%) and 99.40% (n=6, RSD=0.73%), respectively. The results showed that RP-HPLC can be used for the quantification of eupatilin and jaceosidin in the folium of A. argyi. With this method, we found there was no significant difference of jaceosidin content within all the samples collected, but the content of eupatilin was significantly higher in samples from Qichun, Ningbo, Xiangyang and Nanyang, located in the south of Huaihe River compared with these from other areas. Copyright© by the Chinese Pharmaceutical Association.

Phase behavior and formation of o/w nano-emulsion in vegetable oil/ mixture of polyglycerol polyricinoleate and polyglycerin fatty acid ester/water systems.

Science.gov (United States)

Wakisaka, Satoshi; Nakanishi, Masami; Gohtani, Shoichi

2014-01-01

It is reported that mixing polyglycerol polyricinoleate (PGPR) and polyglycerol laurilester has a great emulsifying capacity, and consequently fine oil-in-water (o/w) emulsions can be formed. However, the role of PGPR is not clear. The objective of this research is to investigate the phase behavior of vegetable oil/mixture of PGPR and polyglycerol fatty acid ester/water systems, and to clarify the role of PGPR in making a fine emulsion. Phase diagrams were constructed to elucidate the optimal process for preparing fine emulsions. In all the systems examined in this study, the phases, including the liquid crystal phase (L(c)) and sponge phase (L(3)), spread widely in the phase diagrams. We examined droplet size of the emulsions prepared from each phase and found that o/w nano-emulsions with droplet sizes as small as 50 nm were formed by emulsifying either from a single L(3) phase or a two-phase region, L(c) + L(3). These results indicate that a sponge phase L(3) or liquid crystal phase L(c) or both is necessary to form an o/w nano-emulsion whose average droplet diameter is less than 50 nm for PGPR and polyglycerin fatty acid ester mixtures used as surfactant.

A rapid MCM-41 dispersive micro-solid phase extraction coupled with LC/MS/MS for quantification of ketoconazole and voriconazole in biological fluids.

Science.gov (United States)

Yahaya, Noorfatimah; Sanagi, Mohd Marsin; Abd Aziz, Noorizan; Wan Ibrahim, Wan Aini; Nur, Hadi; Loh, Saw Hong; Kamaruzaman, Sazlinda

2017-02-01

A rapid dispersive micro-solid phase extraction (D-μ-SPE) combined with LC/MS/MS method was developed and validated for the determination of ketoconazole and voriconazole in human urine and plasma samples. Synthesized mesoporous silica MCM-41 was used as sorbent in d-μ-SPE of the azole compounds from biological fluids. Important D-μ-SPE parameters, namely type desorption solvent, extraction time, sample pH, salt addition, desorption time, amount of sorbent and sample volume were optimized. Liquid chromatographic separations were carried out on a Zorbax SB-C 18 column (2.1 × 100 mm, 3.5 μm), using a mobile phase of acetonitrile-0.05% formic acid in 5 mm ammonium acetate buffer (70:30, v/v). A triple quadrupole mass spectrometer with positive ionization mode was used for the determination of target analytes. Under the optimized conditions, the calibration curves showed good linearity in the range of 0.1-10,000 μg/L with satisfactory limit of detection (≤0.06 μg/L) and limit of quantitation (≤0.3 μg/L). The proposed method also showed acceptable intra- and inter-day precisions for ketoconazole and voriconazole from urine and human plasma with RSD ≤16.5% and good relative recoveries in the range 84.3-114.8%. The MCM-41-D-μ-SPE method proved to be rapid and simple and requires a small volume of organic solvent (200 μL); thus it is advantageous for routine drug analysis. Copyright © 2016 John Wiley & Sons, Ltd.

A Review and Modern Approach to LC Ladder Synthesis

Directory of Open Access Journals (Sweden)

Alexander J. Casson

2011-01-01

Full Text Available Ultra low power circuits require robust and reliable operation despite the unavoidable use of low currents and the weak inversion transistor operation region. For analogue domain filtering doubly terminated LC ladder based filter topologies are thus highly desirable as they have very low sensitivities to component values: non-exact component values have a minimal effect on the realised transfer function. However, not all transfer functions are suitable for implementation via a LC ladder prototype, and even when the transfer function is suitable the synthesis procedure is not trivial. The modern circuit designer can thus benefit from an updated treatment of this synthesis procedure. This paper presents a methodology for the design of doubly terminated LC ladder structures making use of the symbolic maths engines in programs such as MATLAB and MAPLE. The methodology is explained through the detailed synthesis of an example 7th order bandpass filter transfer function for use in electroencephalogram (EEG analysis.

Sensitivity of GC-EI/MS, GC-EI/MS/MS, LC-ESI/MS/MS, LC-Ag(+) CIS/MS/MS, and GC-ESI/MS/MS for analysis of anabolic steroids in doping control.

Science.gov (United States)

Cha, Eunju; Kim, Sohee; Kim, Ho Jun; Lee, Kang Mi; Kim, Ki Hun; Kwon, Oh-Seung; Lee, Jaeick

2015-01-01

This study compared the sensitivity of various separation and ionization methods, including gas chromatography with an electron ionization source (GC-EI), liquid chromatography with an electrospray ionization source (LC-ESI), and liquid chromatography with a silver ion coordination ion spray source (LC-Ag(+) CIS), coupled to a mass spectrometer (MS) for steroid analysis. Chromatographic conditions, mass spectrometric transitions, and ion source parameters were optimized. The majority of steroids in GC-EI/MS/MS and LC-Ag(+) CIS/MS/MS analysis showed higher sensitivities than those obtained with other analytical methods. The limits of detection (LODs) of 65 steroids by GC-EI/MS/MS, 68 steroids by LC-Ag(+) CIS/MS/MS, 56 steroids by GC-EI/MS, 54 steroids by LC-ESI/MS/MS, and 27 steroids by GC-ESI/MS/MS were below cut-off value of 2.0 ng/mL. LODs of steroids that formed protonated ions in LC-ESI/MS/MS analysis were all lower than the cut-off value. Several steroids such as unconjugated C3-hydroxyl with C17-hydroxyl structure showed higher sensitivities in GC-EI/MS/MS analysis relative to those obtained using the LC-based methods. The steroids containing 4, 9, 11-triene structures showed relatively poor sensitivities in GC-EI/MS and GC-ESI/MS/MS analysis. The results of this study provide information that may be useful for selecting suitable analytical methods for confirmatory analysis of steroids. Copyright © 2015 John Wiley & Sons, Ltd.

Acyl and silyl group effects in reactivity-based one-pot glycosylation: synthesis of embryonic stem cell surface carbohydrates Lc4 and IV(2)Fuc-Lc4.

Science.gov (United States)

Hsu, Yun; Lu, Xin-An; Zulueta, Medel Manuel L; Tsai, Chih-Ming; Lin, Kuo-I; Hung, Shang-Cheng; Wong, Chi-Huey

2012-03-14

Relative reactivity evaluations showed the graded arming of toluenyl thioglucosides by variously positioned silyl groups but not by their acyl counterparts. These findings were applied in reactivity-based one-pot assembly of linker-attached Lc(4) and IV(2)Fuc-Lc(4), which are components of human embryonic stem cell surface. The sugar-galectin-1 binding was also examined.

Development and Validation of LC Method for the Determination of Famciclovir in Pharmaceutical Formulation Using an Experimental Design

Directory of Open Access Journals (Sweden)

Srinivas Vishnumulaka