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Sample records for c18 column hplc

  1. Vantagens e desvantagens das colunas C18 e C30 para a separação de carotenóides por CLAE Advantages and disadvantages of C18 and C30 columns for HPLC separation of carotenoids

    Directory of Open Access Journals (Sweden)

    Itaciara Larroza Nunes

    2006-12-01

    performance liquid chromatography (HPLC using C18 (monomeric, 4 mm, 300 x 3.9 mm and C30 (polymeric 3 mm, 250 x 4.6 mm columns and many different mobile phases, with either isocratic or gradient elution. The carotenoids were identified by their spectral characteristics and co-chromatography with standards. The best chromatographic conditions were achieved with the C30 column, temperature set at 33 ºC and as mobile phase an isocratic elution of methanol (0.1% triethylamine/tert-butyl methyl ether (50:50 to separate lycopene isomers and (95:5 for lutein and zeaxanthin, both at 1 mL/min. However, for quantitative analysis, it is necessary to evaluate the peak area repeatability on the C30 column. In addition, the monomeric C18 column can be employed for separation of lutein and zeaxanthin.

  2. Cheap C18-modified silica monolith particles as HPLC stationary phase of good separation efficiency

    Energy Technology Data Exchange (ETDEWEB)

    Ali, Ashraf; Ali, Faiz; Cheong, Woo Jo [Dept. of of Chemistry, Inha University, Incheon (Korea, Republic of)

    2015-06-15

    The columns packed with particles have a high efficiency but they are accompanied with a high column back pressure due to lower permeability, while the monolithic columns have a high permeability but they result in inferior separation efficiency for the analysis of small molecules in HPLC. In our laboratory,we have been using the pseudo-monolithic silica particles with C-18 ligand or polystyrene film. The column to column reproducibility was evaluated based on three columns made of three different batches of silica monolith particles, and better than 4.5% in N, and 1.6% in retention time were observed. The day to day reproducibility of a single column for three consecutive days was found better than 1.5% both in N and retention time. The van Deemter plots were derived for awide range of flow rates. The trends of van Deemter plots were similar to those of common patterns and the optimal flow rate was found to be 25 μL/min.

  3. The effect of re-dissolution solvents and HPLC columns on the analysis of mycosporine-like amino acids in the eulittoral macroalgae and

    National Research Council Canada - National Science Library

    Karsten, Ulf; Escoubeyrou, Karine; Charles, François

    2009-01-01

    .... Therefore pure methanol, distilled water and HPLC eluent were evaluated as re-dissolution solvent for dried and extracts, which were subsequently analyzed on three reversed-phase C8 and C18 HPLC columns...

  4. Separation analysis of macrolide antibiotics with good performance on a positively charged C18HCE column.

    Science.gov (United States)

    Wei, Jie; Shen, Aijin; Yan, Jingyu; Jin, Gaowa; Yang, Bingcheng; Guo, Zhimou; Zhang, Feifang; Liang, Xinmiao

    2016-03-01

    The separation of basic macrolide antibiotics suffers from peak tailing and poor efficiency on traditional silica-based reversed-phase liquid chromatography columns. In this work, a C18HCE column with positively charged surface was applied to the separation of macrolides. Compared with an Acquity BEH C18 column, the C18HCE column exhibited superior performance in the aspect of peak shape and separation efficiency. The screening of mobile phase additives including formic acid, acetic acid and ammonium formate indicated that formic acid was preferable for providing symmetrical peak shapes. Moreover, the influence of formic acid content was investigated. Analysis speed and mass spectrometry compatibility were also taken into account when optimizing the separation conditions for liquid chromatography coupled with tandem mass spectrometry. The developed method was successfully utilized for the determination of macrolide residues in a honey sample. Azithromycin was chosen as the internal standard for the quantitation of spiramycin and tilmicosin, while roxithromycin was used for erythromycin, tylosin, clarithromycin, josamycin and acetylisovaleryltylosin. Good correlation coefficients (r(2) > 0.9938) for all macrolides were obtained. The intra-day and inter-day recoveries were 73.7-134.7% and 80.7-119.7% with relative standard deviations of 2.5-8.0% and 3.9-16.1%, respectively. Outstanding sensitivity with limits of quantitation (S/N ≥ 10) of 0.02-1 μg/kg and limits of detection (S/N ≥ 3) of 0.01-0.5 μg/kg were achieved. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. A chemometric approach to determine the phenolic compounds in different barley samples by two different stationary phases: a comparison between C18 and pentafluorophenyl core shell columns.

    Science.gov (United States)

    Gómez-Caravaca, Ana María; Verardo, Vito; Berardinelli, Annachiara; Marconi, Emanuele; Caboni, Maria Fiorenza

    2014-08-15

    Barley (Hordeum vulgare L.) is a cereal crop that has been cultivated since ancient times. However, its interest as nutritional food and as food ingredient is relatively new. Thus, in this study, the phenolic compounds of eighteen different varieties of barley (4 waxy and 14 non-waxy) grown under the same agronomic conditions in the same experimental field have been determined by HPLC-DAD-MS. Two new methodologies were developed using new generation superficially porous HPLC columns with different stationary phases: C18 and pentafluorophenyl (PFP). Twelve free phenolic compounds and eight bound phenolic compounds could be identified in barley samples in less than 22min. The study of different method parameters showed that C18 column was more suitable for the analysis of phenolic compounds of barley. Hierarchical cluster analysis (HCA) was conducted in order to assess the different ability of the two different core shell HPLC columns in the discrimination between "waxy" and "non-waxy" varieties, and only HCA of C18 column could separate waxy and non-waxy genotypes. Significant differences in the content of phenolic compounds between waxy and non-waxy samples were found, being waxy barley samples the ones which presented higher content of free and bound phenolic compounds. Once the best discriminant HPLC column was established, principal component analysis (PCA) was applied and it was able to discriminate between "waxy" and "non-waxy" varieties; however it discriminated the barley samples based only in free phenolic compounds. Because of that, partial least squares discriminant analysis (PLS-DA) and Artificial Neural Networks (ANN) were carried out. PLS-DA and ANN permitted the classification of waxy and non-waxy genotypes from both free and bound phenolic compounds. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Improved HPLC column-switching determination of Coenzyme Q and Vitamin E in plasma.

    Science.gov (United States)

    Bompadre, Stefano; Tulipani, Sara; Romandini, Stefania; Giorgetti, Raffaele; Battino, Maurizio

    2008-01-01

    A novel isocratic modified column-switching HPLC method for automated quantitative analysis of Vitamin E and Coenzyme Q, in the reduced and oxidized form, is described. Many column-switching HPLC methods are found in the literature, also for determining antioxidant substances, but we developed a different system of column-switching. An empty column, 5 cm long, was connected to the switching valve, before the sample loop and the extraction column. The sample loop was connected directly after the empty column. The inserted column, containing about 1.4 ml of the extraction eluent simulated a gradient elution, enhancing sensitivity and resolution. When switching the columns, the empty column is placed right before the extraction column and acts as a static mixer for the extraction phase and the incoming analytical phase. Samples were cleaned from interfering compounds by transfer onto a extraction-column, using a C-8 silica. Separation was performed onto an analytical column C18 3 m icrom, 150 mm x 4.6 mm at a flow rate of 1.0 ml/min with 20 mmol/l lithium perchlorate/perchloric acid, pH3.0 in Ethanol as analytical eluent. Detection was performed with a ESA Coulochem 5100 A model. The method was found to be suitable for automated analysis of Coenzyme Q, reduced and oxidized form, and Vitamin E in serum.

  7. Purification of bufadienolides from the skin of Bufo bufo gargarizans Cantor with positively charged C18 column.

    Science.gov (United States)

    Li, Xiaolong; Guo, Zhimou; Wang, Chaoran; Shen, Aijin; Liu, Yanfang; Zhang, Xiuli; Zhao, Weijie; Liang, Xinmiao

    2014-04-01

    As a kind of promising anticancer compounds, the preparation of bufadienolides is a hot study spot. However, due to the complexity of biological sample, the purification of bufadienolides from a crude sample (toad skin) is a tough work. In this paper, we reported a new way based on positively charged C18 material (XCharge C18) to quickly separate and purify bufadienolides from toad skin. By this method, the different ionic feature of the amino acid conjugated bufadienolides (AACBs) and the free form bufadienolides (AAUBs) was firstly utilized to obtain distinct separation selectivity on the XCharge C18 column. Additionally, the peak tailing problem of AACBs on conventional C18 was resolved and better resolutions were achieved on the XCharge C18, thus, two kinds of bufadienolides on one column were successfully purified respectively. Taking F13 as an example, the method was validated by liquid chromatography-mass spectrometry (LC-MS), and then 4 AACBs as well as 4 AAUBs were simultaneously purified by preparative XCharge C18. In addition, the application of this method in other fractions was also validated. The results suggested that the developed method is a practical and promising tool for efficient separation and purification of bufadienolides from toad skin. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. HPLC-CUPRAC post-column derivatization method for the determination of antioxidants: a performance comparison between porous silica and core-shell column packing.

    Science.gov (United States)

    Haque, Syed A; Cañete, Socrates Jose P

    2018-01-01

    An HPLC method employing a post-column derivatization strategy using the cupric reducing antioxidant capacity reagent (CUPRAC reagent) for the determining antioxidants in plant-based materials leverages the separation capability of regular HPLC approaches while allowing for detection specificity for antioxidants. Three different column types, namely core-shell and porous silica including two chemically different core-shell materials (namely phenyl-hexyl and C18), were evaluated to assess potential improvements that could be attained by changing from a porous silica matrix to a core-shell matrix. Tea extracts were used as sample matrices for the evaluation specifically looking at catechin and epigallocatechin gallate (EGCG). Both the C18 and phenyl-hexyl core-shell columns showed better performance compared to the C18 porous silica one in terms of separation, peak shape, and retention time. Among the two core-shell materials, the phenyl-hexyl column showed better resolving power compared to the C18 column. The CUPRAC post-column derivatization method can be improved using core-shell columns and suitable for quantifying antioxidants, exemplified by catechin and EGCG, in tea samples.

  9. Repeatability in column preparation of a reversed-phase C18 monolith and its application to separation of tocopherol homologues.

    Science.gov (United States)

    Kositarat, Sirichai; Smith, Norman William; Nacapricha, Duangjai; Wilairat, Prapin; Chaisuwan, Patcharin

    2011-06-15

    This work investigated the repeatability of column preparation for a reversed-phase C18 monolith, namely stearyl methacrylate-co-ethylene glycol dimethacrylate (SMA-EDMA). The columns were thermally polymerised using three commonly available heating devices (GC oven, hot air oven and water bath) and their chromatographic performance evaluated using micro-liquid chromatography for separation of five test compounds. Precision in terms of %RSD of retention times were 9.0, 6.5, and 12.5 using GC oven, hot air oven and water bath, respectively. Between-batch precision for the hot air oven (n=3 days) was less than 10.4% for retention time. The SMA-EDMA monolith was applied to the separation of tocopherol homologues by capillary electrochromatography. Usually tocopherol homologues cannot be completely separated by conventional reversed-phase C8- or C18-packed bed or C18-silica based monolithic columns. Polymer monolith has been shown to give remarkable selectivity towards the tocopherols compared to the conventional microparticulate phase and silica based monolith. Successful separation of the tocopherol isomers was achieved on the SMA-EDMA monolith without any column modification. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Synthesis and applications of monolithic HPLC columns

    Science.gov (United States)

    Liang, Chengdu

    Silica and carbon monolithic columns were synthesized and modified for liquid chromatography applications. Column configurations and cladding techniques were investigated in detail. Three novel approaches have been developed for the synthesis of bimodal porous rods. Out of these three methods, gel-casting was adopted for the synthesis of silica monoliths with ordered mesopores and uniform macropores; the use of colloidal templates and dual phase separation has been successfully implemented for the synthesis of carbon monoliths with well-controlled meso- and macro- porosities. The formation of mesopores in carbon materials has been further studied in the microphase separation of block copolymers. Electrochemical modification of carbon monoliths was discovered to be an efficient method for converting covalently bonded functionalities to carbon monoliths. N,N'-diethylaminobenzene has been attached to carbon surface for the separation of proteins and protein digests. The performances of carbon-based monolithic columns were studied intensely through frontal analysis and Van Deemter plot. Temperature and pressure effects were also investigated in carbon-based columns. The density of bonding on the modified carbon monoliths was characterized by thermogravimetric analysis.

  11. A hybrid FIA/HPLC system incorporating monolithic column chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Adcock, Jacqui L. [School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 (Australia); Francis, Paul S. [School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 (Australia)], E-mail: psf@deakin.edu.au; Agg, Kent M. [School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 (Australia); Marshall, Graham D. [GlobalFIA, Fox Island, WA 98333 (United States); Barnett, Neil W. [School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 (Australia)

    2007-09-26

    We have combined the generation of solvent gradients using milliGAT pumps, chromatographic separations with monolithic columns and chemiluminescence detection in an instrument manifold that approaches the automation and separation efficiency of HPLC, whilst maintaining the positive attributes of flow injection analysis (FIA), such as manifold versatility, speed of analysis and portability. As preliminary demonstrations of this hybrid FIA/HPLC system, we have determined six opiate alkaloids (morphine, pseudomorphine, codeine, oripavine, ethylmorphine and thebaine) and four biogenic amines (vanilmandelic acid, serotonin, 5-hydroxyindole-3-acetic acid and homovanillic acid) in human urine, using tris(2,2'-bipyridyl)ruthenium(III) and acidic potassium permanganate chemiluminescence detection.

  12. HPLC determination of capsaicinoids with cross-linked C18 column and buffer-free eluent.

    Science.gov (United States)

    Daood, Hussein G; Halasz, Gábor; Palotás, Gábor; Palotás, Gabriella; Bodai, Zsolt; Helyes, Lajos

    2015-01-01

    A simple and efficient high-performance liquid chromatographic method was developed and validated for the separation and determination of capsaicin and its major dihydro- and homoderivatives in spice paprika products in 20 min with fluorescent and 35 min with mass-spectrometric detection. The separation was performed on reversed-phase chromatographic adsorbent of cross-linked endcapping with eluent consisting of 1:1 acetonitrile-water or acetonitrile-0.1% formic acid under isocratic conditions. Excellent separation of all the major and minor capsaicinoids with resolution index between 1.08 and 7.34 was achieved. The detection and quantification limits of capsaicinoids in standard material solutions were between 2 and 10 ng/mL. The lowest detectable amount of capsaicin, with fluorescent detection, was found to be capsaicinoids could be distinguished from the non-capsaicinoids compounds appeared on liquid chromatography-fluorescence profile of extract from drastically processed paprika by applying mass spectroscopic detection. Hungarian spice paprika were evaluated as mild to very hot (capsaicinoid content: 334-1,660 µg/g) and chili products as very or extremely hot products (1,543-2,818 µg/g). © The Author [2014]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  13. The effect of re-dissolution solvents and HPLC columns on the analysis of mycosporine-like amino acids in the eulittoral macroalgae Prasiola crispa and Porphyra umbilicalis

    Science.gov (United States)

    Karsten, Ulf; Escoubeyrou, Karine; Charles, François

    2009-09-01

    Many macroalgal species that are regularly exposed to high solar radiation such as the eulittoral green alga Prasiola crispa and the red alga Porphyra umbilicalis synthesize and accumulate high concentrations of mycosporine-like amino acids (MAAs) as UV-sunscreen compounds. These substances are typically extracted with a widely used standard protocol following quantification by various high performance liquid chromatography (HPLC) techniques. However, further preparation steps prior to HPLC analysis as well as different HPLC column types have not been systematically checked regarding separation quality and reproducibility. Therefore pure methanol, distilled water and HPLC eluent were evaluated as re-dissolution solvent for dried Prasiola and Porphyra extracts, which were subsequently analyzed on three reversed-phase C8 and C18 HPLC columns. The data indicate that distilled water and the HPLC eluent gave almost identical peak patterns and MAA contents on the C8 and C18 columns. In contrast, the application of the widely used methanol led to double peaks or even the loss of specific peaks as well as to a strong decline in total MAA amounts ranging from about 35% of the maximum in P. crispa to 80% of the maximum in P. umbilicalis. Consequently, methanol should be avoided as re-dissolution solvent for the HPLC sample preparation. An improved protocol for the MAA analysis in macroalgae in combination with a reliable C18 column is suggested.

  14. Preliminary Study of High Resolution HPLC Analytical Method for Sedimentary Pigments Based on Coupled C8 Columns

    Science.gov (United States)

    Yao, P.; Yu, Z.; Deng, C.; Liu, S.; Zhao, J.

    2008-05-01

    The pigments in marine water columns can provide accurate estimates of community composition and abundance of phytoplankton. In addition, the sedimentary pigments, especially the derivatives of chlorophyll such as pyrophaeophytins, pyrophaeophorbides and steryl chlorin esters (SCEs) formed during early diagenesis can also provide information on the primary producer community and the changes in paleoproductivity. Accordingly, analysis of pigments and their derivatives is of great importance for oceanography, limnology and geochemistry. Many methods have been developed for the separation of chlorophylls, carotenoids and their derivatives derived from phytoplankton and water column samples using high-performance liquid chromatography (HPLC). Methods widely cited in the literatures include those developed by Wright et al. (1991) and Zapata et al. (2000). Both methods use reversed-phase columns, but C18 column was employed in Wright et al. (1991) and C8 column in Zapata et al. (2000). However, evident coelutions are observed in published works. This will particularly cause problematic identification and quantification in dealing with sedimentary pigments which are highly complex and often display a broad range in polarity. Clearly, it is necessary to improve the separation of the complex pigments if the information carried by the pigments is to be used fully. Coupled C18 columns were used in the HPLC method developed by Airs et al. (2001) for the analysis of complex pigment distributions. Improved chromatographic resolution, more pigment components and novel bacteriochlorophyll derivatives were obtained by this method. It indicates a new road for HPLC method development. C8 column has shorter carbon chains than that of C18 column and can provide less retention of apolar compounds which is of particular advantaged to hydrophobic chlorophyll a, b and their derivatives. That is one of the reasons why the C8 method developed by Zapata et al. (2000) is admittedly better than

  15. [Determination of triterpenoic acids in fruits of Ziziphus jujuba using HPLC-MS with polymeric ODS column].

    Science.gov (United States)

    Zhang, Yong; Zhou, An; Xie, Xiao-Mei

    2013-03-01

    A simple and sensitive method has been developed to simultaneously determine betunilic acid, oleanolic acid and ursolic acid in the fruits of Ziziphus jujuba from different regions by HPLC-MS. This HPLC assay was performed on PAH polymeric C18 bonded stationary phase column with mobile phase contained acetonitrile-water (90: 10) and with negative ESI detection mode. The developed approach was characterized by short time consumption for chromatographic separation, high sensitivity and good reliability so as to meet the requirements for rapid analysis of large-batch fruits of Z. jujuba from different habitats.

  16. Fast and Universal HPLC Method for Determination of Permethrin in Formulations Using 1.8-µm Particle-Packed Column and Performance Comparison with Other Column Types

    Science.gov (United States)

    Shishovska, Maja A.; Stefova, Marina T.

    2012-01-01

    An HPLC method has been developed for the fast separation and quantification of permethrin using C18 column packed with 1.8 µm particles. The method is specific with good resolution to degradation products and to other present components. It has acceptable validation results. The run time is 4.5 min (or may be within 1.6 min is rapid resolution mode) with an organic solvent consumption of 3.6 mL per run. The method has been applied to samples of formulations for various uses: mattress cleaner, shampoo, and veterinary powder. The performance of the applied column is compared with other common column types. The relationships between linear velocity of the mobile phase (u) and resolution factor (Rs), back-pressure (ΔP), and efficiency (H) are presented. The experimental data shows the advantages of 1.8-µm particle columns to be a significant reduction in solvent consumption (by factor of 4.4 and 1.5) and a reduction in run-time (by factor 4.7 and 1.5), and the weaknesses are a high back-pressure and lower efficiency. Finally, it has been shown that use of 1.8-µm particle packed columns with conventional HPLC systems is possible, but with limitations in mobile phase flow-rate. PMID:22291055

  17. Reversed Phase Column HPLC-ICP-MS Conditions for Arsenic Speciation Analysis of Rice Flour.

    Science.gov (United States)

    Narukawa, Tomohiro; Matsumoto, Eri; Nishimura, Tsutomu; Hioki, Akiharu

    2015-01-01

    New measurement conditions for arsenic speciation analysis of rice flour were developed using HPLC-ICP-MS equipped with a reversed phase ODS column. Eight arsenic species, namely, arsenite [As(III)], arsenate [As(V)], monomethylarsonic acid (MMAA), dimethylarsinic acid (DMAA), trimethylarsine oxide (TMAO), tetramethylarsonium (TeMA), arsenobetaine (AsB) and arsenocholine (AsC), were separated and determined under the proposed conditions. In particular, As(III) and MMAA and DMAA and AsB were completely separated using a newly proposed eluent containing ammonium dihydrogen phosphate. Importantly, the sensitivity changes, in particular those of As(V) and As(III) caused by coexisting elements and by complex matrix composition, which had been problematical in previously reported methods, were eliminated. The new eluent can be applied to C8, C18 and C30 ODS columns with the same effectiveness and with excellent repeatability. The proposed analytical method was successfully applied to extracts of rice flour certified reference materials.

  18. A rapid and efficient preparation of [{sup 123}I]radiopharmaceuticals using a small HPLC Rocket[reg] column

    Energy Technology Data Exchange (ETDEWEB)

    Katsifis, Andrew [Radiopharmaceuticals Division R and D, Australian Nuclear Science and Technology, Organisation, Menai, NSW 2234, Sydney (Australia)]. E-mail: akx@ansto.gov.au; Papazian, Vahan [Radiopharmaceuticals Division R and D, Australian Nuclear Science and Technology, Organisation, Menai, NSW 2234, Sydney (Australia); Jackson, Timothy [Radiopharmaceuticals Division R and D, Australian Nuclear Science and Technology, Organisation, Menai, NSW 2234, Sydney (Australia); Loc' h, Christian [Radiopharmaceuticals Division R and D, Australian Nuclear Science and Technology, Organisation, Menai, NSW 2234, Sydney (Australia)

    2006-01-01

    A simplified method for the rapid and efficient preparation of [{sup 123}I]radiopharmaceuticals is described. Three radiopharmaceuticals, [{sup 123}I]{beta}-CIT, [{sup 123}I]MIBG and [{sup 123}I]clioquinol, were synthesised and purified as model compounds. The radiotracers were labelled with iodine-123 using electrophilic oxidative conditions and purified by a compact semi-preparative reverse phase column (C-18, 3 {mu}m, 7x53 mm, Alltima Rocket[reg, Alltech] using aqueous-ethanol as HPLC solvents that were directly used for radiopharmaceutical formulation. The radiochemical purity of the radioiodinated tracers as assessed by analytical HPLC was higher than 99% with specific activity higher than 3 GBq/nmol. The total preparation time of a radiotracer ranged from 40 to 60 min and, starting from 3.7 GBq of iodine-123, more than 2.5 GBq of formulated radiopharmaceuticals were available for clinical investigations.

  19. Application of narrow-bore HPLC columns in rapid determination of sildenafil citrate in its pharmaceutical dosage forms.

    Science.gov (United States)

    Ghodsi, Razieh; Kobarfard, Farzad; Tabatabai, Sayyed Abbas

    2012-01-01

    A special type of silica-based columns has been recently introduced into the market which is called narrow-bore columns. They have lower internal volume than the standard high-performance liquid chromatography (HPLC) columns and thus reduce the solvent consumption by almost 80%. A simple, accurate and environmentally friendly reversed phase- HPLC (RP-HPLC method) which could be used in fast and high throughput analyses has been developed for the purpose of determining the sildenafil in bulk and pharmaceutical dosage forms, using narrow-bore C18 column (50 × 3.2 mm, 5 µm particle size) in isocratic mode, with mobile phase comprising of buffer (pH = 3) and acetonitrile in the ratio of 75:25 v/v. The flow rate was 0.7 mL/min and the detection was monitored through Ultraviolet detector (UV detector) at 292 nm. Clonazepam was used as the internal standard and the run time was 4 min. The proposed method has permitted the quantification of sildenafil over the linearity in the range of 30-4000 ng/mL and its percentage recovery was found to be 99-105%. Limit of quantitation (LOQ) is determined as 30 ng/mL. The intra-day and inter-day precisions were found 1.2-2.2% and 1.56-3.4% respectively. The solvent consumption was 2.8 mL per sample of which ca 0.7 mL was acetonitrile. This study shows that the application of narrow-bore column instead of the conventional reversed phase column in HPLC analyses has the advantages of shorter run time and less organic solvent consumption. This method is highly sensitive with excellent recoveries and precision and there is no need for special column and pre-column or post-column treatment of the sample. Moreover, the method is free from interference by common additives and excipients, suggesting applications in routine quality control analyses.

  20. Rapid trace level determination of sulfonamide residues in honey with online extraction using short C-18 column by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Sajid, Muhammad; Na, Na; Safdar, Muhammad; Lu, Xin; Ma, Lin; He, Lan; Ouyang, Jin

    2013-11-01

    A sensitive and inexpensive quantification method with online extraction using a short C-18 column for sulfonamide residues in honey by high performance liquid chromatography with fluorescence detector was developed and validated. In sample preparation, acid hydrolysis was used to break the N-glycoside bond between the honey sugar and sulfonamide drugs and derivatization of sulfonamide residues with fluorescamine was conducted at pH 3.5 using a citrate buffer (0.5M) in the honey matrix. The chromatography was carried out on Zorbax Extended C-18 (250mm×4.6mm; 5μm) column, using a mixture of acetonitrile and an acetate buffer (pH 4.50, 20mM) as a mobile phase. A Zorbax Extended C-18 (12mm×4.6mm; 5μm) column was used for online extraction of fifteen sulfonamide residues from honey sample with the help of a two position valve. The limit of quantification of sulfonamide residues in honey was less than 3ngg(-1), and the percentage recovery of study compounds in spiked honey sample was from 80% for sulfacetamide to 100% of sulfachloropyridazine. The developed method has excellent linearity for all studied sulfonamides with a correlation coefficient 0.993. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. New validated high-performance liquid chromatographic method for simultaneous analysis of ten flavonoid aglycones in plant extracts using a C18 fused-core column and acetonitrile-tetrahydrofuran gradient.

    Science.gov (United States)

    Olszewska, Monika A

    2012-09-01

    An HPLC method of high resolution has been developed and validated for the simultaneous determination of ten prominent flavonoid aglycones in plant materials using a fused-core C18-silica column (Ascentis® Express, 4.6 mm × 150 mm, 2.7 μm). The separation was accomplished with an acetonitrile-tetrahydrofuran gradient elution at a flow rate of 1 mL/min and temperature of 30°C. UV spectrophotometric detection was employed at 370 nm for flavonols (quercetin [QU], myricetin [MY], isorhamnetin [IS], kaempferol [KA], sexangularetin [SX], and limocitrin [LM]) and 340 nm for flavones (apigenin [AP], acacetin [AC], chrysoeriol [CH], and luteolin [LU]). The high resolution of critical pairs QU/LU (10.50), QU/CH (3.40), AP/CH (2.51), SX/LM (2.30), and IS/KA (2.70) was achieved within 30.3 min. The observed column back pressure was less than 4300 psi, thus acceptable for conventional HPLC equipment. The method was sensitive enough having LODs of 0.115-0.525 ng and good linearity (r > 0.9999) over the test range. The precision values, expressed as RSD values, were Sorbus species. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Comparing monolithic and fused core HPLC columns for fast chromatographic analysis of fat-soluble vitamins.

    Science.gov (United States)

    Kurdi, Said El; Muaileq, Dina Abu; Alhazmi, Hassan A; Bratty, Mohammed Al; Deeb, Sami El

    2017-06-27

    HPLC stationary phases of monolithic and fused core type can be used to achieve fast chromatographic separation as an alternative to UPLC. In this study, monolithic and fused core stationary phases are compared for fast separation of four fat-soluble vitamins. Three new methods on the first and second generation monolithic silica RP-18e columns and a fused core pentafluoro-phenyl propyl column were developed. Application of three fused core columns offered comparable separations of retinyl palmitate, DL-α-tocopheryl acetate, cholecalciferol and menadione in terms of elution speed and separation efficiency. Separation was achieved in approx. 5 min with good resolution (Rs > 5) and precision (RSD ≤ 0.6 %). Monolithic columns showed, however, a higher number of theoretical plates, better precision and lower column backpressure than the fused core column. The three developed methods were successfully applied to separate and quantitate fat-soluble vitamins in commercial products.

  3. Characterization of insulin-loaded liposome using column-switching HPLC.

    Science.gov (United States)

    Ohnishi, Naozumi; Tanaka, Shota; Tahara, Kohei; Takeuchi, Hirofumi

    2015-02-20

    We evaluated the drug-encapsulation state of insulin (INS)-loaded liposome using a novel column-switching HPLC system that can automatically separate unloaded drug from encapsulated drug by hydrophobic interaction. When the INS-loaded liposome was dispersed in water (pH 7.4), the encapsulation efficiency (EE) obtained by the column-switching HPLC system was consistent with that obtained by a conventional ultracentrifugation method. However, the INS-loaded liposome dispersed in 0.1% acetic acid (pH 3.3) showed disagreement between the EEs obtained by both methods. Considering the results of particle size, zeta potential, and transmission electron microscope (TEM) observations, we hypothesized that the column-switching HPLC method was able to distinguish INS adsorbed onto the liposome surface from the encapsulated INS, although an ultracentrifugation method precipitated the adsorbed INS onto the liposome surface along with the encapsulated INS. Therefore, the novel column-switching HPLC system may be a more accurate and useful technique for characterization and optimization of the INS-loaded liposome formulation. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. A rapid HPLC column switching method for sample preparation and determination of β-carotene in food supplements.

    Science.gov (United States)

    Brabcová, Ivana; Hlaváčková, Markéta; Satínský, Dalibor; Solich, Petr

    2013-11-15

    A simple and automated HPLC column-switching method with rapid sample pretreatment has been developed for quantitative determination of β-carotene in food supplements. Commercially samples of food supplements were dissolved in chloroform with help of saponification with 1M solution of sodium hydroxide in ultrasound bath. A 20-min sample dissolution/extraction step was necessary before chromatography analysis to transfer β-carotene from solid state of food supplements preparations (capsules,tablets) to chloroform solution. Sample volume - 3μL of chloroform phase was directly injected into the HPLC system. Next on-line sample clean-up was achieved on the pretreatment precolumn Chromolith Guard Cartridge RP-18e (Merck), 10×4.6mm, with a washing mobile phase (methanol:water, 92:8, (v/v)) at a flow rate of 1.5mL/min. Valve switch to analytical column was set at 2.5min in a back-flush mode. After column switching to the analytical column Ascentis Express C-18, 30×4.6mm, particle size 2.7μm (Sigma Aldrich), the separation and determination of β-carotene in food supplements was performed using a mobile phase consisting of 100% methanol, column temperature at 60°C and flow rate 1.5mL/min. The detector was set at 450nm. Under the optimum chromatographic conditions standard calibration curve was measured with good linearity - correlation coefficient for β-carotene (r(2)=0.999014; n=6) between the peak areas and concentration of β-carotene 20-200μg/mL. Accuracy of the method defined as a mean recovery was in the range 96.66-102.40%. The intraday method precision was satisfactory at three concentration levels 20, 125 and 200μg/mL and relative standard deviations were in the range 0.90-1.02%. The chromatography method has shown high sample throughput during column-switching pretreatment process and analysis in one step in short time (6min) of the whole chromatographic analysis. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Separation of Alkyne Enantiomers by Chiral Column HPLC Analysis of Their Cobalt-Complexes

    Directory of Open Access Journals (Sweden)

    Qiaoyun Liu

    2017-03-01

    Full Text Available Separation of the enantiomers of new chiral alkynes in strategic syntheses and bioorthogonal studies is always problematic. The chiral column high-performance liquid chromatography (HPLC method in general could not be directly used to resolve such substrates, since the differentiation of the alkyne segment with the other alkane/alkene segment is not significant in the stationary phase, and the alkyne group is not a good UV chromophore. Usually, a pre-column derivatization reaction with a tedious workup procedure is needed. Making use of easily-prepared stable alkyne-cobalt-complexes, we developed a simple and general method by analyzing the in situ generated cobalt-complex of chiral alkynes using chiral column HPLC. This new method is especially suitable for the alkynes without chromophores and other derivable groups.

  6. Design and Prototype of an Automated Column-Switching HPLC System for Radiometabolite Analysis

    Directory of Open Access Journals (Sweden)

    Neil Vasdev

    2016-08-01

    Full Text Available Column-switching high performance liquid chromatography (HPLC is extensively used for the critical analysis of radiolabeled ligands and their metabolites in plasma. However, the lack of streamlined apparatus and consequently varying protocols remain as a challenge among positron emission tomography laboratories. We report here the prototype apparatus and implementation of a fully automated and simplified column-switching procedure to allow for the easy and automated determination of radioligands and their metabolites in up to 5 mL of plasma. The system has been used with conventional UV and coincidence radiation detectors, as well as with a single quadrupole mass spectrometer.

  7. Design and Prototype of an Automated Column-Switching HPLC System for Radiometabolite Analysis.

    Science.gov (United States)

    Vasdev, Neil; Collier, Thomas Lee

    2016-08-17

    Column-switching high performance liquid chromatography (HPLC) is extensively used for the critical analysis of radiolabeled ligands and their metabolites in plasma. However, the lack of streamlined apparatus and consequently varying protocols remain as a challenge among positron emission tomography laboratories. We report here the prototype apparatus and implementation of a fully automated and simplified column-switching procedure to allow for the easy and automated determination of radioligands and their metabolites in up to 5 mL of plasma. The system has been used with conventional UV and coincidence radiation detectors, as well as with a single quadrupole mass spectrometer.

  8. Preparation and Characterization of a Polymeric Monolithic Column for Use in High-Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    Bindis, Michael P.; Bretz, Stacey Lowery; Danielson, Neil D.

    2011-01-01

    The high-performance liquid chromatography (HPLC) experiment, most often done in the undergraduate analytical instrumentation laboratory course, generally illustrates reversed-phase chromatography using a commercial C[subscript]18 silica column. To avoid the expense of periodic column replacement and introduce a choice of columns with different…

  9. Adaptation of Glass Columns For Clean-up in RP-HPLC Determination of Aflatoxins With Post-column Derivatisation With Bromine and Fluorescence Detection

    National Research Council Canada - National Science Library

    Owino, Victor Ochieng

    2009-01-01

    .... This study aimed to evaluate the possibility of using cheap florisil-packed glass columns instead of relatively complex and expensive pre-packed cartridges for afltoxin clean-up prior to HPLC analysis...

  10. [Determination of acrylamide in processed foods by column-switching HPLC with UV detection].

    Science.gov (United States)

    Terada, Hisaya; Tamura, Yukio

    2003-12-01

    A simple and convenient analytical method for the determination of acrylamide in processed foods was established. Acrylamide was extracted with water in an ultrasonic bath. The extract was passed through an OASIS HLB cartridge and the eluate was injected into the HPLC system using a column-switching technique. The HPLC system consisted of two pumps, two 6-port-2-position valves, two columns and a UV detector. At first, the sample solution was chromatographed on an ODS column with a mobile phase of water, then the flow of the mobile phase was switched using a 6-port-2-position valve, and the acrylamide peak fraction was introduced into an aqueous gel permeation column (analytical column). The fraction was chromatographed again on the analytical column with a mobile phase of water, and the eluate was monitored with a UV detector (205 nm). The recoveries of acrylamide from potato chips, fried potato, croquette and instant noodle fortified at levels of 50 to 1,000 micrograms/kg were 93.1 to 101.5% and the coefficient of variation was 1.5 to 5.2%. The detection limit corresponded to 10 micrograms/kg in processed foods. Forty-six samples, potato chips (11), fried potato (10), croquette (20) and instant noodle (5), were analyzed by this method. The acrylamide level was 67-4,499 micrograms/kg for potato chips, 125-1,183 micrograms/kg for fried potato, nd-255 micrograms/kg for croquette and nd-151 micrograms/kg for instant noodle.

  11. Determination of Bovine Lactoferrin in Food by HPLC with a Heparin Affinity Column for Sample Preparation.

    Science.gov (United States)

    Zhang, Yin; Lou, Fei; Wu, Wei; Dong, Xin; Ren, Jia; Shen, Qiuguang

    2017-01-01

    An HPLC method was developed for the quantitative determination of bovine lactoferrin (bLF) in sterilized milk, modified milk, fermented milk, infant formula, adult formula, rice cereal, vitamin function drink, and protein powder products. bLF was first extracted with a phosphate buffer (pH 8), underwent cleanup in a heparin affinity column, and was detected by HPLC with a C4 column and diode-array detector at a wavelength of 280 nm. The proposed method provided a linear detection range of 10.0-1000 μg/mL with an LOD of 0.6 mg/100 g in liquid samples and 3 mg/100 g in solid samples and an LOQ of 2 mg/100 g in liquid samples and 10 mg/100 g in solid samples. In addition, the method showed good recovery for various samples, ranging from 76 to 96%. The method had several remarkable advantages, including ease of handling, high sensitivity and accuracy, good reproducibility, and low-cost detection. Based on the distinctive properties presented here, we believe the proposed HPLC assay holds great promise for the oversight and detection of bLF in testing organizations, dairy enterprises, and regulatory authorities.

  12. Rapid determination of three alkaloids from Lotus Plumule in human serum using an HPLC-DAD method with a short monolithic column.

    Science.gov (United States)

    Du, Hui; Ren, Jing; Wang, Sicen

    2011-12-01

    An HPLC-DAD with a short monolithic column method was used for rapid, sensitive and simultaneous determination of liensinine, isoliensinine and neferine in human serum. Chromatographic separation was achieved on an RP-HPLC chromolithic column (50mm×4.6mm i.d.), with a mobile phase of methanol-water-triethylamine-acetic acid (70:30:0.2:0.05), at a flow rate of 1mlmin(-1). The calibration curves of three alkaloids exhibit good linear relationship in the concentration range of 0.5-80μgml(-1). The limits of detection (LODs) of three alkaloids were 20, 20 and 25ngml(-1), respectively, and the separation time was 4min. A comparative study has been performed with a normal C18 packed column (150mm×4.6mm i.d., 5μm). The separation time was 16min and the LODs of three alkaloids were 56, 56 and 65ngml(-1), respectively. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. A review of post-column photochemical reaction systems coupled to electrochemical detection in HPLC

    Energy Technology Data Exchange (ETDEWEB)

    Fedorowski, Jennifer [Department of Chemistry and Biochemistry, University of Maryland, Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250 (United States); LaCourse, William R., E-mail: lacourse@umbc.edu [Department of Chemistry and Biochemistry, University of Maryland, Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250 (United States)

    2010-01-04

    Post-column photochemical reaction systems have developed into a common approach for enhancing conventional methods of detection in HPLC. Photochemical reactions as a means of 'derivatization' have a significant number of advantages over chemical reaction-based methods, and a significant effort has been demonstrated to develop an efficient photochemical reactor. When coupled to electrochemical (EC) detection, the technique allows for the sensitive and selective determination of a variety of compounds (e.g., organic nitro explosives, beta-lactam antibiotics, sulfur-containing antibiotics, pesticides and insecticides). This review will focus on developments and methods using post-column photochemical reaction systems followed by EC detection in liquid chromatography. Papers are presented in chronological order to emphasize the evolution of the approach and continued importance of the application.

  14. C18 core-shell column with in-series absorbance and fluorescence detection for simultaneous monitoring of changes in stilbenoid and proanthocyanidin concentrations during grape cane storage.

    Science.gov (United States)

    Sáez, Vania; Gayoso, Camila; Riquelme, Sebastián; Pérez, Julie; Vergara, Carola; Mardones, Claudia; von Baer, Dietrich

    2017-12-29

    Grape canes, the residues from the annual pruning of vines, contain high levels of inducible (E)-resveratrol and also oligomeric stilbenoids and proanthocyanidins. These two families of phenolic compounds are bioactive, but to quantify them in a single chromatographic run using only ultraviolet detection is a difficult task. To overcome this limitation, a chromatographic method was developed using a core shell column for separation, an ultraviolet-visible diode array detector (DAD) and a fluorescence (FL) detector connected in series for quantification, with an electrospray ionization interface (ESI) and a triple quadrupole mass spectrometric detector (MS/MS) added for identification of the analytes. The proanthocyanidins (+)-catechin, (-)-epicatechin, procyanidins B1, B2, and C1, an unknown dimer and trimer, two prodelphinidin dimers, and monogallate procyanidin dimers were detected in the tested grape cane samples. The stilbenoids detected were (E)-resveratrol, (E)-piceatannol, (E)-piceid, (E)-ε-viniferin, vitisin B, a glycosylated monomer, three oxidized dimers, an unknown dimer and a tetramer, pallidol, hopeaphenol, (E)-δ-viniferin, and (E)-ω-viniferin. However, this method required 60min for each analysis. A faster and more efficient method for quantitative analysis was developed based on HPLC-DAD-FL, reducing the time required to 24min for the simultaneous quantification of proanthocyanidins and stilbenoids in Cabernet Sauvignon, Pinot Noir, and Tintorera grape canes stored at controlled temperatures and relativity humidities for 134days after pruning. To the best of our knowledge, this is the first time a prodelphinidin dimer has been quantified in grape canes. The incorporation of fluorescence detection in series with DAD not only allowed the quantification of proanthocyanidins, it also improved the detectability of some minor stilbenoids present in the canes, such as (E)-piceid. The (E)-resveratrol and (E)-piceatannol levels increased significantly

  15. Simultaneous column chromatographic extraction and purification of abscisic acid in peanut plants for direct HPLC analysis.

    Science.gov (United States)

    Zhang, Ya-Wen; Fan, Wei-Wei; Li, Hui; Ni, He; Han, Han-Bing; Li, Hai-Hang

    2015-10-01

    Abscisic acid (ABA), a universal signaling molecule, plays important roles in regulating plant growth, development and stress responses. The low contents and complex components in plants make it difficult to be accurately analyzed. A novel one-step sample preparation method for ABA in plants was developed. Fresh peanut (Arachis hypogaea) plant materials were fixed by oven-drying, microwave drying, boiling or Carnoy's fixative, and loaded onto a mini-preparing column. After washed the impurities, ABA was eluted with a small amount of solvent. ABA in plant materials was completely extracted and purified in 2mL solution and directly analyzed by HPLC, with a 99.3% recovery rate. Multiple samples can be simultaneously prepared. Analyses using this method indicated that the endogenous ABA in oven-dried peanut leaves increased 20.2-fold from 1.01 to 20.37μgg(-1) dry weight within 12h and then decreased in 30% polyethylene glycol 6000 treated plants, and increased 3.34-fold from 0.85 to 2.84μgg(-1) dry weight in 5 days and then decreased in soil drought treated plants. The method combined the column chromatographic extraction and solid-phase separation technologies in one step and can completely extracted plant endogenous ABA in a purified and highly concentrated form for direct HPLC analysis. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Solid phase extraction for multiresidue analysis of anabolic steroids and related substances from calf urine using C18 and alumina columns

    NARCIS (Netherlands)

    Koole, A; Franke, JP; de Zeeuw, RA

    1999-01-01

    A solid phase extraction method for anabolic steroids and related substances in calf urine is reported, that is suitable as a screening method for illegal growth promoters. Two types of sorbent were used: a reversed phase C18 material and a polar alumina material. After overnight enzymatic

  17. Method for determination of aflatoxin M₁ in cheese and butter by HPLC using an immunoaffinity column.

    Science.gov (United States)

    Sakuma, Hisako; Kamata, Yoichi; Sugita-Konishi, Yoshiko; Kawakami, Hiroshi

    2011-01-01

    A rapid, sensitive convenient method for determination of aflatoxin M₁ (AFM₁) in cheese and butter by HPLC was developed and validated. The method employs a safe extraction solution (mixture of acetonitrile, methanol and water) and an immunoaffinity column (IAC) for clean-up. Compared with the widely used method employing chloroform and a Florisil column, the IAC method has a short analytical time and there are no interference peaks. The limits of quantification (LOQ) of the IAC method were 0.12 and 0.14 µg/kg, while those of the Florisil column method were 0.47 and 0.23 µg/kg in cheese and buffer, respectively. The recovery and relative standard deviation (RSD) for cheese (spiked at 0.5 µg/kg) in the IAC method were 92% and 7%, respectively, while for the Florisil column method the corresponding values were 76% and 10%. The recovery and RSD for butter (spiked at 0.5 µg/kg) in the IAC method were 97% and 9%, and those in the Florisil method were 74% and 9%, respectively. In the IAC method, the values of in-house precision (n=2, day=5) of cheese and butter (spiked at 0.5 µg/kg) were 9% and 13%, respectively. The IAC method is superior to the Florisil column method in terms of safety, ease of handling, sensitivity and reliability. A survey of AFM₁ contamination in imported cheese and butter in Japan was conducted by the IAC method. AFM₁ was not detected in 60 samples of cheese and 30 samples of butter.

  18. Rapid Determination of the Monosaccharide Composition and Contents in Tea Polysaccharides from Yingshuang Green Tea by Pre-Column Derivatization HPLC

    Directory of Open Access Journals (Sweden)

    Yujie Ai

    2016-01-01

    Full Text Available A pre-column derivatization high-performance liquid chromatography (HPLC method was developed and optimized to characterize and quantify the monosaccharides present in tea polysaccharides (TPS isolated from Yingshuang green tea. TPS sample was hydrolyzed with trifluoroacetic acid, subjected to pre-column derivatization using 1-phenyl-3-methyl-5-pyrazolone (PMP, and separated on an Agilent TC-C18 column (4.6 mm × 250 mm, 5 μm with UV detection at 250 nm. A mixture of ten PMP derivatives of standard monosaccharides (mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose, arabinose, and fucose could be baseline separated within 20 min. Moreover, quantitative analysis of the component monosaccharides in Yingshuang green tea TPS was achieved, indicating the TPS consisted of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose, and arabinose in the molar contents of 0.72, 0.78, 0.89, 0.13, 0.15, 0.36, 0.39, 0.36, 0.36, and 0.38 μM, respectively. Recovery efficiency for component monosaccharides from TPS ranged from 93.6 to 102.4% with RSD values lower than 2.5%. In conclusion, pre-column derivatization HPLC provides a rapid, reproducible, accurate, and quantitative method for analysis of the monosaccharide composition and contents in TPS, which may help to further explore the relationship between TPS monosaccharides isolated from different tea varieties and their biological activity.

  19. HPLC method for rapidly following biodiesel fuel transesterification reaction progress using a core-shell column

    Energy Technology Data Exchange (ETDEWEB)

    Allen, Samuel J.; Ott, Lisa S. [California State University, Chico, CA (United States)

    2012-07-15

    There are a wide and growing variety of feedstocks for biodiesel fuel. Most commonly, these feedstocks contain triglycerides which are transesterified into the fatty acid alkyl esters (FAAEs) which comprise biodiesel fuel. While the tranesterification reaction itself is simple, monitoring the reaction progress and reaction products is not. Gas chromatography-mass spectrometry is useful for assessing the FAAE products, but does not directly address either the tri-, di-, or monoglycerides present from incomplete transesterification or the free fatty acids which may also be present. Analysis of the biodiesel reaction mixture is complicated by the solubility and physical property differences among the components of the tranesterification reaction mixture. In this contribution, we present a simple, rapid HPLC method which allows for monitoring all of the main components in a biodiesel fuel transesterification reaction, with specific emphasis on the ability to monitor the reaction as a function of time. The utilization of a relatively new, core-shell stationary phase for the HPLC column allows for efficient separation of peaks with short elution times, saving both time and solvent. (orig.)

  20. HPLC method with monolithic column for simultaneous determination of irbesartan and hydrochlorothiazide in tablets

    Directory of Open Access Journals (Sweden)

    Alanazi Amer M.

    2014-06-01

    Full Text Available A simple, sensitive and accurate HPLC method with high throughput has been developed and validated for the simultaneous determination of irbesartan (IRB and hydrochlorothiazide (HCT in combined pharmaceutical dosage forms. The proposed method employed, for the first time, a monolithic column in the analysis. Optimal chromatographic separation of the analytes was achieved on Chromolith® Performance RP-18e column using a mobile phase consisting of phosphate buffer (pH 4/acetonitrile (50:50, V/V pumped isocratically at a flow rate of 1.0 mL min-1. The eluted analytes were monitored with a UV detector set at 270 nm. Under the optimum chromatographic conditions, linear relationship with a good correlation coefficient (R ≥ 0.9997 was found between the peak area and the corresponding concentrations of both IRB and HCT in the ranges of 10-200 and 1-20 ng mL-1. The limits of detection were 2.34 and 0.03 ng mL-1 for IRB and HCT, respectively. The intra- and inter-assay precisions were satisfactory as the RSD values did not exceed 3 %. The accuracy of the proposed method was > 97 %. The proposed method had high throughput as the analysis involved a simple procedure and a very short run- -time of < 3 min. The results demonstrated that the method is applicable in the quality control of combined pharmaceutical tablets containing IRB and HCT

  1. Use of small diameter column particles to enhance HPLC determination of histamine and other biogenic amines in seafood

    DEFF Research Database (Denmark)

    Simat, Vida; Dalgaard, Paw

    2011-01-01

    for determination of biogenic amines in lean canned tuna and fatty frozen herring The modified methods using smaller column particles of 1 8 mu m or 3 mu m allowed biogenic amines to be separated and quantified faster (23-59%) and with less eluent consumption (59-62%) than classical HPLC methods Backpressures were...

  2. Influence of batch-to-batch reproducibility of Luna C-18(2) packing material, nature of column wall material, and column diameter on the liquid chromatographic analysis of basic analytes

    NARCIS (Netherlands)

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

    This paper discusses aspects arising on transferring liquid chromatography (LC) methods developed on conventional size columns to micro LC, i.e. the influence of batch-to-batch reproducibility of packing material, the nature of the column wall material, and the column inner diameter. it was shown

  3. Surface Characterization of Some Novel Bonded Phase Packing Materials for HPLC Columns Using MAS-NMR Spectroscopy

    Directory of Open Access Journals (Sweden)

    Jude Abia

    2015-03-01

    Full Text Available Information on the surface properties of three novel chemically bonded phase packing materials for High performance liquid chromatography (HPLC were obtained using spectra obtained by solid state cross-polarization (CP magic-angle spinning (MAS nuclear magnetic resonance (NMR spectroscopic experiments for the 29Si, and 13C nuclei. These packing materials were: Cogent bidentate C18 bonded to type-C silica, hybrid packing materials XTerra MS C18, and XBridge Prep. C18. The spectra obtained using cross-polarization magic angle spinning (CP-MAS on the Cogent bidentate C18 bonded to type-C silica show the surface to be densely populated with hydride groups (Si-H, with a relative surface coverage exceeding 80%. The hybrid packing materials XTerra and XBridge gave spectra that reveal the silicon atoms to be bonded to organic moieties embedded in the molecular structure of these materials with over 90% of the alkyl silicon atoms found within the completely condensed silicon environments. The hydrolytic stability of these materials were investigated in acidic aqueous solutions at pHs of 7.0 and 3.0, and it was found that while the samples of XTerra and XBridge were not affected by hydrolysis at this pH range, the sample of Cogent lost a significant proportion of its Si-H groups after five days of treatment in acidic aqueous solution.

  4. Absorbance detector based on a deep UV light emitting diode for narrow-column HPLC.

    Science.gov (United States)

    Bui, Duy Anh; Bomastyk, Benjamin; Hauser, Peter C

    2013-10-01

    A detector for miniaturized HPLC based on deep UV emitting diodes and UV photodiodes was constructed. The measurement is accomplished by the transverse passage of the radiation from the light-emitting diode (LED) through fused-silica tubing with an internal diameter of 250 μm. The optical cell allows flexible alignment of the LED, tubing, and photodiode for optimization of the light throughput and has an aperture to block stray light. A beam splitter was employed to direct part of the emitted light to a reference photodiode and the Lambert-Beer law was emulated with a log-ratio amplifier circuitry. The detector was tested with two LEDs with emission bands at 280 and 255 nm and showed noise levels as low as 0.25 and 0.22 mAU, respectively. The photometric device was employed successfully in separations using a column of 1 mm inner diameter in isocratic as well as gradient elution. Good linearities over three orders of magnitude in concentration were achieved, and the precision of the measurements was better than 1% in all cases. Detection down to the low micromolar range was possible. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Determination of catecholamines and related compounds in mouse urine using column-switching HPLC.

    Science.gov (United States)

    Kanamori, Takahiro; Funatsu, Takashi; Tsunoda, Makoto

    2016-04-21

    We have developed an analytical method for the determination of catecholamines and related compounds in mouse urine by column-switching HPLC. Selective extraction of the catechol compounds was performed using a precolumn modified with phenylboronic acid, which has a pH dependent affinity for the catechol structures. The pretreatment buffer, which facilitated binding of the catechols to the precolumn, was optimized to ensure high analyte recoveries and good peak shapes. We found that using the same acetonitrile content in the pretreatment buffer and hydrophilic interaction liquid chromatography mobile phase was necessary to improve peak shapes. Eight catechol compounds were selectively extracted and separated using 100 mmol L(-1) ammonium formate/acetonitrile (20/80 v/v, pH 8.0) for the extraction step, and 20 mmol L(-1) ammonium formate (pH 2.5)/acetonitrile (20/80 v/v) for elution and separation. Native fluorescence of the separated catechol compounds was monitored, and the limits of detection, corresponding to a signal to noise ratio of 3, were 9-58 nmol L(-1). Five catechol compounds (dopamine, epinephrine, norepinephrine, 3,4-dihydroxyphenylglycol, and 3,4-dihydroxymandelic acid) were successfully quantified in mouse urine. Intra- and inter-day precisions were less than 10%, and performance was superior to that afforded by manual sample pretreatment.

  6. Rapid simultaneous determination of eperisone, tolperisone, and tizanidine in human serum by using a MonoSpin® C18 extraction column and liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Miura, Naoya; Saito, Takeshi; Taira, Takayuki; Yamagiwa, Takeshi; Morita, Sein; Inokuchi, Sadaki

    2014-01-01

    A method was developed for rapid toxicological analysis of eperisone, tolperisone, and tizanidine in human serum using a MonoSpin® C18 extraction column and LC/MS/MS. The method was validated for LOD, linearity, precision, and extraction recovery. This method was rapid with an LOD of 0.5 ng/mL, linearity range 1-500.0 ng/mL (r2 = 0.999), and RSD value below 14.6%. Extraction recovery from the sample was greater than 98.6, 98.8, and 88.5% for eperisone, tolperisone, and tizanidine, respectively. Results showed that combination of the MonoSpin C18 extraction column and LC/MS/MS is a simple and rapid method for the analysis of these three analytes, and a method is described for simultaneous quantitative determination of the analytes in human serum by LC/MSIMS. This method was used to determine the serum levels of eperisone in a patient with eperisone poisoning, and could be successfully applied for screening analyses in clinical cases other than poisoning.

  7. HPLC enantioseparation of racemic bupropion, baclofen and etodolac: modification of conventional ligand exchange approach by pre-column formation of chiral ligand exchange complexes.

    Science.gov (United States)

    Singh, Manisha; Bhushan, Ravi

    2016-11-01

    Separation of racemic mixture of (RS)-bupropion, (RS)-baclofen and (RS)-etodolac, commonly marketed racemic drugs, has been achieved by modifying the conventional ligand exchange approach. The Cu(II) complexes were first prepared with a few l-amino acids, namely, l-proline, l-histidine, l-phenylalanine and l-tryptophan, and to these was introduced a mixture of the enantiomer pair of (RS)-bupropion, or (RS)-baclofen or (RS)-etodolac. As a result, formation of a pair of diastereomeric complexes occurred by 'chiral ligand exchange' via the competition between the chelating l-amino acid and each of the two enantiomers from a given pair. The diastereomeric mixture formed in the pre-column process was loaded onto HPLC column. Thus, both the phases during chromatographic separation process were achiral (i.e. neither the stationary phase had any chiral structural feature of its own nor did the mobile phase have any chiral additive). Separation of diastereomers was successful using a C18 column and a binary mixture of MeCN and TEAP buffer of pH 4.0 (60:40, v/v) as mobile phase at a flow rate of 1 mL/min and UV detection at 230 nm for (RS)-Bup, 220 nm for (RS)-Bac and 223 nm for (RS)-Etd. Baseline separation of the two enantiomers was obtained with a resolution of 6.63 in <15 min. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  8. Simultaneous determination of iodide and iodate in soil solution samples by HPLC with electrochemical detection and post-column reaction method

    Energy Technology Data Exchange (ETDEWEB)

    Takeda, Akira; Takaku, Yuichi; Hisamatsu, Shun' ichi [Department of Radioecology, Institute for Environmental Sciences, Aomori 039-3212 (Japan); Tsukada, Hirofumi [Department of Radioecology, Institute for Environmental Sciences, Aomori 039-3212 (Japan); Institute of Environmental Radioactivity, Fukushima University, Fukushima 960-1196 (Japan)

    2014-07-01

    Iodine-129 (half-life 1.6 x 10{sup 7} y) discharged into the atmosphere from nuclear facilities (e.g., a nuclear fuel reprocessing plant) is partly deposited on land and introduced into soil. Stable iodine ({sup 127}I) can be used as a natural analogue to predict the long-term behavior of {sup 129}I in the terrestrial environment. Iodine in soil mainly exists as I{sup -}, IO{sub 3}{sup -}, and organic iodine. Because the mobilities of these species in soil are quite different, iodine speciation in soil solution is a key for predicting the behavior of iodine in soil. We developed a new speciation method suitable for routine analysis of many soil solution samples, and successfully applied the method to real samples. The method involves determining the concentration of total iodine and then separately measuring the I{sup -} and IO{sub 3}{sup -} concentrations with an HPLC system. The HPLC system (Nano-space SI-2; Shiseido, Tokyo, Japan) consisted of a UV/Vis spectrometer and an electrochemical (amperometric) detector (50 mV Ag/AgCl). Two reverse-phase columns (2.0 x 50 mm Capcel Pak DD C8 and 2.0 x 250 mm Capcel Pak MGII C18; Shiseido) were serially connected, and a switching valve was set between them. I{sup -} and IO{sub 3}{sup -} in the sample solution were separated from each other in the DD C8 column. IO{sub 3}{sup -} eluted first from the column, while I{sup -} was retained. After IO{sub 3}{sup -} was further separated from other halogen acids with the C18 column, IO{sub 3}{sup -} was reacted with KBr and o-dianisidine in a thermos-reactor (90 deg. C), and absorption at 450 nm was measured with the UV/Vis spectrometer. The concentration of I{sup -} eluted from the first column was determined with the electrochemical detector. To determine the concentration of total iodine in the sample solution, organic iodine was decomposed by UV irradiation (UV digester 705; Metrohm AG, Herisau, Switzerland) for 30 min at 20 deg. C. The iodine in the solution was reduced to I

  9. Separation of chlorophylls and carotenoids from marine phytoplankton: a new HPLC method using a reversed phase C8 column and pyridine-containing mobile phases

    National Research Council Canada - National Science Library

    Manuel Zapata; Francisco Rodríguez; José L. Garrido

    2000-01-01

    A high-performance liquid chromatographic (HPLC) method based on a reversed-phase C8 column and pyridine-containing mobile phases was developed for the simultaneous separation of chlorophylls and carotenoids...

  10. Determination of blood urea using cation exchange column solid phase extraction combined with HPLC

    National Research Council Canada - National Science Library

    Hideharu SHINTANI; Takashi INOUE

    1994-01-01

    ...°C with detection at 200 nm. By comparing ultrafiltrated blood urea and native blood urea, the blood urea from the HPLC chromatogram was found to be completely free of blood admixtures and almost free of blood protein...

  11. Pico-HPLC system integrating an equal inner diameter femtopipette into a 900 nm I.D. porous layer open tubular column.

    Science.gov (United States)

    Li, Ruonan; Shao, Yunlong; Yu, Yanmin; Wang, Xiayan; Guo, Guangsheng

    2017-04-06

    A picoflow high performance liquid chromatography (pico-HPLC) system was developed, which could directly pipette femtoliter samples using a separation column tip driven by an electroosmotic pump. Amino acid enantiomers were separated in the 900 nm I.D. porous layer open tubular column at a flow rate of 13.50 pL min(-1).

  12. Measurement uncertainty assessment of magnesium trisilicate column for determination of Sudan colorants in food by HPLC using C8 column.

    Science.gov (United States)

    Chen, Ying; He, Chao; Cheng, Jing-Jun; Huang, Wen-Yao; Shao, Sheng-Wen; Jiang, Ya-Ping; Dai, Ling-Feng; Liu, Jia-Fa; Song, Yi

    2016-10-01

    This study aimed to conduct measurement uncertainty assessment of a new method for determination of Sudan colorants (Sudan I, II, III and IV) in food by high performance liquid chromatography (HPLC). Samples were extracted with organic solvents (hexane, 20% acetone) and first purified by magnesium trisilicate (2MgO·3SiO2). The Sudan colorants (Sudan I-IV) were also initially separated on C8 by gradient elution using acetonitrile and 0.1% (v/v) formic acid aqueous solution as the mobile phases and detected with diode-array detector (DAD). The uncertainty of mathematical model of Sudan I, II, III and IV is based on EURACHEM guidelines. The sources and components of uncertainty were calculated. The experiment gave a good linear relationship over the concentration from 0.4 to 4.0 μg/mL and spiked recoveries were from 74.0% to 97.5%. The limits of determination (LOD) were 48, 61, 36, 58 μg/kg for the four analytes, respectively. The total uncertainty of Sudan colorants (Sudan I, II, III and IV) was 810±30.8, 790±28.4, 750±27.0, 730±50.0 μg/kg, respectively. The recovery uncertainty was the most significant factor contributing to the total uncertainty. The developed method is simple, rapid, and highly sensitive. It can be used for the determination of trace Sudan dyes in food samples. The sources of uncertainty have been identified and uncertainty components have been simplified and considered.

  13. Study on the Alkaloids in Tibetan Medicine Aconitum pendulum Busch by HPLC-MSn Combined with Column Chromatography.

    Science.gov (United States)

    Wang, Beibei; Dong, Jie; Ji, Jiaojiao; Yuan, Jiang; Wang, Jiali; Wu, Jiarui; Tan, Peng; Liu, Yonggang

    2016-01-01

    A rapid, convenient and effective identification method of alkaloids was established and an attempt on isolating and analyzing the alkaloids in Aconitum pendulum Busch was conducted successfully. In this article, four high-content components including deoxyaconitine, benzoylaconine, aconine and neoline were isolated by using column chromatography. HPLC-MS(n)was employed to deduce the regulations of fragmentation of diterpenoid alkaloids which displayed a characteristic behavior of loss of CO(28u), CH3COOH(60u), CH3OH(32u), H2O(18u) and C6H5COOH(122u). Then, according to fragmentation regulation of mass spectrometry, 42 alkaloids were found inA. pendulum Among them, 38 compounds were identified and 29 alkaloids were reported for the first time for this herb. Therefore, this means that HPLC-MS(n)combined with column chromatography could work as an effective and reliable tool for rapid identification of the chemical components of herbal medicine. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

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

    African Journals Online (AJOL)

    ... rapid, cost-effective and accurate reverse phase-high performance liquid chromatography (RP-HPLC) method for the determination of aprepitant (APT) in capsule dosage form. Methods: The method developed for the determination of APT in capsule formulation involved using RP-HPLC which incorporated a C18 column ...

  15. Adaptation of glass columns for clean-up RP-HPLC determination of ...

    African Journals Online (AJOL)

    SD 0.1), 10.7 (SD 0.3) and 9.1 (SD 0.3) minutes, respectively for AFB1, AFB2, AFG1 and AFG2. Florisil powder that had been dried for 12 hours at 100 oC, then hydrated with water at 5% (w/v) before packing on glass columns gave slightly ...

  16. Isolation and identification of arctiin and arctigenin in leaves of burdock (Arctium lappa L.) by polyamide column chromatography in combination with HPLC-ESI/MS.

    Science.gov (United States)

    Liu, Shiming; Chen, Kaoshan; Schliemann, Willibald; Strack, Dieter

    2005-01-01

    A simple method involving polyamide column chromatography in combination with HPLC-PAD and HPLC-ESI/MS for isolating and identifying two kinds of lignans, arctiin and arctigenin, in the leaves of burdock (Arctium lappa L.) has been established. After extraction of burdock leaves with 80% methanol, the aqueous phase of crude extracts was partitioned between water and chloroform and the aqueous phase was fractionated on a polyamide glass column. The fraction, eluting with 100% methanol, was concentrated and gave a white precipitate at 4 degrees C from which two main compounds were purified by semi-preparative HPLC. In comparison with the UV and ESI-MS spectra and the HPLC retention time of authentic standards, the compounds were determined to be arctiin and arctigenin. The extraction/separation technique was validated using an internal standard method.

  17. Pre-Column Derivatization HPLC Procedure for the Quantitation of Aluminium Chlorohydrate in Antiperspirant Creams Using Quercetin as Chromogenic Reagent.

    Science.gov (United States)

    Kalogria, Eleni; Varvaresou, Athanasia; Papageorgiou, Spyridon; Protopapa, Evaggelia; Tsaknis, Ioannis; Matikas, Alexios; Panderi, Irene

    2014-01-01

    This article describes the development and validation of a selective high-performance liquid chromatography method that allows, after liquid-liquid extraction and pre-column derivatization reaction with quercetin, the quantification of aluminium chlorohydrate in antiperspirant creams. Chromatographic separation was achieved on an XTerra MS C18 analytical column (150 × 3.0 mm i.d., particle size 5 μm) using a mobile phase of acetonitrile:water (15:85, v/v) containing 0.08 % trifluoroacetic acid at a flow rate of 0.30 mL min -1 . Ultraviolet spectrophotometric detection at 415 nm was used. The assay was linear over a concentration range of 3.7-30.6 μg mL -1 for aluminium with a limit of quantitation of 3.74 μg mL -1 . Quality control samples (4.4, 17.1 and 30.6 μg mL -1 ) in five replicates from five different runs of analysis demonstrated intra-assay precision (% coefficient of variation creams containing 11.0, 13.0 and 16.0 % (w/w) aluminium chlorohydrate, respectively.

  18. Molecularly imprinted polydopamine nano-layer on the pore surface of porous particles for protein capture in HPLC column.

    Science.gov (United States)

    Nematollahzadeh, Ali; Shojaei, Akbar; Abdekhodaie, Mohammad J; Sellergren, Börje

    2013-08-15

    Bio-inspired Human Serum Albumin (HSA) imprinted polydopamine nano-layer was produced through oxidative polymerization of dopamine on the pore surface of HSA modified porous silica particles. The coating thickness was controlled by the reaction time and thereby varied within 0-12 nm. The samples were characterized by elemental analysis, FT-IR, DSC, SEM, TEM, TGA, physisorption and thermoporometry. The characterization confirmed the success of evolution and deposition of polydopamine layer on the silica pore surface. Batch rebinding experiment showed that the molecularly imprinted polymer (MIP) with 8.7 nm coating thickness, in comparison with the thinner and thicker coatings, displays the highest uptake of the target protein. The chromatographic evaluation of the materials packed in HPLC columns showed that the HSA imprinted polydopamine offers good mechanical stability and retains practically all the target protein from an HSA solution or human plasma. Affinity of the imprinting column was examined by using Bovine Serum Albumin (BSA) and Immunoglobulin G (IgG) as competitive proteins. The results showed that the template, HSA, was the most adsorbed protein by the imprinted polydopamine layer. Crown Copyright © 2013. Published by Elsevier Inc. All rights reserved.

  19. [HPLC Characteristic Fingerprint of "Boju" Chrysanthemum morifolium].

    Science.gov (United States)

    Yu, Nian-jun; Yu, Jiao; Zheng, Wei; Cao, Yong; Zheng, Tai-hua; Wang, Yun-qin

    2015-03-01

    To provide a scientific basis for quality control of "Boju" Chrysanthemum morifolium by establishing a HPLC characteristic fingerprint. The HPLC analysis was performed on a Spursil C18 chromatographic column (250 mm x 4. 6 mm, 5 µm), and the mobile phase was acetonitrile-0. 1% phosphoric acid in a gradient mode with the flow rate of 1. 0 mL/min. The detection wavelength was 325 nm and the temperature of column was 30 °C. The common pattern of HPLC characteristic chromatographic profile was established. There were 16 common peaks, five of which were identified in the pattern. The similarities of 10 batches of "Boju" Chrysanthemum monrfolium were evaluated, and all of them were greater than 0. 900. This analysis method of HPLC characteristic chromatographic fingerprint is simple and reproducible, and it can provide a scientific basis for identification and quality evaluation of "Boju" Chrysanthemum monfolium.

  20. Stability-Indicating HPLC Method for the Simultaneous ...

    African Journals Online (AJOL)

    Methods: A stability indicating method for the simultaneous estimation of valsartan and ezetimibe in combined tablet formulation using a RP-HPLC was developed and validated as per ICH guidelines using a symmetry C18 column with a mobile phase comprising phosphate buffer and acetonitrile (58:42 v/v, pH 3.15) with a ...

  1. A Stability Indicating HPLC Method for the Determination of ...

    African Journals Online (AJOL)

    Purpose: The present study was undertaken to develop a validated, rapid, simple and economic stability indicating reverse phase HPLC method for estimating meloxicam (MLX) in bulk and commercial preparations. Method: Reversed phase chromatographic analysis was performed on a C18 Hi Q Sil column with ...

  2. HPLC analysis of closed, open, and reflex eye tear proteins

    OpenAIRE

    Sitaramamma T; Shivaji S; Rao Gullapalli

    1998-01-01

    Changes in the closed, open and reflex eye tear proteins of normal subjects were compared and analysed. Tear proteins were resolved by high-performance liquid chromatography (HPLC) utilising both gel filtration (P-300 SW) and reverse-phase (C-18) columns and the HPLC fractions were further analysed by sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE) under reducing and non-reducing conditions. The protein composition of the closed-eye tear was significantly different fro...

  3. Multi residue method using coupled-column HPLC and GC-MS for the determination of anabolic compounds in samples of urine

    NARCIS (Netherlands)

    Herbold HA; Sterk SS; Stephany RW; Ginkel LA van; ARO

    1998-01-01

    This report describes a multi-residue method for the detection and identification of residues of anabolic compounds. The method is based on automated sample preparation (Solid Phase Extraction (SPE)) and coupled-column High Performance Liquid Chromatography (HPLC) and Gas Chromatography-Mass

  4. A new selective pre-column ninhydrin-based derivatization for a RP-HPLC determination of plasma asymmetric dimethyl-L-arginine (ADMA) by fluorescence detection.

    Science.gov (United States)

    Sotgia, Salvatore; Zinellu, Angelo; Pinna, Gerard Aime; Deiana, Luca; Carru, Ciriaco

    2008-05-01

    We report a new selective and direct pre-column ninhydrin-based derivatization reaction for determination of plasma ADMA levels. This original derivatization procedure matched to a validated and rapid RP-HPLC method can be a useful alternative to other assays in which time consuming and expensive extraction and/or purification steps are required.

  5. Rapid determination of oxindole alkaloids in cat's claw by HPLC using ionic liquid-based microwave-assisted extraction and silica monolithic column.

    Science.gov (United States)

    Chang, Chih-Wei; Yeh, Yu-Ying; Chang, Li-Ching; Hsu, Mei-Chich; Wu, Yu-Tse

    2017-08-01

    Cat's claw is a large woody vine with hook-like thorns, and has been traditionally used to treat inflammatory disorders in South and Central America. In this study, a rapid, validated high-performance liquid chromatographic (HPLC) method using a silica monolithic column was developed for the simultaneous determination of oxindole alkaloids, namely rhynchophylline, pteropodine, isomitraphylline and isopteropodine, in cat's claw. The ionic liquid-based microwave-assisted extraction (ILMAE), considered as an environmentally friendly and powerful tool, was first applied in the extraction of oxindole alkaloids. To optimize the HPLC method, the stationary phases, pH values of mobile phase and flow rates were investigated. The validated HPLC method using a Monolithic RP18e column (100 × 4.6 mm) enables these analytes to be separated almost twice as fast as with a conventional particulate column (~16 vs ~30 min) with limits of quantification and detection of 0.5 and 0.15 μg/mL, respectively. The ILMAE conditions were optimized by the Taguchi orthogonal array design. In comparison with conventional water boiling extraction, ILMAE offers almost four times higher yields within an extremely short extraction time. The developed HPLC coupled with ILMAE method could be efficient and practical for rapid determination of oxindole alkaloids in cat's claw. Copyright © 2016 John Wiley & Sons, Ltd.

  6. A porous graphitized carbon column HPLC method for the quantification of paracetamol, pseudoephedrine, and chlorpheniramine in a pharmaceutical formulation.

    Science.gov (United States)

    Kalogria, Eleni; Koupparis, Michael; Panderi, Irene

    2010-01-01

    A simple, rapid, and stability-indicating HPLC method has been developed, fully validated, and applied to the quantification of paracetamol, pseudoephedrine hydrochloride, and chlorpheniramine maleate in a pharmaceutical formulation, using hydrochlorothiazide as an internal standard. Chromatographic separation was achieved isocratically on an RP porous graphitized carbon analytical column (125 x 2.1 mm id, particle size 5 microm) using 5.0 mM ammonium acetate-acetonitrile (35 + 65, v/v) mobile phase at a flow rate of 0.50 mL/min. UV spectrophotometric detection at 220 nm was used. The method had linear calibration curves over the range of 30-70 microg/mL for paracetamol, 1.8-4.2 microg/mL for pseudoephedrine hydrochloride, and 120-280 ng/mL for chlorpheniramine maleate. The intraday and interday RSD values were less than 3.2% for all compounds, while the relative error was less than 2.9%. Accelerated stability studies performed under various stress conditions proved the selectivity of the method. The developed method was applied successfully to QC and content uniformity tests of commercial tablets.

  7. Development of an HPLC Method with an ODS Column to Determine Low Levels of Aspartame Diastereomers in Aspartame.

    Directory of Open Access Journals (Sweden)

    Takashi Ohtsuki

    Full Text Available α-L-Aspartyl-D-phenylalanine methyl ester (L, D-APM and α-D-aspartyl-L-phenylalanine methyl ester (D, L-APM are diastereomers of aspartame (N-L-α-Aspartyl-L-phenylalanine-1-methyl ester, L, L-APM. The Joint FAO/WHO Expert Committee on Food Additives has set 0.04 wt% as the maximum permitted level of the sum of L, D-APM and D, L-APM in commercially available L, L-APM. In this study, we developed and validated a simple high-performance liquid chromatography (HPLC method using an ODS column to determine L, D-APM and D, L-APM in L, L-APM. The limits of detection and quantification, respectively, of L, D-APM and D, L-APM were found to be 0.0012 wt% and 0.004 wt%. This method gave excellent accuracy, repeatability, and reproducibility in a recovery test performed on five different days. Moreover, the method was successfully applied to the determination of these diastereomers in commercial L, L-APM samples. Thus, the developed method is a simple, useful, and practical tool for determining L, D-APM and D, L-APM levels in L, L-APM.

  8. Improvement of accuracy for the quantitation of selenoproteins in post-column isotope dilution technique with HPLC ICP/MS

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Soo Jin; Pak, Yong Nam [Dept. of Chemistry Education, Korea National University of Education, Cheongju (Korea, Republic of)

    2016-11-15

    Post-column isotope dilution (PCID) or species-unspecific isotope dilution (ID) is a very useful quantitative technique in chromatography hyphenated with inductively coupled plasma–mass spectrometer (ICP/MS). In quantitative analysis, the calibration curve method is the most common one. However, it is not an absolute technique for the determination of concentration for the samples. ID method is the absolute one for the quantitative measurement technique, which means the quantity could be determined by the fundamental unit such as weight. To use the technique, it needs some requirements and there are several restrictions especially in the application for high-performance liquid chromatography (HPLC). The impurities in eluting solvent were in question. Thus, different purity of AA was examined and the result is shown in Figure 3. Figure 3(a) is the one for low (98%) and Figure 3(b) is high (99.999%) purity AA. The dotted area is for the eluting solvent. When low purity of 98% AA was introduced, the background was increasing while it remained the same for high-purity AA.

  9. An automated HPLC method for the fractionation of polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans in fish tissue on a porous graphitic carbon column

    Science.gov (United States)

    Echols, Kathy R.; Gale, Robert W.; Tillitt, Donald E.; Schwartz, Ted R.; O'Laughlin, Jerome

    1997-01-01

    The Ah (aryl-hydrocarbon) hydroxylase-receptor active polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were fractionated by an automated high-performance liquid chromatography (HPLC) system using the Hypercarb™ porous graphitic carbon (PGC) column. This commercially available column was used to fractionate the di-, mono-, and non-ortho PCBs into three fractions for gas chromatography (GC)/electron capture detection analysis, and a fourth fraction containing the PCDDs/PCDFs for GC/mass spectrometry analysis. The recoveries of the PCBs ranged from 68 to 96%, and recoveries of the PCDDs/PCDFs ranged from 74 to 123%. The PGC column has the advantage of faster separations (110 min versus 446 min) and less solvent use (275 ml versus 1,100 ml) compared with automated fractionation of these compounds on activated carbon (PX-21), while still affording good separation of the classes. The PGC column may have an advantage over the pyrenyl-based HPLC method because it has a greater loading capacity (400 μg total PCBs versus 250 μg). Overall, the PGC is a standard column that provides reproducible fractionation of PCDD/PCDFs and PCBs for analytical measurement in environmental samples.

  10. The Human HPLC Column

    Science.gov (United States)

    Frantz, Kyle

    2007-01-01

    Initiatives in education reform emphasize inquiry-based active learning and real-world relevance to increase science literacy nationwide. Active teaching and learning approaches yield rapid intellectual development and may increase interest and motivation to learn science. Incorporating the topic of drug use with neuroscience, biology, psychology,…

  11. DETERMINATION AND VALIDATION OF MEBHYDROLINE NAPADISYLATE IN TABLETS BY HPLC

    Directory of Open Access Journals (Sweden)

    Lestyo Wulandari

    2010-06-01

    Full Text Available An accurate and sensitive HPLC method has been developed for the determination of mebhydroline napadisylate in the tablet. The Chromatography was performed on a reversed phase C-18 column, using a mobile phase of acetonitrile : ammonia 25% (80 : 20 v/v at ambient temperature 25±5 °C and UV detection operates at 320 nm in an overall analysis time of about 15 min, based on peak area. This HPLC method is selective, precise, and accurate and can be used for routine analysis of pharmaceutical preparation in industrial quality-control laboratories.   Keywords : HPLC, mebhydroline napadisylate, validation

  12. Lipophilicity of porphyrins and their retention in IAM, C8-C18 and HILIC chromatographic systems.

    Science.gov (United States)

    Essaid, D; Chaminade, P; Maillard, Ph; Kasselouri, A

    2015-10-10

    Porphyrins are a class of photosensitizers used in photodynamic therapy (PDT). Understanding the interaction of porphyrins with membrane cells components is important in order to improve this therapy. Many analytical methods can be used for this aim. High performance liquid chromatography (HPLC) was used for the separation of porphyrins on RP and HILIC stationary phases as well as on a biomimetic membrane IAM phase. Twenty-six tetraphenyl porphyrins (TPP) were successfully separated on an IAM column, a C18 Gravity RP column, a C8 Gravity RP column, a PolarTec RP column and a HILIC column. Stationary phases were chosen as the most appropriate to cover the study of different types of interactions. Elution was performed with a 45 min linear gradient. Obtained gradient retention times were converted to gradient chromatography hydrophobicity index (CHI) and to an apparent retention factor (kapp). The partition coefficients (logP) of the 26 compounds were measured in a 2-octanol/PBS system and estimated in silico. Correlation between kapp values was studied. Moreover, a multivariate analysis was performed to explain columns relationships. Obtained results show that porphyrins are separated mainly according to hydrophobic interactions that are relative to their structure (sugar number and the disposition around the porphyrin macrocycle). Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Development and validation of a RP-HPLC method for assay of ...

    African Journals Online (AJOL)

    Purpose: To develop and validate a novel reverse phase high performance liquid chromatographic (RP-HPLC) method for the quantification of atorvastatin in thermosensitive hydrogel-based nanocrystal formulation. Method: Chromatographic identification was achieved on C18 (5 μm) column using acetonitrile and 0.025 M ...

  14. On-column deracemization of an atropisomeric biphenyl by quinine-based stationary phase and determination of rotational energy barrier by enantioselective stopped-flow HPLC and CEC.

    Science.gov (United States)

    Tobler, E; Lämmerhofer, M; Mancini, G; Lindner, W

    2001-01-01

    The reversible enantiomerization of axially chiral 2'-dodecyloxy-6-nitrobiphenyl-2-carboxylic acid was studied in the presence of a brush type chiral stationary phase based on O-(tert-butylcarbamoyl) quinine as chiral selector unit by stopped-flow high-performance liquid chromatography (sfHPLC) and capillary electrochromatography (sfCEC). After initial separation of the enantiomers in the first section of the column, the flow was stopped and the resolved species allowed to enantiomerize on-column. From this conversion, which could be determined from the enantiomeric ratios at different enantiomerization times, kinetic rate constants were calculated. By sfHPLC at a constant temperature of 15 degrees C, kinetic rate constants in the presence of the CSP were found to be 4.1 x 10(-5) s(-1) and 2.2 x 10(-5) s(-1) for the (-) and (+)-enantiomers, respectively, corresponding to half-lives of 279 and 530 min. Thus, apparent activation energies of enantiomerization were calculated to be 93.0 and 94.6 kJ mol(-1) for the (-) and (+)-enantiomers. On the macroscopic level, the apparent difference of rotational energy barriers and kinetic rate constants for both enantiomers is reflected as deracemization. For example, starting from a racemic mixture, an enantiomeric excess (ee) of 14% was seen in the stopped-flow HPLC experiment described after an enantiomerization time of 220 min at 15 degrees C, and a maximal ee of 17% can be approximated after infinite enantiomerization time. There is good agreement between HPLC and CEC results as well as their experimental errors, confirming that the new sfCEC technique may be a valuable and convenient tool to study interconversion processes. Copyright 2001 Wiley-Liss, Inc.

  15. Titania as a chemo-affinity support for the column-switching HPLC analysis of phosphopeptides: application to the characterization of phosphorylation sites in proteins by combination with protease digestion and electrospray ionization mass spectrometry.

    Science.gov (United States)

    Sano, Akira; Nakamura, Hiroshi

    2004-05-01

    A method for the determination of phosphorylation sites in phosphoproteins based on column-switching high-performance liquid chromatography (HPLC) has been developed. The HPLC system consisted of a titania precolumn for the selective adsorption of phosphopeptides, an anion-exchange analytical column and a UV detector (215 nm). Rabbit muscle phosphorylase a (RPa) and porcine stomach pepsin (PSP) were tested as model phosphoproteins. After protease digestion, the resulting phosphopeptides were successfully isolated by column-switching HPLC. The phosphopeptide fractions were analyzed by electrospray ionization mass spectrometry with a positive or negative ion mode after purification by reversed-phase HPLC. Pseudo-molecular ion peaks corresponding to Gln-Ile-Ser(p)-Val-Arg (MW 681.7) and Glu-Ala-Thr-Ser(p)-Gln-Glu-Leu (MW 856.8) were detected from the tryptic digest of RPa and chymotryptic digest of PSP, respectively, which agreed with the theoretically expected phosphopeptide fragments.

  16. Rapid optimization of the post-column fluorogenic ninhydrin reaction for the HPLC-based determination of bradykinin and related fragments.

    Science.gov (United States)

    Wimalasena, R; Audus, K L; Stobaugh, J F

    2003-01-01

    A flow injection analysis scheme is demonstrated for the rapid optimization of reagent concentrations, flow rates, delay time and temperature using the guanidino moiety specific fluorogenic ninhydrin reaction. Using the amino acid arginine, non-arginine containing peptides, and the arginine-containing peptides, bradykinin and related fragments, specificity is demonstrated. These results serve to extend previous descriptions of the post-column reaction by offering a time efficient approach for the optimization of newly assembled post-column reactors using this chemistry. The reactor is subsequently added to a gradient elution HPLC system with the separation of bradykinin and bradykinin fragments demonstrated. Detection sensitivity in the high femtomole-low picomole mass range was achieved for these substances. Copyright 2003 John Wiley & Sons, Ltd.

  17. [HPLC fingerprint of Calendula officinalis flower].

    Science.gov (United States)

    Xing, Zhan-Fen; Cheng, Hong-Da; Zhang, Ping-Ping; Gong, Lei; Ma, Li-Ya

    2014-07-01

    To establish an HPLC fingerprint of Calendula officinalis flower for its quality control. Hypersil ODS C18 column (250 mm x 4.6 mm, 5 μm) was used with acetonitrile and water as mobile phase in a gradient mode at the flow rate of 1.0 mL/min. The detection wavelength was 220 nm and the temperature of column was set at 35 degrees C. The similarity was analyzed with the Estimating System of Similarity on the Chinese Medicine Fingerprint Chromatogram. The HPLC fingerprint of Calendula officinalis flower containing eleven peaks was set up. The similarity of Calendula officinalis flower from different habitats was greater than 0.90. This method is easy and reliable, which can be used to judge the habitat and control the quality of Calendula officinalis flower.

  18. Development of a simple column-switching high-performance liquid chromatography (HPLC) method for rapid and simultaneous routine serum monitoring of lamotrigine, oxcarbazepine and 10-monohydroxycarbazepine (MHD).

    Science.gov (United States)

    Greiner, Christine; Haen, Ekkehard

    2007-07-01

    Using isocratic column-switching high-performance liquid chromatography (HPLC) we established a group method for automated quantitative analysis of the antiepileptic drugs lamotrigine, oxcarbazepine and its metabolite 10-monohydroxycarbazepine (MHD) that are also used in psychiatry as mood stabilizers. Samples were cleaned from interfering proteins and lipids by transfer onto a pre-column, using a PerfectBond C-8 material, with 8% acetonitrile in water as a pre-column eluent. Separation was performed by elution onto the analytical column (Betasil C6 5 microm, 250 mm x 4.6 mm) at a flow rate of 1.0 ml/min with potassium dihydrogenphosphate buffer (20 mmol/l, pH3.0)/acetonitrile (70/30; v/v) as analytical eluent. UV-spectrophotometric detection was set to 215 nm for all three compounds. The analytical run was finished within 18 min. Detection limit was 30 ng/ml for lamotrigine, 35 ng/ml for oxcarbazepine and 25 ng/ml for 10-monohydroxycarbazepine. The method was found to be suitable for automated analysis of serum samples of patients treated with lamotrigine and oxcarbazepine.

  19. Analysis of sterigmatocystin in cereals, animal feed, seeds, beer and cheese by immunoaffinity column clean-up and HPLC and LC-MS/MS quantification.

    Science.gov (United States)

    Marley, Elaine; Brown, Phyllis; Mackie, Jennifer; Donnelly, Carol; Wilcox, Joyce; Pietri, Amedeo; Macdonald, Susan

    2015-01-01

    A method is reported for the analysis of sterigmatocystin in various food and feed matrices using a commercial sterigmatocystin immunoaffinity column (IAC) for sample clean-up prior to HPLC analysis by UV with mass spectrometric detection (LC-MS/MS). Cereals (wheat, oats, rye, maize and rice), sunflower seeds and animal feed were spiked with sterigmatocystin at levels from 0.75 to 50 µg kg(-1) to establish method performance. Using acetonitrile/water extraction followed by IAC clean-up, and analysis by HPLC with detection at 325 nm, recoveries ranged from 68% to 106%, with repeatability from 4.2% to 17.5%. The limit of quantification with UV detection in these matrices was 1.5 µg kg(-1). For the analysis of beer and cheese the sample preparation prior to IAC clean-up was changed to accommodate the different properties of the matrix, prior to analysis by LC-MS/MS. For beer and cheese spiked at 5.0 µg kg(-1) the recoveries were 94% and 104%, and precision (RSDs) were 1.9% and 2.9% respectively. The limits of quantification by LC-MS/MS in beer and cheese were 0.02 and 0.6 µg kg(-1) respectively. The sterigmatocystin IAC was demonstrated to provide an efficient clean-up of various matrices to enable this mycotoxin to be determined by either HPLC with UV detection or LC-MS/MS.

  20. Application of an online post-column derivatization HPLC-DPPH assay to detect compounds responsible for antioxidant activity in Sonchus oleraceus L. leaf extracts.

    Science.gov (United States)

    Ou, Zong-Quan; Schmierer, David M; Rades, Thomas; Larsen, Lesley; McDowell, Arlene

    2013-02-01

    To use an online assay to identify key antioxidants in Sonchus oleraceus leaf extracts and to investigate the effect of leaf position and extraction conditions on antioxidant concentration and activity. Separation of phytochemicals and simultaneous assessment of antioxidant activity were performed online using HPLC and post-column reaction with a free-radical reagent (2, 2-diphenylpicrylhydrazyl, DPPH). Active compounds were identified using nuclear magnetic resonance spectroscopy and mass spectrometry. We applied the online HPLC-DPPH radical assay to evaluate antioxidants in leaves from different positions on the plant and to assess the effect of pre-treatment of leaves with liquid N(2) before grinding, extraction time, extraction temperature and method of concentrating extracts. Key antioxidants identified in S. oleraceus leaf extracts were caftaric acid, chlorogenic acid and chicoric acid. Middle leaves contained the highest total amount of the three key antioxidant compounds, consisting mainly of chicoric acid. Pre-treatment with liquid N(2), increasing the extraction temperature and time and freeze-drying the extract did not enhance the yield of the key antioxidants. The online HPLC-DPPH radical assay was validated as a useful screening tool for investigating individual antioxidants in leaf extracts. Optimized extraction conditions were middle leaves pre-treated with liquid N(2), extraction at 25°C for 0.5 h and solvent removal by rotary evaporation. © 2012 The Authors. JPP © 2012. Royal Pharmaceutical Society.

  1. Analysis of amino acid composition in proteins of animal tissues and foods as pre-column o-phthaldialdehyde derivatives by HPLC with fluorescence detection.

    Science.gov (United States)

    Dai, Zhaolai; Wu, Zhenlong; Jia, Sichao; Wu, Guoyao

    2014-08-01

    Studies of protein nutrition and biochemistry require reliable methods for analysis of amino acid (AA) composition in polypeptides of animal tissues and foods. Proteins are hydrolyzed by 6M HCl (110°C for 24h), 4.2M NaOH (105°C for 20 h), or proteases. Analytical techniques that require high-performance liquid chromatography (HPLC) include pre-column derivatization with 4-chloro-7-nitrobenzofurazan, 9-fluorenyl methylchloroformate, phenylisothiocyanate, naphthalene-2,3-dicarboxaldehyde, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, and o-phthaldialdehyde (OPA). OPA reacts with primary AA (except cysteine or cystine) in the presence of 2-mercaptoethanol or 3-mercaptopropionic acid to form a highly fluorescent adduct. OPA also reacts with 4-amino-1-butanol and 4-aminobutane-1,3-diol produced from oxidation of proline and 4-hydroxyproline, respectively, in the presence of chloramine-T plus sodium borohydride at 60°C, or with S-carboxymethyl-cysteine formed from cysteine and iodoacetic acid at 25°C. Fluorescence of OPA derivatives is monitored at excitation and emission wavelengths of 340 and 455 nm, respectively. Detection limits are 50 fmol for AA. This technique offers the following advantages: simple procedures for preparation of samples, reagents, and mobile-phase solutions; rapid pre-column formation of OPA-AA derivatives and their efficient separation at room temperature (e.g., 20-25°C); high sensitivity of detection; easy automation on the HPLC apparatus; few interfering side reactions; a stable chromatography baseline for accurate integration of peak areas; and rapid regeneration of guard and analytical columns. Thus, the OPA method provides a useful tool to determine AA composition in proteins of animal tissues (e.g., skeletal muscle, liver, intestine, placenta, brain, and body homogenates) and foods (e.g., milk, corn grain, meat, and soybean meal). Copyright © 2014 Elsevier B.V. All rights reserved.

  2. A Validated Stability-Indicating HPLC Method for Routine Analysis of an Injectable Lincomycin and Spectinomycin Formulation

    OpenAIRE

    Abualhasan,Murad; Batrawi, Nidal; SUTCLIFFE, Oliver; ZAID, Abdel

    2012-01-01

    Lincomycin and spectinomycin combination therapy is widely used in veterinary medicine for the treatment of gastrointestinal and respiratory infections caused by lincomycin- and spectinomycin-sensitive microorganisms. A simple, reverse phase HPLC method for the analysis of samples of an injectable lincomycin and spectinomycin preparation containing a mixture of inactive excipients has been developed. The HPLC was carried out using the RP-C18 (250 mm × 4.0 mm, 5 μm) column, with the gradient m...

  3. [HPLC fingerprint of ethyl acetate extraction of Saxifraga stolonifera].

    Science.gov (United States)

    Zhou, Mei; Chen, Hua-Guo; Xian, Chun; Huang, Zhi-Jin; Zhou, Xin

    2013-04-01

    To establish an HPLC fingerprint of ethyl acetate extraction of Saxifraga stolonifera. The HPLC analysis was performed on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column with isocratic elution of acetonitrile-0.05% phosphoric acid at a flow rate of 1.0 mL x min(-1). The detection wavelength was set at 256 nm and the column temperature was set at 30 degrees C. The HPLC fingerprint of ethyl acetate extraction of S. stolonifera has been established. There were fifteen common peaks, seven of which were identified by reference substances. The RSD of relative retention time was less than 3% in the precision and repeated tests. Eleven samples from different area can be distinguished from their fingerprints. This method is reasonable and reliable and can be used for quality control of S. stolonifera.

  4. Caution to HPLC analysis of tricarbonyl technetium radiopharmaceuticals: an example of changing constitution of complexes in column.

    Science.gov (United States)

    Chen, Xiangji; Guo, Yunhang; Liu, Boli

    2007-03-12

    Radio-HPLC is a powerful tool for analyzing radioactive species in radiopharmaceutical chemistry. In this paper, we found an example that the commonly used eluting solvent, acetonitrile, could coordinate with the popular radiopharmaceutical nuclides, technetium-99m, during chromatography. [(99m)Tc(CO)(3)(H(2)O)(3)](+) and [Re(CO)(3)(H(2)O)(3)](+) showed quite different retention time when they were eluted using acetonitrile/water as mobile phase. However, they almost demonstrated the same retention time when they were eluted using methanol/water as mobile phase. Further analysis showed that both [(99m)Tc(CO)(3)(H(2)O)(3)](+) and [Re(CO)(3)(H(2)O)(3)](+) could be changed into [(99m)Tc(CO)(3)(CH(3)CN)(3)](+) and [Re(CO)(3)(CH(3)CN)(x)(H(2)O)(3-x)](+) during the separation, respectively. Some former works mistook the [(99m)Tc(CO)(3)(CH(3)CN)(3)](+) for [(99m)Tc(CO)(3)(H(2)O)(3)](+) when using acetonitrile and water in analysis. Quality control of the radiopharmaceuticals containing metal complex should be careful since HPLC solvent could replace some liable ligand molecules.

  5. Identification and quantification of genipin and geniposide from Genipa americana L. by HPLC-DAD using a fused-core column

    Directory of Open Access Journals (Sweden)

    Grazielle NÁTHIA-NEVES

    2018-01-01

    Full Text Available Abstract In this work, it was developed a fast, simple and selective method for quantification of genipin and geniposide from unripe fruits of genipap, which are known as natural colorants, blue and yellow, respectively. The compounds separation was performed in a fused-core C18 column using as mobile phase water (A and acetonitrile (B both acidified with 0.1% formic acid, with the following gradient: 0 min, 99% A; 9 min, 75% A; 10 min, 99% A and 13 min, 99% A. The temperature and flow rate that allowed the best chromatographic performance were 35 °C and 1.5 mL/min, respectively, resulting a total run time of 13 min, including column clean-up and re-equilibration. This short analysis time represents an advantage compared to the methods reported in the literature where the running times are 2-5 times greater. The detection wavelength was set at 240 nm. The method validation was performed based on specificity, linearity, detection and quantification limits, precision and accuracy, according to ICH methodology. Finally, the developed method was suitable for monitoring analysis of those compounds content in vegetable samples.

  6. Evaluating the interactions of organic compounds with multi-walled carbon nanotubes by self-packed HPLC column and linear solvation energy relationship.

    Science.gov (United States)

    Chu, Yingqian; Li, Xuehua; Xie, Hongbin; Fu, Zhiqiang; Yang, Xianhai; Qiao, Xianliang; Cai, Xiyun; Chen, Jingwen

    2013-12-15

    Understanding the interactions between organic pollutants and carbon nanotubes (CNTs) is critical for fate assessment of both CNTs and organic pollutants. In this study, the chromatographic approach was introduced based on CNTs as stationary phase for the evaluation of such interactions. The pristine multi-walled carbon nanotubes (MWCNTs) were packed into columns of high-pressure liquid chromatography (HPLC) and the retention factors (k') were determined to characterize the adsorption affinity of organic compounds onto MWCNTs. Nine compounds were tested. The results showed that their lnk' values followed the order: benzene compounds correlate positively with molecular polarizability (E) significantly, suggesting that the π-/n-electrons-dependent polarizable interactions play a major role for the adsorption. Moreover, the thermodynamic parameters calculated from van't Hoff equations revealed that the interactions between the compounds and MWCNTs were spontaneous and exothermic processes. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Bamboo charcoal as adsorbent for SPE coupled with monolithic column-HPLC for rapid determination of 16 polycyclic aromatic hydrocarbons in water samples.

    Science.gov (United States)

    Ma, Jiping; Li, Mo; Li, Jinhua; Rui, Cuijie; Xin, Yanping; Xue, Qinzhao; Chen, Lingxin

    2011-10-01

    The coupling of solid-phase extraction (SPE) using bamboo charcoal (BC) as an adsorbent with a monolithic column-high performance liquid chromatography (MC-HPLC) method was developed for the high-efficiency enrichment and rapid determination of 16 polycyclic aromatic hydrocarbons (PAHs) in water. Key influence factors, such as the type and the volume of the elution solvent, and the flow rate and the volume of the sample loading, were optimized to obtain a high SPE recovery and extraction efficiency. BC as an SPE adsorbent presented a high extraction efficiency due to its large specific surface area and high adsorption capacity; MC as an HPLC column accelerated the separation within 8 min because of its high porosity, fast mass transfer, and low-pressure resistance. The calibration curves for the PAHs extracted were linear in the range of 0.2-15 µg/L, with the correlation coefficients (r(2)) between 0.9970-0.9999. This method attained good precisions (relative standard deviation, RSD) from 3.5 to 10.9% for the standard PAHs I aqueous solutions at 5 µg/L; the method recoveries ranged in 52.6-121.6% for real spiked river water samples with 0.4 and 4 µg/L. The limits of detection (LODs, S/N = 3) of the method were determined from 11 and 87 ng/L. The developed method was demonstrated to be applicable for the rapid and sensitive determination of 16 PAHs in real environmental water samples.

  8. Evaluating the interactions of organic compounds with multi-walled carbon nanotubes by self-packed HPLC column and linear solvation energy relationship

    Energy Technology Data Exchange (ETDEWEB)

    Chu, Yingqian; Li, Xuehua, E-mail: lixuehua@dlut.edu.cn; Xie, Hongbin; Fu, Zhiqiang; Yang, Xianhai; Qiao, Xianliang; Cai, Xiyun; Chen, Jingwen

    2013-12-15

    Highlights: • The HPLC-based approach using CNTs as stationary phase is introduced. • The interactions of nine compounds to MWCNTs are evaluated by retention factors. • The LSER theory is adopted to probe the interactions of the compounds with MWCNTs. • π-/n-Electrons-dependent polarizable interaction plays a key role for adsorption. -- Abstract: Understanding the interactions between organic pollutants and carbon nanotubes (CNTs) is critical for fate assessment of both CNTs and organic pollutants. In this study, the chromatographic approach was introduced based on CNTs as stationary phase for the evaluation of such interactions. The pristine multi-walled carbon nanotubes (MWCNTs) were packed into columns of high-pressure liquid chromatography (HPLC) and the retention factors (k′) were determined to characterize the adsorption affinity of organic compounds onto MWCNTs. Nine compounds were tested. The results showed that their ln k′ values followed the order: benzene < toluene < phenol < chlorobenzene < bromobenzene < aniline < sulfamethoxazole < sulfadiazine ≈ sulfadimidine. The linear solvation energy relationship (LSER) theory was adopted to correlate ln k′ with the molecular solvatochromic parameters. We found that ln k′ of the studied compounds correlate positively with molecular polarizability (E) significantly, suggesting that the π-/n-electrons-dependent polarizable interactions play a major role for the adsorption. Moreover, the thermodynamic parameters calculated from van’t Hoff equations revealed that the interactions between the compounds and MWCNTs were spontaneous and exothermic processes.

  9. A simple and sensitive HPLC method based on pre-column fluorescence labelling for multiple classes of plant growth regulator determination in food samples.

    Science.gov (United States)

    Li, Guoliang; Liu, Shucheng; Sun, Zhiwei; Xia, Lian; Chen, Guang; You, Jinmao

    2015-03-01

    The determination of trace plant growth regulator (PGR) has received more and more attentions in the field of phytophysiology and food safety. But the simple and sensitive method for simultaneously analysing multiple classes of PGR remains poorly investigated. In this study, a new pre-column fluorescence labelling method using 2-(11H-benzo[a]carbazol-11-yl)-ethyl-4-methylbenzenesulfonate (BCETS) as the labelling reagent has been developed for simultaneous determination of seven PGRs (i.e., indole-3-acetic acid, 3-indolybutyric acid, 3-indolepropionic acid, jasmonic acid, gibberellin A3, 1-naphthylacetic acid and 2-naphthaleneacetic acid) by HPLC with fluorescent detection (FLD). The proposed method offered the LOD of 0.34-0.73 ng/mL for seven PGRs, which were significantly lower than the reported methods. The crude extract without complex pre-treatments and purification was directly labelled by BCETS and analysed by HPLC-FLD, which facilitates the high-throughput sample screening. This method was proven to be inexpensive, simple, selective, sensitive, accurate and reliable for trace PGR determination. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. A nitromethane-based HPLC system alternative to acetonitrile for carotenoid analysis of fruit and vegetables.

    Science.gov (United States)

    Sandmann, Gerhard

    2010-01-01

    Acetonitrile-based HPLC systems are the most commonly used for carotenoid analysis from different plant tissues. Because of the acetonitrile shortage, an HPLC system for the separation of carotenoids on C(18) reversed-phase columns was developed in which an acetonitrile-alcohol-based mobile phase was replaced by nitromethane. This solvent comes closest to acetonitrile with respect to its elutrophic property. Our criterion was to obtain similar separation and retention times for a range of differently structured carotenoids. This was achieved by further increase in the lipophilicity with ethylacetate. For all the carotenoids which we tested, we found co-elution only of β-cryptoxanthin and lycopene. By addition of 1% of water, separation of this pair of carotenoids was also achieved. The final recommended mobile phase consisted of nitromethane : 2-propanol : ethyl acetate : water (79 : 10 : 10 : 1, by volume). On Nucleosil C(18) columns and related ones like Hypersil C(18), we obtained separation of carotenes, hydroxyl, epoxy and keto derivatives, which resembles the excellent separation properties of acetonitrile-based mobile phases on C(18) reversed phase columns. We successfully applied the newly developed HPLC system to the separation of carotenoids from different vegetables and fruit. Copyright © 2010 John Wiley & Sons, Ltd.

  11. Standardization of RP-HPLC methods for the detection of the major peanut allergens Ara h 1, Ara h 2 and Ara h 3

    Science.gov (United States)

    Crude peanut extract (CPE) was analyzed for three major allergens (Ara h 1, h 2, and h 3) using a C12 and a C18 column at two wavelengths (280 and 220 nm) and under different solvent conditions. HPLC profiles were compared for retention time, resolution, and peak heights. CPE samples were spiked wit...

  12. Determination for multiple mycotoxins in agricultural products using HPLC-MS/MS via a multiple antibody immunoaffinity column.

    Science.gov (United States)

    Zhang, Zhaowei; Hu, Xiaofeng; Zhang, Qi; Li, Peiwu

    2016-05-15

    Mycotoxins usually found in agricultural products such as peanut, corn, and wheat, are a serious threat to human health and their detection requires multiplexed and sensitive analysis methods. Herein, a simultaneous determination for aflatoxin B1, B2, G1, G2, ochratoxin A, zearalanone and T-2 toxin was investigated using high performance liquid chromatography coupled with tandem mass spectrometry in a single run via a home-made multiple immunoaffinity column. Four monoclonal antibodies were produced in our lab against aflatoxins, ochratoxin A, zearalanone and T-2 toxin, respectively, then combined as a pool and bound to Sepharose-4B for affinity chromatography. Seven mycotoxins were effectively extracted from the agricultural product samples by using acetonitrile/water/acetic acid (80:19:1, v/v/v) Then, the extraction was cleanup by multiple immunoaffinity column. This method demonstrated a considerable linear range of 0.30-25, 0.12-20, 0.30-20, 0.12-20, 0.60-30, 0.30-25, and 1.2-40μgkg(-1)and lower limits of detection at 0.1, 0.04, 0.1, 0.04, 0.2, 0.1 and 0.4μgkg(-1) for AFB1, AFB2, AFG1, AFG2, OTA, ZEN and T-2, respectively, in comparison with previously reported methods, as well as excellent recoveries. The mIAC capacity for AFB1, AFB2, AFG1, AFG2, OTA, ZEN, and T-2 were 187, 181, 153, 151, 105, 130, 88ng, respectively. It was found that all of the 7 mycotoxins were present in 90 agricultural product samples. The proposed method meets the requirements for rapid sample preparation and highly sensitive identification of multiple mycotoxins in agricultural product and food safety. This method provides a promising alternative with high throughput and high sensitivity for rapid analysis of seven mycotoxins in the monitoring of food safety. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Determination of fluoxetine and norfluoxetine in rat plasma by HPLC with pre-column derivatization and fluorescence detection.

    Science.gov (United States)

    Guo, Xingjie; Fukushima, Takeshi; Li, Famei; Imai, Kazuhiro

    2003-01-01

    Both fluoxetine (FLX) and its N-demethylated metabolite, norfluoxetine (NFLX), have been reported to be potent serotonin-reuptake inhibitors. A sensitive and reliable method that allows simultaneous quantification of their plasma levels would be valuable and was developed in this work. The procedure included extraction of FLX and NFLX from plasma, fluorescence derivatization with 4-(N-chloroformylmethyl-N-methyl) amino-7-nitro-2,1,3-benzoxadiazole (NBD-COCl), separation of the derivatives on an octadecylsilica column with acetonitrile-water (55:45,v/v) as mobile phase and fluorescence detection with emission at 537 nm and excitation at 478 nm. The calibration curves were linear for FLX and NFLX concentration over the range of 10-1000 nM (r = 0.9992 and r = 0.9997) and the limits of quantitation were 10 nM in 100 micro L of plasma. Precision of intra- and inter-day RSD of less than 12% and accuracy of intra- and inter-day RE within -6.0-13% were achieved. The method described was applied to analysis of the plasma samples from rats treated with FLX hydrochloride and to the pharmacokinetic study. Copyright 2002 John Wiley & Sons, Ltd.

  14. Simultaneous determination of four aflatoxins and ochratoxin A in ginger and related products by HPLC with fluorescence detection after immunoaffinity column clean-up and postcolumn photochemical derivatization.

    Science.gov (United States)

    Wen, Jing; Kong, Weijun; Wang, Jian; Yang, Meihua

    2013-12-01

    Ginger, a widely used spice and traditional Chinese medicine, is prone to be contaminated by mycotoxins. A simple, sensitive, and reproducible method based on immunoaffinity column clean-up coupled with HPLC and on-line postcolumn photochemical derivatization with fluorescence detection was developed for the simultaneous determination of aflatoxins (AFs) B1 , B2 , G1 , G2 , and ochratoxin A (OTA) in 25 batches of gingers and related products marketed in China for the first time. The samples were first extracted by ultrasonication with methanol/water (80:20, v/v) and then cleaned up with immunoaffinity columns for analysis. Under the optimized conditions, the LODs and LOQs for the five mycotoxins were 0.03-0.3 and 0.1-0.9 μg/kg, respectively. The average recoveries ranged from 81.3-100.8% for AFs and from 88.6-99.5% for OTA at three spiking levels. Good linearity was observed for the analytes with correlation coefficients all >0.9995. All moldy gingers were contaminated with at least one kind of the five investigated mycotoxins, while none of them were found in normal gingers. Ginger powder samples were contaminated slightly with the contamination levels below the LOQs, while ginger tea bags were mainly contaminated by OTA at 1.05-1.19 μg/kg and ginger black tea bags were mainly contaminated by AFs at 3.37-5.76 μg/kg. All the contamination levels were below the legally allowable limits. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. [RP-HPLC characteristics of dragon's blood].

    Science.gov (United States)

    Gao, Xiu-Li; Jiang, Qian; Wang, Peng-Jiao; Zhang, Min

    2007-10-01

    To study the fingerprint of dragon's blood resina draconis by high performance liquid chromatography. The samples are extracted with methanol and separated on a Eclipse XDB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase of acetonitrile-H2O in gradient mode, and the flow rate was 1.0 mL x min(-1), the detection wavelength was 275 nm and the temperature of column was 40 degrees C. Loureirin B was used as the reference compound. HPLC fingerprint of dragon's blood was established and the similarity of the fingerprint was compared. The method is simple, accurate, and can be used to control the quality of dragon's blood.

  16. Validated HPLC-PDA method for rosmarinic acid quantification in Rosemary

    OpenAIRE

    Bara, María Teresa. F.; Rezende, Kênnia R.; Alves,Suzana F.; Oliveira, Ezequiane M. S.; Chaul, Luiza T.; Conceição, Edmilson C. da; Couto, Renê O.; Paula, José R.

    2011-01-01

    A fast and simple HPLC-PDA method for rosmarinic acid quantification in Rosemary leaves powder was validated. The analyses were performed using a C18 column in isocratic conditions and detection at 254 nm. Performing the system suitability tests, the method showed to be capable of providing data of acceptable quality. Results showed that the method was selective, linear for concentration ranges between 2.5-50 μg.mL–1 , sensitive, precise, accurate and robust. The main advantages of...

  17. Reversed-phase HPLC retention data in correlation studies with in lipophilicity molecular descriptors of carotenoids

    OpenAIRE

    Podunavac-Kuzmanović Sanja O.; Jevrić Lidija R.; Tepić Aleksandra N.; Šumić Zdravko

    2013-01-01

    Influence of chemical structure on the lipophilicity of isolated free carotenoids from paprika oleoresin has been studied by QSRR approach (Quantitative structure-retention relationship). The chromatographic behavior of these compounds was investigated by using reversed phase high-pressure liquid chromatography (RP HPLC). Retention mechanism has been determined using the following mobile phase: acetone -water, on reversed - phase column (SB-C18). A variety ...

  18. RP-HPLC Determination of Raloxifene in Pharmaceuticl Tablets

    Directory of Open Access Journals (Sweden)

    P. Venkata Reddy

    2006-01-01

    Full Text Available A reverse phase HPLC method is developed for the determination of Raloxifene in pharmaceutical dosage forms. Chromatography was carried out on an inertsil C18 column using a mixture of acetonitrile and phosphate buffer (30:70 v/v as the mobile phase at a flow rate of 1 mL/min. Detection was carried out at 290 nm .The retention time of the drug was 10.609 min. The method produced linear responses in the concentration range of 0.5-200 µg/mL of Raloxifene. The method was found to be applicable for determination of the drug in tablets.

  19. Quantitative analysis by HPLC-MS2 of the pyrrolizidine alkaloid adonifoline in Senecio scandens.

    Science.gov (United States)

    Zhang, Fang; Wang, Chang-Hong; Wang, Wan; Chen, Lu-Xin; Ma, Hong-Yan; Zhang, Chao-Feng; Zhang, Mian; Bligh, S W Annie; Wang, Zheng-Tao

    2008-01-01

    A quantitative method using HPLC-MS(2) has been developed for the determination of adonifoline, one of the retronecine-type hepatotoxic pyrrolizidine alkaloids in Senecio scandens Buch.-Ham. ex D. Don., a traditional Chinese herb. Using an orthogonal design test, a simple and rapid sample extraction method was developed. HPLC analysis was conducted using a C(18) column as stationary phase and a mixture of acetonitrile and aqueous formic acid as mobile phase. Good linearity for adonifoline was found in the concentration range 0.12-4.18 microg/mL, and the HPLC-MS/MS method was shown to be appropriate, in terms of sensitivity, precision and reproducibility. The quantities of adonifoline in extracts of 18 plant samples from different collection sources and from different parts (flowers, leaves, thick stems, slim stems and roots) of S. scandens were determined using the newly developed HPLC/MS(2) analysis. (c) 2007 John Wiley & Sons, Ltd.

  20. Determination of Arctiin and Arctigenin Contents in Arctium Tomentosum Mill. by HPLC Method

    OpenAIRE

    Xiaoying Zhou; Haoke Zhang; Liang Ge; Haiyan Gong; Shuge Tian

    2011-01-01

    A simple, precise, rapid and accurate, binary-phase high performance liquid chromatographic method has been developed for the determination of arctiin and arctigenin contents in the Arctium tomentosumMill. with short run time. Chromatographic separation was achieved by using HPLC system, consisting of a Shimadzu LC-6AD and Kromasil C18 column (250×4.6 mm, 5 μm, with pre-column), the mobile phase consists of methanol and water (55: 45). Detection wavelength was 280 nm. The speed of flow was 1....

  1. A high-temperature liquid chromatographic reactor approach for investigating the solvolytic stability of a pharmaceutical compound and an investigation of its retention behavior on a C18-modified zirconia stationary phase.

    Science.gov (United States)

    Skrdla, Peter J; Bopra, Angela; Chasse, Tyson; Wang, Tao

    2008-06-09

    The solvolysis kinetics of a developmental active pharmaceutical ingredient (API) were investigated using a high temperature (HT)-HPLC reactor approach to determine whether it might be possible to use the technique to efficiently screen the relative stabilities of typical APIs (particularly those that are stable at the column temperatures achievable on most HPLC systems and over durations of less than 60 min-a reasonable upper limit for typical method run time). It was discovered that the on-column API degradation kinetics better obeyed a second-order model than a first-order one. Employing a newly developed mathematical treatment, the apparent activation energy for the process was determined to be 85.7+/-1.6 kJ/mol; the apparent frequency factor was found to be (3.9+/-0.4)x10(4) s(-1). The retention mechanism of the API on the C18-modified zirconia column (ZirChrom) Diamondbond-C18) was investigated using a van't Hoff analysis. It was discovered that the logarithm of the retention factor (following correction for the gradient elution of the assay method) exhibited a quadratic dependence on the reciprocal of the absolute temperature. While the retention was found to be predominantly enthalpically driven over the majority of temperatures investigated in this study (ranging from 40 to 200 degrees C), a regression fit of the curve predicted a maximum at approximately 20 degrees C, indicative of a transition from predominantly enthalpically controlled retention to a mainly entropically driven mechanism. A table summarizing the thermodynamic retention parameters at each experimental column temperature is provided. Finally, the preliminary application of the HT-HPLC reactor approach to the study of degradation kinetics of other APIs is discussed in terms of some unexpected findings obtained using the zirconia column.

  2. Multi residue method using coupled-column HPLC and GC-MS for the determination of anabolic compounds in samples of urine

    NARCIS (Netherlands)

    Herbold HA; Sterk SS; Stephany RW; Ginkel LA van; ARO

    1998-01-01

    Dit rapport beschrijft een multiresidu methode voor de analyse, detectie en identificatie van residuen van anabolica. De analytische methode is gebaseerd op een geautomatiseerde vaste faseextractie en een High Performance Liquid Chromatography (HPLC) kolomschakelingssysteem. Detectie vindt

  3. [HPLC fingerprinting of total glycosides of Swertia franchetiana].

    Science.gov (United States)

    Tian, Wei; Chen, Zhao-hui; Zhai, Jing; Chen, Li-ren; Li, Yong-min

    2005-05-01

    To establish a sensitive and specific HPLC method for controlling the quality of total glycosides from Swertia franchetiana H. Smith. HPLC method was applied for quality and quantitative assessment of the pharmaceutical extracts from Swertia franchetiana H. Smith. The preparation of sample, the HPLC column, mobile phase, elution mode (isocratic or gradient) and gradient program were optimized in order to obtain HPLC profile. The HPLC system consisted of a SPD-1OAvp pump, SPD-M1OAVP photodiode-array detector (PAD), SIL-10ADVP auto injector. Data were acquired and processed with the CLASS-VP6.1 workstation. HPLC analysis was performed on a Kromasil C18 column (250 mm x 4. 6 mm ID, 5 microm) with methanol and water as mobile phase. The column temperature was set up at 40 degrees C and the flow-rate was 1 mL x min(-1). The reference solution of chemical standards and sample were injected into HPLC system, separately. The HPLC chromatographic fingerprinting of the total glycosides, showing 16 characteristic peaks which were partitioned into three parts: one peak in 0-10 min of retention time, nine peaks containing main 1-7 peaks in 10-15 min of retention time, 6 peaks in 15-30 min of retention time, was established from 10 lots of their products. By comparison of the retention time and the on-line UV spectra and their molecule weights of chemical standards, peak 1-7 were identified as swertiamarin (1), gentiopicroside (2), sweroside (3), isoorientin (4), swertisin (5), isoswertisin (6) and swetianolin (7), respectively. The ratios of peak area between 1-16 were in their extent. Moreover, comparison of the HPLC profiles of the total glycosides, the extracts prepared using another process and the plant indicated that they were closely related to each other. The HPLC profiles and quantitative assessment of the total glycosides from Swertia franchetiana H. Smith with high specificity can be used to control their quality and assure lot to lot consistency.

  4. [Studies on fingerprinting of Flos Buddleja by RP-HPLC].

    Science.gov (United States)

    Han, Peng; Cui, Ya-jun; Guo, Hong-zhu; Guo, De-an

    2004-10-01

    To establish fingerprinting of Flos Buddleja by using RP-HPLC for the quality control. The HPLC condition was as follows: Inertsil ODS-3 C18 analytical column (4.6 mm x 250 mm, 5 microm), gredient eluation with MeCN (0.1% TFA)-H2O (0.1%TFA), flow rate 1.0 mL x min(-1), detection wavelength 254 nm. 10 commercial samples were analyzed to establish a fingerprinting. Among the obtained fingerprinting, most of the detected peaks were separated effectively. The accuracy, repeatability and stability of this method were satisfied. The RSDs of relative retention time and area of aimed peaks which existed in all samples wereless than 5%. Theresults were in accordance with the request of fingerprinting. The established fingerprinting can be used for the quality control of Flos Buddleja.

  5. HPLC analysis of neo-clerodane diterpenoids from Teucrium chamaedrys.

    Science.gov (United States)

    Avula, B; Manyam, R B; Bedir, E; Khan, I A

    2003-07-01

    A simple, rapid analytical method for the quantitative determination of nine neo-clerodane diterpenoids was developed. The neo-clerodane diterpenoids present in the plant material and extracts were separated with an acetonitrile-water gradient at a flow rate of 1 mL per minute. The HPLC separation was performed on a Phenomenex Luna C18(2) (150 x 4.6 mm I.D., particle size 5 microm) reversed phase column with detection at 220 nm. The limit of detection was 0.24-0.90 microg/mL. The relative standard deviation (RSD) values for the determination of neo-clerodane diterpenoids in plant extracts were less than 3.20%. This is the first analytical method developed for qualitative and quantitative analysis of nine neo-clerodane diterpenoids by HPLC with PDA detection.

  6. Retention Models on Core-Shell Columns.

    Science.gov (United States)

    Jandera, Pavel; Hájek, Tomáš; Růžičková, Marie

    2017-07-13

    A thin, active shell layer on core-shell columns provides high efficiency in HPLC at moderately high pressures. We revisited three models of mobile phase effects on retention for core-shell columns in mixed aqueous-organic mobile phases: linear solvent strength and Snyder-Soczewiński two-parameter models and a three-parameter model. For some compounds, two-parameter models show minor deviations from linearity due to neglect of possible minor retention in pure weak solvent, which is compensated for in the three-parameter model, which does not explicitly assume either the adsorption or the partition retention mechanism in normal- or reversed-phase systems. The model retention equation can be formulated as a function of solute retention factors of nonionic compounds in pure organic solvent and in pure water (or aqueous buffer) and of the volume fraction of an either aqueous or organic solvent component in a two-component mobile phase. With core-shell columns, the impervious solid core does not participate in the retention process. Hence, the thermodynamic retention factors, defined as the ratio of the mass of the analyte mass contained in the stationary phase to its mass in the mobile phase in the column, should not include the particle core volume. The values of the thermodynamic factors are lower than the retention factors determined using a convention including the inert core in the stationary phase. However, both conventions produce correct results if consistently used to predict the effects of changing mobile phase composition on retention. We compared three types of core-shell columns with C18-, phenyl-hexyl-, and biphenyl-bonded phases. The core-shell columns with phenyl-hexyl- and biphenyl-bonded ligands provided lower errors in two-parameter model predictions for alkylbenzenes, phenolic acids, and flavonoid compounds in comparison with C18-bonded ligands.

  7. Determination of 7 Kinds of Sulfa Drugs in a Pork Sample by HPLC Using a Biphenyl Core-shell Stationary-phase Column

    National Research Council Canada - National Science Library

    Yoshimi ISHIHARA; Hirotaka SUGITA; Jiro TAKANO; Hideaki KITAMI

    2016-01-01

    ...) with an ultraviolet detector (UV) on a biphenyl core-shell stationary phase column was developed. This analytical method provides high linearity of the calibration curve as well as repeatability...

  8. Development and validation of a RP-HPLC method for the determination of gentamicin sulfate and its related substances in a pharmaceutical cream using a short pentafluorophenyl column and a charged aerosol detector.

    Science.gov (United States)

    Joseph, Arul; Rustum, Abu

    2010-02-05

    Gentamicin sulfate is a potent broad spectrum aminoglycoside antibiotic which is used as an active pharmaceutical ingredient (API) against both Gram-positive and Gram-negative bacteria. A reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated to determine the composition of gentamicin sulfate and to estimate its related substances (without any pre- or post-column derivatization) in a pharmaceutical cream. As gentamicin has a weak UV chromophore, it is not possible to detect low levels of known and unknown related substances of gentamicin using a UV detector. In this method, a Charged Aerosol Detector (CAD) was used to obtain high sensitivity that was necessary for the intended purpose of the method. This method can separate all the analogues of gentamicin including all known and unknown related substances of the API. A short (5cmx4.6mm) pentafluorophenyl HPLC column from Restek (Allure PFP) was used with an ion-pair gradient mobile phase consisting of (A) heptafluorobutyric acid:water:acetonitrile (0.025:95:5, v/v/v) and (B) trifluoroacetic acid:water:acetonitrile (1:95:5, v/v/v).

  9. Determination of spectinomycin hydrochloride and its related substances by HPLC-ELSD and HPLC-MSn.

    Science.gov (United States)

    Wang, Jian; Hu, Xiaojun; Tu, Ying; Ni, Kunyi

    2006-04-13

    A new and simple high-performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) method for the determination of spectinomycin hydrochloride and its related substances was developed. The column was Agilent SB-C(18) (250 mm x 4.6 mm, 5 microm). The mobile phase was 25 mM trifluoroacetic acid. The drift tube temperature was 40 degrees C. The pressure of nebulizing gas was 3.5 bar. Good separation of spectinomycin from main related substances could be achieved. The standard curve was rectilinear in the range of 0.07-3.8 mg/ml (r = 0.9997). Precision was 1.0% (R.S.D.). The limit of detection was 6 microg/ml. The method is simple and rapid, and the results are accurate and reproducible. The HPLC-MS(n) method was used to characterize the structures of impurities contained in the spectinomycin. In positive mode, impurities were elucidated by use of electrospray ion trap mass spectrometry in the multi-stage MS full scan mode. The possible structures of impurities C and D in spectinomycin were deduced based on the HPLC-MS(n) data.

  10. Determination of free salicylic acid in chewing aspirin tablets by HPLC.

    Science.gov (United States)

    Tian, Jun; Chen, Xin-shan; Wang, Rui-dong

    2003-07-01

    To establish a HPLC method for determining the content of free salicylic acid in chewing aspirin tablets. The determination was conducted on a HPLC column (C(18), 150 mm x 4.6 mm x 5 microm) with methanol-water-glacial acetic acid (8.0 5.5 1.0) as the mobile phase and the detection wavelength of 302 nm. The calibration curve was linear within the concentration range of 2.65 to 31.77 microg/ml (r=0.999 97) of salicylic acid. The average recovery rate was 100.21% with relative standard deviation of 0.53% (n=6). HPLC is quick and accurate of determining the content of free salicylic acid for chewing aspirin tablets.

  11. [Comparative study on HPLC fingerprint of ordinary powder and ultrafine powder for Gardenia jasminoides f. longicarpa].

    Science.gov (United States)

    Ju, Ai-Hua; Zhou, Kai; Zhang, Jing; Cai, Li-Juan; Song, Ping-Ping

    2013-07-01

    To establish an HPLC fingerprint of Gardenia jasminoides f. longicarpa and compare the differences between its ordinary powder and ultrafine powder. The analysis was carried out on a Kromasil C18 (250 mm x 4.6 mm, 5 microm) column with gradient elution of acetonitrile-0.4% phosphoric acid at the flow rate of 1.0 ml/min. The wavelength was 240 nm during 0 - 40 min and 440 nm during 40 - 80 min. HPLC fingerprint of Gardenia jasminoides f. longicarpa was established, 23 common peaks were identified,and the similarity of 10 samples was greater than 0.9. Ultrafine grinding did not change the types and number of chemical compositions, but it obviously increased the content of main chemical compositions. The HPLC fingerprint is accurate, reliable and repeatable, which can be used for quality control of Gardenia jasminoides f. longicarpa. Ultrafine grinding can stimulate the release of chemical components of Gardenia jasminoides f. longicarpa.

  12. Simultaneous separation of polar and non-polar mixtures by capillary HPLC based on an ostadecylsilane and taurine derivatized silica continuously packed column.

    Science.gov (United States)

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

    2016-12-01

    A capillary column was prepared by continuously packing ostadecylsilane (ODS) and taurine derivatized silica (TDS) in one column without interface. This continuously packing chromatography (CPC) column is easy to operate, has good stability and shows simultaneously separation of both polar and non-polar compounds. The simultaneous separation of a series of complex samples with highly hydrophobic components (benzene, toluene, ethylbenzene, and PAHs) and highly hydrophilic components (biogenic amines, bases and nucleosides) using this CPC method was investigated. The relative parameters such as the volume fraction of acetonitrile and length of the ODS and the TDS phases were investigated and optimized. The experimental results show that this column combines the advantages of both ODS and TDS stationary phases, and exhibits a reversed phase liquid chromatography (RPLC) mode followed by a hydrophilic interaction liquid chromatography (HILIC) mode when 80% of acetonitrile was used in the mobile phase. The satisfactory results indicate that the CPC method provides an easy way to simultaneously separate polar and non-polar compounds. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. [Studies on fingerprints of Centella asiatica by HPLC].

    Science.gov (United States)

    Lu, Qiang; Li, Wan-hong; Hu, Shi-qiang

    2011-01-01

    To study the HPLC fingerprints and establish a sensitive and specific method for controlling the quality of Centella asiatica. HPLC gradient elution was applied for the fingerprints of Centella asiatica. All 16 samples are collected from different habitats of China. The columni was Alltech C18 column (250 mm x 4.6 mm, 5 microm), mobile phase was acetonitrile-water, flow rate was 1.0 ml/min, wavelength was 205 nm. The fingerprint of Centella asiatica was established, 16 samples of different areas of Centella asiatica were detected. There were 15 common peaks in the HPLC fingerprints of Centella asiatica. By comparison with the reference standards and using LC-ESI-MS(n) to corroborate the structure, 5-10 peaks were identified as madecassoside, asiaticoside, quercetin, kaemperol, madecassic acid and asiatic acid respectively. After calculating the similarity of the HPLC fingerprints of 16 habitants, the similarity of different habitats has been bad quite. The method is accurate, reliable and good repeatability. This chromatographic fingerprint method can be used to controll the quality of Centella asiatica.

  14. Development and Validation of New RP-HPLC Method for the Determination of Dexrazoxane

    OpenAIRE

    Basaveswara Rao, M. V.; Prasanthi, V.; Rao, G. Venkata; Raman, B. V.

    2012-01-01

    A new sensitive, precise, rapid and linear RP-HPLC method was developed and validated for the determination of dexrazoxane in formulations and human serum samples. Good chromatographic separation of dexrazoxane was achieved by using Kromasil C18 column. The system was operated at ambient temperature using a mobile phase consisting of methanol, 5% ortho phosphoric acid, 0.01M ammonium dihydrogen phosphate and tetrahydrofuran, pH 4.2 (10:40:30:20, v/v) isocratically at a flow rate of 1 ml/min. ...

  15. Simultaneous Determination of Lamivudine, Zidovudine and Abacavir in Tablet Dosage Forms by RP HPLC Method

    OpenAIRE

    Kumar, D. Anantha; Rao, G. Srinivasa; Rao, J. V. L. N. Seshagiri

    2010-01-01

    A simple, accurate and reproducible RP-HPLC method has been developed for the simultaneous determination of lamivudine, zidovudine and abacavir in tablet dosage forms. Chromatography was carried out on a HiQ Sil C 18 V column using a mobile phase consisting of 0.01 M potassium dihydrogen ortho-phosphate (pH 3.0) and methanol (55:45 v/v) at a flow rate of 0.8 mL/min. The detection was made at 272 nm and stavudine was used as the internal standard for this study. The retention times for lamivud...

  16. Vitamin D analysis in plasma by high performance liquid chromatography (HPLC) with C30 reversed phase column and UV detection - easy and acetonitrile-free

    DEFF Research Database (Denmark)

    Hymøller, Lone; Jensen, Søren Krogh

    2011-01-01

    Two physiologically important forms of vitamin D exist: vitamin D2 and vitamin D3, which by liver based hydroxylase enzymes are converted to 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3, respectively. These hydroxylated metabolites of vitamin D are measured in plasma to assess the vtamin D status...... of animals and humans. Therefore cheap and reliable analytical methods are very much in demand in nutritional and physiological research. After saponification and extraction of plasma or serum samples the current method uses reverse phase high performance liquid chromatography on a C30 column and with UV...

  17. Development of a pre-column derivatization HPLC method with diode-array detection for the determination of amino sugars in peat and soil humic acids.

    Science.gov (United States)

    Beňo, Erik; Góra, Róbert; Hutta, Milan

    2017-11-22

    The work is focused on the development of a high-performance liquid chromatography method with diode-array detection for the separation and quantitation of the three most abundant amino sugars; d-glucosamine, d-galactosamine, and d-mannosamine. The high-performance liquid chromatography separation was carried out by reversed-phase chromatography on Chromolith Performance RP-18e monolithic column after acid hydrolysis (5 M HCl) and pre-column derivatization of samples using diethyl ethoxymethylenemalonate. Gradient elution and a mobile phase composed of ammonium formate buffer solution (10 mmol L-1 , pH 3.60) and methanol with flow rate of 1.0 mL min-1 were used. The monitoring wavelength was set at 280 nm. The limits of detection and quantitation for analytes ranged from 0.017 to 0.122 mg L-1 and from 0.057 to 0.407 mg L-1 , respectively. The proposed method was successfully applied for the determination of amino sugars in samples of humic acids isolated from different soils and peat. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  18. Quantification of methamphetamine, amphetamine and enantiomers by semi-micro column HPLC with fluorescence detection; applications on abusers' single hair analyses.

    Science.gov (United States)

    Al-Dirbashi, O Y; Kuroda, N; Wada, M; Takahashi, M; Nakashima, K

    2000-08-01

    Achiral and chiral semi-micro column high-performance liquid chromatographic methods with fluorescence detection to determine methamphetamine and amphetamine in human hair are described. These compounds were extracted into 5% trifluoroacetic acid (TFA) in methanol, derivatized with 4-(4,5-diphenyl-1H-imidazol-2-yl)-benzoyl chloride and separated either on a 250 x 1.5 mm i.d. octadecyl-silane (ODS) or a 150 x 2 mm i.d. OD-RH column. Linear calibration curves extending over a wide range of concentration that covers the practical samples were obtained for amphetamine, methamphetamine and their enantiomers (r = 0.999). Resolution values for amphetamine and methamphetamine enantiomers were 3.4 and 1.1, respectively. Intra- and inter-day variations of both the methods were not larger than 8.9% expressed as relative standard deviations (n >/= 5). The limits of detection at a signal-to-noise ratio of 3 obtained by both the methods were in the range of 1.0-4.7 fmol/5 microL injection with the achiral method being more sensitive. Abusers' hair samples were analyzed by the two methods and only the S(+)-enantiomers were found in eight Japanese abusers' hair samples. The achiral method was used to study the concentrations of these compounds in single black and white hair strands of abusers. Copyright 2000 John Wiley & Sons, Ltd.

  19. Comparison of UHPLC and HPLC in Benzodiazepines Analysis of Postmortem Samples

    Science.gov (United States)

    Behnoush, Behnam; Sheikhazadi, Ardeshir; Bazmi, Elham; Fattahi, Akbar; Sheikhazadi, Elham; Saberi Anary, Seyed Hossein

    2015-01-01

    Abstract The aim of this study was to compare system efficiency and analysis duration regarding the solvent consumption and system maintenance in high-pressure liquid chromatography (HPLC) and ultra high-pressure liquid chromatography (UHPLC). In a case–control study, standard solutions of 7 benzodiazepines (BZs) and 73 biological samples such as urine, tissue, stomach content, and bile that screened positive for BZs were analyzed by HPLC and UHPLC in laboratory of forensic toxicology during 2012 to 2013. HPLC analysis was performed using a Knauer by 100-5 C-18 column (250 mm × 4.6 mm) and Knauer photodiode array detector (PAD). UHPLC analysis was performed using Knauer PAD detector with cooling autosampler and Eurospher II 100-3 C-18 column (100 mm × 3 mm) and also 2 pumps. The mean retention time, standard deviation, flow rate, and repeatability of analytical results were compared by using 2 methods. Routine runtimes in HPLC and UHPLC took 40 and 15 minutes, respectively. Changes in mobile phase composition of the 2 methods were not required. Flow rate and solvent consumption in UHPLC decreased. Diazepam and flurazepam were detected more frequently in biological samples. In UHPLC, small particle size and short length of column cause effective separation of BZs in a very short time. Reduced flow rate, solvent consumption, and injection volume cause more efficiency and less analysis costs. Thus, in the detection of BZs, UHPLC is an accurate, sensitive, and fast method with less cost of analysis. PMID:25860209

  20. Solid-phase extraction and HPLC assay of nicotine and cotinine in plasma and brain.

    Science.gov (United States)

    Dawson, Ralph; Messina, S M; Stokes, C; Salyani, S; Alcalay, N; De Fiebre, N C; De Fiebre, C M

    2002-01-01

    The aim of this study was to develop a simple and reliable assay for nicotine (NIC) and its major metabolite, cotinine (COT), in plasma and brain. A method was developed that uses an extraction method compatible with reverse-phase high-performance liquid chromatography (HPLC) separation and ultraviolet (UV) detection. Sequential solid-phase extraction on silica columns followed by extraction using octadecyl (C18) columns resulted in mean percent recovery (n = 5) of 51 +/- 5, 64 +/- 10, and 52 +/- 10% for NIC, COT, and phenylimidazole (PI), respectively, in spiked 1-mL serum samples. Recovery (mean +/- SEM) of the internal standard (PI) from spiked samples of nicotine-injected rats averaged 64.1 +/- 1.5% (n = 138) from plasma, and 20.7+/-0.8% (n = 128) from brain. The limits of detection of NIC in plasma samples were approximately 8 ng per mL, and of COT, 13.6 ng per mL. Further optimization of our extraction method, using slower flow rates and solid-phase extraction on silica columns, followed by C18 column extraction, yielded somewhat better recoveries (38 +/-3%) for 1-mL brain homogenates. Interassay precision (coefficient of variation) was determined on the basis of daily calibrations for 2 months and was found to be 7%, 9%, and 9% for NIC, COT, and PI, respectively, whereas intra-assay variability was 3.9% for both NIC and COT. Limited studies were performed on analytical columns for comparison of retention, resolution, asymmetry, and column capacity. We concluded that a simple two-step solid-phase extraction method, coupled with HPLC separation and UV detection, can be used routinely to measure NIC and COT in biological fluids and tissues.

  1. Comparison of HPLC, UV spectrophotometry and potentiometric titration methods for the determination of lumefantrine in pharmaceutical products.

    Science.gov (United States)

    da Costa César, Isabela; Nogueira, Fernando Henrique Andrade; Pianetti, Gérson Antônio

    2008-09-10

    This paper describes the development and evaluation of a HPLC, UV spectrophotometry and potentiometric titration methods to quantify lumefantrine in raw materials and tablets. HPLC analyses were carried out using a Symmetry C(18) column and a mobile phase composed of methanol and 0.05% trifluoroacetic acid (80:20), with a flow rate of 1.0ml/min and UV detection at 335nm. For the spectrophotometric analyses, methanol was used as solvent and the wavelength of 335nm was selected for the detection. Non-aqueous titration of lumefantrine was carried out using perchloric acid as titrant and glacial acetic acid/acetic anhydride as solvent. The end point was potentiometrically determined. The three evaluated methods showed to be adequate to quantify lumefantrine in raw materials, while HPLC and UV methods presented the most reliable results for the analyses of tablets.

  2. Development and application of HPLC-RI and HPLC-MS/MS based methods for quantification of residual deoxycholate levels in pneumococcal polysaccharides.

    Science.gov (United States)

    Gairola, Sunil; Gautam, Manish; Patil, Dada; Manoj Kumar, Krishna; Shinde, Pravin; Jana, S K; Dhere, Rajeev; Jadhav, Suresh

    2016-11-01

    The analysis of residual sodium deoxycholate (DOC); a detergent of biological origin used in manufacturing of polysaccharide vaccines is challenging due to complex sample matrices and the lack of suitable methods. Here we report, rapid and sensitive high-performance liquid chromatography-refractive index (HPLC-RI) and tandem mass spectrometry (HPLC-MS/MS) methods for estimation of residual DOC in pneumococcal polysaccharides. For HPLC-RI method, separation was achieved using Luna C18 column and mobile phase compositions of acetonitrile: methanol: 20 mM sodium acetate (60:05:35% v/v). For HPLC-MS/MS method, separation was achieved using a Hypersil BDS C18 column with gradient elution of methanol and water (0.1% formic acid). MS/MS method showed linearity (r(2) = 0.997) over the range of 10-320 ng/mL with limits of detection (LOD) and lower limit of quantitation (LOQ) of 3 and 10 ng/mL respectively. Precision (% RSD) and accuracy (% recovery) for both methods were in the range of 0.74-8.29% and 82.33-117.86% respectively. Sample matrices interferences were addressed following novel sample clean-up method based on liquid-liquid extraction. Both methods enabled traceable quantitation of DOC in intermediate and purified pneumococcal polysaccharides of serotypes: 1, 5, 6A, 6B, 7F, 9V, 14, 19A, 19F and 23F. Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  3. [Comparative study of HPLC fingerprint between Zanthoxylum bungeanum and Zanthoxylum schinifolium].

    Science.gov (United States)

    Song, Li; Liu, You-Ping

    2012-01-01

    To establish the HPLC fingerprint of Zanthoxylum bungeanum and Zanthoxylum schinifolium for finding the difference. Samples were extracted with 50% methanol 25 mL by ultrasonic wave and then separated on Hypersil BDS C18 (250 mm x 4.6 mm, 5 microm) column. Gradient elution was carried out with a mobile phase of methanol-water. The detection wavelength was 268 nm, the column temperature was set at 35 degrees C, the flow rate was 1.0 mL/min and the analytic time was 130 min. The software "Similarity Evaluation System for Chromatographic Fingerprint of TCMs" (Version 2004A) was employed to generate the mean chromatogram and carry out the similarity analysis of the samples. SPSS 17.0 was employed to carry out the cluster analysis. Similarity of Z. bungeanum was 0.909 - 0.992 and that of Z. schinifolium was 0.930 - 0.999. There were 27 common peaks in HPLC Fingerprints of Z. bungeanum and 24 in that of Z. schinifolium. Their HPLC standard fingerprints were obvious difference. They belonged to different categories in cluster analysis. The method is simple, accurate and rapid. It could obviously distinguish Z. bungeanum from Z. schinifolium. So it suggests that HPLC fingerprints should be one of the quality control indexes of Zanthoxyli Pericarpium.

  4. [HPLC fingerprint chromatogram of Polygonum multiflorum from Guizhou].

    Science.gov (United States)

    Li, Yan; Wang, Hui-Juan; Lin, Bing; Zhao, Zhi; Zhou, Ying

    2012-12-01

    To establish the fingerprint of Polygonum multiflorum from Guizhou and provide a standard for its quality control. HPLC analysis was performed on Agillent ZABAX-C18 (4.6 mm x 250 mm, 5 microm), gradient eluted composed of acetonitrile-0.4% water solution of phosphoric acid. Column temperature was set at 25 degrees C and the flow rate was 1 mL/min. The detection wavelength was 280 nm and the analysis time was 60 min. 9 common peaks were identified. The RSD of the relative retention time and the relative peak area were less than 3% in analyzing its precision, stability and repeatability of the common peaks, and the similarity of the 16 batches of sample was more than 0.9. The method is simple and reliable, and it can provide a standard and guidance for quality control of Polygonum multiflorum.

  5. HPLC analysis of closed, open, and reflex eye tear proteins

    Directory of Open Access Journals (Sweden)

    Sitaramamma T

    1998-01-01

    Full Text Available Changes in the closed, open and reflex eye tear proteins of normal subjects were compared and analysed. Tear proteins were resolved by high-performance liquid chromatography (HPLC utilising both gel filtration (P-300 SW and reverse-phase (C-18 columns and the HPLC fractions were further analysed by sodium dodecyl sulphate - polyacrylamide gel electrophoresis (SDS-PAGE under reducing and non-reducing conditions. The protein composition of the closed-eye tear was significantly different from that of the open and reflex-eye tear. Secretory IgA (sIgA was the predominant protein in closed eye tears constituting 49% of the total protein compared to 11% in reflex tears, whereas lysozyme was the predominant protein (53% in reflex tears. Levels of lactoferrin, lipocalin and lysozyme were relatively constant in both open and reflex tears. HPLC profiles of the closed-eye tears, upon continuous stimulation of lacrimal glands indicated that sIgA was significantly reduced whereas lactoferrin, lipocalin, and lysozyme were significantly increased. These results indicate that the tear composition upon waking attains that of the open eye within 4 to 5 minutes, and upon continuous stimulation this reflects the reflex-eye tear composition. It also indicates that mechanisms responsible for changes in concentration of constitutive and regulated tear protein with stimulus can be studied successfully using non-invasive methods to collect human tears.

  6. HPLC analysis of closed, open, and reflex eye tear proteins.

    Science.gov (United States)

    Sitaramamma, T; Shivaji, S; Rao, G N

    1998-12-01

    Changes in the closed, open and reflex eye tear proteins of normal subjects were compared and analysed. Tear proteins were resolved by high-performance liquid chromatography (HPLC) utilising both gel filtration (P-300 SW) and reverse-phase (C-18) columns and the HPLC fractions were further analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing and non-reducing conditions. The protein composition of the closed-eye tear was significantly different from that of the open and reflex-eye tear. Secretory IgA (sIgA) was the predominant protein in closed eye tears constituting 49% of the total protein compared to 11% in reflex tears, whereas lysozyme was the predominant protein (53%) in reflex tears. Levels of lactoferrin, lipocalin and lysozyme were relatively constant in both open and reflex tears. HPLC profiles of the closed-eye tears, upon continuous stimulation of lacrimal glands indicated that sIgA was significantly reduced whereas lactoferrin, lipocalin, and lysozyme were significantly increased. These results indicate that the tear composition upon waking attains that of the open eye within 4 to 5 minutes, and upon continuous stimulation this reflects the reflex-eye tear composition. It also indicates that mechanisms responsible for changes in concentration of constitutive and regulated tear protein with stimulus can be studied successfully using non-invasive methods to collect human tears.

  7. [Chromatographic Fingerprinting Study of Zhenyuan Granules Dry Extract by HPLC-DAD and HPLC-MS/MS].

    Science.gov (United States)

    Li, Yuan-yuan; Shan, Jin-feng; Tan, Qing-jie; Wang, Song-lin; Jiang, Jian-lan

    2015-09-01

    To establish a novel, accurate and valid fingerprint method of Zhenyuan granules dry extract by using HPLC-DAD method, to study herbs belonging of fingerprint peaks and to identify some of the chromatographic peaks by HPLC-MS/MS analysis, for providing the basis for scientific evaluation of the quality. The sample solutions were analyzed by an Agilent SB C18 (250 mm x 4.6 mm, 5 µm) column, and gradiently eluted with acetonitrile (containing 0.1% formic acid) and aqueous phase (containing 0.1% formic acid) as the mobile phase. The flow rates were 1.2 mL/min (0~70 min) and 0.8 mL/min (70~150 min); the column temperature was 30 °C; and the detection wavelength was 254 nm. 40 peaks were selected as fingerprint peaks under the optimal chromatographic condition, and the similarity coefficients of 10 batches of Zhenyuan granules dry extract were all greater than 0.98. 27 peaks were tentatively identified with reference to literature data based on their mass spectrometry. The chromatographic fingerprint of Zhenyuan granules is proved to be a reliable method for comprehensive quality control and assessment.

  8. Qualitative and Quantitative Analysis of Lignan Constituents in Caulis Trachelospermi by HPLC-QTOF-MS and HPLC-UV.

    Science.gov (United States)

    Liu, Xiao-Ting; Wang, Xu-Guang; Yang, Yu; Xu, Rui; Meng, Fan-Hua; Yu, Neng-Jiang; Zhao, Yi-Min

    2015-05-05

    A high-performance liquid chromatography coupled with quadrupole tandem time-of-flight mass (HPLC-QTOF-MS) and ultraviolet spectrometry (HPLC-UV) was established for simultaneous qualitative and quantitative analysis of the major chemical constituents in Caulis Trachelospermi, respectively. The analysis was performed on an Agilent Zorbax Eclipse Plus C18 column (4.6 mm×150 mm, 5 μm) using a binary gradient system of water and methanol, with ultraviolet absorption at 230 nm. Based on high-resolution ESI-MS/MS fragmentation behaviors of the reference standards, the characteristic cleavage patterns of lignano-9, 9'-lactones and lignano-8'-hydroxy-9, 9'-lactones were obtained. The results demonstrated that the characteristic fragmentation patterns are valuable for identifying and differentiating lignano-9,9'-lactones and lignano-8'-hydroxy-9,9'-lactones. As such, a total of 25 compounds in Caulis Trachelospermi were unambiguously or tentatively identified via comparisons with reference standards or literature. In addition, 14 dibenzylbutyrolatone lignans were simultaneously quantified in Caulis Trachelospermi by HPLC-UV method. The method is suitable for the qualitative and quantitative analyses of dibenzylbutyrolatone lignans in Caulis Trachelospermi.

  9. Qualitative and Quantitative Analysis of Lignan Constituents in Caulis Trachelospermi by HPLC-QTOF-MS and HPLC-UV

    Directory of Open Access Journals (Sweden)

    Xiao-Ting Liu

    2015-05-01

    Full Text Available A high-performance liquid chromatography coupled with quadrupole tandem time-of-flight mass (HPLC-QTOF-MS and ultraviolet spectrometry (HPLC-UV was established for simultaneous qualitative and quantitative analysis of the major chemical constituents in Caulis Trachelospermi, respectively. The analysis was performed on an Agilent Zorbax Eclipse Plus C18 column (4.6 mm × 150 mm, 5 μm using a binary gradient system of water and methanol, with ultraviolet absorption at 230 nm. Based on high-resolution ESI-MS/MS fragmentation behaviors of the reference standards, the characteristic cleavage patterns of lignano-9, 9'-lactones and lignano-8'-hydroxy-9, 9'-lactones were obtained. The results demonstrated that the characteristic fragmentation patterns are valuable for identifying and differentiating lignano-9,9'-lactones and lignano-8'-hydroxy-9,9'-lactones. As such, a total of 25 compounds in Caulis Trachelospermi were unambiguously or tentatively identified via comparisons with reference standards or literature. In addition, 14 dibenzylbutyrolatone lignans were simultaneously quantified in Caulis Trachelospermi by HPLC-UV method. The method is suitable for the qualitative and quantitative analyses of dibenzylbutyrolatone lignans in Caulis Trachelospermi.

  10. Validation of simultaneous volumetric and HPLC methods for the determination of pridinol mesylate in raw material.

    Science.gov (United States)

    Simionato, Laura D; Ferello, Leonardo; Stamer, Sebastián; Zubata, Patricia D; Segall, Adriana I

    2013-01-01

    Simple, sensitive, and economical simultaneous volumetric and HPLC methods for the determination of pridinol mesylate in raw material have been developed. The volumetric method is based on the reaction of pridinol with sodium lauryl sulphate in diluted sulphuric acid. Dimethyl yellow was used as indicator to detect the end point of the titration in aqueous/organic layer. The HPLC method for the determination of pridinol mesylate employs a reverse phase C18 column at ambient temperature with a mobile phase consisting of acetonitrile: 0.05 M potassium dihydrogen phosphate, pH adjusted to 5.0 (1 : 2, v/v). The flow rate was 0.8 mL/min. Quantitation was achieved with UV detection at 258 nm based on peak area. Both methods were found to be suitable for the quality control of pridinol mesylate in raw material.

  11. A Simple HPLC Bioanalytical Method for the Determination of ...

    African Journals Online (AJOL)

    Purpose: To develop a simple, accurate, and precise high performance chromatography (HPLC) method with spectrophotometric detection for the determination of doxorubicin hydrochloride in rat plasma. Methods: Doxorubicin hydrochloride and daunorubicin hydrochloride (internal standard, IS) were separated on a C18.

  12. Comparison of microbial communities in Lake Tahoe surface sample with Tonga Trench water column samples using High Pressure Liquid Chromatography - Electrospray Ionization - Mass Spectroscopy (HPLC - ESI - MS) and Global Natural Products Social Molecular Network (GNPS)

    Science.gov (United States)

    Belmonte, M. A.

    2015-12-01

    Intact polar lipids (IPLs) are lipids composed of a head group, a glycerol, and a fatty acid chain that make up the lipid bilayer of cell membranes in living cells; and the varying head groups can be indicative of the type of microbes present in the environment (Van Mooy 2010). So by distinguishing and identifying the IPL distribution in an environment one can make inferences about the microbial communities in the said environment. In this study, we used High Pressure Liquid Chromatography-Electrospray Ionization- Mass Spectroscopy (HPLC-ESI-MS) and Global Natural Products Social Molecular Networking (GNPS) to compare the IPL distributions of two oligotrophic environments: surface waters of Lake Tahoe in the Sierra Nevada Mountains, and the water column of the Tonga Trench in the South Pacific. We hypothesized that the similar nutrient dynamics of the two oligotrophic environments would result in similar eukaryotic and prokaryotic communities, which would be reflected in the IPL composition of suspended particulate organic matter (POM). For simplicity we focused on the classes of IPLs most commonly observed in the marine environment: phosphotidylglycerol (PG), phosphotidylethanolamine (PE), diacylglyceryl-trimethyl-homoserine (DGTS), diacylglyceryl-hydroxymethyl-trimethylalanine (DGTA), sulfoquinovosyldiacylglycerol (SQDG), monoglycosyldiacylglycerol (MGDG) and diglycosyldiacylglycerol (DGDG). Our results showed that all of the marine IPLs of interest were present in Lake Tahoe which confirms that there are many of the same microbial communities in the fresh waters of Lake Tahoe and the salt waters Tonga Trench.

  13. High-resolution peptide mapping separations with MS-friendly mobile phases and charge-surface-modified C18.

    Science.gov (United States)

    Lauber, Matthew A; Koza, Stephan M; McCall, Scott A; Alden, Bonnie A; Iraneta, Pamela C; Fountain, Kenneth J

    2013-07-16

    Ionic analytes, such as peptides, can be challenging to separate by reverse-phase chromatography with optimal efficiency. They tend, for instance, to exhibit poor peak shapes, particularly when eluted with mobile phases preferred for electrospray ionization mass spectrometry. We demonstrate that a novel charged-surface C18 stationary phase alleviates some of the challenges associated with reverse-phase peptide separations. This column chemistry, known as CSH (charged-surface hybrid) C18, improves upon an already robust organosilica hybrid stationary phase, BEH (ethylene-bridged hybrid) C18. Based on separations of a nine-peptide standard, CSH C18 was found to exhibit improved loadability, greater peak capacities, and unique selectivity compared to BEH C18. Its performance was also seen to be significantly less dependent on TFA-ion pairing, making it ideal for MS applications where high sensitivity is desired. These performance advantages were evaluated through application to peptide mapping, wherein CSH C18 was found to aid the development of a high-resolution, high-sensitivity LC-UV-MS peptide mapping method for the therapeutic antibody, trastuzumab. From these results, the use of a C18 stationary phase with a charged surface, such as CSH C18, holds significant promise for facilitating challenging peptide analyses.

  14. STABILITY INDICATING HPLC METHOD DEVELOPMENT AND VALIDATION FOR DETERMINATION OF DACLATASVIR IN PURE AND TABLETS DOSAGE FORMS

    OpenAIRE

    Hanaa Saleh, Gamal H. Ragab, Mohammed A. Othman

    2017-01-01

    A sensitive, simple, selective and accurate HPLC method was developed and validated for analysis of antiviral drug Daclatasvir (BMS-790052, DCV) in pure form and in tablet dosage form in the presence of its degradation products. The chromatographic separation achieved by isocratic elution on Hypersil BDS C18, 4.6×150 mm, 5µm column at 25˚c. The mobile phase was a mixture of 0.05M Potassium dihydrogen phosphate (pH-4.5) and acetonitrile in ratio of 50:50 (v/v). The injection volume was 10µl. T...

  15. Determination of related substances in lisinopril and amlodipine tablets by HPLC

    Directory of Open Access Journals (Sweden)

    Dao-Rui Yu

    2016-06-01

    Full Text Available Objective: To establish an HPLC method for determining the related substances in lisinopril and amlodipine tablets. Methods: An Inertsil Thermo BDS HYPERSIL C18 (4.6 mm伊250 mm, 5 μm column was used with the Acetonitrile-water-phosphoric acid (10:90:0.1 as mobile phase A and Acetonitrile-water-phosphoric acid (90:10:0.1 as mobile phase B by gradient elution at the detection wavelength of 215 nm. The flow rate was 1.0 mL/min and the column temperature was 30 ℃. Results: The separation of the impurity peak and peak was good. Besides, all the impurities could be detected effectively. Conclusions: The method is sensitive, accurate and selective. It is suitable for control the related substances in lisinopril and Amlodipine tablets.

  16. [Simultaneous determination of 6 active components in Chrysanthemum morifolium by HPLC].

    Science.gov (United States)

    Qin, Shan; Wen, Xuesen

    2011-06-01

    To develop a HPLC method quantitative method for simultaneous determination of chlorogenic acid, 1, 5-dicaffeoylquinic acid, isochlorogenic acid A, isochlorogenic acid C, luteolin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside in Chrysanthemum morifolium Ramat. A Phenomenex Gemini-NX C18 column (4.6 mm x 250 mm, 5 microm) was used with CH3 OH and 0.4% H3PO4 as mobile phases. The flow rate was 1 mL x min(-1), the column temperature was 25 degrees C, and the detection wavelength was set at 350 nm. The 6 active components were in baseline separation. The linearity of this method was good (r > or = 0.999 7), and the average recoveries were 100.6% - 102.4%, RSD Chrysanthemum.

  17. Analysis of C-glycosyl flavonoids from South American Passiflora species by HPLC-DAD and HPLC-MS.

    Science.gov (United States)

    Zucolotto, Silvana Maria; Fagundes, Carize; Reginatto, Flávio Henrique; Ramos, Freddy A; Castellanos, Leonardo; Duque, Carmenza; Schenkel, Eloir Paulo

    2012-01-01

    Leaves and fruits of Passiflora species are widely used around the world in popular medicine, mainly as sedatives and tranquilisers. C-glycosyl flavonoids are the main components of these species. To investigate the constituent patterns and to develop a chromatographic method for the characterisation of the C-glycosyl flavonoids profile of the extracts of the leaves and the pericarp of South American Passiflora species. The chemical composition of extracts from the leaves and the fruits' pericarp of Passiflora edulis var. flavicarpa, P. edulis var. edulis, Passiflora alata, Passiflora tripartita var. mollissima, Passiflora quadrangularis, Passiflora manicata and Passiflora ligularis was evaluated for the presence of C-glycosyl flavonoids. Two separate HPLC methods were developed suitable for a diode array detector (DAD) and a MS detector. Separation by HPLC-DAD was achieved on a Luna C-18 column, using solvent A (tetrahydrofuran-isopropanol-acetonitrile) and solvent B (H₃PO₄ 0.5%) in an isocratic elution mode. In the HPLC-MS, the components were separated on a Luna RP-18A column by a gradient elution (water-acetonitrile-formic acid). The presence of C-glycosyl flavonoids was identified in leaves and pericarp of P. edulis var. flavicarpa, P. alata, P. edulis var. edulis and P. tripartita var. molissima, but only in leaf extracts of P. quadrangularis and P. manicata and not at all in P. ligularis. The different species and varieties showed different major constituents. The C-glycosyl flavonoids identified more frequently were orientin, isoorientin, vitexin and isovitexin. The methods established are simple and can be used as a tool for the characterisation and quality control of pharmaceutical preparations containing these Passiflora extracts. Copyright © 2011 John Wiley & Sons, Ltd.

  18. Application of RP-HPLC in Detecting Content of Ferulic Acid in Fuke Qianjin Capsule

    Directory of Open Access Journals (Sweden)

    Ming-zhi WANG

    2015-09-01

    Full Text Available Objective: To establish a method for the determination of ferulic acid content in Fuke Qianjin Capsule. Methods: Reversed phase high performance liquid chromatography (RP-HPLC was applied in this study, with the detection conditions as follows: chromatographic column: Boston green ODS·min-1, detection wavelength: 316 nm, and column temperature: 30℃. C18 (250 mm × 4.6 mm, 5 μm, mobile phase: methanol-0.1% phosphoric acid (25:75, flow velocity: 1.0 mLResults: Ferulic acid, whose sample size was 0.017760-0.10656 μg, was in favorable linear relationship with the integral value of peak area, with correlation coefficient r=0.9998. The average sample-injecting recovery rate and degree of precision (RSD were 97.6% (n=6 and 1.6%, respectively. The results of this study also showed that the specificity of RP-HPLC in this study was excellent; negative samples had no interference on chromatographic peak of target substance (ferulic acid; and RSD of accuracy, repeatability, stability and recovery rate were 1.1%, 1.4%, 1.2% and 1.6%, respectively.Conclusion: RP-HPLC is accurate, rapid, stable and convenient, so it can be used as an optimal method for the detection of ferulic acid content in Fuke Qianjin Capsule.

  19. Stability-indicating HPLC method for the determination of darunavir ethanolate.

    Science.gov (United States)

    Reddy, B V Rami; Jyothi, G; Reddy, B S; Raman, N V V S S; Reddy, K Subhash Chander; Rambabu, C

    2013-01-01

    A novel stability-indicating reversed-phase high-performance liquid chromatographic (HPLC) method has been developed for the quantitative determination of darunavir ethanolate, an HIV-1 protease inhibitor. The chromatographic separation was achieved using an X-Bridge C18 (150 × 4.6 mm × 3.5 µm) HPLC column in isocratic mode employing 0.01M ammonium formate (pH.3.0) buffer and acetonitrile in the ratio of 55:45 (v/v) with a flow rate of 1.0 mL/min. The detector wavelength was monitored at 265 nm and the column temperature was maintained at 30°C. Darunavir ethanolate was exposed to thermal, photolytic, acid, base and oxidative stress conditions. Considerable degradation of the drug substance was found to occur under acid, base and oxidative stress conditions. The peak homogeneity data of darunavir ethanolate obtained by photodiode array detection demonstrated the specificity of the method in the presence of degradants. The degradation products were well resolved from primary peak of darunavir, indicating that the method is specific and stability-indicating. The HPLC method was validated as per International Conference on Harmonization guidelines with respect to specificity, precision, linearity, accuracy and robustness. Regression analysis showed a correlation coefficient value greater than 0.999. The accuracy of the method was established based on the recovery obtained for darunavir ethanolate.

  20. [Correlation of HPLC Characteristic Spectra of Vinegar Corydalis Rhizoma Decoction Pieces, Water Decoction and Formula Granules].

    Science.gov (United States)

    Wei, Mei; Du, Lan-zhe; Li, Hui; Zhang, Guang-da; Chen, Xiang-dong

    2015-05-01

    To study the correlation of characteristic spectra of Vinegar Corydalis Rhizoma decoction pieces, water decoction and formula granules by HPLC, and to investigate the transfer of the main chemical constituents between three different forms. The analysis was carried out by a Phenomenex Gemini C18 column (250 mm x 4.6 mm,5 μm) with acetonitrile-1% acetic acid and ammonium acetate buffer solution (pH 6.0) as the mobile phase in a gradient elution mode. The detection wavelength was 280 nm with a flow rate of 0.8 mL /min. The column temperature was 30 degrees C. The characteristic spectra from 11 batches of Vinegar Corydalis Rhizoma decoction pieces, 11 batches of water decoction and 11 batches of formula granules were established respectively. Ten peaks in the HPLC characteristic spectra from 11 batches of formula granules could be tracked in the water decoction, nine peaks in the HPLC characteristic spectra could be tracked in the decoction pieces. In the ten common peaks, four components such as protopine, palnatine chloride, berberine hydrochloride and tetrahydropalmatine were verified. The main chemical components of Vinegar Corydalis Rhizoma decoction pieces, water decoction and formula granules are basically the same, the common component contents have similar proportion.

  1. Development and optimisation of an HPLC-DAD-ESI-Q-ToF method for the determination of phenolic acids and derivatives.

    Directory of Open Access Journals (Sweden)

    Annalaura Restivo

    Full Text Available A method for the HPLC-MS/MS analysis of phenols, including phenolic acids and naphtoquinones, using an amide-embedded phase column was developed and compared to the literature methods based on classical C18 stationary phase columns. RP-Amide is a recently developed polar embedded stationary phase, whose wetting properties mean that up to 100% water can be used as an eluent. The increased retention and selectivity for polar compounds and the possibility of working in 100% water conditions make this column particularly interesting for the HPLC analysis of phenolic acids and derivatives. In this study, the chromatographic separation was optimised on an HPLC-DAD, and was used to separate 13 standard phenolic acids and derivatives. The method was validated on an HPLC-ESI-Q-ToF. The acquisition was performed in negative polarity and MS/MS target mode. Ionisation conditions and acquisition parameters for the Q-ToF detector were investigated by working on collision energies and fragmentor potentials. The performance of the method was fully evaluated on standards. Moreover, several raw materials containing phenols were analysed: walnut, gall, wine, malbec grape, French oak, red henna and propolis. Our method allowed us to characterize the phenolic composition in a wide range of matrices and to highlight possible matrix effects.

  2. Simultaneous HPLC quantitative analysis of active compounds in leaves of Moringa oleifera Lam.

    Science.gov (United States)

    Vongsak, Boonyadist; Sithisarn, Pongtip; Gritsanapan, Wandee

    2014-08-01

    Moringa oleifera Lam. has been used as a traditional medicine for the treatment of numerous diseases. A simultaneous high-performance liquid chromatography (HPLC) analysis was developed and validated for the determination of the contents of crypto-chlorogenic acid, isoquercetin and astragalin, the primary antioxidative compounds, in M. oleifera leaves. HPLC analysis was successfully conducted by using a Hypersil BDS C18 column, eluted with a gradient of methanol-1% acetic acid with a flow rate of 1 mL/min, and detected at 334 nm. Parameters for the validation included linearity, precision, accuracy and limits of detection and quantitation. The developed HPLC method was precise, with relative standard deviation oleifera leaf extracts were 98.50, 98.47 and 98.59%, respectively. The average contents of these compounds in the dried ethanolic extracts of the leaves of M. oleifera collected from different regions of Thailand were 0.081, 0.120 and 0.153% (w/w), respectively. The developed HPLC method was appropriate and practical for the simultaneous analysis of crypto-chlorogenic acid, isoquercetin and astragalin in the leaf extract of M. oleifera. This work is valuable as guidance for the standardization of the leaf extracts and pharmaceutical products of M. oleifera. © The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. 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.

  4. Comparison of monolithic and microparticulate columns for reversed-phase liquid chromatography of tryptic digests of industrial enzymes in cleaning products.

    Science.gov (United States)

    Beneito-Cambra, M; Herrero-Martínez, J M; Ramis-Ramos, G; Lindner, W; Lämmerhofer, M

    2011-10-14

    Enzymes of several classes used in the formulations of cleaning products were characterized by trypsin digestion followed by HPLC with UV detection. A polymeric monolithic column (ProSwift) was used to optimize the separation of both the intact enzymes and their tryptic digests. This column was adequate for the quality control of raw industrial enzyme concentrates. Then, monolithic and microparticulate columns were compared for peptide analysis. Under optimized conditions, the analysis of tryptic digests of enzymes of different classes commonly used in the formulation of cleaning products was carried out. Number of peaks, peak capacity and global resolution were obtained in order to evaluate the chromatographic performance of each column. Particulate shell-core C18 columns (Kinetex, 2.6 μm) showed the best performance, followed by a silica monolithic column (Chromolith RP-18e) and the conventional C18 packings (Gemini, 5 μm or 3 μm). A polymeric monolithic column (ProSwift) gave the worst performances. The proposed method was satisfactorily applied to the characterization of the enzymes present in spiked detergent bases and commercial cleaners. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. Simultaneous Determination of Five Active Components in the Chinese Patent Medicine Niuhuang Jiangya Pill by HPLC-MS/MS.

    Science.gov (United States)

    Xiong, Shan; Lei, Shanshan

    2017-05-01

    Niuhuang Jiangya (NHJY) pill is one of the well-known Chinese patent medicines in China used in the treatment of high blood pressure. The primary purpose of this study was to establish and validate a method using HPLC with tandem MS for the quality evaluation of NHJY pill through simultaneous determination of the following five active components: baicalin, paeoniflorin, astragaloside IV, ferulic acid, and emodin. Chromatographic separation was carried out on a Hypersil GOLD HPLC C18 column (50 × 4.6 mm, 3 μm) with acetonitrile and water as mobile phase and gradient elution at a flow rate of 0.4 mL/min. The method established in this study was selective, linear, precise, and accurate and was successfully applied to evaluate five active components in NHJY pill collected from different production batches, which could be considered a good approach to control the quality of NHJY pill and other related botanical drugs.

  6. A validated stability-indicating HPLC method for routine analysis of an injectable lincomycin and spectinomycin formulation.

    Science.gov (United States)

    Abualhasan, Murad N; Batrawi, Nidal; Sutcliffe, Oliver B; Zaid, Abdel Naser

    2012-01-01

    Lincomycin and spectinomycin combination therapy is widely used in veterinary medicine for the treatment of gastrointestinal and respiratory infections caused by lincomycin- and spectinomycin-sensitive microorganisms. A simple, reverse phase HPLC method for the analysis of samples of an injectable lincomycin and spectinomycin preparation containing a mixture of inactive excipients has been developed. The HPLC was carried out using the RP-C(18) (250 mm × 4.0 mm, 5 μm) column, with the gradient mobile phase consisting of an acetonitrile and phosphate buffer at pH 6; the flow rate of 1 mL/min and ultraviolet detection at 220 nm. This method was validated in accordance with both FDA and ICH guidelines and showed good linearity, accuracy, precision, selectivity, and system suitability results within the acceptance criteria. A stability-indicating study was also carried out and indicated that this method can also be used for purity and degradation evaluation of these formulations.

  7. Simultaneous quantitative determination of cinnamaldehyde and methyl eugenol from stem bark of Cinnamomum zeylanicum Blume using RP-HPLC.

    Science.gov (United States)

    Gursale, Atish; Dighe, Vidya; Parekh, Guarang

    2010-01-01

    A simple, sensitive, and precise reversed-phase high performance liquid chromatographic (HPLC) method has been developed, validated and used for simultaneous quantitative determination of cinnamaldehyde and methyl eugenol from the methanolic extract of dried bark powder of Cinnamomum zeylanicum Blume (family Lauraceae). The ultrasonic extraction method was used for the extraction of these compounds. The reversed-phase HPLC analysis was carried out using a Intersil ODS-3V-C(18) (150 mm x 4.6 mm, 5 microm) column and a mobile phase comprising of methanol-acetonitrile-water in the volume ratio of 35:20:45, delivered at a flow rate of 1.0 cm(3)/min. The detection and quantitation of both the compounds was carried out at 221 nm.

  8. On-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma with a weak ion exchange monolithic column.

    Science.gov (United States)

    Yang, Gengliang; Feng, Sha; Liu, Haiyan; Yin, Junfa; Zhang, Li; Cai, Liping

    2007-07-01

    A weak ion exchange monolithic column prepared by modifying the GMA-MAA-EDMA (glycidyl methacrylate-methacrylic acid-ethylene glycol dimethacrylate) monoliths with ethylenediamine was applied to remove matrix compounds in biological fluid. Using this monolithic column, on-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma samples had been investigated. Chromatography was performed by reversed-phase HPLC on a C(18) column with ultraviolet detection at 225 nm. Results showed that the ion exchange monolithic column could be used for deproteinization and retaining oxacillin and cloxacillin in human urine and plasma, which provided a simple and fast method for assaying drugs in human urine and plasma.

  9. DEVELOPMENT OF A NEW STABILITY INDICATING RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF METFORMIN HYDROCHLORIDE AND TENELIGLIPTIN HYDROBROMIDE AND ITS VALIDATION AS PER ICH GUIDELINES

    OpenAIRE

    Vinutha Kommineni* , K.P.R.Chowdary and S.V.U.M.Prasad

    2017-01-01

    A new stability indicating RP HPLC method has been developed and validated for simultaneous estimation of Metformin Hydrochloride and Teneligliptin in bulk and dosage forms. The method involves separation on YMC C18column(150mm x 4.6mm x5µm particle size). The optimized mobile phase consists of Phosphate buffer (pH 3) and Acetonitrile (80:20v/v) with a flow rate of 0.8ml/min and UV detection at 220mn. Retention time was 2.138min (Metformin Hydrochloride), 2.943min (Teneligliptin), 5.075 Piogl...

  10. Determination of levodopa in human plasma by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS: application to a bioequivalence study

    Directory of Open Access Journals (Sweden)

    Heliana F. Martins

    2013-01-01

    Full Text Available A sensitive, accurate and simple method using HPLC-MS/MS was developed and validated for levodopa quantitation in human plasma. Analysis was achieved on a pursuit® C18 analytical column (5 µm; 150 x 4.6 mm i.d. using a mobile phase (methanol and water , 90:10, v/v containing formic acid 0.5% v/v, after extracting the samples using a simple protein plasma precipitation with perchloric acid. The developed method was validated in accordance with ANVISA guidelines and was successfully applied to a bioequivalence study in 60 healthy volunteers demonstrating the feasibility and reliability of the proposed method.

  11. Development and Validation of a RP-HPLC Method for Estimation of Montelukast Sodium in Bulk and in Tablet Dosage Form

    OpenAIRE

    Singh R; Saini P; Mathur S; Singh G; Lal B

    2010-01-01

    The present work describes a simple, precise and accurate HPLC method for estimation of montelukast sodium in bulk and in tablet dosage form. The separation was achieved by using octadecylsilane column (C18) and acetonitrile:1 mM sodium acetate adjusted to pH 6.3 with acetic acid in proportion of 90:10 v/v as mobile phase, at a flow rate of 1.5 ml/min. Detection was carried out at 285 nm. The retention time of montelukast sodium was found to be 3.4 min. The limit of detection was found 1.31 &...

  12. HPLC analysis of methylxanthines in human breast milk.

    Science.gov (United States)

    Blanchard, J; Weber, C W; Shearer, L E

    1990-12-01

    A sensitive and specific high-performance liquid chromatographic (HPLC) procedure is developed for simultaneously quantitating the levels of caffeine, theophylline, theobromine and paraxanthine in breast milk. The method involved the precipitation of proteins present in the milk samples with a 6% v/v perchloric acid solution containing the internal standard, proxyphylline, followed by centrifugation at 12,800 Xg for 10 minutes. The clear supernatant was then chromatographed on a C18 reversed-phase analytical column at ambient temperature using a wavelength of 272 nm. Samples were eluted from the column at a constant flow rate of 1.5 mL/min using a gradient program in which the concentration of methanol in the mobile phase varied from 0 to 16%. The mean recoveries of the methylxanthines averaged over all the concentrations examined were generally excellent and ranged from 96.3 +/- 5.4% for caffeine to 102.3 +/- 8.9% for paraxanthine. The assay precision was very good and the peaks of interest were extremely well resolved. The method is recommended for assessing the total caffeine and dimethylxanthine load to which the nursing infant is exposed in mothers ingesting typical amounts of caffeine.

  13. Determining trace amounts and the origin of formaldehyde impurity in Neisseria meningitidis A/C/Y/W-135-DT conjugate vaccine formulated in isotonic aqueous 1× PBS by improved C18-UPLC method.

    Science.gov (United States)

    Gudlavalleti, Seshu K; Crawford, Erika N; Tran, Nhi N; Orten, Dana J; Harder, Jeffery D; Reddy, Jeeri R

    2015-03-25

    The ability to accurately measure and report trace amounts of residual formaldehyde impurity in a vaccine product is not only critical in the product release but also a regulatory requirement. In many bacterial or viral vaccine manufacturing procedures, formaldehyde is used either at a live culture inactivation step or at a protein de-toxification step or at both. Reported here is a validated and improved C18-UPLC method (developed based on previously published C-8 HPLC method) to determine the traces of formaldehyde process impurity in a liquid form Neisseria meningitidis A/C/Y/W-135-DT conjugate vaccine formulated in isotonic aqueous 1× PBS. UPLC C-18 column and the conditions described distinctly resolved the 2,4-DNPH-HCHO adduct from the un-reacted 2,4-DNPH as detected by TUV detector at 360 nm. This method was shown to be compatible with PBS formulation and extremely sensitive (with a quantitation limit of 0.05 ppm) and aided to determine formaldehyde contamination sources by evaluating the in-process materials as a track-down analysis. Final nanogram levels of formaldehyde in the formulated single dose vialed vaccine mainly originated from the diphtheria toxoid carrier protein used in the production of the conjugate vaccine, whereas relative contribution from polysaccharide API was minimal. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Simultaneous Determination of Eight Hypotensive Drugs of Various Chemical Groups in Pharmaceutical Preparations by HPLC-DAD.

    Science.gov (United States)

    Stolarczyk, Mariusz; Hubicka, Urszula; Żuromska-Witek, Barbara; Krzek, Jan

    2015-01-01

    A new sensitive, simple, rapid, and precise HPLC method with diode array detection has been developed for separation and simultaneous determination of hydrochlorothiazide, furosemide, torasemide, losartane, quinapril, valsartan, spironolactone, and canrenone in combined pharmaceutical dosage forms. The chromatographic analysis of the tested drugs was performed on an ACE C18, 100 Å, 250×4.6 mm, 5 μm particle size column with 0.0.05 M phosphate buffer (pH=3.00)-acetonitrile-methanol (30+20+50 v/v/v) mobile phase at a flow rate of 1.0 mL/min. The column was thermostatted at 25°C. UV detection was performed at 230 nm. Analysis time was 10 min. The elaborated method meets the acceptance criteria for specificity, linearity, sensitivity, accuracy, and precision. The proposed method was successfully applied for the determination of the studied drugs in the selected combined dosage forms.

  15. Development and Validation of a RP-HPLC Method for Determination of Nimodipine in Sustained Release Tablets

    Directory of Open Access Journals (Sweden)

    Xiaojun Shang

    2013-01-01

    Full Text Available A rapid, sensitive, and reproducible reverse phase high performance liquid chromatographic (RP-HPLC method with UV detector for the determination of nimodipine in sustained release tablets was developed. The method involved using a SinoChoom ODS-BP C18 reversed phase column (5 μm, 4.6 mm × 200 mm and mobile phase consisting of methanol-acetonitrile-water (35 : 38 : 27, v/v. The flow rate is 1.0 mL/min, the UV detector was operated at 237 nm, and the column was maintained at 25°C. The method was validated according to official compendia guidelines. The calibration curve of nimodipine for RP-HPLC method was linear over the range of 10–100 μg/mL. The retention time was found at 7.50 min for nimodipine. The variation for interday and intraday assay was found to be less than 0.72%. The proposed RP-HPLC was proved to be suitable for the determination of nimodipine in sustained release tablets.

  16. [Species differentiation and quality assessment of Ziziphus jujuba var. spinosa based on the HPLC fingerprint and quantitative analysis].

    Science.gov (United States)

    Zhang, Cui-Ying; Wang, Jie; Guo, Li-Li; Cheng, Hui-Ping; Cui, Han-Ming; Wu, Ping; Dong, Yu; Huang, Shi-Jing

    2012-11-01

    To establish an HPLC method of a characteristical chemical fingerprint analysis in combination with simultaneous determination of four bioactive components for species differentiation and quality assessment of Ziziphus jujuba var. spinosa. The chromatographic separation was performed on an Agilent TC-C18 BDS (250 mm x 4.6 mm, 5 microm) column. The mobile phase consisted of acetonitrile and water in a linear gradient elution procedure. The evaporator tube temperature of ELSD was set at 110.5 degrees C with the nebulizing gas flow rate of 3.1 mL/min and the flow rate of mobile phase was 1.0 mL/min. The column was maintained at 30 degrees C. The injection volume was 20 microL. HPLC methodology for both chemical fingerprint analysis and quantitative determination of four active ingredients were validated, respectively. According to the contents of the four ingredients and the chemical fingerprints of Ziziphus jujuba var. spinosa using principal component analysis, Ziziphus jujuba var. spinosa was different from the fake derived from the seeds of Ziziphus mauritiana. The developed HPLC method is exclusive and repetitive for the species identification and quality evaluation of Ziziphus jujuba var. spinosa.

  17. Determination of Diphenylamine in Gunshot Residue by HPLC-MS/MS

    Directory of Open Access Journals (Sweden)

    Hongcheng Mei

    2016-01-01

    Full Text Available A high performance liquid chromatography tandem mass spectrometry/mass spectrometry (HPLC-MS/MS protocol was developed for the determination of diphenylamine (DPA. Four productions of DPA were selected for qualitative assay and the peak area of the main product ion for quantitation. By means of separation using an Agilent Extend-C18 column (CA, USA (150 mm × 4.6 mm, 5 μm with methanol-water (90:10 as the mobile phase, DPA was detected by electrospray ionization (ESI tandem mass spectrometry in positive mode. The linearity of the peak area versus concentration ranged 5-500 ng/mL, r 2 = 0.9978. The limit of detection (S/N =3 of this method was 0.3 ng/mL. This method is applicable for the determination of DPA in gunshot residue.

  18. A Validated RP-HPLC Method for theDetermination of Impurities in Montelukast Sodium

    Directory of Open Access Journals (Sweden)

    N. Rashmitha

    2010-01-01

    Full Text Available The present paper describes the development of a reverse phase chromatographic (RPLC method for montelukast sodium in the presence of its impurities and degradation products generated from forced degradation studies. The drug substance was subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal degradation. The degradation of montelukast sodium was observed under acid and oxidative environment. The drug was found to be stable in other stress conditions studied. Successful separation of the drug from the process impurities and degradation products formed under stress conditions were achieved on an Atlantis dC18 (250 x 4.6 mm 5 μm column. The gradient LC method employs solution A and solution B as mobile phase. The solution A contains aqueous 0.1% OPA and solution B contains a mixture of water, acetonitrile (5:95 v/v. The HPLC method was developed and validated with respect to linearity, accuracy, precision, specificity and ruggedness.

  19. Development and Validation of HPLC-PDA Assay method of Frangula emodin

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    Deborah Duca

    2016-03-01

    Full Text Available Frangula emodin, (1,3,8-trihydroxy-6-methyl-anthraquinone, is one of the anthraquinone derivatives found abundantly in the roots and bark of a number of plant families traditionally used to treat constipation and haemorrhoids. The present study describes the development and subsequent validation of a specific Assay HPLC method for emodin. The separation was achieved on a Waters Symmetry C18, 4.6 × 250 mm, 5 μm particle size, column at a temperature of 35 °C, with UV detection at 287 and 436 nm. An isocratic elution mode consisting of 0.1% formic acid and 0.01% trifluoroacetic acid as the aqueous mobile phase, and methanol was used. The method was successfully and statistically validated for linearity, range, precision, accuracy, specificity and solution stability.

  20. RP - HPLC method for the estimation of Tamsulosin Hydrochloride in Tablet Dosage Form.

    Science.gov (United States)

    Kumari, Richa; Dash, P P; Lal, V K; Mishra, A; Murthy, P N

    2010-11-01

    A rapid and sensitive reverse phase RP-HPLC method is proposed for the estimation of tamsulosin hydrochloride in tablets. Tamsulosin hydrochloride was chromatographed on a reverse phase C18 column with a mobile phase consisting of acetonitrile and water in the ratio of 50:50 v/v. The mobile phase was pumped at a flow rate of 1.5 ml/min. The eluents were monitored at 214 nm. The retention time of the drug was 1.7 min. With this method, linearity was observed between area under curve and concentration of tamsulosin hydrochloride in the injected solution, in the range of 5 to 100 μg/ml. The method was found to be applicable for analysis of the drug in tablets. The results were validated statistically.

  1. A Validated RP – HPLC Method for Simultaneous Estimation of Cefixime and Cloxacillin in Tablets

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    G. Rathinavel

    2008-01-01

    Full Text Available This paper presents a RP-HPLC method for the simultaneous estimation of cefixime and cloxacillin in tablets. The process was carried out on C18 column (5 μm, 25 cm × 4.6 mm, i.d using phosphate buffer (pH 5.0, acetonitrile and methanol in the ratio 80:17:3 respectively as a mobile phase at a flow rate of 2mL/min. Wavelength was fixed at 225 nm. The retention time of cefixime and cloxacillin was found to be 5.657 and 6.200 min, respectively. The developed method is rapid and sensitive and it can be used for estimation of combination of these drugs in tablets.

  2. A Validated HPLC Method for the Determination of Vanillyl Butyl Ether in Cosmetic Preparations

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    Francisco Ríos

    2017-02-01

    Full Text Available A specific HPLC (High-Performance Liquid Chromatography method has been developed and validated for the determination of vanillyl butyl ether in cosmetic products. The extraction procedure with an isopropanol water 1:1 mixture is described. The method uses a RP-C-18 column with isocratic elution and an ultraviolet (UV detector. The mobile phase consists of a mixture of acetonitrile and buffer (Na2HPO4 20 mM in water (30:70 v/v with a variable flow rate. The method was validated with respect to accuracy, precision (repeatability and reproducibility, specificity and linearity. The procedure described here is simple, selective and reliable for routine quality control analysis and stability tests of commercially available cosmetic products.

  3. Determination of Arctiin and Arctigenin Contents in Arctium Tomentosum Mill. by HPLC Method

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    Xiaoying Zhou

    2011-01-01

    Full Text Available A simple, precise, rapid and accurate, binary-phase high performance liquid chromatographic method has been developed for the determination of arctiin and arctigenin contents in the Arctium tomentosumMill. with short run time. Chromatographic separation was achieved by using HPLC system, consisting of a Shimadzu LC-6AD and Kromasil C18 column (250×4.6 mm, 5 μm, with pre-column, the mobile phase consists of methanol and water (55: 45. Detection wavelength was 280 nm. The speed of flow was 1.0 mL/min. The specimen handing quantity was 10 μL. The arctiin’s linearity range was 1.575∼4.725 μg (r=0.9995. The arctigenin’s linearity range was 0.613, 3.063 μg (r = 0.9998 and the linear relationship was accurate. The average recovery (n=5 of arctiin and arctigenin were 101.55% (RSD=2.23% 101.63% (RSD =1.49 % respectively. The contents of arctiin and arctigenin in Arctium tomentosum Mill. were 10.69 mg/g and 0.15 mg/g, respectively. Therefore, the developed HPLC method can be applied to both in vitro studies of arctiin and arctigenin formulations as well as drug estimation in biological samples.

  4. [Determination of oleanolic acid in fufang zhenqin feining capsules by RP-HPLC].

    Science.gov (United States)

    Rong, Lu-Ping; Chen, Yi-Shen; Liao, Hua-Wei; Wan, Zheng-Lan

    2013-08-01

    To investigate the optimal technology of extraction of fufang zhenqin feining capsules and establish a method for determination its oleanolic acid. Orthogonal test was employed for selecting the optimum extraction technology by the index of the content of oleanolic acid in extraction. HPLC was performed on a C18 (250 mm x 4.6 mm, 5 microm) column using methanol-0.3% triethylamine solution (80: 20) as the mobile phase at a flow rate of 1 ml/min. The detection wavelength was set at 210 nm. The column temperature was 30 degrees C and the injection size was 25 microL. The optimum extraction technology was as follows: the concertration of alcohol was 70%, the ratio of solid/material was 8: 1, extracting 3 times with half an hour for each time. The linear range of oleanolic acid was 59.68 - 596.8 microg/ml (r = 0.9997), the average recovery was 99.79% (RSD = 0.5%). The considerable extraction rate of active components in the drugs is achieved by applying the selected technology, and the simple method is fit for production. The established HPLC method is simple, accurate, special and can be used for the quality control of fufang zhenqin feining capsules.

  5. Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC

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    Cheng Bai

    2007-01-01

    Full Text Available High-performance liquid chromatography (HPLC analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and quantitative analysis of asparagine and citrulline using the Platinum EPS reverse-phase C18 column (Alltech Associates. Asparagine alone is eluted earlier than citrulline alone, but when both of them are present in biological samples they may co-elute. HPLC retention times for asparagine and citrulline were influenced by other ureides in the mixture. We found that at various asparagines and citrulline ratios [= 3:1, 1:1, and 1:3; corresponding to 75:25, 50:50, and 25:75 (μMol ml–1/μMol ml–1], the resulting peak exhibited different retention times. Adjustment of ureide ratios as internal standards enables peak identification and quantification. Both chemicals were quantified in xylem sap samples of pecan [Carya illinoinensis (Wangenh. K. Koch] trees. Analysis revealed that tree nickel nutrition status affects relative concentrations of Urea Cycle intermediates, asparagine and citrulline, present in sap. Consequently, we concluded that the HPLC methods are presented to enable qualitative and quantitative analysis of these metabolically important ureides.

  6. Simultaneous quantitative determination of five alkaloids in Catharanthus roseus by HPLC-ESI-MS/MS.

    Science.gov (United States)

    Zhang, Lin; Gai, Qing-Hui; Zu, Yuan-Gang; Yang, Lei; Ma, Yu-Liang; Liu, Yang

    2014-10-01

    To establish a method to simultaneously determine the main five alkaloids of Catharanthus roseus for trace samples, a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) analysis method was developed. The five Catharanthus alkaloids, vinblastine, vincristine, vinleurosine, vindoline, and catharanthine were chromatographically separated on a C18 HPLC column. The mobile phase was methanol-15 nmol·L(-1) ammonium acetate containing 0.02% formic acid (65 : 35, V/V). The quantification of these alkaloids was based on the Multiple Reaction Monitoring (MRM) mode. This method was validated, and the results achieved the aims of the study. The intra- and inter-day precision and accuracy of the five alkaloids were within 1.2%-11.5% (RSD%) and -10.9%-10.5% (RE%). The recovery rates of the five alkaloids of samples were from 79.9% to 91.5%. The five analytes were stable at room temperature for 2 h, at 4 °C for 12 h, and at -20 °C for two weeks. The developed method was applied successfully to determine the content of the five alkaloids in three plant parts of three batches of C. roseus with a minute amount collected from three regions of China. The HPLC-ESI-MS/MS method can be used for the simultaneous determination of five important alkaloids in trace C. roseus samples. Copyright © 2014 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.

  7. Spectrophotometric and HPLC determination of deflazacort in pharmaceutical dosage forms

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    Amarilis Scremin

    2010-06-01

    Full Text Available Deflazacort (DFZ is a glucocorticoid used as an anti-inflammatory and immunosuppressant drug. No official methods are available for DFZ determination in pharmaceutical formulations. The objective of this study was to develop, validate and compare spectrophotometric (UV and colorimetric and high-performance liquid chromatography (HPLC methods, for the quantitative determination of DFZ in tablets and oral suspension. For the UV method, ethanol was used as the solvent, with detection at 244 nm. The colorimetric method was based on the redox reaction with blue tetrazolium in alkaline medium, with detection at 524 nm. The method by HPLC was carried out using a C18 column, mobile phase consisting of acetonitrile:water (80:20, v/v with a flow rate of 1.0 mL min-1 and detection at 244 nm. The methods proved linear (r > 0.999, precise (RSD 97%. Statistical analysis of the results indicated that the UV and HPLC methods were statistically equivalent, while the values obtained for the colorimetric method differed significantly from the other methods.O deflazacorte (DFZ é um fármaco glicocorticóide usado como antiinflamatório e imunossupressor. Métodos oficiais não estão disponíveis para a determinação de DFZ em formas farmacêuticas. Este estudo teve como objetivo desenvolver, validar e comparar métodos por espectrofotometria (UV e colorimetria e cromatografia líquida de alta eficiência (CLAE, na determinação quantitativa de DFZ em comprimidos e suspensão oral. O método por UV utilizou etanol como solvente, com detecção em 244 nm. O método colorimétrico foi baseado na reação de redução com azul de tetrazólio em meio alcalino, com detecção em 524 nm. O método por CLAE utilizou coluna C18; fase móvel constituída de acetonitrila:água (80:20, v/v, com fluxo de 1,0 mL min-1 e detecção em 244 nm. Os métodos foram lineares (r > 0,999; precisos (RSD 97%. As análises estatísticas dos resultados obtidos indicaram que os m

  8. Factors influencing the separation of oligonucleotides using reversed-phase/ion-exchange mixed-mode high performance liquid chromatography columns.

    Science.gov (United States)

    Biba, Mirlinda; Jiang, Eileen; Mao, Bing; Zewge, Daniel; Foley, Joe P; Welch, Christopher J

    2013-08-23

    New mixed-mode columns consisting of reversed-phase and ion-exchange separation modes were evaluated for the analysis of short RNA oligonucleotides (∼20mers). Conventional analysis for these samples typically involves using two complementary methods: strong anion-exchange liquid chromatography (SAX-LC) for separation based on charge, and ion-pair reversed-phase liquid chromatography (IP-RPLC) for separation based on hydrophobicity. Recently introduced mixed-mode high performance liquid chromatography (HPLC) columns combine both reversed-phase and ion-exchange modes, potentially offering a simpler analysis by combining the benefits of both separation modes into a single method. Analysis of a variety of RNA oligonucleotide samples using three different mixed-mode stationary phases showed some distinct benefits for oligonucleotide separation and analysis. When using these mixed-mode columns with typical IP-RPLC mobile phase conditions, such as ammonium acetate or triethylammonium acetate as the primary ion-pair reagent, the separation was mainly based on the IP-RPLC mode. However, when changing the mobile phase conditions to those more typical for SAX-LC, such as salt gradients with NaCl or NaBr, very different separation patterns were observed due to mixed-mode interactions. In addition, the Scherzo SW-C18 and SM-C18 columns with sodium chloride or sodium bromide salt gradients also showed significant improvements in peak shape. Copyright © 2013 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2010-01-01

    We prepared hybrid particle-monolithic polymethacrylate columns for micro-HPLC by in situ polymerization in fused silica capillaries pre-packed with 3-5microm C(18) and aminopropyl silica bonded particles, using polymerization mixtures based on laurylmethacrylate-ethylene dimethacrylate (co)polymers for the reversed-phase (RP) mode and [2-(methacryloyloxy)ethyl]-dimethyl-(3-sulfopropyl) zwitterionic (co)polymers for the hydrophilic interaction (HILIC) mode. The hybrid particle-monolithic columns showed reduced porosity and hold-up volumes, approximately 2-2.5 times lower in comparison to the pure monolithic columns prepared in the whole volume of empty capillaries. The elution volumes of sample compounds are also generally lower in comparison to packed or pure monolithic columns. The efficiency and permeability of the hybrid columns are intermediate in between the properties of the reference pure monolithic and particle-packed columns. The chemistries of the embedded solid particles and of the interparticle monolithic moiety in the hybrid capillary columns contribute to the retention to various degrees, affecting the selectivity of separation. Some hybrid columns provided improved separations of proteins in comparison to the reference particle-packed columns in the reversed-phase mode. Zwitterionic hybrid particle-monolithic columns show dual mode retention HILIC/RP behaviour depending on the composition of the mobile phase and allow separations of polar compounds such as phenolic acids in the HILIC mode at lower concentrations of acetonitrile and, often in shorter analysis time in comparison to particle-packed and full-volume monolithic columns. Copyright 2009 Elsevier B.V. All rights reserved.

  10. Simple and reliable methods for the determination of sixteen marker components for quality control of Daochi pill by HPLC coupled with diode array detection.

    Science.gov (United States)

    Lu, Binan; Liu, Yuetao; Yin, Lianhong; Wang, Xiaona; Peng, Jinyong

    2009-01-01

    Traditional Chinese medicine plays a very important role in the healthcare system of China and thus the quality control of medicinal herb products is of paramount concern. To establish a simple and effective high-performance liquid chromatography (HPLC) method to evaluate the quality of Daochi pill. Two HPLC methods were developed for the determination of 16 marker components in Daochi pills. In method A, the analytes were separated on a Lichrosorb C(18) column using a gradient elution of methanol and 1% aqueous phosphoric acid (pH 2.9, adjusted by triethylamine). In method B, the separation was achieved on an Agilent Eclipse Plus C(18) column using a gradient elution of methanol and 3% aqueous phosphoric acid (pH 2.0, adjusted by triethylamine) in a gradient elution mode. Methods were linear over the range 0.27-500 microg/mL (r(2) >or= 0.9995). Accuracy, precision and repeatability were all within the required limits. The mean recoveries measured at three concentrations were higher than 95% with RSD quality evaluation and control of this natural product.

  11. A Comparative Study of Newly Developed HPLC-DAD and UHPLC-UV Assays for the Determination of Posaconazole in Bulk Powder and Suspension Dosage Form

    Directory of Open Access Journals (Sweden)

    Dalia A. Hamdy

    2014-01-01

    Full Text Available Objective. To develop and compare HPLC-DAD and UHPLC-UV assays for the quantitation of posaconazole in bulk powder and suspension dosage form. Methods. Posaconazole linearity range was 5–50 μg/mL for both assays. For HPLC-DAD assay, samples were injected through Zorbax SB-C18 (4.6 × 250 mm, 5 μm column. The gradient elution composed of the mobile phase acetonitrile: 15 mM potassium dihydrogen orthophosphate (30 : 70 to 80 : 20, linear over 7 minutes pumped at 1.5 mL/min. For UHPLC-UV assay, samples were injected through Kinetex-C18 (2.1 × 50 mm, 1.3 μm column. The mobile phase composed of acetonitrile: 15 mM potassium dihydrogen orthophosphate (45 : 55 pumped isocratically at 0.4 mL/min. Detection wavelength was 262 nm in both methods. Results. The run time was 11 and 3 minutes for HPLC-DAD and UHPLC-UV assays, respectively. Both assays were linear (r2>0.999 with CV% and % error of the mean <3%. Limits of detection and quantitation were 0.82 and 2.73 μg/mL for HPLC-DAD and 1.04 and 3.16 μg/mL for UHPLC-UV, respectively. The methods quantitated PSZ in suspension dosage form with no observable interferences. Conclusions. Both assays were proven sensitive and selective according to ICH guidelines. UHPLC-UV assay exhibited some economic and chromatographic separation superiority.

  12. A rapid and fully automatic method for the accurate determination of a wide carbon-chain range of per- and polyfluoroalkyl substances (C4-C18) in human serum.

    Science.gov (United States)

    Gao, Ke; Gao, Yan; Li, Yili; Fu, Jianjie; Zhang, Aiqian

    2016-11-04

    A rapid and fully automatic method for determining 21 per- and polyfluoroalkyl substances (with carbon chains ranging from C4 to C18, including 13 PFCAs, 5 PFSAs, 2 Cl-PFESAs, and PFOSA) in human serum samples was developed. The HPLC parameters, Turboflow column, mobile phase, sample injection volume, loading flow rate, and sample cleanup and elution time were optimized. 25μL serum sample was directly injected into the developed on-line Turboflow SPE HPLC-MS/MS system for analysis after dilution. Matrix effects were corrected due to the matrix removal efficiency of the Turboflow column and sufficient types of internal isotope standards that were used. The established method showed a good linearity (r2>0.99), rapid processing time (20min per sample), satisfactory recoveries (matrix spiked recoveries range from 84.6% to 114%) and precision (intra-day and inter-day RSDs ranged from 1.5% to 9.2% and from 1.1% to 7.0%, respectively). The limits of detection (LODs) of the 21 analyzed PFASs were between 0.008 and 0.19ngmL-1. The LODs of short- and long-chain PFASs, such as PFBA, PFPeA, PFHxDA, and PFODA, were 0.008, 0.022, 0.15 and 0.19ngmL-1, respectively; the spiked recoveries of these PFASs were 101, 105, 87.1, and 85.8%, respectively. Both the LODs and recoveries were better than previous studies. Further, serum PFASs concentrations detected by the presented on-line SPE method were consistent with the traditional off-line SPE method (r: 0.98-0.99), which verified the accuracy and applicability of the present method. The method shows good practical prospects in the analysis of trace per- and polyfluoroalkyl substances in human serum. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. [Qualitative and quantitative analysis of major constituents in Tetrastigma hemsleyanum by HPLC-Q-TOF-MS and UPLC-QqQ-MS].

    Science.gov (United States)

    Xu, Wen; Fu, Zhi-qin; Lin, Jing; Huang, Xue-cheng; Chen, Dan; Yu, Hong-min; Huang, Ze-hao; Fan, Shi-ming

    2014-11-01

    A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.

  14. Comparison of fused-core and conventional particle size columns by LC-MS/MS and UV: application to pharmacokinetic study.

    Science.gov (United States)

    Song, Wei; Pabbisetty, Deepthi; Groeber, Elizabeth A; Steenwyk, Rick C; Fast, Douglas M

    2009-10-15

    The chromatographic performance of fused-core (superficially porous) HPLC packing materials was compared with conventional fully porous particle materials for LC-MS/MS analysis of two pharmaceuticals in rat plasma. Two commercially available antidepressants, imipramine and desipramine, were assayed using a conventional analytical C(18) column (5 microm, 2.0 mm x 30 mm) and a fused-core C(18) column (2.7 microm, 2.1 mm x 30 mm). Retention time, column efficiency, pressure drop, resolution, and loading capacity were compared under the same operating conditions. The fused-core column demonstrated reduced assay time by 34% and 2-3-fold increased efficiency (N). Loading capacity up to 25 microl of extract injected on column showed no peak distortion. The registered back-pressure from a flow rate of 1.0 ml/min did not exceed 3400 psi making it compatible with standard HPLC equipment (typically rated to 6000 psi). Two mobile phases were examined, and morpholine as an organic base modifier yielded a 2-5-fold increase in S/N near the limit of detection over triethylamine. The 2.7 microm fused-core column was applied to the analysis of imipramine and desipramine in extracted, protein precipitated rat plasma by LC-MS/MS. The calibration curves were linear in the concentration range of 0.5-1000 ng/ml for both imipramine and desipramine. Intra-run precisions (%CV) and accuracies (%bias) were within +/-7.8% and +/-7.3% at three QC levels and within 14.7% and 14.4% at the LOQ level for both analytes. Following a single method qualification run, the method was applied to the quantitation of pharmacokinetic study samples after oral administration of imipramine to male rats.

  15. A simple method for simultaneous determination of basic dyes encountered in food preparations by reversed-phase HPLC.

    Science.gov (United States)

    Dixit, Sumita; Khanna, Subhash K; Das, Mukul

    2011-01-01

    The present method utilizes a simple pretreatment step, cleanup on polyamide SPE cartridges, and HPLC resolution on reversed-phase C18 for the detection of the three basic nonpermitted dyes encountered in food matrixes. Polyamide cartridges were chosen because both acidic and basic dyes can be cleaned up due to their amphoteric nature. Analysis was performed on a reversed-phase C18 micro-Bondapak column using the isocratic mixture of acetonitrile-sodium acetate with a flow rate of 1.5 mL/min and a programmable lambda(max) specific visible detection to monitor colors, achieving higher sensitivity and expanded scope to test multicolor blends. All the colors showed linearity with the regression coefficient, from 0.9983 to 0.9995. The LOD and LOQ ranged between 0.107 and 0.754 mg/L and 0.371 and 2.27 mg/L or mg/kg, respectively. The intraday and interday precision gave good RSDs, and percentage recoveries in different food matrixes ranged from 75 to 96.5%. The study demonstrates that the use of a combination of a simple SPE cleanup and HPLC resolution with UV-Vis end point detection was successful in screening the presence of these three basic nonpermitted dyes individually or in blend, in a variety of food matrixes.

  16. Oxidative stability of structured lipids containing C18:0, C18:1, C18:2, C18:3 or CLA in sn 2-position - as bulk lipids and in milk drinks

    DEFF Research Database (Denmark)

    Timm Heinrich, Maike; Nielsen, Nina Skall; Xu, Xuebing

    2004-01-01

    In this study, we compared the oxidative stability of a specific structured lipid (SL) containing conjugated linoleic acid (CLA) in the sn2-position with SL containing other C18 fatty acids of different degree of unsaturation (stearic, oleic, linoleic or linolenic acid). SL was produced by enzyma......In this study, we compared the oxidative stability of a specific structured lipid (SL) containing conjugated linoleic acid (CLA) in the sn2-position with SL containing other C18 fatty acids of different degree of unsaturation (stearic, oleic, linoleic or linolenic acid). SL was produced...... by enzymatic interesterification with caprylic acid. Oxidative stability was compared in the five lipids themselves and in milk drinks containing 5% of the different SL. During storage, samples were taken for chemical and physical analyses. Moreover, sensory assessments were performed on milk drinks....... The oxidative stability of our SL was very different when comparing (a) bulk lipids and milk drink and (b) the five different batches of each product. SL based on oleic acid was the most unstable as bulk lipid, while SL based on linoleic acid was the most unstable in milk drink. SL based on CLA was the second...

  17. Modeling of chromatographic lipophilicity of food synthetic dyes estimated on different columns.

    Science.gov (United States)

    Sârbu, Costel; Casoni, Dorina; Kot-Wasik, Agata; Wasik, Andrzej; Namieśnik, Jacek

    2010-08-01

    The retention behavior of some food synthetic dyes has been studied by RP-HPLC on chemically bonded C18, C8, C16 and CN stationary phases. Using methanol-ammonium acetate (0.08 mol/L, pH=6.76) as mobile phase, a linear behavior of retention parameters throughout the methanol fraction variance was obtained in all cases (r>0.99). The patterns of chromatographic behavior of the compounds illustrate high similarities between the C18, C8 and C16 columns, respectively. Highly significant correlations were obtained between experimental lipophilicity indices log k(w) and phi(0) estimated on C18 and C8 stationary phases and some computed log P-values. An extensive investigation made for quantitative structure-property (lipophilicity) relationships of studied dyes, using descriptors from Dragon software, multiple linear regression and genetic algorithm, revealed that the molecular descriptors appearing in the best models combine 2-D and 3-D aspects of the molecular structure. The most significant descriptors can be classified as radial distribution function, GETAWAY (autocorrelation), 3D-MoRSE signal, Burden eigenvalues and edge adjacency descriptors.

  18. Content determination of obeticholic acid tablets by HPLC

    Directory of Open Access Journals (Sweden)

    Yunxia LU

    2017-04-01

    Full Text Available In order to establish a method for the content determination of obeticholic acid tablets, HPLC-UV method is adopted. The determination is performed on Agilent HC-C18 column(250 mm×4.6 mm,5 μm) with mobile phase acetonitrile (002% formic acid-water (0.02% formic acid = 60∶40 (V/V at the flow rate of 1.0 mL/min. The detection wavelength is 195 nm, the column temperature is 30 ℃ and the volume is 100 μL. The result shows that there is a good liner relationship between the mass concentration of obeticholic acid in the range of 0.200 26~1.001 30 mg/mL and the peak area, and r=0.999 9. The RSD of precision, stability and reproducible tests are all less than 1.0%. The average recovery rate is 99.64% , and RSD is 0.58%(n=9. The method is simple, exclusive, accurate and durable, and can be used for the content determination of obeticholic acid tablets.

  19. Identification of Rhodiola species by using RP-HPLC*

    Science.gov (United States)

    Wang, Qiang; Ruan, Xiao; Jin, Zhi-hua; Yan, Qi-chuan; Tu, Shan-jun

    2005-01-01

    An approach was established using RP-HPLC (reversed-phase high-performance liquid chromatography) to identify ten species of Rhodiola, R. coccinea A. Bor, R. junggarica C.Y. Yang et N.R. Cui spn., R. heterodonta A. Bor, R. linearifolia A. Bor, R. pamiro alaiucm A. Bor, R. kaschgarica A. Bor, R. litwinowii A. Bor, R. gelida schrenk, R. rosea L. and R. quadrifide Fisch et Mey collected from the Tianshan Mountains areas of China. Chromatograms of alcohol-soluble proteins, generated from these ten Rhodiola spp. were compared. Each chromatogram of alcohol-soluble proteins came from a single seed of one wild species 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 22%–55% solvent B with a flow rate of 1 ml/min and a run time of 67 min, the chromatography gave optimum separation of Rhodiola alcohol-soluble proteins. Chromatogram of each species was different and could be used to identify those species. Cluster analysis of genetic similarity coefficients of 37% to 60% showed a medium degree of genetic diversity among the species in these eco-areas. Cluster analysis showed that the ten species of Rhodiola can be divided into four clusters and yielded the general and unique biochemical markers of these species. RP-HPLC was shown to be a rapid, repeatable and reliable method for Rhodiola species identification and analysis of genetic diversity. PMID:15909330

  20. Lipstick dermatitis due to C18 aliphatic compounds.

    Science.gov (United States)

    Hayakawa, R; Matsunaga, K; Suzuki, M; Arima, Y; Ohkido, Y

    1987-04-01

    An 18-year-old girl developed cheilitis. She had a past history of lip cream dermatitis, but the cause was not found. Patch tests with 2 lipsticks were strongly positive. Tests with the ingredients were positive to 2 aliphatic compounds, glyceryl diisostearate and diisostearyl malate. Impurities in the materials were suspected as the cause. Analysis by gas chromatography detected 3 chemicals in glyceryl diisostearate and 1 in diisostearyl malate as impurities. Patch testing with the impurities and glyceryl monoisostearate 0.01% pet in glyceryl diisostearate and isostearyl alcohol 0.25% pet in diisostearyl malate were strongly positive. The characteristics common to the 2 chemicals were liquidity at room temperature, branched C18 aliphatic compound and primary alcohol. Chemicals lacking any of the above 3 features did not react.

  1. Determination of Dyclonine Hydrochloride by a HPLC Method and Camphor and Menthol by a GC Method in Compound Lotion

    Directory of Open Access Journals (Sweden)

    Suying Ma

    2013-01-01

    Full Text Available A high performance liquid chromatographic (HPLC method with UV detector for the determination of dyclonine hydrochloride and a gas chromatography (GC method with flame ionization detector (FID for the determination of camphor and menthol in lotion were developed. The developed HPLC method involved using a SinoChoom ODS-BP C18 reversed-phase column (5 μm, 4.6 mm × 200 mm and mobile phase consisting of acetonitrile : water : triethylamine in a ratio of 45 : 55 : 1.0; pH was adjusted to 3.5 with glacial acetic acid. The developed GC method for determination of camphor and menthol involved using an Agilent 19091J-413 capillary chromatographic column (30 m × 320 μm × 0.25 μm. The two methods were validated according to official compendia guidelines. The calibration of dyclonine hydrochloride for HPLC method was linear over the range of 20–200 μg/mL. The retention time was found at 6.0 min for dyclonine hydrochloride. The calibration of camphor and menthol of GC method was linear over the range of 10–2000 μg/mL. The retention time was found at 2.9 min for camphor and 3.05 min for menthol. The proposed HPLC and GC methods were proved to be suitable for the determination of dyclonine hydrochloride, camphor, and menthol in lotion.

  2. A Simple HPLC Bioanalytical Method for the Determination of ...

    African Journals Online (AJOL)

    Purpose: To develop a simple, accurate, and precise high performance chromatography (HPLC) method with spectrophotometric detection for the determination of doxorubicin hydrochloride in rat plasma. Methods: Doxorubicin hydrochloride and daunorubicin hydrochloride (internal standard, IS) were separated on a C18 ...

  3. Analysis and theoretical modeling of 18O enriched carbon dioxide spectrum by CRDS near 1.35 μm: (II) 16O13C18O, 16O13C17O, 12C18O2, 17O12C18O, 12C17O2, 13C18O2 and 17O13C18O

    Science.gov (United States)

    Karlovets, E. V.; Campargue, A.; Kassi, S.; Tashkun, S. A.; Perevalov, V. I.

    2017-04-01

    This contribution is the second part of the analysis of the room temperature absorption spectrum of 18O enriched carbon dioxide by very high sensitivity Cavity Ring Down spectroscopy between 6977 and 7918 cm-1 (1.43-1.26 μm). Overall, more than 8600 lines belonging to 166 bands of eleven carbon dioxide isotopologues were rovibrationnally assigned. In a first part (Kassi et al. J Quant Spectrosc Radiat Transfer 187 (2017) 414-425, http://dx.doi.org/10.1016/j.jqsrt.2016.09.002), the results relative to mono-substituted isotopologues, 16O12C18O, 16O12C17O, 12C16O2 and 13C16O2, were presented. This second contribution is devoted to the multiply-substituted isotopologues or clumped isotopologues of particular importance in geochemistry: 16O13C18O, 16O13C17O, 12C18O2, 17O12C18O, 12C17O2, 13C18O2 and 17O13C18O. On the basis of the predictions of effective Hamiltonian models, a total of 3195 transitions belonging to 73 bands were rovibrationnally assigned for these seven species. Among the 73 observed bands, 55 are newly reported. All the identified bands correspond to ΔP=10 and 11 series of transitions, where P= 2V1+V2+3V3 is the polyad number (Vi are vibrational quantum numbers). The accurate spectroscopic parameters of 70 bands have been determined from the standard band-by-band analysis. Global fits of the measured line intensities of the ΔP=10 series of transitions of 17O12C18O and 16O13C18O and of the ΔP=11 series of transitions of 12C18O2, 17O12C18O, 16O13C18O and 13C18O2 were performed to determine the corresponding sets of the effective dipole moment parameters.

  4. Simultaneous determination of eight catechins and four theaflavins in green, black and oolong tea using new HPLC-MS-MS method.

    Science.gov (United States)

    Tao, Wuqun; Zhou, Zhiguang; Zhao, Bin; Wei, Tongyu

    2016-11-30

    A simple, rapid and accurate analytical method was developed for the analysis of eight tea catechins and four theaflavins simultaneously in three types of tea (green, black and oolong tea), using ultra high performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (HPLC-MS-MS) in multiple-reaction monitoring (MRM) mode. This method using HPLC-MS-MS in positive mode was performed on a CAPCELL PAK C18 MGIII (2.0mm×100mm, 3μm) column (Shiseido) with the mobile phase consisting of 0.1% aqueous formic acid (A) and methanol (B) in gradient elution at a flow rate of 0.3mLmin-1, and the column temperature set at 30°C. The optimized HPLC-MS-MS methodology is selective and specific, and was validated for eight catechins and four theaflavins widely reported in different teas. Satisfactory linearity was achieved in linear range (0.02-5μgmL-1 for catechins and 0.02-20μgmL-1 for theaflavins) and fine determination coefficient (r2>0.9935). The recoveries ranged from 65% to 115% with the RSD ranging from 2.4% to 6.7%. The methodology was used to evaluate the target polyphenols concentration in three types of tea samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Rapid screening of tetrodotoxin in urine and plasma of patients with puffer fish poisoning by HPLC with creatinine correction.

    Science.gov (United States)

    Yu, Chung-Him; Yu, Chun-Fai; Tam, Sidney; Yu, Peter Hoi-Fu

    2010-01-01

    A rapid and simple detection method for tetrodotoxin (TTX) in urine and plasma of patients with puffer fish poisoning was developed using commercially pre-packed solid-phase extraction (SPE) cartridges (C18 and weak cation exchange columns) and subsequent analyses by HPLC with UV detection. The detection limit of the standard TTX, TTX-spiked urine and plasma samples were all 10 ng/ml and the average TTX recovery in urine and plasma samples after SPE were 90.3 +/- 4.0 and 87.1 +/- 2.9%, respectively. It was noticed that the creatinine-adjusted urinary TTX levels obtained within the first 24 h of presentation apparently correlated much better with the severity of poisoning than the urinary TTX concentration without adjusting for variations in concomitant creatinine excretion.

  6. Chromatographic fingerprint analysis and simultaneous determination of eight lignans in Justicia procumbens and its compound preparation by HPLC-DAD.

    Science.gov (United States)

    Wang, Linan; Pan, Jianyu; Yang, Meihua; Wu, Jun; Yang, Junshan

    2011-03-01

    HPLC fingerprints were developed for the quality evaluation of Justicia procumbens and its compound preparation, Jian-er syrup, together with the simultaneous quantification of eight arylnaphthalide lignans (6'-hydroxy justicidin B, 6'-hydroxy justicidin A, 6'-hydroxy justicidin C, justicidin B, chinensinaphthol methyl ether, justicidin C, taiwanin C, and neojusticin A). Samples were separated with a Shiseido Capcell Pak C(18) reversed-phase column (250×4.6 mm id, 5 μm) using acetonitrile and water as the mobile phase. The column temperature was maintained at 35°C and the wavelength of detector was set at 256 nm. For fingerprint analysis, 17 peaks were selected as the characteristic peaks for the evaluation of the similarities among different J. procumbens samples collected in different places. The structures of lignans were confirmed by diagnostic fragments in the positive ESI-MS(n) . The new method was successfully applied for the chromatographic fingerprint analysis and simultaneous determination of eight lignans in its compound preparation, Jian-er syrup. All the results indicated that HPLC fingerprint assay in combination with multi-marker determination afforded a useful method for the quality control of J. procumbens and its compound preparation, Jian-er syrup. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Multivariate development and validation of a stability-indicating HPLC method for the determination of glimepiride in tablets.

    Science.gov (United States)

    Bonfilio, Rudy; Peres, Carolina; Salgado, Hérida R N; De Araújo, Magali B; Tarley, César R T

    2013-01-01

    This paper describes the multivariate development of a stability-indicating HPLC method for the quantification of glimepiride in pharmaceutical tablets. Full factorial design, Doehlert design, and response-surface methodology were used in conjunction with the desirability function approach. This procedure allowed the adequate separation of glimepiride from all degradant peaks in a short analysis time (about 9 min). This HPLC method uses potassium phosphate buffer (pH 6.5; 27.5 mmol/L)-methanol (34 + 66, v/v) mobile phase at a flow rate of 1.0 mL/min and UV detection at 228 nm. A Waters Symmetry C18 column (250 x 4.6 mm, 5.0 pm) at controlled room temperature (25 degrees C) was used as the stationary phase. The method was validated according to International Conference on Harmonization guidelines and demonstrated linearity from 2 to 40 mg/L glimepiride, selectivity, precision, accuracy, and robustness. The LOD and LOQ were 0.315 and 1.050 mg/L, respectively. The multivariate strategy adopted in this work can be successfully applied in routine laboratories because of its fast optimization without the additional cost of columns or equipment.

  8. A new validated HPLC method for the determination of sulforaphane: application to study pharmacokinetics of sulforaphane in rats.

    Science.gov (United States)

    Ong, Chau; Elbarbry, Fawzy

    2016-07-01

    A simple, accurate and reproducible high-performance liquid chromatography (HPLC) method has been developed and validated for the quantification of sulforaphane (SF) in rat plasma. The method involves a simple liquid-liquid extraction procedure to extract both SF and 7-hyrdoxycoumarin, the internal standard. The chromatographic analysis was achieved on a Shimadzu LC 20A HPLC system equipped with a Zorbax Eclipse XDB C18 column and an isocratic mobile phase consisting of 10 mm KH2 PO4 (pH 4.5) and acetonitrile HPLC grade (40:60, v/v) run at a flow rate of 1 mL/min for 10 min. The UV detection wavelength was set at 202 nm. The method exhibited good linearity (R(2) > 0.999) over the assayed concentration range (0.05-2 μg/mL) and demonstrated good intra- and inter-day precision and accuracy (relative standard deviations and the deviation from predicted values were <15%). This method was also successfully applied for studying the pharmacokinetics of SF in spontaneously hypertensive rats following single oral dietary doses of SF. The pharmacokinetics of SF show linear behavior at the dose range investigated in this study. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  9. Development and validation of a stability-indicating RP–HPLC method for estimation of atazanavir sulfate in bulk

    Directory of Open Access Journals (Sweden)

    S. Dey

    2017-04-01

    Full Text Available A stability-indicating reverse phase–high performance liquid chromatography (RP–HPLC method was developed and validated for the determination of atazanavir sulfate in tablet dosage forms using C18 column Phenomenix (250 mm×4.6 mm, 5 μm with a mobile phase consisting of 900 mL of HPLC grade methanol and 100 mL of water of HPLC grade. The pH was adjusted to 3.55 with acetic acid. The mobile phase was sonicated for 10 min and filtered through a 0.45 μm membrane filter at a flow rate of 0.5 mL/min. The detection was carried out at 249 nm and retention time of atazanavir sulfate was found to be 8.323 min. Linearity was observed from 10 to 90 μg/mL (coefficient of determination R2 was 0.999 with equation, y=23.427x+37.732. Atazanavir sulfate was subjected to stress conditions including acidic, alkaline, oxidation, photolysis and thermal degradation, and the results showed that it was more sensitive towards acidic degradation. The method was validated as per ICH guidelines.

  10. Validated Stability Indicating RP-HPLC Method for Simultaneous Estimation of Codeine Phosphate and Chlorpheniramine Maleate from Their Combined Liquid Dosage Form

    Directory of Open Access Journals (Sweden)

    Ramakrishna Kommana

    2013-01-01

    Full Text Available The present paper describes the development of quick stability indicating RP-HPLC method for the simultaneous estimation of codeine phosphate and chlorpheniramine maleate in the presence of its degradation products, generated from forced degradation studies. The developed method separates codeine phosphate and chlorpheniramine maleate in impurities/degradation products. Codeine phosphate and chlorpheniramine maleate and their combination drug product were exposed to acid, base, oxidation, dry heat, and photolytic stress conditions, and the stressed samples were analysed by proposed method. The proposed HPLC method utilizes the Shimadzu HPLC system on a Phenomenex C18 column (, 5 μ using a mixture of 1% o-phosphoric acid in water : acetonitrile : methanol (78 : 10 : 12 mobile phase with pH adjusted to 3.0 in an isocratic elution mode at a flow rate of 1 mL/min, at 23°C with a load of 20 μL. The detection was carried out at 254 nm. The retention time of codeine phosphate and chlorpheniramine maleate was found to be around 3.47 min and 9.45 min, respectively. The method has been validated with respect to linearity, robustness, precision, accuracy, limit of detection (LOD, and limit of quantification (LOQ. The developed validated stability indicating HPLC method was found to be simple, accurate, and reproducible for the determination of instability of these drugs in bulk and commercial products.

  11. Greener liquid chromatography using a guard column with micellar mobile phase for separation of some pharmaceuticals and determination of parabens.

    Science.gov (United States)

    Youngvises, Napaporn; Chaida, Thanatcha; Khonyoung, Supada; Kuppithayanant, Nattawan; Tiyapongpattana, Warawut; Itharat, Arunporn; Jakmunee, Jaroon

    2013-03-15

    In this research, a greener chromatography employing a short column, Zorbax SB C18 cartridge (12.5 × 4.6 mm, 5 μm) commonly used as a guard column in a reverse phase high performance liquid chromatography (RP-HPLC), was utilized as the analytical column in conjunction with a more eco-friendly micellar mobile phase of sodium dodecyl sulfate (SDS) for separation tertiary mixtures of local anesthetics and antihistamines; and binary mixture of colds drugs; and quaternary mixture of some parabens with different separation conditions. The chromatographic behavior of these analytes was studied to demonstrate separation efficiency of this guard column in a micellar mobile phase. Moreover, this column and SDS mobile phase was exploited for determination of parabens in 64 samples of cosmetic product, both those that were produced locally in the community and those that were commercially manufactured. Linear calibration graphs of the parabens as detected at 254 nm were obtained in the range of 1-100 μmol L(-1) with R(2)>0.9990. Percentage recoveries were 92.4-109.2 with %RSD<3, and the limit of detection and quantitation were 0.04-0.10 and 0.20-0.80 μmol L(-1), respectively. This analytical system is not only greener but also faster and employing simpler sample preparation than a conventional liquid chromatographic system. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Determination of Tramadol in human plasma by HPLC with fluorescence detection

    Directory of Open Access Journals (Sweden)

    Daniela Baconi

    2016-04-01

    Full Text Available Tramadol is a centrally acting analgesic, atypical opioid, and although it is generally considered as a medicinal drug with a low potential for dependence, there is growing evidence of tramadol abuse in some countries. The ultraviolet detection is not suitable for analysis of tramadol in plasma, due to the lack of sensitivity and selectivity. However, it was shown that tramadol has a weak fluorescence, and the latest techniques for determination of tramadol in plasma include liquid chromatographic methods with fluorescence detection (FL. The objective of the paper was to develop a HPLC-FL method applicable for quantification of tramadol in human plasma.The separation was achieved by reverse phase HPLC method, using as stationary phase C18 – Kromasil® column and a mobile phase consisted of acetonitrile:0.1% formic acid (20:80. The fluorescence detection has been applied with λex/em= 280/310 nm. A solid phase extraction procedure using C18 cartridge was carried out. The linearity of the method has been demonstrated in the range of both therapeutic and toxic plasma tramadol levels (concentrations of 0.100 – 1µg/mL. The selectivity, precision, and accuracy of the method have been demonstrated. The limit of detection (LOD= 0.010 µg/mL and the limit of quantification (LOQ = 0.100 µg/mL have been established.The proposed method can be used to assess tramadol levels in human plasma in pharmacokinetic studies, as well as in overdose cases. The utility of the method for the quantification of therapeutic levels of tramadol has been shown on the plasma samples from the patients with tramadol treatment as analgesic (doses ranging from 100 mg to 400 mg/day.The developed method is rapid, using simple experimental conditions and an accurate and short extraction procedure.

  13. Simultaneous determination of olanzapine and fluoxetine hydrochloride in capsules by spectrophotometry, TLC-spectrodensitometry and HPLC.

    Science.gov (United States)

    Tantawy, Mahmoud A; Hassan, Nagiba Y; Elragehy, Nariman A; Abdelkawy, Mohamed

    2013-03-01

    This paper describes sensitive, accurate and precise spectrophotometric, TLC-spectrodensitometric and high performance liquid chromatographic (HPLC) methods for simultaneous determination of olanzapine and fluoxetine HCl. Two spectrophotometric methods were developed, namely; first derivative (D (1)) and derivative ratio (DD (1)) methods. The TLC method employed aluminum TLC plates precoated with silica gel GF254 as the stationary phase and methanol:toluene:ammonia (7:3:0.1, by volume) as the mobile phase, where the chromatogram was scanned at 235 nm. The developed HPLC method used a reversed phase C18 column with isocratic elution. The mobile phase composed of phosphate buffer pH 4.0:acetonitrile:triethylamine (53:47:0.03, by volume) at flow rate of 1.0 mL min(-1). Quantitation was achieved with UV detection at 235 nm. The methods were validated according to the International Conference on Harmonization (ICH) guidelines. The selectivity of the proposed methods was tested using laboratory-prepared mixtures. The developed methods were successfully applied for the determination of olanzapine and fluoxetine HCl in bulk powder and combined capsule dosage form.

  14. Simultaneous determination of olanzapine and fluoxetine hydrochloride in capsules by spectrophotometry, TLC-spectrodensitometry and HPLC

    Directory of Open Access Journals (Sweden)

    Mahmoud A. Tantawy

    2013-03-01

    Full Text Available This paper describes sensitive, accurate and precise spectrophotometric, TLC-spectrodensitometric and high performance liquid chromatographic (HPLC methods for simultaneous determination of olanzapine and fluoxetine HCl. Two spectrophotometric methods were developed, namely; first derivative (D1 and derivative ratio (DD1 methods. The TLC method employed aluminum TLC plates precoated with silica gel GF254 as the stationary phase and methanol:toluene:ammonia (7:3:0.1, by volume as the mobile phase, where the chromatogram was scanned at 235 nm. The developed HPLC method used a reversed phase C18 column with isocratic elution. The mobile phase composed of phosphate buffer pH 4.0:acetonitrile:triethylamine (53:47:0.03, by volume at flow rate of 1.0 mL min−1. Quantitation was achieved with UV detection at 235 nm. The methods were validated according to the International Conference on Harmonization (ICH guidelines. The selectivity of the proposed methods was tested using laboratory-prepared mixtures. The developed methods were successfully applied for the determination of olanzapine and fluoxetine HCl in bulk powder and combined capsule dosage form.

  15. HPLC Quantitative Analysis of Main Stilbenes and Flavones in Different Parts of Matteuccia struthiopteris

    Directory of Open Access Journals (Sweden)

    Shubei Li

    2013-01-01

    Full Text Available A simple and accurate HPLC-UV method was developed for the simultaneous quantitative analysis of main stilbenes and flavones in different parts (fronds, rhizomes, and frond bases of M. struthiopteris. The chromatographic separation was performed on a Kromasil C18 column (4.6 mm × 250 mm, 5 μm with the mobile phase of MeOH-H2O (including 0.1% phosphoric acid using a gradient elution at the flow rate of 1.0 mL min−1 and UV detection at 295 nm. The method was validated by specificity, linearity, accuracy (recovery, and precision tests (repeatability, intra- and interday. For all the six compounds, the linear regression coefficients ranged from 0.9958 to 0.9998 within the test ranges; intra- and interday precisions were <2% and the mean recoveries ranged from 98.09 to 103.56%. The amount of these compounds in the frond bases was almost the same as in the rhizomes but much higher than that in the fronds. The results indicate that the HPLC method developed was appropriate for the analysis of the six compounds in different parts (fronds, rhizomes, and frond bases of M. struthiopteris.

  16. Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine.

    Science.gov (United States)

    Pavlović, Bojana; Cvijetić, Nataša; Dragačević, Luka; Ivković, Branka; Vujić, Zorica; Kuntić, Vesna

    2016-01-01

    One of the most commonly used surfactants in the production of split virus influenza vaccine is nonionic surfactant Triton X-100. After splitting of the virus is accomplished, Triton X-100 is removed from the vaccine by subsequent production steps. Because of toxicity of Triton X-100, which remains in the vaccine in residual amounts, a sufficiently sensitive method for its detection and quantification needs to be defined. Two methods for determination of Triton X-100 residuals were developed: the UV-spectrophotometry and HPLC methods. For both methods, preparation of vaccine samples and removal of proteins and virus particles were crucial: samples were treated with methanol (1:1) and then centrifuged at 25 000 × g for 30 min. After such treatment, the majority of vaccine components that interfered in the UV region were removed, and diluted samples could be directly measured. The chromatographic system included C18 column, step methanol gradient, and detection at 225 nm with a single peak of Triton X-100 at 12.6 min. Both methods were validated and gave satisfactory results for accuracy, precision, specificity, linearity, and robustness. LOQ was slightly lower for the HPLC method. Hence, it was shown that both methods are suitable for analysis of residual amounts of Triton X-100, with the advantages of the UV method being its simplicity and availability in most laboratories.

  17. [Spectrum-effect Relationship Between Total Antioxidant Activity and HPLC Fingerprint of Arctium lappa Root].

    Science.gov (United States)

    Wang, Xiao-juan; Jiang, Lin

    2014-12-01

    To explore the spectrum-effect relationship between the HPLC fingerprint of Arctium lappa root methanol extract and the total antioxidant activity. The experiment was carried out with Gemini C18 110A (250 mm x 4.6 mm, 5 µm) column using methanol-0.04% phosphoric acid as gradient mobile phase at the flow rate of 1.0 mL/min, detection wavelength of 320 nm. The total antioxidant activity was determined by measuring the absorbance of each sample after being reacted with ammonium molybdate reagent. The spectrum-effect relationship was investigated using canonical correlation analysis (CCA). The spectrum-effect relationship between the HPLC fingerprint of Arctium lappa root methanol extract and the total antioxidant activity were established, the similarity of fingerprint of all samples was above 0.9. Peaks 1, 6, 9, 12 and 14 were principle components of Arctium lappa root for the total antioxidant activity. This method contributes to the fast comprehensive evaluation of quality of Arctium lappa root.

  18. UPLC and HPLC of caffeoyl esters in wild and cultivated Arctium lappa L.

    Science.gov (United States)

    Haghi, Ghasem; Hatami, Alireza; Mehran, Mehdi

    2013-05-01

    Analytical methods including ultra-performance liquid chromatography (UPLC) and high-performance liquid chromatography (HPLC) with photodiode array (PDA) detector were developed for the analysis of caffeoylquinic acid derivatives in seeds, leaves and roots of Arctium lappa L. Separation was performed on C(18) column utilising 5% (v/v) acetic acid in water and acetonitrile at 330 nm. Both methodologies were validated in terms of linearity, precision, and recovery. The results showed that the major advantages of UPLC, over HPLC were the fast analysis, narrow peaks, high sensitivity, and reduction of solvent consumption. Subsequently the methods were applied for the identification and quantification of chlorogenic acid (5-CQA) and 1,5-dicaffeoylquinic acid (1,5-DCQA) as main compounds in samples. The total phenolic content of samples ranged from 3.93 to 14.13 g of 5-CQA equivalent/100g dry weight (DW). There was a significant variability from 89 to 571 mg/100g for 5-CQA and 48 to 486 mg/100g for 1,5-DCQA in dry material. Copyright © 2012 Elsevier Ltd. All rights reserved.

  19. HPLC and chemometric assisted spectrophotometric methods for simultaneous determination of diprophylline, phenobarbitone and papaverine hydrochloride.

    Science.gov (United States)

    El-Gindy, Alaa

    2005-09-01

    Three methods are developed for the simultaneous determination of diprophylline (DP), phenobarbitone (PH) and papaverine hydrochloride (PP). The chromatographic method depends on a high performance liquid chromatographic (HPLC) separation on a reversed-phase C18 column with a mobile phase consisting of 0.02 M potassium dihydrogen phosphate, pH 3.5--acetonitrile (55:45 v/v). Quantitation was achieved with UV detection at 210 nm based on peak area. The other two chemometric methods applied were principal component regression (PCR) and partial least squares (PLS-1). These approaches were successfully applied to quantify the three drugs in the mixture using the information included in the UV absorption spectra of appropriate solutions in the range 215-245 nm with the intervals Delta lambda = 0.2 nm. The calibration PCR and PLS-1 models were evaluated by internal validation (prediction of compounds in its own designed training set of calibration), by cross-validation (obtaining statistical parameters that show the efficiency for a calibration fit model) and by external validation over laboratory-prepared mixtures and pharmaceutical preparations. The PCR and PLS-1 methods require neither any separation step, nor any priori graphical treatment of the overlapping spectra of the three drugs in a mixture. The results of PCR and PLS-1 methods were compared with HPLC method obtained in pharmaceutical formulation and a good agreement was found.

  20. Column-to-column packing variation of disposable pre-packed columns for protein chromatography.

    Science.gov (United States)

    Schweiger, Susanne; Hinterberger, Stephan; Jungbauer, Alois

    2017-12-08

    In the biopharmaceutical industry, pre-packed columns are the standard for process development, but they must be qualified before use in experimental studies to confirm the required performance of the packed bed. Column qualification is commonly done by pulse response experiments and depends highly on the experimental testing conditions. Additionally, the peak analysis method, the variation in the 3D packing structure of the bed, and the measurement precision of the workstation influence the outcome of qualification runs. While a full body of literature on these factors is available for HPLC columns, no comparable studies exist for preparative columns for protein chromatography. We quantified the influence of these parameters for commercially available pre-packed and self-packed columns of disposable and non-disposable design. Pulse response experiments were performed on 105 preparative chromatography columns with volumes of 0.2-20ml. The analyte acetone was studied at six different superficial velocities (30, 60, 100, 150, 250 and 500cm/h). The column-to-column packing variation between disposable pre-packed columns of different diameter-length combinations varied by 10-15%, which was acceptable for the intended use. The column-to-column variation cannot be explained by the packing density, but is interpreted as a difference in particle arrangement in the column. Since it was possible to determine differences in the column-to-column performance, we concluded that the columns were well-packed. The measurement precision of the chromatography workstation was independent of the column volume and was in a range of±0.01ml for the first peak moment and±0.007 ml 2 for the second moment. The measurement precision must be considered for small columns in the range of 2ml or less. The efficiency of disposable pre-packed columns was equal or better than that of self-packed columns. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

  1. HPLC determination of selected incestisides in cosmetic

    OpenAIRE

    Kameníčková, Daniela

    2013-01-01

    Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Daniela Kameníčková Supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of Diploma Thesis: HPLC determination of selected incestisides in cosmetic Active ingredients fenoxycarb and permethrin were determined in cosmetic anti- parasitic product Arpalit® Neo shampoo against parasites with bamboo extract. Analysis was performed by HPLC using RP-Amide column 100 x 3 mm with a particl...

  2. Determination of 3 ingredients in Anshenbao granules by HPLC-ELSD

    Directory of Open Access Journals (Sweden)

    Lili LI

    2018-02-01

    Full Text Available In order to improve the quality control standard of Anshenbao granules, a high performance liquid chromatography (HPLC method is developed for the determination of three active ingredients including Jujuboside A, Scopoletin and Quercetin in Anshenbao granules. The analysis is performed on a Agilent Zorbox SB C18 column(4.6 mm×150 mm,5 μm with a gradient mobile phase of methanol-0.5% glacial acetic acid at a flow rate of 1.0 mL/min. The evaporative light scattering detector(ELSD is used, the column temperature is 35 ℃, and the injection is 10 μL. The three active ingredients are separated completely under the given chromatogram system with a good linearity (r=0.995 6~0.998 0 of peak area and the injection. The results of precision, repeatability and stability experiments can meet the requirement, and the mean recoveries of each active ingredients are all in the range of 94.74%~96.08%. The analytical method is simple, accurate and sensitive, and it can be used for determination of active ingredients in Anshenbao granules. It is of practical significance to improve the quality standard of Anshenbao and improve the quality control of medicine.

  3. [HPLC fingerprints of tibetan medicinal herb "songdi" (Saxifraga umbellulata var. pectinata)].

    Science.gov (United States)

    Fei, Yao; Zhong, Guo-Yue; Jiang, Wei

    2014-07-01

    The research was carried out to establish HPLC fingerprints of Tibetan medicinal herb "Songdi" (Saxifraga umbellulata var. pectinata), and to provide reference for identification an quality control of it. It was performed on an Amethyst-C18-P (4.6 mm x 250 mm, 5 microm) column with the mobile phase of methanol-0.4% formic acid in a linear gradient mode at a flow rate of 1.0 mL x min(-1). The column temperature was 30 degrees C, and the detection wavelength was set at 254 nm. The software for chromatographic fingerprint was applied to analyse the pattern analysis, the common peaks and similarity. Cluster analysis was done based on the common peaks data of 33 samples from different plant species and sources by SPSS software. Ten common chromatographic peaks were identified by fingerprint, showing a low similarity in constituent and variety. Flavonoids and saponins were the principal components. The number and area of peaks were affected by the collection sources and method. The high similarity are showed by the samples derived from the same area with high accuracy and high purity. The method is so simple, exclusive, stable and high repeatable that it can provide reference for identification and quality assessment of "Songdi" (S. umbellulata var. pectinata).

  4. [Simultaneous determination of five sesquiterpene lactones in Carpesium abrotanoides by HPLC].

    Science.gov (United States)

    Liu, Min; Zhou, Xiao-jiang

    2015-05-01

    An HPLC method was established to simultaneously determine the five sesquiterpene lactones, carabrone, carabrol, 2-desoxy-4-epi-pulchellin, telekinand 11(13)-dehydroivaxillin in Carpesium abrotanoides. Samples were analyzed on a kromasil C18 column (4.6 mm x 250 mm, 5 µm); mobile phase: A was acetonitrile, B was water, with gradient elution; flow speed: 1.0 mL · min(-1); detection was carried out using a photodiode array detector at 211 nm; temperatureof column: 30 °C. The five sesquiterpene lactones were well separated with good linear correlations in the range of 0.270-2.700 (r = 0.999 7 ), 1.336-13.360 (r = 0.999 6), 0.258-2.580 (r = 0.999 7), 0.238-2.380 (r = 0.999 9), 0.490-4.900 µg (r = 0.999 9) for carabrone, carabrol, 2-desoxy-4-epi-pulchellin, telekin and 11 (13) -dehydroivaxillin. The average recoveries of these five sesquiterpene lactoneswere 96.78%-98.41% (RSD 1.3%-2.9%). The method was simple, repeatable and stable, which could be used for quality control of C. abrotanoide.

  5. Simultaneous determination of aceclofenac, paracetamol, and chlorzoxazone by RP-HPLC in pharmaceutical dosage form.

    Science.gov (United States)

    Shaikh, K A; Devkhile, A B

    2008-08-01

    A simple, rapid, and precise reversed-phase liquid chromatographic method is developed for simultaneous determination of paracetamol, aceclofenac, and chlorzoxazone in their ternary mixtures of commercial pharmaceutical preparation. This method uses a Zorbax SB C18, 250 x 4.6 mm, 5 microm analytical column. Mobile phase is acetonitrile and buffer (40:60, v/v), buffer containing 50 mM orthophosporic acid; pH of the buffer is adjusted to 6 with 10% w/v sodium hydroxide solution. The instrumental settings are at a flow rate of 1 mL/min; the column temperature is 25 degrees C, and detector wavelength is 270 nm. The sample concentrations are measured on weight basis to avoid the internal standard. The method is validated and shown to be linear. The correlation coefficients for paracetamol, aceclofenac, and chlorzoxazone are 0.9981, 0.9990, and 0.9986, respectively. The recovery values for paracetamol, aceclofenac, and chlorzoxazone ranged from 100.7-101.4%, 100.4-101.0%, and 100.5-101.3%, respectively. The relative standard deviation for six replicates is always less than 2%. This HPLC method is successfully applied to the simultaneous quantitative analysis of the title drugs in tablets and can be applied for assay and dissolution test of tablets for the estimation of paracetamol, aceclofenac, and chlorozoxazone in their commercial samples.

  6. Creative columns

    National Research Council Canada - National Science Library

    Hoff, Diane

    2017-01-01

      Here, Hoff presents creative columns. As her seventh-graders began learning about ancient Greece in social studies, in art they observed ancient Greek architecture, paying attention to the orders of Pork, Ionic, and Corinthian...

  7. [Study and comparison on HPLC fingerprints of flavonoids of frequently used Chinese materia medica in citrus].

    Science.gov (United States)

    Chen, Yonggang; Lin, Li

    2011-10-01

    To establish the HPLC fingerprint of flavonoids of the six clinical frequently used Chinese materia medica for regulating Qi flow,such as Citri grandis, C. grands, Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride, Aurantii Fructus, and Aurantii Fructus Immaturus from Citrus, and analysis differences in the fingerprints to provide scientific basis for profile-effect research and clinical reasonable use. HPLC was performed on a C18 column with methanol-water (with acetic acid), to establish HPLC fingerprints of the six kinds of medicinal herbs on the same chromatograph condition. The six frequently used Chinese materia medica were divided into naringin type and hesperidin type according to the method of phytochemotaxonomy. Based on the retention time of chromatograph peaks, C. grandis and C. grands had fifteen common peaks; Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride, Aurantii Fructus and Aurantii Fructus Immaturus had ten common peaks. All herbs had five common peaks. Compared with mutual model, the holistic similarity of chromatograms of C. grandis and C. grands was in the range of 0.9285 - 0.9962. The degree of similarity was high. For Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride and Aurantii Fructus Immaturus, it was in the range of 0.9221 - 0.9973 and high. But the similarity of Aurantii Fructus was only in 0.4547 - 0.7733 with the mutual model. The established fingerprints of flavonoids of the six common traditional Chinese medicines can be used to compare the differences intuitively. Meanwhile, the peak height and peak areas of characteristic peaks are different remarkably, but whether it is connected with the different function of regulating Qi flow of the six medical materials in clinical use, is still needed to be studied.

  8. HPLC Determination of Taurine in Sports Drinks

    Science.gov (United States)

    Orth, Dale L.

    2001-06-01

    The amino acid taurine (2-aminoethanesulfonic acid) is present as a nutritional supplement in many sports drinks. An experiment, suitable for a junior-senior level instrumental analysis course, is described to measure the amount of taurine in these sports drinks. A pre-column derivatization with Sanger's reagent, 2,4-dinitrofluorobenzene, is followed by an HPLC separation utilizing a gradient elution, and detection at 360 nm.

  9. Application of HPLC and MALDI-TOF MS for studying as-synthesized ligand-protected gold nanoclusters products.

    Science.gov (United States)

    Zhang, Yan; Shuang, Shaomin; Dong, Chuan; Lo, Chung Keung; Paau, Man Chin; Choi, Martin M F

    2009-02-15

    Samples of polydisperse gold nanoclusters (AuNCs) protected with monolayers of N-acetyl-L-cysteine (NAC) have been chromatographically separated by a C18 column (4.6 mm x 250 mm) using a gradient elution program with a mobile phase of methanol (MeOH)/water containing tetrabutylammonium fluoride (Bu(4)N(+)F(-)) and sodium chloride. The effects of Bu(4)N(+)F(-) and MeOH on the separation have been investigated in detail. In conjunction with absorbance-based, fluorescence, and electrochemical detectors, the elution order of these water-soluble AuNCs is confirmed according to their core size in an ascending order. The onset oxidation potential closely follows the core size of AuNC. The separated fractions from high-performance liquid chromatography (HPLC) were collected and analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry to determine the number of Au atoms in the fractions. The sizes of AuNCs in some selected HPLC fractions were also assessed by transmission electron microscopy and high-resolution transmission electron microscopy. Photoluminescence spectra of the fractions show that the luminescent shift in the visible/near-infrared region does not follow with the core size of AuNC. More importantly, the proposed HPLC methodology has been successfully applied to analyze various polydisperse AuNC products synthesized from different gold-to-ligand mole ratios (Au/NAC) and reaction temperatures. The results confirm that larger Au/NAC and higher temperature will produce larger core size AuNCs products with narrower dispersity. In addition, AuNC samples obtained from various synthesis reaction times were analyzed by our HPLC methodology, demonstrating that the reaction's behavior follows the nucleation-growth-disintegration process.

  10. A validated HPLC-UV method and optimization of sample preparation technique for norepinephrine and serotonin in mouse brain.

    Science.gov (United States)

    Thomas, Jaya; Khanam, Razia; Vohora, Divya

    2015-01-01

    Norepinephrine and serotonin are two important neurotransmitters whose variations in brain are reported to be associated with many common neuropsychiatric disorders. Yet, relevant literature on estimation of monoamines in biological samples using HPLC-UV is limited. The present study involves the development of a simultaneous HPLC-UV method for estimation of norepinephrine and serotonin along with optimization of the sample preparation technique. Chromatographic separation was achieved by injecting 20 µL of the sample after extraction into Quaternary pump HPLC equipped with C18 column using 0.05% formic acid and acetonitrile (90:10, v/v) as the mobile phase with 1 mL min(-1) flow rate. The developed method was validated as per the ICH guidelines in terms of linearity, accuracy, repeatability, precision, and robustness. The method showed a wide range of linearity (50-4000 and 31.25-4000 ng mL(-1) for norepinephrine and serotonin, respectively). The recovery was found to be in the range of 86.04-89.01% and 86.43-89.61% for norepinephrine and serotonin, respectively. The results showed low value of %RSD for repeatability, intra and inter-day precision, and robustness studies. Four different methods were used for the extraction of these neurotransmitters and the best one with maximum recovery was ascertained. Here, we developed and validated a simple, accurate, and reliable method for the estimation of norepinephrine and serotonin in mice brain samples using HPLC-UV. The method was successfully applied to quantify these neurotransmitters in mice brain extracted by optimized sample preparation technique.

  11. CHIMPS: the 13CO/C18O (J = 3 → 2) Heterodyne Inner Milky Way Plane Survey

    Science.gov (United States)

    Rigby, A. J.; Moore, T. J. T.; Plume, R.; Eden, D. J.; Urquhart, J. S.; Thompson, M. A.; Mottram, J. C.; Brunt, C. M.; Butner, H. M.; Dempsey, J. T.; Gibson, S. J.; Hatchell, J.; Jenness, T.; Kuno, N.; Longmore, S. N.; Morgan, L. K.; Polychroni, D.; Thomas, H.; White, G. J.; Zhu, M.

    2016-03-01

    We present the 13CO/C18O (J = 3 → 2) Heterodyne Inner Milky Way Plane Survey (CHIMPS) which has been carried out using the Heterodyne Array Receiver Program on the 15 m James Clerk Maxwell Telescope (JCMT) in Hawaii. The high-resolution spectral survey currently covers |b| ≤ 0.5° and 28° ≲ l ≲ 46°, with an angular resolution of 15 arcsec in 0.5 km s-1 velocity channels. The spectra have a median rms of ˜0.6 K at this resolution, and for optically thin gas at an excitation temperature of 10 K, this sensitivity corresponds to column densities of NH2 ˜ 3 × 1020 cm-2 and NH2 ˜ 4 × 1021 cm-2 for 13CO and C18O, respectively. The molecular gas that CHIMPS traces is at higher column densities and is also more optically thin than in other publicly available CO surveys due to its rarer isotopologues, and thus more representative of the three-dimensional structure of the clouds. The critical density of the J = 3 → 2 transition of CO is ≳104 cm-3 at temperatures of ≤20 K, and so the higher density gas associated with star formation is well traced. These data complement other existing Galactic plane surveys, especially the JCMT Galactic Plane Survey which has similar spatial resolution and column density sensitivity, and the Herschel infrared Galactic Plane Survey. In this paper, we discuss the observations, data reduction and characteristics of the survey, presenting integrated-emission maps for the region covered. Position-velocity diagrams allow comparison with Galactic structure models of the Milky Way, and while we find good agreement with a particular four-arm model, there are some significant deviations.

  12. Automatization for development of HPLC methods.

    Science.gov (United States)

    Pfeffer, M; Windt, H

    2001-01-01

    Within the frame of inprocess analytics of the synthesis of pharmaceutical drugs a lot of HPLC methods are required for checking the quality of intermediates and drug substances. The methods have to be developed in terms of optimal selectivity and low limit of detection, minimum running time and chromatographic robustness. The goal was to shorten the method development process. Therefore, the screening of stationary phases was automated by means of switching modules equipped with 12 HPLC columns. Mobile phase and temperature could be optimized by using Drylab after evaluating chromatograms of gradient elutions performed automatically. The column switching module was applied for more than three dozens of substances, e.g. steroidal intermediates. Resolution (especially of isomeres), peak shape and number of peaks turned out to be the criteria for selection of the appropriate stationary phase. On the basis of the "best" column the composition of the "best" eluent was usually defined rapidly and with less effort. This approach leads to savings in manpower by more than one third. Overnight, impurity profiles of the intermediates were obtained yielding robust HPLC methods with high selectivity and minimized elution time.

  13. A new solid-phase extraction and HPLC method for determination of patulin in apple products and hawthorn juice in China.

    Science.gov (United States)

    Zhou, Yuchun; Kong, Weijun; Li, Yan; Logrieco, Antonio F; Xu, Jun; Yang, Meihua

    2012-03-01

    A new solid-phase extraction (SPE) pretreatment method using a home-made polyvinylpolypyrrolidone-florisil (PVPP-F) column was developed for the analysis of patulin in apple and hawthorn products in China. Fifty samples (25 apple juices, 12 apple jams, and 13 hawthorn juices) were prepared using the new method and then analyzed by high performance liquid chromatography with diode array detection (HPLC-DAD) on an Agela Venusil MP C(18) reversed-phase column (4.6 mm × 250 mm, 5 μm). The cleanup results for all samples using home-made PVPP-F column were compared with those obtained using a MycoSep®228 AflaPat column. The correlation coefficient R (0.9998) fulfilled the requirement of linearity for patulin in the concentration range of 2.5-250 μg/kg. The limits of detection (LODs) and quantification (LOQs) of patulin were 3.99 and 9.64 μg/kg for PVPP-F column, and 3.56 and 8.07 μg/kg for MycoSep®228 AflaPat column, respectively. Samples were spiked with patulin at levels ranging from 25 to 250 μg/kg, and recoveries using PVPP-F and MycoSep®228 AflaPat columns were in the range of 81.9-100.9% and 86.4-103.9%, respectively. Naturally occurring patulin was found in 2 of 25 apple juice samples (8.0%) and 1 of 13 hawthorn juice samples (7.7%) at concentrations ranging from 12.26 to 36.81 μg/kg. The positive results were further confirmed by liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS). © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. A Novel HPLC Method for the Concurrent Analysis and Quantitation of Seven Water-Soluble Vitamins in Biological Fluids (Plasma and Urine: A Validation Study and Application

    Directory of Open Access Journals (Sweden)

    Margherita Grotzkyj Giorgi

    2012-01-01

    Full Text Available An HPLC method was developed and validated for the concurrent detection and quantitation of seven water-soluble vitamins (C, B1, B2, B5, B6, B9, B12 in biological matrices (plasma and urine. Separation was achieved at 30°C on a reversed-phase C18-A column using combined isocratic and linear gradient elution with a mobile phase consisting of 0.01% TFA aqueous and 100% methanol. Total run time was 35 minutes. Detection was performed with diode array set at 280 nm. Each vitamin was quantitatively determined at its maximum wavelength. Spectral comparison was used for peak identification in real samples (24 plasma and urine samples from abstinent alcohol-dependent males. Interday and intraday precision were <4% and <7%, respectively, for all vitamins. Recovery percentages ranged from 93% to 100%.

  15. A Novel HPLC Method for the Concurrent Analysis and Quantitation of Seven Water-Soluble Vitamins in Biological Fluids (Plasma and Urine): A Validation Study and Application

    Science.gov (United States)

    Grotzkyj Giorgi, Margherita; Howland, Kevin; Martin, Colin; Bonner, Adrian B.

    2012-01-01

    An HPLC method was developed and validated for the concurrent detection and quantitation of seven water-soluble vitamins (C, B1, B2, B5, B6, B9, B12) in biological matrices (plasma and urine). Separation was achieved at 30°C on a reversed-phase C18-A column using combined isocratic and linear gradient elution with a mobile phase consisting of 0.01% TFA aqueous and 100% methanol. Total run time was 35 minutes. Detection was performed with diode array set at 280 nm. Each vitamin was quantitatively determined at its maximum wavelength. Spectral comparison was used for peak identification in real samples (24 plasma and urine samples from abstinent alcohol-dependent males). Interday and intraday precision were vitamins. Recovery percentages ranged from 93% to 100%. PMID:22536136

  16. Development and Validation of a Stability-Indicating RP-HPLC Method for the Assay of Pristinamycin in Bulk and Tablet Dosage Form.

    Science.gov (United States)

    Mallikarjuna Rao, Nagasarapu; Gowrisankar, Dannana

    2016-01-01

    Pristinamycin is an antibiotic used mainly in the treatment of Staphylococcus infections. The aim of this study was to develop a rapid and simple stability-indicating RP-HPLC method for the determination of pristinamycin in tablet dosage form. Pristinamycin was eluted on the ACE-5, C18-HL, 250 x 4.6 mm, 5 µm analytical column with a mobile phase consisting of 0.2% orthophosphoric acid and acetonitrile 63:37 v/v, pumped at 1.5 ml/min flow rate. The column was maintained at 40°C and 10 μl of the solutions were injected. UV detection was performed at 206 nm. The procedure separated pristinamycin and its potential degradation products in an overall analysis time of less than 10 min with pristinamycin eluting at about 3 min. The method was validated according to the regulatory guidelines with respect to specificity, precision, accuracy, linearity, and robustness. Forced degradation studies were also performed for pristinamycin bulk drug samples to demonstrate the stability-indicating power of the HPLC method. The % RSD of system precision and method precision was found to be 0.64 and 1.49%, respectively. The procedure provided a linear response over the concentration range 25-150 μg/ml (r = 0.9998). Finally, the applicability of the method was evaluated in the tablet dosage form as well as in stability samples.

  17. Development and Validation of New RP-HPLC Method for Simultaneous Determination of Methyl and Propyl Parabens with Levetiracetam in Pure Form and Pharmaceutical Formulation

    Directory of Open Access Journals (Sweden)

    Soad S. Abd El-Hay

    2016-01-01

    Full Text Available A simple and robust high-performance liquid chromatography (HPLC method is described for the assay for levetiracetam (LTC, methyl paraben (MHB, and propyl paraben (PHB either in their pure form or in commercial Levepsy® syrup. The method is selective and stability indicating and all chromatographic conditions were studied to obtain adequate separation of LTC, MHB, and PHB from their degradation products and from excipients. The HPLC separation was carried out on a RP C18 Hypersil BDS analytical column (150 mm × 4.6 mm ID using gradient elution system. The mobile phase flow rate was 1.5 mLmin−1 and the column temperature was kept at 40°C. Complete separation of the studied components was obtained within a cycle time of 8 min. LTC, MHB, and PHB were eluted at 1.56, 5.86, and 7.85 min, respectively. Detection was carried out at 240 nm using a dual wavelength detector. The method has been validated for linearity, accuracy, precision, specificity, limit of detection, limit of quantitation, robustness, and ruggedness. The proposed method was successfully applied for the determination of LTC in the presence of parabens in Levepsy syrup.

  18. Development and validation of SPE-HPLC method for the determination of carbamazepine and its metabolites carbamazepine epoxide and carbamazepine trans-diol in plasma

    Directory of Open Access Journals (Sweden)

    Spasić Mirjana

    2012-01-01

    Full Text Available SPE-HPLC method has been developed and validated for rapid analysis of carbamazepine and its two metabolites carbamazepine epoxide and carbamazepine trans-diol in human plasma. The analysis was performed using C18 Bakerbond-BDC analytical column (250 mm x 4.6 mm i.d., particle size 5 μm. The optimal conditions for the separation were established with the mobile phase acetonitrile - 10 mM phosphate buffer, pH 7.0 (30:70, v/v at the flow rate of 1.5 mL min-1, temperature 35°C, and UV detection at 210 nm. Total run time was about 8 minutes. SPE procedure for extraction of the analytes from plasma sample was developed using Oasis HLB cartridges and subsequently eluate was injected into the HPLC system for analysis. Afterwards, SPE-HPLC method was subjected to validation. Linearity was obtained over the concentration range of 0.2-25 μg/mL for carbamazepine, carbamazepine epoxide and carbamazepine trans-diol with correlation coefficients higher than 0.995. The method showed good intra-day and inter-day precision with relative standard deviation below 7.96%, while accuracy ranged from 92.09% to 108.5% for all analytes. Finally, the method was successfully applied to analysis of plasma samples of epileptic patients in monotherapy and polytherapy. [Acknowledgments. Projekat Ministarstva nauke Republike Srbije, br. OI 172033].

  19. HPLC-DAD and UV-spectrophotometry for the determination of lychnopholide in nanocapsule dosage form: validation and application to release kinetic study.

    Science.gov (United States)

    Branquinho, Renata Tupinambá; Mosqueira, Vanessa Carla Furtado; Kano, Eunice Kazue; de Souza, Jacqueline; Dorim, Diego Dias Ramos; Saúde-Guimarães, Dênia Antunes; de Lana, Marta

    2014-01-01

    Simple and sensitive methods using high-performance liquid chromatography-diode array detection (HPLC-DAD) and ultraviolet (UV)-spectrophotometry were developed and compared to quantify lychnopholide (LYC) in poly-ε-caprolactone nanocapsules and to study its release kinetics. Both methods were validated concerning their specificity, linearity, limits of detection and quantification, precision, accuracy and stability. HPLC-DAD analyses were conducted using an RP C18 column, isocratic elution with a methanol-water (60:40 v/v) mobile phase at 0.8 mL/min flow rate and detection at 265 nm. The linear response (r(2) > 0.999) was obtained within a concentration range of 2-25 µg/mL using HPLC-DAD and 5-40 µg/mL using spectrophotometry. Intra-day and inter-day precision were obtained with low relative standard deviation values. The accuracy of the methods was within the range 98-101% for HPLC-DAD and from 96-100% for UV-spectrophotometry. Both methods were suitable to be applied for the determination of drug loading percentage (>96%) and encapsulation efficiency (>90%). Furthermore, the sensitivity of HPLC-DAD method allows studies of LYC release/dissolution in sink conditions. LYC presented 100% dissolution after 24 h, whereas only 60% of LYC was released from the nanocapsule dosage form, with no burst effect. The methods fulfilled all validation parameters evaluated for LYC quantification in the polymeric nanocapsules and have proven to be accurate, selective and sensitive in the previously mentioned applications.

  20. Chemometrics-Assisted UV Spectrophotometric and RP-HPLC Methods for the Simultaneous Determination of Tolperisone Hydrochloride and Diclofenac Sodium in their Combined Pharmaceutical Formulation

    Science.gov (United States)

    Gohel, Nikunj Rameshbhai; Patel, Bhavin Kiritbhai; Parmar, Vijaykumar Kunvarji

    2013-01-01

    Chemometrics-assisted UV spectrophotometric and RP-HPLC methods are presented for the simultaneous determination of tolperisone hydrochloride (TOL) and diclofenac sodium (DIC) from their combined pharmaceutical dosage form. Chemometric methods are based on principal component regression and partial least-square regression models. Two sets of standard mixtures, calibration sets, and validation sets were prepared. Both models were optimized to quantify each drug in the mixture using the information included in the UV absorption spectra of the appropriate solution in the range 241–290 nm with the intervals λ = 1 nm at 50 wavelengths. The optimized models were successfully applied to the simultaneous determination of these drugs in synthetic mixture and pharmaceutical formulation. In addition, an HPLC method was developed using a reversed-phase C18 column at ambient temperature with a mobile phase consisting of methanol:acetonitrile:water (60:30:10 v/v/v), pH-adjusted to 3.0, with UV detection at 275 nm. The methods were validated in terms of linearity, accuracy, precision, sensitivity, specificity, and robustness in the range of 3–30 μg/mL for TOL and 1–10 μg/mL for DIC. The robustness of the HPLC method was tested using an experimental design approach. The developed HPLC method, and the PCR and PLS models were used to determine the amount of TOL and DIC in tablets. The data obtained from the PCR and PLS models were not significantly different from those obtained from the HPLC method at 95% confidence limit. PMID:24482768

  1. Development and Validation of HPLC Method for Determination of Crocetin, a constituent of Saffron, in Human Serum Samples

    Directory of Open Access Journals (Sweden)

    Amir Hooshang Mohammadpour

    2013-01-01

    Full Text Available Objective(s:The present study reports the development and validation of a sensitive and rapid extraction method beside high performance liquid chromatographic method for the determination of crocetin in human serum. Materials and Methods:The HPLC method was carried out by using a C18 reversed-phase column and a mobile phase composed of methanol/water/acetic acid (85:14.5:0.5 v/v/v at the flow rate of 0.8 ml/min. The UV detector was set at 423 nm and 13-cis retinoic acid was used as the internal standard. Serum samples were pretreated with solid-phase extraction using Bond Elut C18 (200mg cartridges or with direct precipitation using acetonitrile. Results:The calibration curves were linear over the range of 0.05-1.25 µg/ml for direct precipitation method and 0.5-5 µg/ml for solid-phase extraction. The mean recoveries of crocetin over a concentration range of 0.05-5 µg/ml serum for direct precipitation method and 0.5-5 µg/ml for solid-phase extraction were above 70 % and 60 %, respectively. The intraday coefficients of variation were 0.37- 2.6% for direct precipitation method and 0.64 - 5.43% for solid-phase extraction. The inter day coefficients of variation were 1.69 – 6.03% for direct precipitation method and 5.13-12.74% for solid-phase extraction, respectively. The lower limit of quantification for crocetin was 0.05 µg/ml for direct precipitation method and 0.5 µg/ml for solid-phase extraction. Conclusion: The validated direct precipitation method for HPLC satisfied all of the criteria that were necessary for a bioanalytical method and could reliably quantitate crocetin in human serum for future clinical pharmacokinetic study

  2. Development and Validation of an RP-HPLC Method for Quantitative Estimation of Eslicarbazepine Acetate in Bulk Drug and Tablets.

    Science.gov (United States)

    Singh, M; Kumar, L; Arora, P; Mathur, S C; Saini, P K; Singh, R M; Singh, G N

    2013-11-01

    A convenient, simple, accurate, precise and reproducible RP-HPLC method was developed and validated for the estimation of eslicarbazepine acetate in bulk drug and tablet dosage form. Objective was achieved under optimised chromatographic conditions on Dionex RP-HPLC system with Dionex C18 column (250×4.6 mm, 5 μm particle size) using mobile phase composed of methanol and ammonium acetate (0.005 M) in the ratio of 70:30 v/v. The separation was achieved using an isocratic elution method with a flow rate of 1.0 ml/ min at room temperature. The effluent was monitored at 230 nm using diode array detector. The retention time of eslicarbazepine acetate is found to be 4.9 min and the standard calibration plot was linear over a concentration range of 10-90 μg/ml with r(2)=0.9995. The limit of detection and quantification were found to be 3.144 and 9.52 μg/ml, respectively. The amount of eslicarbazepine acetate in bulk and tablet dosage form was found to be 99.19 and 97.88%, respectively. The method was validated statistically using the percent relative standard deviation and the values are found to be within the limits. The recovery studies were performed and the percentage recoveries were found to be 98.33± 0.5%.

  3. Development and Validation of RP-HPLC Method for Quantitative Estimation of Vinpocetine in Pure and Pharmaceutical Dosage Forms

    Directory of Open Access Journals (Sweden)

    Subrata Bhadra

    2011-01-01

    Full Text Available A simple, precise, specific, and accurate reversed phase high performance liquid chromatographic (RP-HPLC method was developed and validated for determination of vinpocetine in pure and pharmaceutical dosage forms. The different analytical performance parameters such as linearity, accuracy, specificity, precision, and sensitivity (limit of detection and limit of quantitation were determined according to International Conference on Harmonization ICH Q2 (R1 guidelines. RP-HPLC was conducted on Zorbax C18 (150 mm length × 4.6 mm ID, 5 μm column. The mobile phase was consisting of buffer (containing 1.54% w/v ammonium acetate solution and acetonitrile in the ratio (40 : 60, v/v, and the flow rate was maintained at 1.0 mLmin−1. Vinpocetine was monitored using Agilent 1200 series equipped with photo diode array detector (λ = 280 nm. Linearity was observed in concentration range of 160–240 μgmL−1, and correlation coefficient was found excellent (R2 = 0.999. All the system suitability parameters were found within the range. The proposed method is rapid, cost-effective and can be used as a quality-control tool for routine quantitative analysis of vinpocetine in pure and pharmaceutical dosage forms.

  4. An improved HPLC method with fluorescence detection for the determination of pyrene in rat plasma and its pharmacokinetics.

    Science.gov (United States)

    Zhao, Xiaozhi; Wan, Jiangling; Xu, Huibi; Yang, Xiangliang

    2008-12-01

    A high-performance liquid chromatographic method with fluorescence detection (HPLC-FLD) for the determination of pyrene in rat plasma was developed and validated. The method used fluorene as internal standard (IS), following a single-step protein precipitation, the analyte and internal standard were separated on a C18 column with a mobile phase containing methanol-water (90:10, v/v) at a flow rate of 1 ml/min. The analytes were detected by using fluorescence detection at an excitation and emission wavelength of 265 and 394 nm, respectively. Two calibration curves were constructed in the range of 2-100 ng/ml and 0.1-5 microg/ml for pyrene with a lower limit of quantitation (LLOQ) of 2 ng/ml. Both intra-day and inter-day precision were less than 6% except at LLOQ, for which the precision was 10.6 and 9.8, respectively. Accuracy ranged from 98.3 to 103.6%, except at LLOQ, for which the accuracy was about 85%. The recovery ranged from 84.7 to 95.0% at the low, medium and high concentrations. The present HPLC-FLD method was rapid, sensitive, and reliable. The method described herein had been successfully applied for the pharmacokinetic studies in female Wistar rats after administration of 10mg equivalent pyrene/kg dose of solution of pyrene and 1mg equivalent pyrene/kg dose of pyrene-loaded nanoparticle.

  5. 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.

  6. Development of a HPLC-MS/MS method for the simultaneous determination of nifedipine and lidocaine in human plasma.

    Science.gov (United States)

    Grigoriev, Alexander; Nikitina, Alexandra; Yaroshenko, Irina; Sidorova, Alla

    2016-11-30

    The method for simultaneous determination of nifedipine (NIF) and lidocaine (LID) in human plasma by one-step sample preparation has been developed for the first time. Due to the photosensitivity of nifedipine and its low plasma concentrations a precise and reliable method was required. The method involved liquid-liquid extraction (methyl tert-butyl ether, MTBE), and 10μL of the resulting sample was analyzed by HPLC-MS/MS. Chromatographic separation was achieved on an YMC-Triart C18 HPLC column (100×2.0mm; S-5μm 12nm). The mobile phase was methanol:water, 60:40 (v/v) and contained 0.15% acetic acid. The linearity of the method was established in the concentration ranges of 0.5-50ng/mL for NIF and 1.0-500ng/mL for LID. Photodestruction of NIF under ambient light was evaluated. The validated method was successfully applied to analyze human plasma samples after rectal application of the drug (1g) containing 2.0% LID and 0.3% NIF. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Isotope dilution HPLC-MS/MS for simultaneous quantification of acrylamide and 5-hydroxymethylfurfural (HMF) in thermally processed seafood.

    Science.gov (United States)

    Qin, Lei; Zhang, Yu-Ying; Xu, Xian-Bing; Wang, Xu-Song; Liu, Hong-Wei; Zhou, Da-Yong; Zhu, Bei-Wei; Thornton, Megan

    2017-10-01

    The objective of this study was to develop an accurate, robust and rapid HPLC-MS/MS-based method capable of quantifying acrylamide and 5-hydroxymethylfurfural (HMF) simultaneously. A Phenomenex Synergi Fusion-RP C18 column (50mm×2.5mm, 2μm) was used, and the MS/MS instrument was operated in multiple reaction monitoring mode. The isotope dilution method was used to correct for the matrix effect from samples. This method demonstrated low limits of quantification (2.12ng/mL for acrylamide and 4.86ng/mL for HMF) and excellent linearity (R2>0.999). This method also demonstrated excellent quantification accuracy, precision, and recovery (87-110%). Using the method developed, target analytes were quantified in 14 thermally-processed seafood samples. Resulting concentrations ranged from 5.58 to 50.35μg/kg for acrylamide, and from 12.54 to 2276.44μg/kg for HMF. The proposed isotope dilution HPLC-MS/MS method is a valid and rapid technique for simultaneous analysis of acrylamide and HMF in seafood. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Quantitative estimation of (--hinokinin, a trypanosomicidal marker in Piper cubeba, and some of its commercial formulations using HPLC-PDA

    Directory of Open Access Journals (Sweden)

    Kothapalli Haribabu

    2015-04-01

    Full Text Available The fruits of Piper cubeba have been used in Ayurvedic system of medicine for pain, tastelessness, painful urination and mouth diseases. Among its various chemical constituents, (--hinokinin, a trypanosomicidal dibenzylbutyrolactone lignan, is found in significant quantities. For quality evaluation of P. cubeba fruit and its commercial formulations, there is an urgent need to develop an analytical method based on (--hinokinin. For this purpose, an HPLC method was developed using photo diode array detector and Waters HR C18 column with gradient elution consisting of water and acetonitrile. The developed method was validated as per ICH-Q2B guidelines and found to be accurate, precise and linear over a wide range of concentrations (5–300 µg/mL. (--Hinokinin contents were found to be in the range of 0.005–0.109% (m/m in various P. cubeba samples. The developed method was extended to LC–MS for further identification and characterization of (--hinokinin in samples. The developed method is simple, rapid and specific, and can be used as a tool for quality control of P. cubeba fruits and its commercial formulations. Keywords: Piper cubeba, (--Hinokinin, HPLC, Quality control, Herbal formulations

  9. Development and optimization of the SPE procedure for determination of pharmaceuticals in water samples by HPLC-diode array detection.

    Science.gov (United States)

    Mutavdzić Pavlović, Dragana; Babić, Sandra; Dolar, Davor; Asperger, Danijela; Kosutić, Kresimir; Horvat, Alka J M; Kastelan-Macan, Marija

    2010-02-01

    This paper focuses on the investigation of different types of SPE sorbents for the preconcentration of eight veterinary pharmaceuticals from water samples. The pharmaceuticals studied were sulfamethazine, sulfadiazine, sulfaguanidine, trimethoprim, oxytetracycline, enrofloxacin, norfloxacin and penicillin G/procaine. Five different SPE materials (Strata-X, Strata-X-C, Strata SDB-L, Strata C8 and Strata C18) from Phenomenex were compared with Oasis HLB with a view to obtaining the best cartridges for all pharmaceuticals investigated. Extraction efficiency was determined by HPLC with diode array detection (DAD). HPLC-DAD separation and quantification of the selected pharmaceuticals were carried out under gradient elution by a binary mixture of 0.01 M oxalic acid and ACN based on cyano modified column (LiChrosphere 100 CN) from Merck. Strata-X provided the best results in the preconcentration of 100 mL water samples, yielding average pharmaceutical recoveries of higher than 90%, except for sulfaguanidine (76.1%). The developed Strata-X-HLPC-DAD method was validated and applied, for the efficient investigation of reverse osmosis/nanofiltration membranes and for the removal of these eight pharmaceuticals from the production wastewater samples. NF90 and XLE membranes were shown to be the best for the rejection of all investigated pharmaceuticals.

  10. COLUMN TESSELLATIONS

    Directory of Open Access Journals (Sweden)

    Linh Ngoc Nguyen

    2015-06-01

    Full Text Available A new class of non facet-to-facet random tessellations in three-dimensional space is introduced -- the so-called column tessellations. The spatial construction is based on a stationary planar tessellation; each cell of the spatial tessellation is a prism whose base facet is  congruent to a cell of the planar tessellation. Thus intensities, topological and metric mean values of the spatial tessellation can be calculated from suitably chosen parameters of the planar tessellation.

  11. 26 CFR 1.501(c)(18)-1 - Certain funded pension trusts.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Certain funded pension trusts. 1.501(c)(18)-1... pension trusts. (a) In general. Organizations described in section 501(c)(18) are trusts created before June 25, 1959, forming part of a plan for the payment of benefits under a pension plan funded only by...

  12. Stability-Indicating RP-HPLC Method for Simultaneous Estimation of Enrofloxacin and Its Degradation Products in Tablet Dosage Forms

    Directory of Open Access Journals (Sweden)

    V. Ashok Chakravarthy

    2015-01-01

    Full Text Available The present work was the development of a simple, efficient, and reproducible stability-indicating reverse-phase high performance liquid chromatographic (RP-HPLC method for simultaneous determination enrofloxacin (EFX and its degradation products including ethylenediamine impurity, desfluoro impurity, ciprofloxacin impurity, chloro impurity, fluoroquinolonic acid impurity, and decarboxylated impurity in tablet dosage forms. The separation of EFX and its degradation products in tablets was carried out on Kromasil C-18 (250×4.6 mm, 5 μm column using 0.1% (v/v TEA in 10 mM KH2PO4 (pH 2.5 buffer and methanol by linear gradient program. Flow rate was 1.0 mL min−1 with a column temperature of 35°C and detection wavelength was carried out at 278 nm and 254 nm. The forced degradation studies were performed on EFX tablets under acidic, basic, oxidation, thermal, humidity, and photolytic conditions. The degraded products were well resolved from the main active drug and also from known impurities within 65 minutes. The method was validated in terms of specificity, linearity, LOD, LOQ, accuracy, precision, and robustness as per ICH guidelines. The results obtained from the validation experiments prove that the developed method is a stability-indicating method and suitable for routine analysis.

  13. 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.

  14. Screening for pharmaceutically important taxoids in Taxus baccata var. Aurea corr. with CC/SPE/HPLC-PDA procedure.

    Science.gov (United States)

    Mroczek, T; Glowniak, K; Hajnos, M

    2000-12-01

    Needles of 'the golden yew' Taxus baccata var. Aurea Corr. were extracted with methanol followed by pre-purification of the crude extract and column chromatographic (CC) separation on florisil in gradient mode (an increasing concentration of acetone in dichloromethane). The obtained fractions were concentrated and purified on silanized silica gel SPE cartridges and taxoids eluted with 75% methanol were analysed by HPLC-PDA procedure using Waters Symmetry C(18) column with gradient elution. The applied method enabled not only determination of four taxoids commonly occurring in yew extracts (10-deacetylbaccatin III, baccatin III, paclitaxel and cephalomannine), but also, on the basis of chromatographic behaviour and UV spectrum, 10-deacetylated taxoids (10-deacetylpaclitaxel, 10-deacetylcephalomannine, 7-xyloside-10-deacetylpaclitaxel and 10-deacetyltaxol C) could be detected together with 7-epi-10-deacetylpaclitaxel. From the needles of Taxus baccata var. Aurea Corr. the largest amounts isolated were of 10-deacetylbaccatin III, then 10-deacetylpaclitaxel and 7-xyloside-10-deacetylpaclitaxel, all compounds considered to be paclitaxel precursors in semisynthesis. The efficient mechanism of the separation of 10-acetylated taxoids from their 10-deacetylated derivatives on florisil on the basis of electron acceptor-electron donor interactions is discussed. Copyright 2000 John Wiley & Sons, Ltd.

  15. Validated RP-HPLC method for simultaneous determination and quantification of chlorpheniramine maleate, paracetamol and caffeine in tablet formulation.

    Science.gov (United States)

    Acheampong, Akwasi; Gyasi, Wilfred Owusu; Darko, Godfred; Apau, Joseph; Addai-Arhin, Sylvester

    2016-01-01

    Chlorpheniramine maleate-paracetamol-caffeine tablet formulation is one of the common over-the-counter drugs used for the treatment of cold and cough. A reversed-phase high-performance liquid-chromatography method has been successfully developed for the simultaneous determination of chlorpheniramine maleate, paracetamol and caffeine in a drug formulation. The RP-HPLC method employed a Phenomenex C18 reversed phase column (Luna 5µ, 250 × 4.6 mm) with an isocratic mixture of methanol and 0.05 M dibasic phosphate buffer pH 4.0 in the ratio of (30:70; v/v) as the mobile phase. The column temperature was kept at 30 °C. The flow rate was 1.0 mL/min and detection was by means of a UV detector at wavelength of 215 nm. All the active components were successfully eluted with mean retention times of 2.4, 4.2, 7.2 min for chlorpheniramine maleate, paracetamol and caffeine respectively. The method was found to be linear (R(2) > 0.99), precise (RSD paracetamol and caffeine without any interference by excipients.

  16. Qualitative and quantitative analysis of glucosinolates and nucleosides in Radix Isatidis by HPLC and liquid chromatography tandem mass spectrometry

    Directory of Open Access Journals (Sweden)

    Xiuming Wang

    2013-09-01

    Full Text Available Multi-component fingerprinting and quantitation of the glucosinolates and nucleosides in samples of Radix Isatidis have been carried out using high-performance liquid chromatography with diode-array detection and electrospray ionization tandem mass spectrometry (HPLC–DAD–ESI/MS. Five nucleosides together with one glucosinolate were identified by comparing retention times, ultraviolet spectra, mass spectra and/or empirical molecular formulae of reference compounds. Quantitation of these six compounds was carried out simultaneously by HPLC on a Phenomenex Luna C18 column using gradient elution with methanol and water and detection at 254 nm. All calibration curves were linear (r>0.9994 within test ranges. Limits of detection and quantitation were 0.33 ng and 2.50 ng on column, respectively. Intra- and inter-day precision (as relative standard deviation for all analytes was <2.19% with recoveries in the range 99.6%–101.8% at three concentration levels. The validated method was successfully applied to fingerprinting and assay of 25 batches of Radix Isatidis sourced from different geographical regions of China. The method is simple and reliable and has potential value in the quality control of Radix Isatidis.

  17. Efficiency of Polyphenol Extraction from Artificial Honey Using C18 Cartridges and Amberlite® XAD-2 Resin: A Comparative Study

    Directory of Open Access Journals (Sweden)

    Chua Yung An

    2016-01-01

    Full Text Available A comparative study of the extraction efficiency of nine known polyphenols [phenolic acids (benzoic acid, dihydroxybenzoic acid, gallic acid, trans-cinnamic acid, and vanillic acid and flavonoids (naringenin, naringin, quercetin, and rutin] was conducted by deliberately adding the polyphenols to an artificial honey solution and performing solid phase extraction (SPE. Two SPE methods were compared: one using Amberlite XAD-2 resin and another one using a C18 cartridge. A gradient high performance liquid chromatography system with an RP18 column and photodiode array detector was utilized to analyze the extracted polyphenols. The mean percent of recovery from the C18 cartridges was 74.2%, while that from the Amberlite XAD-2 resin was 43.7%. The recoveries of vanillic acid, naringin, and rutin were excellent (>90%; however, gallic acid was not obtained when C18 cartridges were used. Additionally, the reusability of Amberlite XAD-2 resin was investigated, revealing that the mean recovery of polyphenols decreased from 43.7% (1st extraction to 29.3% (3rd extraction. It was concluded that although Amberlite XAD-2 resin yielded a higher number of compounds, C18 cartridges gave a better extraction recovery. The lower recovery seen for the Amberlite XAD-2 resin also cannot be compensated by repeated extractions due to the gradual decrease of extraction recovery when reused.

  18. Quantitative analysis of three chiral pesticide enantiomers by high-performance column liquid chromatography.

    Science.gov (United States)

    Wang, Peng; Liu, Donghui; Gu, Xu; Jiang, Shuren; Zhou, Zhiqiang

    2008-01-01

    Methods for the enantiomeric quantitative determination of 3 chiral pesticides, paclobutrazol, myclobutanil, and uniconazole, and their residues in soil and water are reported. An effective chiral high-performance liquid chromatographic (HPLC)-UV method using an amylose-tris(3,5-dimethylphenylcarbamate; AD) column was developed for resolving the enantiomers and quantitative determination. The enantiomers were identified by a circular dichroism detector. Validation involved complete resolution of each of the 2 enantiomers, plus determination of linearity, precision, and limit of detection (LOD). The pesticide enantiomers were isolated by solvent extraction from soil and C18 solid-phase extraction from water. The 2 enantiomers of the 3 pesticides could be completely separated on the AD column using n-hexane isopropanol mobile phase. The linearity and precision results indicated that the method was reliable for the quantitative analysis of the enantiomers. LODs were 0.025, 0.05, and 0.05 mg/kg for each enantiomer of paclobutrazol, myclobutanil, and uniconazole, respectively. Recovery and precision data showed that the pretreatment procedures were satisfactory for enantiomer extraction and cleanup. This method can be used for optical purity determination of technical material and analysis of environmental residues.

  19. Design and characterization of microfabricated on-chip HPLC columns

    NARCIS (Netherlands)

    de Bruyne, S.

    2013-01-01

    The analysis of liquid mixtures, as e.g. encountered in environmental monitoring, food safety analysis, the development of novel pharmaceutical compounds, etc... is usually carried out using chromatogaphic separatoin techniques. Theoretical calculations have shown that the efficiency of these

  20. Residue analysis of spectinomycin in tissues of chicken and swine by HPLC

    National Research Council Canada - National Science Library

    Hamamoto, Kouko; Mizuno, Yasuharu; Koike, Ryoji; Yamaoka, Ryozo; Takahashi, Toshio; Takahashi, Yoshiyuki

    2003-01-01

    A reversed-phase HPLC method with ultraviolet detection using p-nitrophenyl hydrazine as a pre-column derivatizing reagent was investigated for the determination of the antibiotic spectinomycin (SPCM...

  1. HPLC SEPARATION OF CHIRAL ORGANOPHOSPHORUS PESTICIDES ON POLYSACCHARIDE CHIRAL STATIONARY PHASES

    Science.gov (United States)

    High-performance liquid chromatographic separation of the individual enantiomers of 12 organophosphorus pesticides (OPs) were obtained on polysaccharide chiral HPLC columns using an alkane-alcohol mobile phase. The OP pesticides were crotoxyphos, dialifor, dyfonate, fenamiphos, ...

  2. A Validated HPLC-DAD Method for Simultaneous Determination of Etodolac and Pantoprazole in Rat Plasma

    Directory of Open Access Journals (Sweden)

    Ali S. Abdelhameed

    2014-01-01

    Full Text Available A simple, sensitive, and accurate HPLC-DAD method has been developed and validated for the simultaneous determination of pantoprazole and etodolac in rat plasma as a tool for therapeutic drug monitoring. Optimal chromatographic separation of the analytes was achieved on a Waters Symmetry C18 column using a mobile phase that consisted of phosphate buffer pH~4.0 as eluent A and acetonitrile as eluent B in a ratio of A : B, 55 : 45 v/v for 6 min, pumped isocratically at a flow rate of 0.8 mL min−1. The eluted analytes were monitored using photodiode array detector set to quantify samples at 254 nm. The method was linear with r2=0.9999 for PTZ and r2=0.9995 for ETD at a concentration range of 0.1–15 and 5–50 μgmL−1 for PTZ and ETD, respectively. The limits of detection were found to be 0.033 and 0.918 μgmL−1 for PTZ and ETD, respectively. The method was statistically validated for linearity, accuracy, precision, and selectivity following the International Conference for Harmonization (ICH guidelines. The reproducibility of the method was reliable with the intra- and interday precision (% RSD <7.76% for PTZ and <7.58 % for ETD.

  3. Application of HPLC-DAD Technique for Determination of Phenolic Compounds in Bee Pollen Loads

    Directory of Open Access Journals (Sweden)

    Waś Ewa

    2017-06-01

    Full Text Available A method was elaborated to determine phenolic compounds (vanillin, caffeic, p-coumaric and salicylic acids, and flavonoids: rutin, hesperetin, quercetin, pinocembrin, apigenin, kaempferol, isorhamnetin, chrysin, and acacetin in bee pollen loads using highperformance liquid chromatography with a diode array detector (HPLC-DAD. Phenolic compounds from bee pollen were isolated on Cleanert C18-SPE columns (500 mg/6 mL, Agela Technologies. Polyphenols were identified by comparing the retention times and spectra of compounds found in pollen load samples with the ones of the standard mixture. Quantitative analysis was conducted using the external standard method. In addition, basic validation parameters for the method were determined. For the identified compounds (except for the salicylic acid, satisfactory (≥0.997 linear correlations were obtained. The elaborated method showed high repeatability and inter-laboratory reproducibility. Variability coeffcients of the majority of phenolic compounds did not exceed 10% in conditions of repeatability and inter-laboratory reproducibility, and for the total polyphenolic content they were 1.7 and 5.1%, respectively. The pollen load samples (n = 15 differed in qualitative and quantitative composition of the phenolic compounds. In all the samples, we identified the p-coumaric and salicylic acids and flavonoids rutin, hesperetin, and apigenin nevertheless, these compounds’ contents significantly differed among individual samples. The total phenolic content in the tested samples of pollen loads ranged from 0.653 to 5.966 mg/100 g (on average 2.737 mg/100 g.

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

    Directory of Open Access Journals (Sweden)

    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.

  5. Quantification of β-ecdysone in differents parts of Pfaffia glomerata by HPLC

    Directory of Open Access Journals (Sweden)

    Lara Z. Serra

    2012-12-01

    Full Text Available Pfaffia glomerata (Spreng. Pedersen, Amaranthaceae, is widely distributed in Brazil. Roots are considered as the world's greatest supplier and β-ecdysone is the most important compound extracted from roots of Pfaffia glomerata. So, the aim this study was analyze the presence of β-ecdysone in the inflorescences and stems and compared with the content from roots of Pfaffia glomerata and determine the best extractive method of β-ecdysone this plant. The crude extracts were obtained by Soxhlet method, reflux, maceration, percolation and turbolyse. Compound extracts were quantified by High Performance Liquid Chromatography (HPLC. The analysis were carried out a Phenomenex Column C18, 5 µm, 250x4,6mm, maintened at 30 ºC, gradient system using as mobile phase a mixture of methanol and water, flow rate 1,0 mL and detection at 245 nm. Results showed Soxhlet method with ethanol:water (90:10 v/v presented the higher concentration of β-ecdysone in P. glomerata and inflorescences showed higher amount of this active substance (3,06%, compared with stems (2,37% and roots (1,63%, showing that the inflorescences and plant stems may also be used as a rich source of β-ecdysone.

  6. Quantification of β-ecdysone in differents parts of Pfaffia glomerata by HPLC

    Directory of Open Access Journals (Sweden)

    Lara Z. Serra

    2012-09-01

    Full Text Available Pfaffia glomerata (Spreng. Pedersen, Amaranthaceae, is widely distributed in Brazil. Roots are considered as the world's greatest supplier and β-ecdysone is the most important compound extracted from roots of Pfaffia glomerata. So, the aim this study was analyze the presence of β-ecdysone in the inflorescences and stems and compared with the content from roots of Pfaffia glomerata and determine the best extractive method of β-ecdysone this plant. The crude extracts were obtained by Soxhlet method, reflux, maceration, percolation and turbolyse. Compound extracts were quantified by High Performance Liquid Chromatography (HPLC. The analysis were carried out a Phenomenex Column C18, 5 µm, 250x4,6mm, maintened at 30 ºC, gradient system using as mobile phase a mixture of methanol and water, flow rate 1,0 mL and detection at 245 nm. Results showed Soxhlet method with ethanol:water (90:10 v/v presented the higher concentration of β-ecdysone in P. glomerata and inflorescences showed higher amount of this active substance (3,06%, compared with stems (2,37% and roots (1,63%, showing that the inflorescences and plant stems may also be used as a rich source of β-ecdysone.

  7. Simultaneous HPLC analysis of crebanine, dicentrine, stephanine and tetrahydropalmatine in Stephania venosa

    Directory of Open Access Journals (Sweden)

    Sumet Kongkiatpaiboon

    Full Text Available ABSTRACT Stephania venosa (Blume Spreng., Menispermaceae, has been traditionally used as tonic drug and treatment of various diseases in South East Asian countries. In order to evaluate the quality and standardization of S. venosa roots, the HPLC method for quantification of the content of major components in S. venosa was developed and validated. The chromatographic separation was performed on a Hypersil BDS C18 column using gradient system of 100 mM ammonium acetate in water and methanol with flow rate 1 ml/min. Detection wavelength was set at 210 nm for tetrahydropalmatine, 280 nm for dicentrine and crebanine, and 270 nm for stephanine. The validated method showed good sensitivity, linearity, precision, and accuracy. The suitable solvent that yielded highest alkaloids contents from the matrix was optimized. S. venosa samples collected from various locations were analyzed. The present study provided comprehensive overview of major components in S. venosa. A remarkable variation in the accumulation of alkaloids in each population and the between individual in the same population could be observed. Our results showed the heterogeneity of S. venosa in Thailand which would need a further study for species delimitations.

  8. [Determination of five kinds of restricted preservatives in cosmetics by HPLC].

    Science.gov (United States)

    Lu, Lin; Li, Hui; Gao, Yanhong

    2014-07-01

    In order to detect and monitor preservative component in cosmetic, the method for determination of five kinds of restricted substances benzoic acid, chloroxylenol, o-phenyl phenol and p-chloro-m-cresol chlorhexidine in cosmetics by HPLC was carried out. 10 kinds of commercial cosmetics as test samples, using methanol as the extracting solution, saturated sodium chloride solution as precipitant, and Agilent Eclipse XDB-C18 (250 mm x 4.6 mm, 5 microm) as analytical column methanol -0.1 nol/L sodium dihydrogen phosphate solution (pH 3.0) was as mobile phase eluting at a flow rate 1.0 ml/min, the detective wavelength was 285 nm. The method for determination of chlorhexidine and other four kinds of restricted substances had good linear relationship, the linear correlation coefficients were greater than 0.9990, the limit of detection ( LOD) of 0.8 - 2.2 ng, the average recoveries were between 95.0% -104.0%. The method is accurate and reliable, and it is a useful supplement for "Hygienic Standard for Cosmetics".

  9. Simultaneous determination of 15 nitroimidazoles in cosmetics by HPLC coupled with electrospray ionization- tandem mass spectrometry.

    Science.gov (United States)

    Meng, Xian-Shuang; Bai, Hua; Zhang, Qing; Lv, Qing; Chen, Yun-Xia; Ma, Hui-Juan; Li, Jing-Rui; Ma, Qiang

    2014-01-01

    A sensitive and reliable analytical method based on HPLC/MSIMS has been developed for the simultaneous determination of 15 nitroimidazoles in cosmetics. A diversity of cosmetic samples, including powder, lotion, shampoo, and cream were collected. The samples were ultrasonically extracted with aqueous methanol, and the extracts were then subjected to cleanup bySPE using an Oasis HLB cartridge followed by filtration with a 0.20 pm membrane filter. Afterwards, chromatographic separation was performed on an XSelect CSH C18 column (2.1 x 150 mm, 3.5 pm) maintained at 30°C within 15 min by a gradient of acetonitrile-0.1% aqueous formic acid solution at a flow rate of 0.25 mL/min. The mass spectrometric detection was carried, out using electrospray positive ionization under the multiple reaction monitoring mode. A good linearity was observed over the concentration range from 0.5 to 500 ng/mL. The intraday and interday precisions, which were investigated by determining all target compounds in cosmetics seven times/day and on 7 consecutive days, were below 5.00%. The mean recoveries at three spiked levels ranged from 80.42 to 100.83% with the RSDs from 0.45 to 9.02%. The LOQs were determined to be between 0.01 and 0.1 mg/kg. The method was sufficiently rapid, reliable, and sensitive for the determination of 15 nitroimidazoles in cosmetics.

  10. A Validated RP-HPLC Method for the Estimation of Pizotifen in Pharmaceutical Dosage Form

    Directory of Open Access Journals (Sweden)

    M. V. Basaveswara Rao

    2012-01-01

    Full Text Available A simple, selective, linear, precise, and accurate RP-HPLC method was developed and validated for rapid assay of Pizotifen in pharmaceutical dosage form. Isocratic elution at a flow rate of 1.0 mL/min was employed on Chromosil C18 (250 mm × 4.6 mm, 5 μm column at ambient temperature. The mobile phase consists of methanol : acetonitrile in the ratio of 10 : 90 v/v. The UV detection wavelength was 230 nm, and 20 μL sample was injected. The retention time for Pizotifen was 2.019 min. The percent RSD for accuracy of the method was found to be 0.2603%. The method was validated as per the ICH guidelines. The method can be successfully applied for routine analysis of Pizotifen in the rapid and reliable determination of Pizotifen in pharmaceutical dosage form.

  11. Fast gradient HPLC/MS separation of phenolics in green tea to monitor their degradation.

    Science.gov (United States)

    Šilarová, Petra; Česlová, Lenka; Meloun, Milan

    2017-12-15

    The degradation of catechins and other phenolics in green tea infusions were monitored using fast HPLC/MS separation. The final separation was performed within 2.5min using Ascentis Express C18 column (50mm×2.1mm i.d.) packed with 2μm porous shell particles. Degradation was studied in relation to the temperature of water (70, 80, 90°C) and the standing time of the infusion (up to 6h). Along with chromatographic separation, the antioxidant properties of the infusions were monitored using two spectrophotometric methods. During staying of green tea infusion, the degradation of some catechins probably to gallic acid was observed. Finally, the influence of tea bag storage on antioxidant properties of green tea was evaluated. Rapid degradation of antioxidants after 3weeks was observed. The principal component analysis, factor analysis and discriminant analysis were used for the statistical evaluation of obtained experimental data. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Simultaneous Determination of Four Compounds in a Nelumbo nucifera Seed Embryo by HPLC-DAD

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    Gahee Ryu

    2017-01-01

    Full Text Available Nelumbo nucifera has a variety of biological activities. So it was importantly used as various herbal medicines since traditional times. A simple, fast, and sensitive high-performance liquid chromatographic (HPLC method was developed in this study for efficient quality control of N. nucifera. Four different compounds, including neferine, 1,2,3,4-tetrahydro-1-[(4-hydroxyphenyl methyl]-2-methyl-7-isoquinolinol, 1-hydroxy-2-methylpropene, and 3-(prop-1-enylbenzene-1,2,4,5-tetrol, were simultaneously determined. The four compounds were isolated through a Dionex C18 column by gradient elution with 0.1% TFA-water and methanol. The flow rate was 1.0 mL/min, and the wavelength was detected at 205, 254, 280, and 330 nm. The chromatograms were acquired at 205 nm. The four compounds showed good linear relationships (r2>0.96 over five different concentrations, and an average recovery of the method ranged from 96.27% to 108.78%. Through the analysis validation test and application of the method, the optimized conditions verified that it is efficient to isolate the compounds of N. nucifera seed embryos.

  13. Gradient HPLC method development and validation for Simultaneous estimation of Rosiglitazone and Gliclazide.

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    Uttam Singh Baghel

    2012-10-01

    Full Text Available Objective: The aim of present work was to develop a gradient RP-HPLC method for simultaneous analysis of rosiglitazone and gliclazide, in a tablet dosage form. Method: Chromatographic system was optimized using a hypersil C18 (250mm x 4.6mm, 5毺 m column with potassium dihydrogen phosphate (pH-7.0 and acetonitrile in the ratio of 60:40, as mobile phase, at a flow rate of 1.0 ml/min. Detection was carried out at 225 nm by a SPD-20A prominence UV/Vis detector. Result: Rosiglitazone and gliclazide were eluted with retention times of 17.36 and 7.06 min, respectively. Beer’s Lambert ’s Law was obeyed over the concentration ranges of 5 to 70 毺 g/ml and 2 to 12 毺 g/ml for rosiglitazone and gliclazide, respectively. Conclusion: The high recovery and low coefficients of variation confirm the suitability of the method for simultaneous analysis of both drugs in a tablets dosage form. Statistical analysis proves that the method is sensitive and significant for the analysis of rosiglitazone and gliclazide in pure and in pharmaceutical dosage form without any interference from the excipients. The method was validated in accordance with ICH guidelines. Validation revealed the method is specific, rapid, accurate, precise, reliable, and reproducible.

  14. Simultaneous isocratic HPLC determination of vigabatrin and gabapentin in human plasma by dansyl derivatization.

    Science.gov (United States)

    Krivanek, Peter; Koppatz, Karl; Turnheim, Klaus

    2003-06-01

    A rapid and low-cost assay for simultaneous vigabatrin (VGA) and gabapentin (GBP) determination is described that can be performed with simple HPLC instrumentation. The method involves derivatization of the primary amine group of VGA and GBP with dansyl chloride followed by isocratic separation (column: microBondapak C-18, 10 microm, 300 x 3.9 mm; mobile phase: 50 mmol/L NaH(2)PO(4) in 40% acetonitrile) at 50 degrees C and fluorometric detection (excitation and emission wavelength: 318 and 510 nm, respectively) of the fluorescent product, which is stable for at least 7 days. Correlation coefficients of the calibration curves are >0.999 with a lower limit of detection of 0.3 microg/mL. Between- and within-run coefficients of variation are below 4.5%, and assay time is 15 minutes. This method may be used for therapeutic drug monitoring in the case of GBP and to control patient compliance in the case of VGA.

  15. The Determination of Food Dyes in Vitamins by RP-HPLC.

    Science.gov (United States)

    Šuleková, Monika; Hudák, Alexander; Smrčová, Miroslava

    2016-10-17

    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.

  16. 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.

  17. An HPLC method for the determination of selected amino acids in human embryo culture medium.

    Science.gov (United States)

    Drábková, Petra; Andrlová, Lenka; Kanďár, Roman

    2017-02-01

    A method for the determination of selected amino acids in culture medium using HPLC with fluorescence detection is described. Twenty hours after intra-cytoplasmic sperm injection, one randomly selected zygote was transferred to the culture medium. After incubation (72 h after fertilization), the culture medium in which the embryo was incubated and blank medium was immediately stored at -80°C. Filtered medium samples were derivatized with ortho-phthalaldehyde (naphthalene-2,3-dicarboxaldehyde), forming highly fluorescent amino acids derivatives. Reverse-phase columns (LichroCART, Purospher STAR RP18e or Ascentis Express C18 ) were used for the separation. The derivatives were analyzed by gradient elution with a mobile phase containing ethanol and sodium dihydrogen phosphate. The analytical performance of this method is satisfactory for all amino acids; the intra-assay coefficients of variation were amino acids before and after human embryo cultivation were observed. After embryo incubation, the levels of all amino acids in the medium were increased, apart from aspartate and asparagine. After the cultivation of some embryos, amino acids which were not part of the medium were detected. Low amino acids turnover was observed in some embryos. Copyright © 2016 John Wiley & Sons, Ltd.

  18. [Determination of dimethyl fumarate in bakery food by d-SPE-HPLC-PDA].

    Science.gov (United States)

    Yang, Jie; Luo, Mengtian; Feng, Di; Miao, Hong; Song, Shufeng; Zhao, Yunfeng

    2015-05-01

    To establish a simple and rapid pretreatment method with dispersive solid phase extraction ( d-SPE) by HPLC for determination of dimethyl fumarate in bakery foods. Dimethyl fumarate in samples was ultrasonically extracted by methanol, and cleaned up with d-SPE. Then, it was separated on C18 chromatographic column (4.6 mm x 25 mm, 5 μm) with a mixture of methanol--0.03 mol/L sodium acetate and 0.008 mol/L tetrabutyl ammonium bromide (40: 60, V/V) as mobile phase. The photodiode array detector was used in the determination under λ = 220 nm. In the linear range of 0.1 -25 μg/ml, the correlation coefficients was r > 0.999, and the average recoveries of the spiked samples were in the range of 82.8% - 107.5% with relative standard deviations (RSD) in the range of 3.30% - 7.30% (n = 6). The limit of detection ( LOD) was 0.4 mg/kg, and the limit of quantification was 1.0 mg/kg. The method is simple, rapid, sensitive and accurate, and suitable for determine dimethyl fumarate in bakery foods.

  19. Reversed-phase HPLC retention data in correlation studies with in lipophilicity molecular descriptors of carotenoids

    Directory of Open Access Journals (Sweden)

    Podunavac-Kuzmanović Sanja O.

    2013-01-01

    Full Text Available Influence of chemical structure on the lipophilicity of isolated free carotenoids from paprika oleoresin has been studied by QSRR approach (Quantitative structure-retention relationship. The chromatographic behavior of these compounds was investigated by using reversed phase high-pressure liquid chromatography (RP HPLC. Retention mechanism has been determined using the following mobile phase: acetone -water, on reversed - phase column (SB-C18. A variety of lipophilicity parameters (logP were calculated by use of different software products. On the basis of correlations, the nonlinear structure-activity models were derived between the retention constants, tr (retention time of investigation compounds, and logP values. Five high quality QSRR models were found to have a good predictive ability and close agreement between experimental and predicted values. The study showed that the retention constants can be used as a measure of lipophilicity of investigated compounds at a high significant level. [Projekat Ministarstva nauke Republike Srbije, br. 172012, br. 172014 i br. 31055

  20. Development and Validation of Acyclovir HPLC External Standard Method in Human Plasma: Application to Pharmacokinetic Studies

    Directory of Open Access Journals (Sweden)

    Selvadurai Muralidharan

    2014-01-01

    Full Text Available A simple, rapid, and selective RP-HPLC method was developed for the estimation of acyclovir in human plasma. The method involves a simple protein precipitation technique. Chromatographic separation was carried out on a reverse phase C18 column using mixture of 5 mM ammonium acetate (pH 4.0 and acetonitrile (40 : 60, v/v at a flow rate of 1.0 mL/min with UV detection at 290 nm. The retention time of acyclovir was 4.12 minutes. The method was validated and found to be linear in the range of 25.0–150.0 ng/mL. Validation studies were achieved by using the fundamental parameters, including accuracy, precision, selectivity, sensitivity, linearity and range, stability studies, limit of detection (LOD, and limit of quantitation (LOQ. It shows recovery at 91.0% which is more precise and accurate compared to the other method. These results indicated that the bioanalytical method was linear, precise, and accurate. The new bioanalytical method was successfully applied to a pharmacokinetic linearity study in human plasma.

  1. Development of a Rapid and Sensitive HPLC Assay Method for Lenalidomide Capsules and Its Related Substances

    Directory of Open Access Journals (Sweden)

    L. Maheshwara Reddy

    2012-01-01

    Full Text Available A chromatographic method was established for the determination of lenalidomide and related substances in 10 mg and 5 mg capsules using Sunfire C-18(250×4.6 mm ID, 5 μm HPCL column with 85:15 v/v ratio of mobile phases A (mixture of phosphoric acid buffer and 1-octane sulphonic acid sodium salt and B(55: 45 v/v ratio of methanol and acetonitrile at 40°C and 210 nm wave length. The degradation studies were performed using 0.1N HCl, 0.1 N NaOH, 1% v/v hydrogen peroxide, humidity, UV at 254 nm, Sun light, and heat to 60°C. No significant degradation of lenalidomide was observed. However, the slight degradation was observed in presence of NaOH. The developed HPLC method gave the peaks purity angle was less their threshold angle, indicating it to be suitable for stability studies. It was validated with respect to linearity, accuracy, precision, ruggedness, and robustness.

  2. Rapid and sensitive method for determination of withaferin-A in human plasma by HPLC.

    Science.gov (United States)

    Patial, Pankaj; Gota, Vikram

    2011-02-01

    To develop and validate a rapid and sensitive high-performance liquid chromatographic method for determination of withaferin-A in human plasma. Withaferin-A, the active molecule of a traditional Indian herb, has demonstrated several biological activities in preclinical models. A validated bioassay is not available for its pharmacokinetic evaluation. The chromatographic system used a reverse-phase C18 column with UV-visible detection at 225 nm. The mobile phase consisted of water and acetonitrile applied in a gradient flow. Withaferin-A was extracted by simple protein-precipitation technique. The calibration curve was linear in the concentration range of 0.05-1.6 µg/ml. The method has the desired sensitivity to detect the plasma concentration range of withaferin-A that is likely to show biological activity based on in vitro data. This is the first HPLC method ever described for the estimation of withaferin-A in human plasma which could be applied for pharmacokinetic studies.

  3. Simultaneous determination of fluoxetine, citalopram, paroxetine, venlafaxine in plasma by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-MS/ESI).

    Science.gov (United States)

    Juan, He; Zhiling, Zhou; Huande, Li

    2005-06-05

    Fluoxetine, citalopram, paroxetine and venlafaxine have been widely used in the treatment of depression. However, no study has been conducted to determine the four drugs simultaneously by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-MS/ESI). To establish a new, rapid and sensitive HPLC-MS/ESI method for simultaneous determination and screening in human plasma of the four most commonly prescribed nontricyclic antidepressants: fluoxetine, citalopram, paroxetine and venlafaxine. The analytes in plasma were extracted by solid-phase-extraction column after samples had been alkalinized. The HPLC separation of the analytes was performed on a MACHEREY-NAGEL C(18) (250 mmx4.6 mm, 5 microm, Germany) column, using water (formic acid 0.6 per thousand, ammonium acetate: 30 mmol/l)-acetonitrile (35:65, v/v) as mobile phase, with a flow-rate of 0.85 ml/min. The compounds were ionized in the electrospray ionization (ESI) ion source of the mass spectrometer and were detected in the selected ion recording (SIR) mode. The calibration curves were linear in the 5.0-1000.0 ng/ml range for all compounds, all of them with coefficients of determination above 0.9900. The average extraction recoveries for all the four analytes were above 73.2%. The methodology recoveries were higher than 95.0%. The limits of detection (LODs) were 0.5, 0.3, 0.3 and 0.1 ng/ml for fluoxetine, citalopram, paroxetine and venlafaxine, respectively. The intra- and inter-day variation coefficients were less than 15.0%. The method is accurate, sensitive and simple for routine therapeutic drug monitoring (TDM) as well as toxicologic screening, and for the study of the pharmacokinetics and metabolism of the four drugs.

  4. Development and Validation of Stability Indicating HPTLC and HPLC Methods for Simultaneous Determination of Telmisartan and Atorvastatin in Their Formulations

    Directory of Open Access Journals (Sweden)

    Kaliappan Ilango

    2013-01-01

    Full Text Available The present study describes development and subsequent validation of stability indicating HPLC and HPTLC methods for simultaneous estimation of Telmisartan (TLM and Atorvastatin (ATV in their combined formulation. The proposed RP-HPLC method utilizes a Phenomenex Luna C18 column using acetonitrile: 0.025 M ammonium acetate (38 : 52%, v/v as mobile phase (pH 3.8, flow rate of 1.0 mL/min. Quantification was achieved with UV detection at 281 nm over concentration range of 12 to 72 μg/mL for TLM and 3 to 18 μg/mL for ATV respectively. In HPTLC, separations were performed on silica gel 60 F254 using toluene-methanol-ethyl acetate-acetic acid (5 : 1 : 1 : 0.3, v/v as mobile phase. The compact bands of TLM and ATV at 0.37 ± 0.02 and 0.63 ± 0.01 respectively were scanned at 279 nm. Linear regression analysis revealed linearity in the range of 40 to 240 ng/band for TLM and 10 to 60 ng/band for ATV respectively. For both the methods, dosage form was exposed to thermal, photolytic, acid, alkali and oxidative stress. The methods distinctly separated the drugs and degradation products even in actual samples. In conclusion, the proposed HPLC and HPTLC methods were appropriate for routine quantification of TLM and ATV in tablet formulation.

  5. Identification and determination of the major constituents in traditional Chinese medicine Longdan Xiegan Pill by HPLC-DAD-ESI-MS

    Directory of Open Access Journals (Sweden)

    Hui Liu

    2011-02-01

    Full Text Available A novel and sensitive HPLC-UV method has been developed for the simultaneous determination of twelve major compounds in Longdan Xiegan Pill. The chemical profile of the twelve compounds, including geniposidic acid (1, geniposide(2, gentiopicroside(3, liquiritin(4, crocin(5, baicalin(6, wogonoside(7, baicalein(8, glycyrrhizic acid (9, wogonin (10, oroxylin A (11 and aristolochic acid A (12, was acquired using high-performance liquid chromatography-diode array detector coupled with an electrospray tandem mass spectrometer (HPLC-DAD-ESI-MS. The analysis was performed on a Dikma Platisil ODS C18 column (250 mm × 4. 6 mm, 5 μm with a gradient solvent system of acetonitrile-0. 1% aqueous formic acid. The validation was carried out and the linearities (r > 0. 9996, repeatability (RSD<1. 8%, intra- and inter-day precision (RSD< 1. 3%, and recoveries (ranging from 96. 6% to 103. 4% were acceptable. The limits of detection (LOD of these compounds ranged from 0.29 to 4. 17 ng. Aristolochic acid A, which is the toxic ingredient, was not detected in all the batches of Longdan Xiegan Pill. Furthermore, hierarchical cluster analysis was used to evaluate the variation of the herbal prescription. The proposed method is simple, effective and suitable for the quality control of this traditional Chinese medicine (TCM. Keywords: Longdan Xiegan Pill, high-performance liquid chromatography-diode array detector coupled with an electrospray tandem mass spectrometer (HPLC-DAD-ESI-MS, qualitative evaluation, aristolochic acid A, hierarchical cluster analysis

  6. HPLC assay of acetylsalicylic acid, paracetamol, caffeine and phenobarbital in tablets.

    Science.gov (United States)

    Franeta, J T; Agbaba, D; Eric, S; Pavkov, S; Aleksic, M; Vladimirov, S

    2002-09-01

    This paper present a HPLC method for simultaneous determination of acetylsalicylic acid, paracetamol, caffeine and phenobarbital in tablets, using chromatographic system consisting a Bio Rad 18 01 solvent pump, Rheodine 71 25 injector and Bio Rad 18 01 UV-Vis Detector. Separation was achieved using Bio SiL HL C18, 5 microm, 250 x 4.6 mm column. Mixture of acetonitrile-water (25:75 v/v) adjusted to pH 2.5 with phosphoric acid was used as a mobile phase at a flow rate of 2.0 ml min(-1). UV detection was at 207 nm range 0.01 AUFS. Under the same conditions it was possible to determine the level of salicylic acid. The chromatographic parameters such as retention times, capacity factor, peak asymmetry, selectivity factor and resolution factor was determined. The validation parameters: linearity (r > 0.998), intra-day precision (RSD: 0.36-1.89%) and inter-day precision (RSD: 0.58-2.18%), sensitivity (LOD: 9 x 10(-5)-1.7 x 10(-4) mg ml(-1) and LOQ: 2.5 x 10(-4)-5.6 x 10(-4) mg ml(-1)), accuracy (recoveries: 98.35-99.14%) and reproducibility (recovery values: 98.74-102.08% for acetylsalicylic acid, 99.93-102.11% for paracetamol, 98.25-102.12% for caffeine and 98.15-102.3% for phenobarbital) (RSD: 1.21-1.85%) were found to be satisfactory. The proposed HPLC method has been applied for the determination of acetylsalicylic acid, paracetamol, caffeine and phenobarbital in Malophenum tablets. The obtained RSD values were within 0.99-1.21%. The developed method is rapid and sensitive and therefore suitable for routine control of these drugs in dosage form.

  7. Robust HPLC-MS/MS method for levofloxacin and ciprofloxacin determination in human prostate tissue.

    Science.gov (United States)

    Szerkus, O; Jacyna, J; Gibas, A; Sieczkowski, M; Siluk, D; Matuszewski, M; Kaliszan, R; Markuszewski, M J

    2017-01-05

    Fluoroquinolones are the drugs of choice in the prevention of bacterial infections after transrectal ultrasound guided prostate biopsy. In order to improve assessment of antibacterial efficacy in the target tissue a simple, selective, rapid and robust HPLC-ESI-MS/MS method for the determination of levofloxacin and ciprofloxacin concentrations in human prostate bioptates was developed and validated. Preparation procedure for prostate samples (10mg) was carried out using homogenization and filtration steps. Analyses were performed within 3.5min using RP C18 column in the isocratic elution mode with mobile phase composed of a mixture of 0.1% formic acid aqueous solution and 0.1% formic acid methanol solution (v/v; 79:21). The method was linear between 0.3μg/g and 15μg/g for levofloxacin and ciprofloxacin with coefficient of correlation (r) ≥0.999. The limit of detection and the limit of quantification for levofloxacin were 0.06μg/g and 0.2μg/g and for ciprofloxacin were 0.04μg/g and 0.13μg/g, respectively. Average concentrations (±SD) of levofloxacin and ciprofloxacin obtained from patients tissue were 5.4±2.2μg/g and 3.9±1.5μg/g, respectively. Additionally, during validation procedure a novel, experimental design approach was applied for the robustness study. For evaluation of analytical method robustness, Plackett-Burman design was employed and for sample preparation method robustness Fractional Factorial design was used. The developed and validated method was successfully applied to examine prostate tissue samples obtained from patients enrolled into a clinical study. Up to now, there has been no other HPLC-ESI-MS/MS method reported for the simultaneous determination of levofloxacin and ciprofloxacin in human prostatic tissue. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Validation of an HPLC method for the simultaneous determination of eletriptan and UK 120.413

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    LJILJANA ZIVANOVIC

    2006-11-01

    Full Text Available Arapid and sensitive RPHPLCmethod was developed for the routine control analysis of eletriptan hydrobromide and its organic impurity UK 120.413 in Relpax® tablets. The chromatography was performed at 20 °Cusing a C18 XTerraTM (5 m, 150 × 4,6 mm column at a flow rate 1.0 ml/min. The drug and its impurity were detected at 225 nm. The mobile phase consisted of TEA (1 % – methanol (67.2:32.8 v/v, the pH of which was adjusted to 6.8 with 85 % orthophosphoric acid. Quantification was accomplished by the internal standard method. The developed RP HPLC method was validated by testing: accuracy, precision, repeatibility, specificity, detection limit, quantification limit, linearity, robustness and sensitivity. High linearity of the analytical procedure was confirmed over the concentration range of 0.05 – 1.00 mg/ml for eletriptan hydrobromide and from 0.10 – 1.50 µg/ml for UK 120.413, with correlation coefficients greater than r = 0.995. The low value of the RSD expressed the good repeatability and precision of the method. Experimental design and a response surface method were used to test robustness of the analytical procedure and to evaluate the effect of variation of the method parameters, namely the mobile phase composition, pH and temperature. They showed small deviations from the method setting. The good recovery and low RSD confirm the suitability of the proposed RP HPLC method for the routine determination of eletriptan hydrobromide and its impurity UK 120.413 in Relpax® tables.

  9. ICH guidelines-compliant HPLC-UV method for pharmaceutical ...

    African Journals Online (AJOL)

    ICH guidelines-compliant HPLC-UV method for pharmaceutical quality control and therapeutic drug monitoring of the multi-targeted tyrosine kinase inhibitor pazopanib. ... Oxamniquine (OXA) was used as internal standard (IS). The analytical column used for the separation was Nucleosil CN with dimensions (i.d. 250 × 4.6 ...

  10. Higher content of C18:1 trans fatty acids in early human milk fat of Roma breast-feeding women.

    Science.gov (United States)

    Marhol, P; Dlouhý, P; Rambousková, J; Pokorný, R; Wiererová, O; Hrncírová, D; Procházka, B; Andel, M

    2007-01-01

    The purpose of our study was to determine the content of trans fatty acids in early human breast milk as an indicator of dietary exposure in a sample of Roma breast-feeding women and in a sample of women from the general Czech population. We collected samples of early human milk from 43 Prague women from the general population and 21 Roma women. After lipid extraction, the fatty acids were converted into methyl esters (FAMEs). Finally, gas chromatography with flame ionization detector (GC-FID) analysis on a CP-Sil 88 column was used to determine C18:1 trans monoenic fatty acid levels and total trans isomers fatty acid levels in human milk. A significantly higher content of C18:1 trans fatty acid isomers was detected in human milk fat from Roma mothers than in women of the general population (2.73 vs. 2.09%, p bad eating habits. (c) 2007 S. Karger AG, Basel.

  11. Theoretische en practische aspecten van het gebruik van micro-HPLC

    NARCIS (Netherlands)

    de Fluiter P; Jansen EHJM

    1992-01-01

    A practical and theoretical approach for the implementation of micro high performance liquid chromatography (HPLC) is described. A new simple and rapid test procedure was developed in wich a HPLC system can be validated for its suitability for micro-bore columns. It appeared that the detector

  12. Highly sensitive determination of 2,4,6-trinitrotoluene and related byproducts using a diol functionalized column for high performance liquid chromatography.

    Directory of Open Access Journals (Sweden)

    Burcu Gumuscu

    Full Text Available In this work, a new detection method for complete separation of 2,4,6-trinitrotoluene (TNT; 2,4-dinitrotoluene (2,4-DNT; 2,6-dinitrotoluene (2,6-DNT; 2-aminodinitrotoluene (2-ADNT and 4-aminodinitrotoluene (4-ADNT molecules in high-performance liquid-chromatography (HPLC with UV sensor has been developed using diol column. This approach improves on cost, time, and sensitivity over the existing methods, providing a simple and effective alternative. Total analysis time was less than 13 minutes including column re-equilibration between runs, in which water and acetonitrile were used as gradient elution solvents. Under optimized conditions, the minimum resolution between 2,4-DNT and 2,6-DNT peaks was 2.06. The recovery rates for spiked environmental samples were between 95-98%. The detection limits for diol column ranged from 0.78 to 1.17 µg/L for TNT and its byproducts. While the solvent consumption was 26.4 mL/min for two-phase EPA and 30 mL/min for EPA 8330 methods, it was only 8.8 mL/min for diol column. The resolution was improved up to 49% respect to two-phase EPA and EPA 8330 methods. When compared to C-18 and phenyl-3 columns, solvent usage was reduced up to 64% using diol column and resolution was enhanced approximately two-fold. The sensitivity of diol column was afforded by the hydroxyl groups on polyol layer, joining the formation of charge-transfer complexes with nitroaromatic compounds according to acceptor-donor interactions. Having compliance with current requirements, the proposed method demonstrates sensitive and robust separation.

  13. Agarose-chitosan-C18film micro-solid phase extraction combined with high performance liquid chromatography for the determination of phenanthrene and pyrene in chrysanthemum tea samples.

    Science.gov (United States)

    Ng, Nyuk Ting; Sanagi, Mohd Marsin; Wan Ibrahim, Wan Nazihah; Wan Ibrahim, Wan Aini

    2017-05-01

    Agarose-chitosan-immobilized octadecylsilyl-silica (C 18 ) film micro-solid phase extraction (μSPE) was developed and applied for the determination of phenanthrene (PHE) and pyrene (PYR) in chrysanthemum tea samples using high performance liquid chromatography-ultraviolet detection (HPLC-UV). The film of blended agarose and chitosan allows good dispersion of C 18 , prevents the leaching of C 18 during application and enhances the film mechanical stability. Important μSPE parameters were optimized including amount of sorbent loading, extraction time, desorption solvent and desorption time. The matrix match calibration curves showed good linearity (r⩾0.994) over a concentration range of 1-500ppb. Under the optimized conditions, the proposed method showed good limits of detection (0.549-0.673ppb), good analyte recoveries (100.8-105.99%) and good reproducibilities (RSDs⩽13.53%, n=3) with preconcentration factors of 4 and 72 for PHE and PYR, respectively. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Direct 13C NMR Detection in HPLC Hyphenation Mode

    DEFF Research Database (Denmark)

    Wubshet, Sileshi Gizachew; Johansen, Kenneth; Nyberg, Nils

    2012-01-01

    is indubitable in simplifying structural elucidations. In the current study, we demonstrated direct (13)C NMR detection of triterpenoids from a Ganoderma lucidum extract in hyphenation mode. The combined advantage of a cryogenically cooled probe, miniaturization, and multiple trapping enabled the first reported......Solid phase extraction (SPE) was introduced as a crucial step in the HPLC-SPE-NMR technique to enable online analyte enrichment from which proton-detected NMR experiments on submicrogram amounts from complex mixtures were possible. However, the significance of direct-detected (13)C NMR experiments...... application of HPLC-SPE-NMR analysis using direct-detected (13)C NMR spectra. HPLC column loading, accumulative SPE trappings, and the effect of different elution solvents were evaluated and optimized. A column loading of approximately 600 mug of a prefractionated triterpenoid mixture, six trappings...

  15. Development of a high-performance liquid chromatography method based on a core-shell column approach for the rapid determination of multiclass polyphenols in grape pomaces.

    Science.gov (United States)

    Fontana, Ariel R; Antoniolli, Andrea; Bottini, Rubén

    2016-02-01

    A rapid and economically affordable reverse-phase chromatographic approach based on a core-shell column with high-performance liquid chromatography multi-wavelength detector (HPLC-MWD) is proposed for the quantification and quality control of multiclass polyphenols (PPs). The separation of 20 relevant polyphenols from grape pomace extracts (GPEs) was achieved in less than 12 min by using a Kinetex C18 column (3.0 mm × 100 mm, 2.6 μm) with a gradient system of ultrapure water (0.1% formic acid) and acetonitrile, a temperature of 35 °C and a flow rate of 0.8 mL min(-1). The maximum backpressure reached was 327 bar, meaning the developed method is adequate for standard HPLC instruments. The applicability of the method was demonstrated by the determination of PPs in GPEs of different red grape varieties. Cabernet Sauvignon GPE showed the highest content of studied PPs (9804.2 μg g(-1)GPE) followed by Bonarda GPE (7302.0 μg g(-1)GPE). Besides the methodological development for a high throughput routine quality control of GPEs, this is the first report of PPs content for Bonarda and Aspirant Bouchet GPE, so the results add knowledge for these grape varieties cultivated in Argentina. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Development and validation of RP-HPLC and UV-spectrophotometric methods for rapid simultaneous estimation of amlodipine and benazepril in pure and fixed dose combination

    Directory of Open Access Journals (Sweden)

    Abhi Kavathia

    2017-05-01

    Full Text Available High-performance liquid chromatographic (HPLC and UV spectrophotometric methods were developed and validated for the quantitative determination of amlodipine besylate (AM and benazepril hydrochloride (BZ. Different analytical performance parameters such as linearity, precision, accuracy, specificity, limit of detection (LOD and limit of quantification (LOQ were determined according to International Conference on Harmonization ICH Q2B guidelines. The RP-HPLC method was developed by the isocratic technique on a reversed-phase Shodex C-18 5e column. The retention time for AM and BZ was 4.43 min and 5.70 min respectively. The UV spectrophotometric determinations were performed at 237 nm and 366 nm for AM and at 237 nm for BZ. Correlation between absorbance of AM at 237 nm and 366 nm was established and based on developed correlation equation estimation of BZ at 237 nm was carried out. The linearity of the calibration curves for each analyte in the desired concentration range was good (r2 > 0.999 by both the HPLC and UV methods. The method showed good reproducibility and recovery with percent relative standard deviation less than 5%. Moreover, the accuracy and precision obtained with HPLC co-related well with the UV method which implied that UV spectroscopy can be a cheap, reliable and less time consuming alternative for chromatographic analysis. The proposed methods are highly sensitive, precise and accurate and hence successfully applied for determining the assay and in vitro dissolution of a marketed formulation.

  17. Development of a validated HPLC method for the determination of sennoside A and B, two major constituents of Cassia obovata Coll.

    Directory of Open Access Journals (Sweden)

    Ghassemi-Dehkordi Nasrollah

    2014-04-01

    Full Text Available Introduction: Cassia obovata Coll is the only Senna species which grows wild in Iran. In the present study, an optimised reverse High Performance Liquid Chromatography (HPLC validated method was established for quantification of sennosides A and B, the major constituents of C. obovata with a simple and accurate method. Methods: HPLC analysis was done using Waters 515 pump on a Nova-Pak C18 (3.9 × 150 mm. Millennium software was used for the determination of the sennoside A and B in Cassia species and processing the information. The method was validated according to USP 32 requirements. Results: The solvent impact on the selectivity factor and partition coefficient parameters evaluated. Using a conventional RP-18 L1 column, 3.9 × 150 mm, the mobile phase was selected after several trials with different mixtures of water and acetonitrile. Sennosides A and B were determined using the external standard calibration method. Using USP 35-NF 30, the LOD and LOQ were calculated. The reliability of the HPLC-method for analysis of sennoside A + B was validated through its linearity, reproducibility, repeatability, and recovery. Fina1ly ethanol:water (1:1 extracts of Cassia obovata and Cassia angustifolia were standardized by assay of sennoside A and B through above HPLC validated method. Conclusion: Through the above method, determination of sennosides in Cassia species are completely possible. Moreover, through comparing the results, even though sennosides are rich in Cassia angustifolia but, the results shows that C. obovata could be considered as an alternative source for sennosides A and B.

  18. Simultaneous Determination of Florfenicol and Diclazuril in Compound Powder by RP-HPLC-UV Method

    Directory of Open Access Journals (Sweden)

    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.

  19. An optimized high-performance liquid chromatography (HPLC method for benzoylmesaconine determination in Radix Aconiti Lateralis Preparata (Fuzi, aconite roots and its products

    Directory of Open Access Journals (Sweden)

    Xu Hongxi

    2008-05-01

    Full Text Available Abstract Background Benzoylmesaconine (BMA is the main Aconitum alkaloid in Radix Aconiti Lateralis Preparata (Fuzi, aconite roots with potent pharmacological activities, such as analgesia and anti-inflammation. The present study developed a simple and reliable method using BMA as a marker compound for the quality control of processed aconite roots and their products. Methods After extraction, a high-performance liquid chromatography (HPLC determination of BMA was conducted on a RP-C18 column by gradient elution with acetonitrile and aqueous phase, containing 0.1% phosphoric acid adjusted with triethylamine to pH 3.0. Results A distinct peak profile was obtained and separation of BMA was achieved. Method validation showed that the relative standard deviations (RSDs of the precision of BMA in all intra-day and inter-day assays were less than 1.36%, and that the average recovery rate was 96.95%. Quantitative analysis of BMA showed that the content of BMA varied significantly in processed aconite roots and their products. Conclusion This HPLC method using BMA as a marker compound is applicable to the quality control of processed aconite roots and their products.

  20. Simultaneous determination of cucurbitacin IIa and cucurbitacin IIb of Hemsleya amabilis by HPLC-MS/MS and their pharmacokinetic study in normal and indomethacin-induced rats.

    Science.gov (United States)

    Wang, Shougang; Guan, Xiaohui; Zhong, Xiuhong; Yang, Zhiping; Huang, Weili; Jia, Baoyang; Cui, Tao

    2016-10-01

    A selective and sensitive HPLC-MS/MS method was developed for the simultaneous determination of cucurbitacin IIa (cuIIa) and cucurbitacin IIb (cuIIb), the major bioactive cucurbitacins of Hemsleya amabilis, in rat plasma using euphadienol as internal standard (IS). After liquid-liquid extraction with dichloromethane, separation was achieved on a Syncronis HPLC C18 column (150 mm × 4.6 mm, 5 μm) using an isocratic mobile phase system consisting of acetonitrile-water (85:15, v/v) at a flow rate of 0.6 mL/min with a split ratio of 1:2. Detection was performed on a TSQ Quantum Ultra mass spectrometer equipped with an positive-ion electrospray ionization source. The lower limits of quantification (LLOQs) were 0.25 and 0.15 ng/mL for cuIIa and cuIIb, respectively. The intra- and inter-day precision was plasma were all >87.1%. The method was fully validated and applied to compare the pharmacokinetic profiles of the two cucurbitacins in rat plasma after oral administration of H. amabilis extract between normal and indomethacin-induced rats. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  1. Simultaneous determination of four iridoid and secoiridoid glycosides and comparative analysis of Radix Gentianae Macrophyllae and their related substitutes by HPLC.

    Science.gov (United States)

    Cao, Xiao-Yan; Wang, Zhe-Zhi

    2010-01-01

    Radix Gentianae Macrophyllae, a traditional Chinese medicine, has been frequently used to dispel rheumatism and ease pain. There are four species of Gentiana (G. macrophylla, G. straminea, G. dahurica and G. crassicaulis) recorded as herbal drugs in the Chinese Pharmacopoeia and two other Gentiana species (G. officinalis and G. siphonantha) are often used as substitutes. Currently, the LC fingerprint comparison among different species and evidence for the equivalent application of these herbs are lacking. To develop an HPLC method for the simultaneous determination of four iridoid and secoiridoid glycosides and a comparative study of six species of Gentiana. HPLC analysis was performed on a C(18) column (Phenomenex, 150 x 4.6 mm, 5 microm particle size) with gradient elution using 0.4% aqueous phosphoric acid and methanol at 242 nm. The proposed method was precise, accurate and sensitive enough for simultaneous quantitative evaluation of four iridoid and secoiridoid glycosides (loganic acid, swertiamarin, gentiopicroside and sweroside) in the six species of Gentiana. Contents of the four marker compounds varied from each other even among the samples from the same species and the LC chromatograms of the six species of Gentiana showed high similarities. he close similarity of LC chromatograms and chemical composition of the four genuine Gentiana species explain their popular usage as Radix Gentianae Macrophyllae in Chinese medicine. By comparing the four genuine Gentiana species, it is suggested that the two substitutes could be used as Radix Gentianae Macrophyllae to relieve the scarcity of resources.

  2. Preliminary investigation of the possibility for implementation of modified pharmacopoeial HPLC methods for quality control of metronidazole and ciprofloxacin in medicinal products used in veterinary medicine

    Directory of Open Access Journals (Sweden)

    Marjan Piponski

    2015-03-01

    Full Text Available Quality control of veterinary medicine products containing two different frequently used antibiotics metronidazole and ciprofloxacin hydrochloride, was considered and performed, using modified pharmacopoeial HPLC methods. Three different HPLC systems were used: Varian ProStar, Perkin Elmer Series and UPLC Shimadzu Prominence XR. The chromatographic columns used were LiChropher RP Select B 75 mm x 4 mm with 5 μm particles and Discovery C18 100 mm x 4,6 mm with 5 μm particles. Chromatographic methods used for both analytes were compendial, with minor modifications made for experimental purposes. Minor modifications of the pharmacopoeia prescribed chromatographic conditions, in both cases, led to better chromatographic parameters, good resolution and shorter analysis times. Optimized methods can be used for: determination of metronidazole in gel formulation, for its simultaneous quantification with preservatives present in the formulation and even for identification and quantification of its specified impurity, 2-methyl-5-nitroimidazole; determination of ciprofloxacin hydrochloride in film coated tablets and eye drops and identification and quantification of its specified impurities. These slightly modified and optimized pharmacopoeial methods for quality control of metronidazole and ciprofloxacin dosage forms used in veterinary medicine can be successfully applied in laboratories for quality control of veterinary medicines.

  3. Screening of preservatives by HPLC-PDA-ESI/MS: A focus on both allowed and recently forbidden compounds in the new EU cosmetics regulation.

    Science.gov (United States)

    Lecce, Raffaele; Regazzoni, Luca; Mustazza, Carlo; Incarnato, Giampaolo; Porrà, Rita; Panusa, Alessia

    2016-06-05

    Commission regulation (EU) No 358/2014 amending the new regulation (EC) No 1223/2009 on cosmetics has prohibited the use of isopropyl-, isobutyl-, phenyl-, benzyl- and pentylparaben. Furthermore, Commission regulation (EU) No 1004/2014 has lowered the maximum permitted concentration of butyl- and propylparaben in cosmetics and it has also banned them in leave-on products designed for application on the nappy area of children under three years of age. A HPLC-PDA-ESI/MS method has been developed herein for the detection of seventeen preservatives, both the most utilised and the recently forbidden by the new EU regulations. The separation of these compounds, including benzoic acid and its derivatives in a 1.10 - 3.04 log Pow range, has been performed with a gradient elution on a Symmetry(®) C18 column (250×4.6mm i.d., particle size 5μm) with water and acetonitrile (0.1% formic acid) as mobile phase. Quantification has been carried out by HPLC-PDA. The method has been validated and successfully applied to the analysis of a large number of cosmetics with different functions like rinse-off and leave-on, or composition like skin, hair, face and oral products. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. A HPLC method to evaluate the influence of photostabilizers on cosmetic formulations containing UV-filters and vitamins A and E.

    Science.gov (United States)

    Gaspar, Lorena Rigo; Campos, Patricia Maria Berardo Gonçalves Maia

    2010-09-15

    This paper reports a simple and reliable HPLC method to evaluate the influence of two currently available photostabilizers on cosmetic formulations containing combined UV-filters and vitamins A and E. Vitamins and UV-filters, widely encountered in products of daily use have to be routinely evaluated since photoinstability can lead to reductions in their efficacy and safety. UV-irradiated formulation samples were submitted to a procedure that included a reliable, precise and specific HPLC method employing a C18 column and detection at 325 and 235 nm. Methanol, isopropanol and water were the mobile phases in gradient elution. The method precision was between 0.28 and 5.07. The photostabilizers studied [diethylhexyl 2,6-naphthalate (DEHN) and benzotriazolyl dodecyl p-cresol (BTDC)], influenced the stability of octyl methoxycinnamate (OMC) associated with vitamins A and E. BTDC was considered the best photostabilizer to vitamins and OMC when the UV-filters were combined with both vitamins A and E. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  5. HPLC-MS method for the simultaneous quantification of the new HIV protease inhibitor darunavir, and 11 other antiretroviral agents in plasma of HIV-infected patients.

    Science.gov (United States)

    D'Avolio, Antonio; Siccardi, Marco; Sciandra, Mauro; Baietto, Lorena; Lorena, Baietto; Bonora, Stefano; Trentini, Laura; Di Perri, Giovanni

    2007-11-15

    A new method using high performance liquid chromatography coupled with electrospray mass spectrometry (HPLC-MS) was developed and validated, for the quantification of plasma concentration of the new protease inhibitors darunavir (DRV) and other 11 antiretroviral agents (ritonavir, amprenavir, atazanavir, lopinavir, saquinavir, indinavir, nelfinavir and its metabolite M-8, nevirapine, efavirenz and tipranavir). A simple protein precipitation extraction procedure was applied on 50 microl of plasma aliquots and chromatographic separation of drugs and Internal Standard (quinoxaline) was achieved with a gradient (acetonitrile and water with formic acid 0.05%) on an C-18 reverse phase analytical column with 25 min of analytical run. Calibration curves were optimised according to expected ranges of drug concentrations in patients, and correlation coefficient (r2) was higher than 0.998 for all analytes. Mean intra- and inter-day precision (relative standard deviation %) for all compounds were 8.4 and 8.3%, respectively, and mean accuracy (% of deviation from nominal level) was 3.9%. Extraction recovery ranged within 93 and 105% for all drugs analysed. This novel HPLC-MS methodology allows a specific, sensitive and reliable determination of DRV and 11 other antiretrovirals. In our hand, it was used to measure DRV and ritonavir plasma concentration in HIV-positive patients, and it is now successfully applied for routine therapeutic drug monitoring and pharmacokinetics studies.

  6. Simultaneous quantification of sulforaphene and sulforaphane by reverse phase HPLC and their content in Raphanus sativus L. var. caudatus Alef extracts.

    Science.gov (United States)

    Sangthong, Sarita; Weerapreeyakul, Natthida

    2016-06-15

    A simple, rapid and precise HPLC assay was developed for the well-known anti-cancer isothiocyanates-sulforaphene (SE) and sulforaphane (SF). The analytical system comprised RP-C18 column with isocratic 5% THF-95% water. High resolution was obtained (and eluted) of two distinct HPLC peaks of similar structures SE and SF analytes (at 23.01±0.02 and 25.65±0.03 min, respectively). The respective LOD vs. LOQ for SE and SF was 0.34 and 0.36 μg/ml vs. 1.02 and 1.08 μg/ml. This assay had the best linearity and accuracy. The recoveries were in the range of 96.83-101.17%. SF and SE were quantified in the pod of Raphanus sativus L. var. caudatus Alef extracts (2253.05±246.18 and 111.94±16.49 μg/g in the crude extract, respectively), while only SE was detected in the stem (1105.14±243.10 μg/g crude extract), as SF was lower than the detection limit. The validated method thus minimized and expedited simultaneous SE and SF analysis. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Development and Validation of a Stability-Indicating HPLC Method for the Simultaneous Determination of Florfenicol and Flunixin Meglumine Combination in an Injectable Solution.

    Science.gov (United States)

    Batrawi, Nidal; Naseef, Hani; Al-Rimawi, Fuad

    2017-01-01

    The combination of the powerful antimicrobial agent florfenicol and the nonsteroidal anti-inflammatory flunixin meglumine is used for the treatment of bovine respiratory disease (BRD) and control of BRD-associated pyrexia, in beef and nonlactating dairy cattle. This study describes the development and validation of an HPLC-UV method for the simultaneous determination of florfenicol and flunixin, in an injectable preparation with a mixture of excipients. The proposed RP-HPLC method was developed by a reversed phase- (RP-) C18e (250 mm × 4.6 mm, 5  μ m) column at room temperature, with an isocratic mobile phase of acetonitrile and water mixture, and pH was adjusted to 2.8 using diluted phosphoric acid, a flow rate of 1.0 mL/min, and ultraviolet detection at 268 nm. The stability-indicating method was developed by exposing the drugs to stress conditions of acid and base hydrolysis, oxidation, photodegradation, and thermal degradation; the obtained degraded products were successfully separated from the APIs. This method was validated in accordance with FDA and ICH guidelines and showed excellent linearity, accuracy, precision, specificity, robustness, LOD, LOQ, and system suitability results within the acceptance criteria.

  8. Development and Validation of a Stability-Indicating HPLC Method for the Simultaneous Determination of Florfenicol and Flunixin Meglumine Combination in an Injectable Solution

    Directory of Open Access Journals (Sweden)

    Nidal Batrawi

    2017-01-01

    Full Text Available The combination of the powerful antimicrobial agent florfenicol and the nonsteroidal anti-inflammatory flunixin meglumine is used for the treatment of bovine respiratory disease (BRD and control of BRD-associated pyrexia, in beef and nonlactating dairy cattle. This study describes the development and validation of an HPLC-UV method for the simultaneous determination of florfenicol and flunixin, in an injectable preparation with a mixture of excipients. The proposed RP-HPLC method was developed by a reversed phase- (RP- C18e (250 mm × 4.6 mm, 5 μm column at room temperature, with an isocratic mobile phase of acetonitrile and water mixture, and pH was adjusted to 2.8 using diluted phosphoric acid, a flow rate of 1.0 mL/min, and ultraviolet detection at 268 nm. The stability-indicating method was developed by exposing the drugs to stress conditions of acid and base hydrolysis, oxidation, photodegradation, and thermal degradation; the obtained degraded products were successfully separated from the APIs. This method was validated in accordance with FDA and ICH guidelines and showed excellent linearity, accuracy, precision, specificity, robustness, LOD, LOQ, and system suitability results within the acceptance criteria.

  9. ICH guidance in practice: validated stability-indicating HPLC method for simultaneous determination of ampicillin and cloxacillin in combination drug products.

    Science.gov (United States)

    Kumar, Vijay; Bhutani, Hemant; Singh, Saranjit

    2007-01-17

    Ampicillin and cloxacillin were degraded together under different stress test conditions prescribed by International Conference on Harmonization. The samples so generated were used to develop a stability-indicating high performance liquid chromatographic (HPLC) method for the two drugs. The drugs were well separated from degradation products using a reversed-phase (C-18) column and a mobile phase comprising of acetonitrile:phosphate buffer (pH 5.0), which was delivered initially in the ratio of 15:85 (v/v) for 1 min, then changed to 30:70 (v/v) for next 14 min, and finally equilibrated back to 15:85 (v/v) from 15 to 20 min. Other HPLC parameters were: flow rate, 1 ml/min; detection wavelength, 225 nm; and injection volume, 5 microl. The method was validated for linearity, precision, accuracy, specificity and selectivity. It was also compared with the assay procedures given in British Pharmacopoeia for individual drugs. Similar results were obtained, indicating that the proposed single method allowed selective analysis of both ampicillin and cloxacillin, in the presence of their degradation products formed under a variety of stress conditions. The developed procedure was also applicable to the determination of instability of the drugs in commercial products.

  10. Simple Reversed-Phase HPLC Method with Spectrophotometric Detection for Measuring Acetaminophen-Protein Adducts in Rat Liver Samples

    Directory of Open Access Journals (Sweden)

    Miteshkumar Acharya

    2012-01-01

    Full Text Available A simple reversed-phase HPLC method for measuring hepatic levels of acetaminophen- (APAP- protein adduct following an overdose of APAP was developed. An aliquot of liver homogenate in phosphate-buffered saline pH 7.4 (PBS was placed on a Nanosep centrifugal device, which was centrifuged to obtain a protein residue. This residue was incubated with a solution of p-aminobenzoic acid (PABA, the internal standard, and bacterial protease in PBS, transferred to a Nanosep centrifugal device, and centrifuged. A 100 μL portion of the filtrate was analyzed on a YMC-Pack ODS-AMQ C18 column, using 100 mM potassium dihydrogen phosphate-methanol-acetic acid (100 : 0.6 : 0.1 as the mobile phase, a flow rate of 1 mL/min, and photometric detection at 254 nm. PABA and APAP-cystein-S-yl (APAP-Cys eluted at ~14.7 min and 22.7 min, respectively. Method linearity, based on on-column concentrations of APAP-Cys, was observed over the range 0.078–40 μg. Recoveries of APAP-Cys from spiked blank liver homogenates ranged from ~83% to 91%. Limits of detection and of quantification of APAP-Cys, based on column concentrations, were 0.06 μg and 0.14 μg, respectively. RSD values for interday and intraday analyses of a blank liver homogenate spiked with APAP-Cyst at three levels were, in all cases, ≤1.0% and <1.5%, respectively. The proposed method was found appropriate for comparing the antidotal properties of N-acetylcysteine and taurine in a rat model of APAP poisoning.

  11. A Validated RP-HPLC Method for the Determination of Recombinant Human Insulin in Bulk and Pharmaceutical Dosage Form

    Directory of Open Access Journals (Sweden)

    B. A. Moussa

    2010-01-01

    Full Text Available A modified RP-HPLC method was developed for the quantitative determination of recombinant human insulin in bulk and pharmaceutical dosage form with reduced retention time. Study of the effects of the column temperature, pH of the mobile phase and presence of vial additives (phenol and m-cresol, or impurities (A-21 Disamido on the accuracy of the assay were assessed. Separation was achieved using a Hypersil BDS C-18 column and the mobile phase was composed of solution A (aqueous solution of 28.3 anhydrous Na2SO4g/L, pH 2.3 and solution B (28.5 g anhydrous Na2SO4 g/L in 50:50 mixture of water and acetonitrile, pH 2.3 in a ratio 48:52 (v/v at 45–50 °C. The column temperature was 40 °C, the flow rate was 1 mL/min and detection was performed at 216 nm. The procedures were validated according to international conference on harmonization (ICH guidelines. Recovery study was done applying standard addition technique for further validation of the procedure. The retention time of recombinant human insulin was 19.7 min as compared to 29 min obtained by the reference method. Analytical conditions fluctuations or presence of vial additives or impurities did not show any significant effect on the accuracy of the method. The prepared standard insulin solution in 0.01 N HCl was found to be stable for 5 days. Statistical comparison showed no significant difference between the described method and reference method regarding the accuracy and precision. The modified method can be applied for routine quality control applications for determination of recombinant human insulin.

  12. Isolation, cytotoxicity evaluation and HPLC-quantification of the chemical constituents from Prangos pabularia.

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    Saleem Farooq

    Full Text Available Phytochemical analysis of the dichloromethane:methanol (1:1 extract of root parts of Prangos pabularia led to the isolation of twelve cytotoxic constituents, viz., 6-hydroxycoumarin (1, 7-hydroxycoumarin (2, heraclenol-glycoside (3, xanthotoxol (4, heraclenol (5, oxypeucedanin hydrate (6, 8-((3,3-dimethyloxiran-2-ylmethyl-7-methoxy-2H-chromen-2-one (7, oxypeucedanin hydrate monoacetate (8, xanthotoxin (9, 4-((2-hydroxy-3-methylbut-3-en-1-yloxy-7H-furo[3,2-g]chromen-7-one (10, imperatorin (11 and osthol (12. The isolates were identified using spectral techniques in the light of literature. 3-(4,5-dimethyl thiazol-2yl-2,5-diphenyltetrazolium bromide (MTT cytotoxicity screening of the isolated constituents was carried out against six human cancer cell lines including lung (A549 and NCI-H322, epidermoid carcinoma (A431, melanoma (A375, prostate (PC-3 and Colon (HCT-116 cell lines. Osthol (12 exhibited the highest cytotoxicity with IC50 values of 3.2, 6.2, 10.9, 14.5, 24.8, and 30.2 µM against epidermoid carcinoma (A431, melanoma (A375, lung (NCI-H322, lung (A549, prostate (PC-3 and colon (HCT-116 cell lines respectively. Epidermoid carcinoma cell line A431 was sensitive to most of the compounds followed by lung (A549 cancer cell line. Finally a simple and reliable HPLC method was developed (RP-HPLC-DAD and validated for the simultaneous quantification of these cytotoxic constituents in Prangos pabularia. The extract was analyzed using a reversed-phase Agilent ZORBAX eclipse plus column C18 (4.6×250 mm, 5 µm at 250 nm wavelength using a gradient water-methanol solvent system at a flow rate of 0.8 ml/min. The RP-HPLC method is validated in terms of recovery, linearity, accuracy and precision (intra and inter-day validation. This method, because of shorter analysis time, makes it valuable for the commercial quality control of Prangos pabularia extracts and its future pharmaceutical preparations.

  13. Retinoid quantification by HPLC/MS(n)

    Science.gov (United States)

    McCaffery, Peter; Evans, James; Koul, Omanand; Volpert, Amy; Reid, Kevin; Ullman, M. David

    2002-01-01

    Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane-isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane-dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids.

  14. [Determination of aloin in aloes by HPLC].

    Science.gov (United States)

    Cao, Hong; Liu, Yun

    2003-04-01

    To describe a simple and rapid isocratic reversed-phase high performance liquid chromatography method for the baseline separation, identification and assay of aloin in aloes. The analytical column was a ZORBAX SB-C18(4.6 mm x 250 mm) filled with a 5 microns stationary phase. The mobile phase consisted of acetonitrile-water (25:75); the flow-rate was 1 mL.min-1. The injection volume was 10 microL. The DAD detector was set at 355 nm. The calibration curve was linear over the range of 0.17-5.9 micrograms (r = 0.9999). The average recovery of the method was 98.6%, RSD 1.32% (n = 6). The results showed that this method was reliable and accurate. The method was applied to eleven Cape and East African aloes of different origin.

  15. Development and validation of a high-performance liquid chromatography method with post-column derivatization for the detection of aflatoxins in cereals and grains.

    Science.gov (United States)

    Asghar, Muhammad Asif; Iqbal, Javed; Ahmed, Aftab; Khan, Mobeen Ahmed; Shamsuddin, Zuzzer Ali; Jamil, Khalid

    2016-06-01

    A novel, reliable and rapid high-performance liquid chromatography (HPLC) method with post-column derivatization was developed and validated. The HPLC method was used for the simultaneous determination of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in various cereals and grains. Samples were extracted with 80:20 (v/v) methanol:water and purified using C18 (40-63 μm) solid-phase extraction cartridges. AFs were separated using a LiChroCART-RP-18 (5 μm, 250 × 4.0 mm(2)) column. The mobile phase consisted of methanol:acetonitrile:buffer (17.5:17.5:65 v/v) (pH 7.4) delivered at the flow rate of 1.0 mL min(-1) The fluorescence of each AF was detected at λex = 365 nm and λem = 435 nm. All four AFs were properly resolved within the total run time of 20 min. The established method was extensively validated as a final verification of the method development by the evaluation of selectivity (AFB1, AFB2, AFG1 and AFG2), linearity (R(2) ≥ 0.9994), precision (average SD ≤ 2.79), accuracy (relative mean error ≤ -5.51), robustness (p developed method was applied for the analysis of AFs in 45 samples comprising rice (n = 20), wheat (n = 15) and maize (n = 10). The results showed that 65% of rice, 20% of wheat and 80% of maize samples were found contaminated with AFs. Thus, according to the achieved results, it is suggested that the newly developed HPLC method could be effectively applied for the routine analysis of the AFs in different cereals and grains. © The Author(s) 2014.

  16. 26 CFR 31.3306(c)(18)-1 - Services of certain nonresident aliens.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Services of certain nonresident aliens. 31.3306...) § 31.3306(c)(18)-1 Services of certain nonresident aliens. (a) (1) Services performed after 1961 by a nonresident alien individual who is temporarily present in the United States as a nonimmigrant under...

  17. Comparison of monolithic silica and polymethacrylate capillary columns for LC.

    Science.gov (United States)

    Moravcová, Dana; Jandera, Pavel; Urban, Jiri; Planeta, Josef

    2004-07-01

    Organic polymer monolithic capillary columns were prepared in fused-silica capillaries by radical co-polymerization of ethylene dimethacrylate and butyl methacrylate monomers with azobisisobutyronitrile as initiator of the polymerization reaction in the presence of various amounts of porogenic solvent mixtures and different concentration ratios of monomers and 1-propanol, 1,4-butanediol, and water. The chromatographic properties of the organic polymer monolithic columns were compared with those of commercial silica-based particulate and monolithic capillary and analytical HPLC columns. The tests included the determination of H-u curves, column permeabilities, pore distribution by inversed-SEC measurements, methylene and polar selectivities, and polar interactions with naphthalenesulphonic acid test samples. Organic polymer monolithic capillary columns show similar retention behaviour to chemically bonded alkyl silica columns for compounds with different polarities characterized by interaction indices, Ix, but have lower methylene selectivities and do not show polar interactions with sulphonic acids. The commercial capillary and analytical silica gel-based monolithic columns showed similar selectivities and provided symmetrical peaks, indicating no significant surface heterogeneities. To allow accurate characterization of the properties of capillary monolithic columns, the experimental data should be corrected for extra-column contributions. With 0.3 mm ID capillary columns, corrections for extra-column volume contributions are sufficient, but to obtain true information on the efficiency of 0.1 mm ID capillary columns, the experimental bandwidths should be corrected for extra-column contributions to peak broadening.

  18. Reversed-phase liquid chromatography of radiolabeled peptides using a C18 guard-PAK precolumn system

    Energy Technology Data Exchange (ETDEWEB)

    Carriere, P.D.; Bennett, H.P. (McGill Univ., Montreal, Quebec (Canada))

    1989-03-01

    In order to avoid radioactive contamination of high-performance liquid chromatography columns and injectors, we have investigated the use of a Guard-PAK precolumn system for the chromatography of ({sup 125}I) labeled peptides. Two gonadotropin-releasing hormone analogs: (1) (D-Ala6-des-Gly10)-GnRH (GnRH-(Ala6)) and (2) (D-Ser(TBu)6-des-Gly10)-GnRH (GnRH-(Ser6)) and rat prolactin (r-PRL) were radiolabeled with {sup 125}I and subjected to reversed-phase liquid chromatography using a C18 Guard-PAK precolumn system. Major peak fractions of purified ({sup 125}I)GnRH-(Ala6), ({sup 125}I)GnRH-(Ser6), and ({sup 125}I)r-PRL eluted at 24%, 28%, and 55% acetonitrile, respectively. Purified ({sup 125}I)GnRH analogs showed specific high affinity binding to rat anterior pituitary gland membranes (specific activity: 1500-1700 Ci/mmol). Purified ({sup 125}I)r-PRL showed high affinity binding to r-PRL antibody by RIA (specific activity: 70-75 microCi/micrograms). This rapid and efficient chromatographic method should be useful in the separation of a wide range of radiolabeled protein and peptide molecules.

  19. Desarrollo y validación de un método analítico (HPLC RP para la determinación de teofilina en plasma

    Directory of Open Access Journals (Sweden)

    Luisa Fernanda Ponce D´léon

    2011-03-01

    Full Text Available Theophylline is a drug widely known for treating asthma and sorne other chronic respiratory diseases. At present time,the blood levels of drugs can be related with eficacy and side effects. There are around 10 drug products ofprogrammedrelease theophyIline forms, commercially available in Colombia. Most of the problems found in the utilization are caused by the non advisable interchange of theophyIline drug products, without any bioequivalency data for them. This study proposes a specific and validated analytical methodology for theophylline in biological fluid s including blood, which is useful for bioavailability and bioequivalencystudies, rutinary monitoring of theophylline blood levels and in forensic chemistry. This metodology has the advantage. touse a mobile phase les s contaminant and cheaper than others previously used. Besides, it makes easier the cleaning of the equipment and extend the useful life of the column. The chromatographic separation system by HPLC-RP, with spectrophotometric detection at 266 nm, ineludes an octadecylsilane C-18 column, and methanol/water as mobile phase.

  20. Design of Experiment (DOE) Utilization to Develop a Simple and Robust Reversed-Phase HPLC Technique for Related Substances' Estimation of Omeprazole Formulations.

    Science.gov (United States)

    Manranjan, Vayeda Chintan; Yadav, Devendra Singh; Jogia, Hitesh Amrutlal; Chauhan, Praful Lalitkumar

    2013-01-01

    A simple, fast, and sensitive reversed-phase HPLC method with UV detection was developed for the quantitation of omeprazole and its eleven related compounds (impurities) in pharmaceutical formulation using the Thermo Accucore C-18 (50 mm × 4.6 mm, 2.6 μm) column. The separation among all the compounds was achieved with a flow rate of 0.8 mL min(-1) employing a gradient program of mobile phase A [0.08 M glycine buffer pH 9.0: acetonitrile; 95:05 (v/v)] and mobile phase B [acetonitrile: methanol; 65:35 (v/v)]. The chromatographic detection was carried out at a wavelength of 305 nm. The method was validated for specificity, linearity, and recovery. The huskiness of the method was determined prior to validation using the Design of Experiments (DOE). The ANOVA analysis of DOE with a 95% confidence interval (CI) confirmed the buffer pH of mobile phase A (p <0.0001) and column temperature (p<0.0001) as significant Critical Method Parameters (CMPs).

  1. An on-line coupling of nanofibrous extraction with column-switching high performance liquid chromatography - A case study on the determination of bisphenol A in environmental water samples.

    Science.gov (United States)

    Háková, Martina; Chocholoušová Havlíková, Lucie; Chvojka, Jiří; Solich, Petr; Šatínský, Dalibor

    2018-02-01

    Polyamide 6 nanofiber polymers were used as modern sorbents for on-line solid phase extraction (SPE) coupled with liquid chromatography. The on-line SPE system was tested for the determination of bisphenol A in river water samples. Polyamide nanofibers were prepared using needleless electrospinning, inserted into a mini-column cartridge (5 × 4.6mm) and coupled with HPLC. The effect of column packing and the amount of polyamide 6 on extraction efficiency was tested and the packing process was optimized. The proposed method was performed using a 50-µL sample injection followed by an on-line nanofibrous extraction procedure. The influence of the washing mobile phase on the retention of bisphenol A during the extraction procedure was evaluated. Ascentis® Express C18 (10cm × 4.6mm) core-shell column was used as an analytical column. Fluorescence detection wavelengths (λex = 225nm and λem = 320nm) were used for identification and quantification of Bisphenol A in river waters. The linearity was tested in the range from 2 to 500µgL-1 (using nine calibration points). The limits of detection and quantification were 0.6 and 2µgL-1, respectively. The developed method was successfully used for the determination of bisphenol A in various samples of river waters in the Czech Republic (The Ohře, Labe, Nisa, Úpa, and Opava Rivers). Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Highly efficient solid phase supported radiosynthesis of [11 C]PiB using tC18 cartridge as a "3-in-1" production entity.

    Science.gov (United States)

    Boudjemeline, Mehdi; Hopewell, Robert; Rochon, Pierre-Luc; Jolly, Dean; Hammami, Iness; Villeneuve, Sylvia; Kostikov, Alexey

    2017-10-05

    Pittsburgh compound B ([11 C]PiB) is the gold standard positron emission tomography (PET) tracer for the in vivo imaging of amyloid plaques. Currently, it is synthesized by either solution chemistry or using a "dry loop" approach followed by HPLC purification within 30 minutes starting from [11 C]CO2 . Here, we report a novel, highly efficient solid phase supported carbon-11 radiolabeling procedure using commercially available disposable tC18 cartridge as a "3-in-1" entity: reactor, purifier, and solvent replacement system. [11 C]PiB is synthesized by passing gaseous [11 C]CH3 OTf through a tC18 cartridge preloaded with a solution of precursor. Successive elution with aqueous ethanol solutions allows for nearly quantitative separation of the reaction mixture to provide chemically and radiochemically pure PET tracer. [11 C]PiB suitable for human injection is produced within 10 minutes starting from [11 C]CH3 OTf (20 min from [11 C]CO2 ) in 22% isolated yield not corrected for decay and molar activity of 190 GBq/μmol using 0.2 mg of precursor. This technique reduces the amount of precursor and other supplies, avoids use of preparative HPLC and toxic solvents, and decreases the time between consecutive production batches. Solid phase supported technique can facilitate [11 C]PiB production compliant with Good Manufacturing Practice (GMP) and improve synthesis reliability. Copyright © 2017 John Wiley & Sons, Ltd.

  3. Development and validation of a HPLC method for the quantification of three flavonoids in a crude extract of Dimorphandra gardneriana

    Directory of Open Access Journals (Sweden)

    Leonardo P. Landim

    2012-09-01

    Full Text Available A method for separation and quantification of three flavonoids by reverse-phase high performance liquid chromatography (HPLC was developed and validated. Flavonoids present in a crude methanolic extract of the inner bark of Dimorphandra gardneriana Tul., Fabaceae, were analyzed. Rutin, isoquercitrin and quercetin were used as calibration standards. The analysis was performed using a Thermo Scientific Hypersil C18 column (250 x 4.0 mm i.d., 5 μm particle size, as stationary phase, with a flow rate of 0.8 mL/min and detection at a wavelength of 356 nm. The proposed method was validated by resolution RE No. 899/2003 of the National Health Surveillance Agency. In this study, an excellent linearity was obtained with r higher than 0.99. Besides, the chromatographic peaks showed good resolution. With other validation data, including precision, specificity, accuracy and robustness, this method demonstrated good reliability and sensitivity, and can be conveniently used for the quantification of rutin, isoquercitrin and quercetin in crude methanolic extract of D. gardneriana pods. Furthermore, there are the advantages of easy sample preparation and short time between each injection.

  4. A Rapid and Sensitive HPLC-Fluorescence Method for Determination of Mirtazapine and Its two Major Metabolites in Human Plasma.

    Science.gov (United States)

    Lavasani, Hoda; Giorgi, Mario; Sheikholeslami, Behjat; Hedayati, Mohammadhasan; Rouini, Mohammad Reza

    2014-01-01

    A rapid and sensitive HPLC method has been developed for the quantification of mirtazapine (MRZ), a noradrenergic and specific serotonergic inhibitor antidepressant (NaSSA) and its two major metabolites N-desmethyl mirtazapine (NDM) and 8-hydroxymirtazapine (8-OHM) in human plasma. The separation was achieved using Chromolith C18 column and a mobile phase of acetonitrile: phosphate buffer (pH = 3, 20:80, v/v) in isocratic mode at a flow rate of 2 mL/min. A fluorescence detector was set at 290 and 350 nm for excitation and emission, respectively. Zolpidem was used as the internal standard. Liquid-liquid extraction was applied for sample clean up. All analytes were eluted in less than 5 minutes with LOQ of 1 ng/mL for MRZ and 2 ng/mL for both NDM and 8-OHM. The developed method was successfully applied to quantify MRZ and its metabolites in plasma of a healthy volunteer.

  5. Simultaneous determination of bioactive compounds in Piper nigrum L. and a species comparison study using HPLC-PDA.

    Science.gov (United States)

    Rao, Vidadala Rama Subba; Raju, Sagi Satyanarayana; Sarma, Vanka Umamaheswara; Sabine, Fouriner; Babu, Kothapalli Hari; Babu, Katragadda Suresh; Rao, Janaswamy Madhusudana

    2011-08-01

    Piper nigrum L. is a traditional medicine widely used in India for illnesses such as constipation, diarrhoea, earache, gangrene, heart disease, hernia, hoarseness, indigestion, insect bites, insomnia, joint pain, liver problems, lung disease, oral abscesses, sunburn, tooth decay and toothaches. In this study, six bioactive compounds, namely piperine (1), pellitorine (2), guineensine (3), pipnoohine (4), trichostachine (5) and piperonal (6) were quantified in different extracts of P. nigrum L. and compared with those of P. longum L. and P. chaba Hunter. To evaluate the quality of P. nigrum, a simple, accurate and precise HPLC-PDA method was developed for the simultaneous determination of the above-mentioned six compounds. The separation was achieved by Phenomenex Luna RP C(18) column (150 × 4.6 mm, 5 µm, Phenomenex Inc, CA, USA) with a binary gradient solvent system of water-acetonitrile, at a flow rate of 1.0 mL min(-1) and detected at 210, 232, 262 and 343 nm. All six calibration curves showed good linearity (R (2) > 0.9966). The method was reproducible with intra- and inter-day variations of less than 2% and 5%, respectively. The results demonstrated that this method is simple, reliable and suitable for the quality control of these plants.

  6. Simultaneous determination of hyoscine N-butyl bromide and paracetamol in their binary mixture by RP-HPLC method

    Directory of Open Access Journals (Sweden)

    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.

  7. Derivative- Ratio Spectrophotometric, Chemometric and HPLC Validated methods for Simultaneous Determination of Amlodipine and Atorvastatin in Combined Dosage Form

    Directory of Open Access Journals (Sweden)

    Ola Moustafa Abdallah

    2011-04-01

    Full Text Available Three methods were developed for simultaneous determination of amlodipine and atorvastatin without previous separation. The first method depends on first derivative of the ratios spectra by measurements of the amplitudes at 228 and 245 nm for amlodipine using 25 μg/mL of atorvastatin as a divisor and at 284 and 295 nm for atorvastatin using 80 μg/mL of amlodipine as a divisor. Calibration graphs were established in the range of 10-100 μg/mL and 2.5-30 μg/mL for amlodipine and atorvastatin, respectively. The second method describes the use of multivariate spectophotometric calibration for the simultaneous determination of the analyzed binary mixture, where the resolution is accomplished by using partial least squares (PLS regression analysis. In the third method (HPLC, separation was performed by using C18 reversed phase column and a mobile phase of acetonitrile: 0.05 M KH2PO4 (60:40v/v adjusted by phosphoric acid to pH 3.5 at flow rate of 1 mL/min. All proposed methods were extensively validated and the results obtained by adopting the three methods were statistically analyzed and compared with those obtained from a reported method.

  8. Simultaneous quantification of citalopram, clozapine, fluoxetine, norfluoxetine, maprotiline, desmethylmaprotiline and trazodone in human serum by HPLC analysis.

    Science.gov (United States)

    Waschgler, R; Hubmann, M R; Conca, A; Moll, W; König, P

    2002-12-01

    We describe an analytical procedure for the simultaneous quantification of citalopram (seropram), clozapine (leponex), fluoxetine (fluctine), norfluoxetine, maprotiline (ludiomil), desmethylmaprotiline and trazodone (trittico) in human serum within a period of 11.5 minutes using reversed phase HPLC. After 2 liquid/liquid extractions in the sample preparation phase, the drugs and metabolites were separated on a C18 column using a mobile phase consisting of acetonitrile/buffer (30/70, v:v) at 70 degrees C, a flow rate of 1.5 m/min and haloperidol as internal standard. Absorption and native fluorescence signals of the eluted compounds were detected simultaneously at 260 nm and 227/300 nm (excitation/emission), respectively. The calibration ranges for citalopram, clozapine, fluoxetine, norfluoxetine, maprotiline, and desmethylmaprotiline ranged from 50-400 microg/l and for trazodone from 50-3,200 microg/l. The CVs varied between 0.6% and 5.5% (within-run) and between 3.2% and 7.1% (between-run). Recoveries were > 90% for all pharmaceuticals. We noticed no interferences from several commonly used drugs.

  9. HPLC quantification of two isomeric triterpenic acids isolated from Mitracarpus scaber and antimicrobial activity on Dermatophilus congolensis.

    Science.gov (United States)

    Gbaguidi, Fernand; Accrombessi, Georges; Moudachirou, Mansourou; Quetin-Leclercq, Joëlle

    2005-10-04

    Oleanolic (OA) and ursolic acids (UA) were isolated for the first time from the alcoholic extract of Mitracarpus scaber possessing antimicrobial effects on Dermatophilus congolensis. These two triterpenic acids were also active (MIC 15 microg/ml) on this causative agent of dermatophilosis in African animals. To quantify OA and UA in M. scaber, a new, simple and rapid high-performance liquid chromatography (HPLC) method compatible with MS detection was developed and validated. The mobile phase acetonitrile:H2O (85:15, v/v) was pumped through a C18 octadecylsilyl silica column at a flow rate of 0.6 ml/min and the eluate was monitored at 215 nm. The calibration curves constructed between 0.5 and 10 microg/ml showed linear relationships with good R2 values. The developed method was precise and reproducible with relative standard deviations (RSD) for these two active constituents between 0.22-2.06% (intraday) and 1.61-3.72% (interday) for concentrations from 0.5 to 6 microg/ml. Limits of detection and quantification were, respectively, 0.2 and 0.5 microg/ml.

  10. 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

  11. Stability-indicating RP-HPLC method for simultaneous estimation of levosalbutamol sulfate and theophylline in combined dosage form

    Directory of Open Access Journals (Sweden)

    Sagar Suman Panda

    2013-09-01

    Full Text Available A novel, simple, accurate and precise RP-HPLC method for simultaneous determination of levosalbutamol sulfate and theophylline has been developed and validated. Separation was achieved on a Phenomenex; C18 column (250 mm × 4.6 mm i.d., 5 µm using methanol: 10 mM TBAHS(tetrabutyl ammonium hydrogen sulfate (50:50, v/v as mobile phase at flow rate of 1.0 mL.min-1. The UV detection wavelength was 274 nm. The linearity is obeyed over a concentration range of 0.5-150 µg.mL-1 with correlation coefficient of 0.999 for both the drugs. The proposed method was validated by determining accuracy, precision, stability and system suitability parameters. The method was found to be robust. Specificity of the method was determined by subjecting the drugs to various stress conditions like acid, alkali, oxidation, thermal and photolytic degradation. The method was used successfully for the simultaneous determination of levosalbutamol sulfate and theophylline in syrup dosage form.

  12. Extraction and determination of trace amounts of chlorpromazine in biological fluids using magnetic solid phase extraction followed by HPLC

    Directory of Open Access Journals (Sweden)

    Yadollah Yamini

    2014-08-01

    Full Text Available A simple, rapid and sensitive method termed as magnetic solid phase extraction (MSPE combined with high-performance liquid chromatography-ultraviolet detector (HPLC-UV has been proposed for the determination of trace amounts of chlorpromazine (CPZ in water, urine and plasma samples. The separation and determination was performed on a C18 column under the optimal chromatographic conditions. Several factors influencing the extraction efficiency of CPZ, such as pH, surfactant and adsorbent amounts, ionic strength, extraction time, sample volume and desorption conditions, were studied and optimized. Under the optimal MSPE conditions, the extraction percentage of CPZ was 74%, 27% and 16% in water, urine and plasma samples, respectively. The limits of detection (LODs of the proposed approach were 0.1, 5.0 and 10 ng/mL in water, urine and plasma samples, respectively. The relative standard deviations (RSDs based on five replicate determinations at 10 ng/mL level of CPZ was 1.2%. Good linear behaviors over the investigated concentration ranges (0.25–300 ng/mL with good coefficient of determination, R2>0.9998, were obtained. Good spike recoveries with relative errors less than 9.0% were obtained when applying the proposed method to water, urine and plasma samples. Keywords: Chlorpromazine, Magnetic solid phase extraction, Mixed-hemimicelles

  13. Determination of itopride hydrochloride in human plasma by RP-HPLC with fluorescence detection and its use in bioequivalence study.

    Science.gov (United States)

    Ma, Jing; Yuan, Li-Hua; Ding, Mei-Juan; Zhang, Jun; Zhang, Qing; Xu, Qun-Wei; Zhou, Xue-Min

    2009-03-01

    A sensitive, selective and simple method using a precipitation of protein with 10% perchloric acid, followed by high-performance liquid chromatography (HPLC) with fluorescence detection was developed for the determination of itopride hydrochloride in human plasma, using levofloxacin as the internal standard (IS). Chromatographic separation was obtained within 7.0 min using a reverse phase Hypersil BDS C(18) (250 mm x 4.6 mm, 5 microm) column and an isocratic mobile phase, constituting of a mixture of 0.1 mol/l ammonium acetate-methanol (30:70, v/v) flowing at 1.1 ml/min. The excitation and emission wavelengths were set at 304 and 344 nm, respectively. The method was validated over the concentration range of 5 ng/ml to 1000.0 ng/ml. The lower limit of quantitation (LLOQ) was 5 ng/ml. The extractive recovery of itopride hydrochloride from the biological matrix was more than 80.77%. The intra-day accuracy of the drug containing serum samples was more than 82.94% with a precision of 2.81-4.37%. The inter-day accuracy was 82.91% or more, with a precision of 6.89-9.54%. The limit we have used (70-143%) is based on the local regulatory authority (SFDA). The developed method was validated and successfully applied to bioequivalence studies of itopride hydrochloride in healthy male volunteers.

  14. HPLC and chemometrics-assisted UV-spectroscopy methods for the simultaneous determination of ambroxol and doxycycline in capsule

    Science.gov (United States)

    Hadad, Ghada M.; El-Gindy, Alaa; Mahmoud, Waleed M. M.

    2008-08-01

    High-performance liquid chromatography (HPLC) and multivariate spectrophotometric methods are described for the simultaneous determination of ambroxol hydrochloride (AM) and doxycycline (DX) in combined pharmaceutical capsules. The chromatographic separation was achieved on reversed-phase C 18 analytical column with a mobile phase consisting of a mixture of 20 mM potassium dihydrogen phosphate, pH 6-acetonitrile in ratio of (1:1, v/v) and UV detection at 245 nm. Also, the resolution has been accomplished by using numerical spectrophotometric methods as classical least squares (CLS), principal component regression (PCR) and partial least squares (PLS-1) applied to the UV spectra of the mixture and graphical spectrophotometric method as first derivative of the ratio spectra ( 1DD) method. Analytical figures of merit (FOM), such as sensitivity, selectivity, analytical sensitivity, limit of quantitation and limit of detection were determined for CLS, PLS-1 and PCR methods. The proposed methods were validated and successfully applied for the analysis of pharmaceutical formulation and laboratory-prepared mixtures containing the two component combination.

  15. Quantitative Detection of Ambroxol in Human Plasma Using HPLC-APCI-MS/MS: Application to a Pharmacokinetic Study.

    Science.gov (United States)

    Mao, Zhengsheng; Wang, Xin; DI, Xin; Liu, Yangdan; Zang, Yanan; Ma, Dongke; Liu, Youping; DI, Xin

    2017-01-01

    In this study, a rapid and reliable high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of ambroxol in human plasma was developed and validated using palmatine as an internal standard (IS). Ambroxol and IS were extracted from 200 μL of human plasma via a simple protein precipitation preparation. Chromatographic separation was achieved on a Platisil C18 column (150 × 4.6 mm, 5 μm) using methanol-0.01% formic acid (70:30, v/v) as the mobile phase at a flow rate of 0.6 mL/min under an isocratic condition. The MS acquisition m/z 379 → 264 for ambroxol and 352 → 336 for IS was performed by atmospheric-pressure chemical ionization (APCI) mass spectrometry in selected reaction monitoring mode. The calibration curve for ambroxol was linear over the concentration range of 2.500 - 180.0 ng/mL. The matrix effects of ambroxol ranged from 104.6 to 112.7%. This fully validated method was successfully applied to a pharmacokinetic study of ambroxol in humans after oral administration of ambroxol at a single dose of 75 mg.

  16. Development and validation of a selective HPLC-UV method for thymol determination in skin permeation experiments.

    Science.gov (United States)

    Angelo, Tamara; Pires, Felipe Q; Gelfuso, Guilherme M; da Silva, Joyce K R; Gratieri, Tais; Cunha-Filho, Marcílio S S

    2016-06-01

    Thymol is a natural monoterpene, whose antioxidant and antimicrobial properties suggest a potential use in topical formulations. A simple, precise and selective HPLC method for thymol determination in skin penetration studies was developed and validated in this paper. Separation was achieved with a RP-C18 column, mobile phase comprised of acetonitrile:water (35:65v/v), flow rate of 1.5mL/min, oven temperature at 40°C, injection volume of 30μL and UV detection at 278nm. The validation procedure certified the method was selective for thymol determination even when extracted from skin matrix extracts. It was also linear in a range from 0.5 to 15.0μg/mL, robust, precise and accurate, with recovery rates from the skin layers higher than 90%. Limits of detection and quantification were 0.05 and 0.14μg/mL, respectively. The method showed, therefore, to be adequate for use in further skin permeation studies employing thymol topical formulations. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Dual Wavelength RP-HPLC Method for Simultaneous Determination of Two Antispasmodic Drugs: An Application in Pharmaceutical and Human Serum

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    Najmul Hasan

    2013-01-01

    Full Text Available A reverse phase stability indicating HPLC method for simultaneous determination of two antispasmodic drugs in pharmaceutical parenteral dosage forms (injectable and in serum has been developed and validated. Mobile phase ingredients consist of Acetonitrile : buffer : sulfuric acid 0.1 M (50 : 50 : 0.3 v/v/v, at flow rate 1.0 mL/min using a Hibar μBondapak ODS C18 column monitored at dual wavelength of 266 nm and 205 nm for phloroglucinol and trimethylphloroglucinol, respectively. The drugs were subjected to stress conditions of hydrolysis (oxidation, base, acid, and thermal degradation. Oxidation degraded the molecule drastically while there was not so much significant effect of other stress conditions. The calibration curve was linear with a correlation coefficient of 0.9999 and 0.9992 for PG and TMP, respectively. The drug recoveries fall in the range of 98.56% and 101.24% with 10 pg/mL and 33 pg/mL limit of detection and limit of quantification for both phloroglucinol and trimethylphloroglucinol. The method was validated in accordance with ICH guidelines and was applied successfully to quantify the amount of trimethylphloroglucinol and phloroglucinol in bulk, injectable form and physiological fluid. Forced degradation studies proved the stability indicating abilities of the method.

  18. RP-HPLC Method Development and Validation for Determination of Eptifibatide Acetate in Bulk Drug Substance and Pharmaceutical Dosage Forms.

    Science.gov (United States)

    Bavand Savadkouhi, Maryam; Vahidi, Hossein; Ayatollahi, Abdul Majid; Hooshfar, Shirin; Kobarfard, Farzad

    2017-01-01

    A new, rapid, economical and isocratic reverse phase high performance liquid chromatography (RP-HPLC) method was developed for the determination of eptifibatide acetate, a small synthetic antiplatelet peptide, in bulk drug substance and pharmaceutical dosage forms. The developed method was validated as per of ICH guidelines. The chromatographic separation was achieved isocratically on C18 column (150 x 4.60 mm i.d., 5 µM particle size) at ambient temperature using acetonitrile (ACN), water and trifluoroacetic acid (TFA) as mobile phase at flow rate of 1 mL/min and UV detection at 275 nm. Eptifibatide acetate exhibited linearity over the concentration range of 0.15-2 mg/mL (r2=0.997) with limit of detection of 0.15 mg/mL The accuracy of the method was 96.4-103.8%. The intra-day and inter-day precision were between 0.052% and 0.598%, respectively. The present successfully validated method with excellent selectivity, linearity, sensitivity, precision and accuracy was applicable for the assay of eptifibatide acetate in bulk drug substance and pharmaceutical dosage forms.

  19. RP-HPLC determination of phenylalkanoids and monoterpenoids in Rhodiola rosea and identification by LC-ESI-TOF.

    Science.gov (United States)

    Avula, Bharathi; Wang, Yan-Hong; Ali, Zulfiqar; Smillie, Troy J; Filion, Vicky; Cuerrier, Alain; Arnason, John T; Khan, Ikhlas A

    2009-08-01

    An HPLC method permitting the simultaneous determination of fourteen analytes (phenylalkanoids and monoterpenoids) from the roots of Rhodiola rosea was developed. A separation was achieved within 35 min using C(18) column material and a water-acetonitrile mobile phase, both containing a 0.05% phosphoric acid gradient system and a temperature of 53 degrees C. The method was validated for linearity, repeatability, limits of detection and limits of quantification. The limits of detection and limits of quantification of 14 phenylalkanoids and monoterpenoids were found to be 0.20-1.0 and 0.5-3.5 microg/mL, respectively. The wavelengths used for quantification of phenylalkanoids and monoterpenoids with a diode array detector were 205, 220 and 251 nm. The method was used to analyze the roots of two species of Rhodiola and commercial extracts of R. rosea and provides preliminary evidence of phytochemical differences between North American and Eurasian populations of R. rosea. LC-mass spectrometry coupled with electrospray ionization (ESI) interface method is described for the identification of phenylalkanoids and monoterpenoids in various Rhodiola samples. This method involved the use of the [M + H](+), [M + NH(4)](+) and [M + Na](+) ions in the positive ion mode with extractive ion monitoring (EIM). (c) 2009 John Wiley & Sons, Ltd.

  20. Development and validation of an HPLC method for the determination of epicatechin in Maytenus ilicifolia (Schrad. Planch., Celastraceae

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    Gisely Cristiny Lopes

    2010-09-01

    Full Text Available A simple, reproducible and efficient high-performance liquid chromatography (HPLC method was developed. Water (0.05% TFA:acetonitrile (0.05% TFA was used as the mobile phase in a gradient system for the determination of epicatechin (EP in leaves of Maytenus ilicifolia (Schrad. Planch. The analysis was performed using an RP C-18 column (5 µm as the stationary phase, with a flow rate of 0.8 mL/min, at a wavelength of 210 nm for detection and determination. The main validation parameters of the method were also determined. The calibration curve was found to be linear, with a range of 10-120 µg/mL (EP. The correlation coefficient of the linear regression analysis was within 0.9988, and the detection and quantification limits were 28.61 and 86.77 µg/mL, respectively. The content of EP was successfully determined, with satisfactory reproducibility and recovery. Recovery of the EP was 99.32%. The method was successfully applied to the determination of epicatechin in leaves of M. ilicifolia. The interlaboratorial evaluation showed the reproducibility of the method with a relative standard deviation of 14.62%.

  1. Development and Validation of RP-HPLC Method for Simultaneous Estimation of Aspirin and Esomeprazole Magnesium in Tablet Dosage Form

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    Dipali Patel

    2013-01-01

    Full Text Available A simple, specific, precise, and accurate reversed-phase HPLC method was developed and validated for simultaneous estimation of aspirin and esomeprazole magnesium in tablet dosage forms. The separation was achieved by HyperChrom ODS-BP C18 column (200 mm × 4.6 mm; 5.0 μm using acetonitrile: methanol: 0.05 M phosphate buffer at pH 3 adjusted with orthophosphoric acid (25 : 25 : 50, v/v as eluent, at a flow rate of 1 mL/min. Detection was carried out at wavelength 230 nm. The retention times of aspirin and esomeprazole magnesium were 4.29 min and 6.09 min, respectively. The linearity was established over the concentration ranges of 10–70 μg/mL and 10–30 μg/mL with correlation coefficients (r2 0.9986 and 0.9973 for aspirin and esomeprazole magnesium, respectively. The mean recoveries were found to be in the ranges of 99.80–100.57% and 99.70–100.83% for aspirin and esomeprazole magnesium, respectively. The proposed method has been validated as per ICH guidelines and successfully applied to the estimation of aspirin and esomeprazole magnesium in their combined tablet dosage form.

  2. HPLC method for urinary theobromine determination: Effect of consumption of cocoa products on theobromine urinary excretion in children.

    Science.gov (United States)

    Rodriguez, Adrian; Costa-Bauza, Antonia; Saez-Torres, Concepcion; Rodrigo, Dolores; Grases, Felix

    2015-11-01

    To validate a simple method of urinary theobromine determination, to assess urinary theobromine levels in 80 healthy children and to relate these levels to consumption of cocoa products. Urine samples were diluted, directly injected into an HPLC system, separated by gradient elution on a C18 column, and detected by UV spectrometry. The method was validated for linearity, limits of detection and quantification, imprecision, accuracy, recovery and interferences. The proposed method was used to assess 12-h day and 12-h night urinary theobromine excretion by 80 healthy children, divided into four groups based on consumption of cocoa products. In addition, urinary excretion of magnesium and oxalate, also present in cocoa, was measured in these four groups. The method was linear to a theobromine concentration of 278μmol/L (50mg/L). LOD and LOQ for urine samples, diluted 1:5 (vol/vol) with water, were 1.1 and 3.6μmol/L respectively. Within-run and between-run imprecisions (CV) were each Theobromine excretion levels were significantly higher in healthy children with higher consumption of cocoa products (ptheobromine determination with 100% recovery, without sample pretreatment. Urinary theobromine levels in healthy children were directly related to their consumption of cocoa products. Copyright © 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  3. Development and Validation of an HPLC Method for the Simultaneous Determination of Fipronil, Chlorfenapyr, and Pyriproxyfen in Insecticide Formulations.

    Science.gov (United States)

    Hafeez, Anum; Tawab, Iffat Abdul; Iqbal, Sajid

    2016-09-01

    In this study, the analytical method development and validation of an HPLC assay for simultaneous determination of fipronil, chlorfenapyr, and pyriproxyfen in formulation products is described. On the basis of solubility and chromatographic separation with good resolution, acetonitrile-water (80 + 20) was selected as the mobile phase in isocratic mode with a flow rate of 1 mL/min. Chromatographic separations were performed on a Beckman C18 analytical column (4.6 mm × 15 cm, 5 μm particle size; Musa Jee & Sons, Karachi, Pakistan). The retention times for fipronil, chlorfenapyr, and pyriproxyfen were 3.70, 8.61 and 10.09 min, respectively. Calibration curves of all studied insecticides were linear in the concentration range of 20 to 800 μg/mL, with R(2) > 0.997. The LODs of fipronil, chlorfenapyr, and pyriproxyfen were 15.1, 13.3, and 20.0 μg/mL, respectively, whereas the LOQs were 45.9, 40.3, and 60.6 μg/mL. Interday precision was RSD, % pyriproxyfen in their formulations.

  4. Simultaneous Determination of Four Active Ingredients in Sargentodoxa cuneata by HPLC Coupled with Evaporative Light Scattering Detection

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    Di-Hua Li

    2016-01-01

    Full Text Available A HPLC coupled with evaporative light scattering detection method had been developed for the simultaneous determination of 3,4-dihydroxyphenylethyl alcohol glycoside, salidroside, chlorogenic acid, and liriodendrin in the stem of Sargentodoxa cuneata. With a C18 column, the analysis was performed using acetonitrile and 0.2% formic acid aqueous solution as mobile phase in gradient program at a flow rate of 0.9 mL/min. The optimum drift tube temperature of evaporative light scattering detection was at 105°C with the air flow rate of 2.5 L/min. The calibration curves showed good linearity during the test ranges. This method was validated for limits of detection and quantification, precision, and reproducibility. The recoveries were within the range of 96.39%–104.64%. The relative standard deviations of intraday and interday precision were less than 2.90% and 3.30%, respectively. The developed method can be successfully used to quantify the four analytes in the stem of Sargentodoxa cuneata from various regions in China.

  5. Sequential Injection Chromatography with an Ultra-short Monolithic Column for the Low-Pressure Separation of α-Tocopherol and γ-Oryzanol in Vegetable Oils and Nutrition Supplements.

    Science.gov (United States)

    Thaithet, Sujitra; Kradtap Hartwell, Supaporn; Lapanantnoppakhun, Somchai

    2017-01-01

    A low-pressure separation procedure of α-tocopherol and γ-oryzanol was developed based on a sequential injection chromatography (SIC) system coupled with an ultra-short (5 mm) C-18 monolithic column, as a lower cost and more compact alternative to the HPLC system. A green sample preparation, dilution with a small amount of hexane followed by liquid-liquid extraction with 80% ethanol, was proposed. Very good separation resolution (Rs = 3.26), a satisfactory separation time (10 min) and a total run time including column equilibration (16 min) were achieved. The linear working range was found to be 0.4 - 40 μg with R2 being more than 0.99. The detection limits of both analytes were 0.28 μg with the repeatability within 5% RSD (n = 7). Quantitative analyses of the two analytes in vegetable oil and nutrition supplement samples, using the proposed SIC method, agree well with the results from HPLC.

  6. Ultrasound-assisted extraction of azadirachtin from dried entire fruits of Azadirachta indica A. Juss. (Meliaceae) and its determination by a validated HPLC-PDA method.

    Science.gov (United States)

    de Paula, Joelma Abadia Marciano; Brito, Lucas Ferreira; Caetano, Karen Lorena Ferreira Neves; de Morais Rodrigues, Mariana Cristina; Borges, Leonardo Luiz; da Conceição, Edemilson Cardoso

    2016-01-01

    Azadirachta indica A. Juss., also known as neem, is a Meliaceae family tree from India. It is globally known for the insecticidal properties of its limonoid tetranortriterpenoid derivatives, such as azadirachtin. This work aimed to optimize the azadirachtin ultrasound-assisted extraction (UAE) and validate the HPLC-PDA analytical method for the measurement of this marker in neem dried fruit extracts. Box-Behnken design and response surface methodology (RSM) were used to investigate the effect of process variables on the UAE. Three independent variables, including ethanol concentration (%, w/w), temperature (°C), and material-to-solvent ratio (gmL(-1)), were studied. The azadirachtin content (µgmL(-1)), i.e., dependent variable, was quantified by the HPLC-PDA analytical method. Isocratic reversed-phase chromatography was performed using acetonitrile/water (40:60), a flow of 1.0mLmin(-1), detection at 214nm, and C18 column (250×4.6mm(2), 5µm). The primary validation parameters were determined according to ICH guidelines and Brazilian legislation. The results demonstrated that the optimal UAE condition was obtained with ethanol concentration range of 75-80% (w/w), temperature of 30°C, and material-to-solvent ratio of 0.55gmL(-1). The HPLC-PDA analytical method proved to be simple, selective, linear, precise, accurate and robust. The experimental values of azadirachtin content under optimal UAE conditions were in good agreement with the RSM predicted values and were superior to the azadirachtin content of percolated extract. Such findings suggest that UAE is a more efficient extractive process in addition to being simple, fast, and inexpensive. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. HPLC, TLC, and first-derivative spectrophotometry stability-indicating methods for the determination of tropisetron in the presence of its acid degradates.

    Science.gov (United States)

    Abdel-Fattah, Laila S; El-Sherif, Zeinab A; Kilani, Khadiga M; El-Haddad, Dalia A

    2010-01-01

    Three stability-indicating assay methods were developed for the determination of tropisetron in a pharmaceutical dosage form in the presence of its degradation products. The proposed techniques are HPLC, TLC, and first-derivative spectrophotometry (1D). Acid degradation was carried out, and the degradation products were separated by TLC and identified by IR, NMR, and MS techniques. The HPLC method was based on determination of tropisetron in the presence of its acid-induced degradation product on an RP Nucleosil C18 column using methanol-water-acetonitrile-trimethylamine (65 + 20 + 15 + 0.2, v/v/v/v) mobile phase and UV detection at 285 nm. The TLC method was based on the separation of tropisetron and its acid-induced degradation products, followed by densitometric measurement of the intact spot at 285 nm. The separation was carried out on silica gel 60 F254 aluminum sheets using methanol-glacial acetic acid (22 + 3, v/v) mobile phase. The 1D method was based on the measurement of first-derivative amplitudes of tropisetron in H2O at the zero-crossing point of its acid-induced degradation product at 271.9 nm. Linearity, accuracy, and precision were found to be acceptable over concentration ranges of 40-240 microg/mL, 1-10 microg/spot, and 6-36 micro/mL for the HPLC, TLC, and 1D methods, respectively. The suggested methods were successfully applied for the determination of the drug in bulk powder, laboratory-prepared mixtures, and a commercial sample.

  8. Revival of physostigmine - a novel HPLC assay for simultaneous determination of physostigmine and its metabolite eseroline designed for a pharmacokinetic study of septic patients.

    Science.gov (United States)

    Pinder, Nadine; Zimmermann, Johannes B; Hofer, Stefan; Brenner, Thorsten; Weigand, Markus A; Gubbe, Ute; Hoppe-Tichy, Torsten; Swoboda, Stefanie

    2015-07-01

    Physostigmine, commonly used as an antidote in anticholinergic poisoning, is reported to have additional pharmacological effects, such as activation of the cholinergic anti-inflammatory pathway in sepsis models. Due to the narrow therapeutic range of physostigmine and its metabolite eseroline, however, the plasma concentrations of these substances need to be determined so as to understand their effect and ensure safety in the treatment of septic patients. To determine physostigmine and its metabolite eseroline, a rapid and sensitive high performance liquid chromatography (HPLC) method has been developed and validated. Spiked plasma samples were cleaned up and concentrated using a simple liquid-liquid extraction (LLE) procedure with N-methylphysostigmine as internal standard. Separation was achieved using reversed-phase HPLC on a Kinetex C18 column with gradient elution and fluorescence detection (254 nm excitation/355 nm emission). LLE produced clean extracts and a mean recovery of 80.3% for eseroline and 84.9% for physostigmine. The HPLC assay revealed a limit of detection (LOD) of 0.025 ng/mL and a lower limit of quantification (LLOQ) of 0.05 ng/mL for both analytes. Linearity was observed at 0.05-10.0 ng/mL (r²>0.999). Intra- and inter-day precision ranged from 0.7% to 6.6%, and intra- and inter-day accuracy 97.5%-110.0%. The presented method is useful for human drug level monitoring of physostigmine and eseroline in accordance with current guidelines. Remarkably low plasma concentrations can be quantified after LLE with gradient elution and fluorescence detection, making this a suitable method for pharmacokinetic studies in a clinical setting.

  9. Application of RP–HPLC method in dissolution testing and statistical evaluation by NASSAM for simultaneous estimation of tertiary combined dosages forms

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    Yogesh Upadhyay

    2015-10-01

    Full Text Available A dissolution method with robust high performance liquid chromatographic (HPLC analysis for immediate release tablet formulation was developed and validated to meet the requirement as per International Conference on Harmonization (ICH and United States Food and Drug Administration (USFDA guidelines. The method involved the use of Agilent ZORBAX Eclipse XDB C18 column, and temperature was maintained at 30 °C. After optimization, the mobile phase was selected as phosphate buffer (KH2PO4, 30 mM : ACN (60:40, v/v with pH 3.0, and retention time Rt was found as 3.24, 4.16, and 2.55 min for paracetamol (PCM, chlorpheniramine maleate (CPM and phenylephrine hydrochloride (PH respectively at 265 nm and at a flow rate of 1 mL/min. The relative standard deviation (%RSD for 6 replicate measurements was found to be less than 2%. Furthermore net analyte signal standard addition method (NASSAM with spectrophotometer was performed for standard and liquid oral suspension. On the basis of selectivity, sensitivity and accuracy analysis, it was confirmed that this novel method could be useful for simultaneous estimation of the given drug combinations. Two-way analysis of variance (ANOVA was applied for evaluating the statistical difference between the assay results obtained via both NASSAM and RP–HPLC methods and ultimately no significant difference was found between both the methods. All the methods and results were acceptable and confirmed that the method was suitable for intended use. Keywords: Dissolution, RP–HPLC, Net analyte signal standard addition method, Two-way ANOVA

  10. Development of a validated HPLC method for the separation and analysis of a Bromazepam, Medazepam and Midazolam mixture

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    Hasan al-Hawasli

    2012-12-01

    Full Text Available The purpose of this work was to develop a rapid, sensitive and validated HPLC method for the separation and analysis of a Bromazepam, Medazepam and Midazolam mixture. The three benzodiazepine compounds were separated on a reversed-phase C18 column at 50 °C using a mobile phase containing 25% acetonitrile, 45% methanol and 30% ammonium acetate (0.05 M. The pH was adjusted to pH=9 by the addition of ammonia solution (35%, w/w. The samples were detected using a UV detector at 240 nm. The validation study of the method included the effect of temperature, flow rate, ratio of the components of the mobile phase and the pH of the mobile phase on the efficiency of separation. The linear range of Bromazepam and Midazolam was between 0.12 and 0.18 mg/mL, while that of Medazepam was between 0.08 and 0.12 mg/mL. The relative standard deviation for precision was less than 2%. The linearity, selectivity, accuracy and robustness of the developed method showed acceptable values. The method was applied to the analysis of the samples of raw material of the three compounds under study, and the percentage of recoveries was 99.89%±1.06. It was also applied to the analysis of samples of pharmaceutical preparations of those compounds and spiked serum samples. Recoveries from serum samples ranged between 91.5% and 99.0%. The developed method is suitable for quality control of Bromazepam, Medazepam and Midazolam in their mixtures and in pharmaceutical preparations (tablets, capsules, ampoules. It can also be used to determine their concentrations in serum. Keywords: Benzodiazepine, Bromazepam, Medazepam, Midazolam, HPLC, Serum

  11. Validation of HPLC-UV method for determination of minor glycosides contained in Stevia rebaudiana Bertoni leaves.

    Science.gov (United States)

    Aranda-González, Irma; Moguel-Ordoñez, Yolanda; Betancur-Ancona, David

    2015-05-01

    Leaves of Stevia rebaudiana contain glycosides with sweetness and biological activity. However besides the major glycosides, there are other glycosides within extracts that may contribute to its activity, and therefore it is important to quantify them. In this work, an isocratic HPLC method was validated for determination of dulcoside A, steviolbioside, rebaudioside C and rebaudioside B. An HPLC method was performed using a C18 column (250 × 4.6 mm, particle size 5 µm) and a UV detector set at 210 nm. The mobile phase consisted of a 32:68 (v/v) mixture of acetonitrile and sodium phosphate buffer (10 mmol/L, pH 2.6), set to a flow rate of 1.0 mL/min. The calculated parameters were: sensitivity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy and precision. The calibration curves were linear over the working range 25-150 µg/mL, with coefficient of correlation of ≥0.99 and coefficient of determination of ≥0.98. The LOD was 5.68-8.81 µg/mL, while the LOQ was 17.21-26.69 µg/mL. The percentage recoveries of fortified samples were 100 ± 10% and precision, relative standard deviation, was <10%. The method validation showed accuracy, linearity and precision; therefore this method can be applied for quantitative analysis of minor steviol glycosides in S. rebaudiana leaves. Copyright © 2014 John Wiley & Sons, Ltd.

  12. Determination of oleanolic acid, ursolic acid and amygdalin in the flower of Eriobotrya japonica Lindl. by HPLC.

    Science.gov (United States)

    Zhou, Chunhua; Chen, Kunsong; Sun, Chongde; Chen, Qingjun; Zhang, Wangshu; Li, Xian

    2007-07-01

    Simple and accurate HPLC methods were developed for the determination of oleanolic acid (OA), ursolic acid (UA) and amygdalin in loquat (Eriobotrya japonica Lindl.) flower, which is commonly used for the treatment of various diseases as a traditional Chinese medicine. HPLC assay was performed on a reversed-phase C(18) column and all three compounds were detected at 210 nm with a flow rate of 1.0 mL/min. The mobile phase consisted of methanol (A) and 0.03 mol/L phosphate buffer (pH 2.8) (B) with a ratio of 88:12 (A:B, v/v) for simultaneous detection of OA and UA, and 25:75 (A:B, v/v) for detection of amygdalin. The established methods showed good precision and accuracy with overall intra-day and inter-day variation of 0.99-3.55 and 1.05-4.05%, respectively, and overall recoveries of 97.37-99.32% for the three compounds. Application of these methods to determine the OA, UA and amygdalin contents in loquat flower showed that cultivar had a minor effect on the contents of all three compounds, with average amounts of 0.38-0.51 mg OA/g dry weight (DW), 2.15-2.68 mg UA/g DW and 1.23-1.56 mg amygdalin/g DW among five loquat cultivars tested. However, developmental stages and flower tissues showed significant effect on the contents of all three bioactive components. Copyright 2007 John Wiley & Sons, Ltd.

  13. Quantification of endogenous brassinosteroids in plant by on-line two-dimensional microscale solid phase extraction-on column derivatization coupled with high performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Wu, Qian; Wu, Dapeng; Shen, Zheng; Duan, Chunfeng; Guan, Yafeng

    2013-07-05

    An on-line two-dimensional microscale solid phase extraction (2DμSPE)-on column derivatization (OCD)-high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) method was developed for quantification of brassinosteroids (BRs) in plant tissues. Five BRs with widest distribution in plant species and high bioactivity (24-epibrassinolide, 24-epicastasterone, 6-deoxo-24-epicastasterone, teasterone and typhastero) were selected as target analytes. 2DμSPE column packed sequentially with phenyl boronic acid silica sorbent (the first dimension) and C18 silica sorbent (the second dimension) was used to selectively extract and enrich BRs by 110-146 times. OCD was carried out on the second dimension of 2DμSPE column with m-aminophenylboronic acid (m-APBA) as a derivatization reagent, enhancing the sensitivity of MS/MS to BRs by 13-8437 times. It was also found that pre-trap of derivatization reagent on the C18 section of 2DμSPE column could increase reaction efficiency by 3-10 times. The whole process time of the on-line system was less than 30min. The detection limits of the method for five BRs were between 1.4 and 6.6pg with RSDs less than 10%. Endogeneous BRs in tomato leaves were analyzed by using this method. Owing to the high selectivity of this on-line 2DμSPE system, BRs in plant extracts could be quantified using matrix-free standard calibration method with relative recoveries in the range of 80-124%. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Numerical method for the estimation of column radial heterogeneity and of the actual column efficiency from tailing peak profiles.

    Science.gov (United States)

    Miyabe, Kanji; Guiochon, Georges

    2011-01-01

    It is probably impossible to prepare high-performance liquid chromatography (HPLC) columns that have a completely homogeneous packing structure. Many reports in the literature show that the radial distributions of the mobile phase flow velocity and the local column efficiency are not flat, even in columns considered as good. A degree of radial heterogeneity seems to be a common property of all HPLC columns and an important source of peak tailing, which prevents the derivation of accurate information on chromatographic behavior from a straightforward analysis of elution peak profiles. This work reports on a numerical method developed to derive from recorded peak profiles the column efficiency at the column center, the degree of column radial heterogeneity, and the polynomial function that best represents the radial distributions of the flow velocity and the column efficiency. This numerical method was applied to two concrete examples of tailing peak profiles previously described. It was demonstrated that this numerical method is effective to estimate important parameters characterizing the radial heterogeneity of chromatographic columns.

  15. Evaluation of different column chemistries for fast urinary metabolic profiling.

    Science.gov (United States)

    Kloos, Dick-Paul; Lingeman, Henk; Niessen, Wilfried M A; Deelder, André M; Giera, Martin; Mayboroda, Oleg A

    2013-05-15

    Fast analytical methodologies are mandatory for large scale metabolic profiling. Here, we present a thorough evaluation of different column chemistries in combination with different mobile phases for fast LC-MS urinary metabolic profiling. Three porous HILIC materials were investigated, next to core-shell C18-, XB-C18- and PFP-RPLC material. The performance of the selected column chemistries was tested in a non-targeted manner with pooled urine samples and in a targeted manner with a set of 54 common urinary metabolites. In order to evaluate the differential behaviour of the tested columns in a targeted manner, we applied a peak scoring algorithm. This algorithm takes into account several quality criteria such as retention time, dead time, peak height and peak shape. In general, HILIC columns generate more retention for polar metabolites. Our results show that the diol-HILIC column outperforms the RPLC columns. However, because of their opposite nature, comprehensive behaviour is observed as well, which was shown by investigating gender differences in a small urinary sample set. All applied column chemistries enabled sufficient peak capacity within a short gradient time. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Quantification of Organic Acids in Fermented Shrimp Waste by HPLC

    Directory of Open Access Journals (Sweden)

    Dalia Isabel Sánchez-Machado

    2008-01-01

    Full Text Available This work describes a simple, rapid, and reliable HPLC method for the determination of organic acids in fermented shrimp waste. Lactic, acetic and citric acids were quantified by HPLC with UV detection, on a 250×4.6 mm Extrasil ODS 5-μm column, mobile phase was ultrapure water adjusted with metaphosphoric acid to pH=2.1, flow rate 0.6 mL/min, column temperature 30 °C, and detection wavelength 210 nm. Under these conditions, the recovery (97.5 % and the method repeatability (RSD=6.2 % for lactic acid were of satisfying quality. Organic acids can preserve the quality and nutritive value of fermented shrimp waste.

  17. The quantitative impact of the mesopore size on the mass transfer mechanism of the new 1.9 μm fully porous Titan-C18 particles II--analysis of biomolecules.

    Science.gov (United States)

    Gritti, Fabrice; Guiochon, Georges

    2015-05-01

    The kinetic performances of 3.0 × 100 mm columns packed with 1.9 μm Titan-C18 particles with average mesopore sizes of 80 Å and 120 Å were investigated quantitatively for the analysis of biomolecules. Large mesopores are expected to speed up the rate of diffusivity of high-molecular-weight compounds across the stationary phase and to generate higher plate counts at high velocities. The mass transfer mechanism of bradykinin acetate salt (1060 Da) and insulin (5733 Da) was determined over a range of flow rates from 0.025 to 1.0 mL/min. The pore diffusivities of these two biomolecules were accurately measured from the peak parking method. Even though the gain in column efficiency was not found significant for small molecules such as valerophenone (162 Da), enlarging the average pore size from 80 to 120 Å induces a measurable diminution of the reduced plate height, h, of bradykinin (from 17 to 11 or -35% at a reduced velocity of 50) and a significant reduction for insulin (from 43 to 12 or -72% at a reduced velocity of 90). Remarkably, while the increase of the column efficiency for bradykinin is consistent with a faster diffusivity of bradykinin across the 120 Å Titan-C18 particles, the higher column efficiencies measured for insulin are mostly due to a faster absorption kinetics into the 120 Å than that into the 80 Å Titan-C18 particles. This result is supported by the fact that the effective pore diffusivity of insulin is even slightly smaller across the 120 Å than that across the 80 Å 1.9μm Titan-C18 particles. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Light-scattering detection of phospholipids resolved by HPLC.

    Science.gov (United States)

    Descalzo, A M; Insani, E M; Pensel, N A

    2003-09-01

    An improved method for the analysis of phospholipids by normal-phase HPLC is described. Addition of methanol and acetonitrile to a gradient based on 2-propanol/hexane/water promoted a rapid separation of major classes of bovine surfactant phospholipids (PL) by using a conventional silica column. The use of an ELSD permitted an accurate analysis of a mixture of PL. Calibration curves were linear within the range of 5-40 microg with detection limits below 1 microg for PE and PC, and CV ranged from 0.6 to 9.6%. PL present in surfactant homogenates were separated by a solid-phase extraction (SPE) procedure before HPLC analysis. This methodology gave a recovery of 95% and combined SPE-HPLC and quantification of biological PL within a 30-min run. The use of ELSD detection of the eluted compounds was precise, linear, and sensitive.

  19. Moessbauer study of C18N/Fe Langmuir-Blodgett layers

    Energy Technology Data Exchange (ETDEWEB)

    Kuzmann, Erno [Institute of Chemistry, Eoetvoes Lorand University (Hungary); Telegdi, Judit [Institute of Nanochemistry and Catalysis, Chemical Research Center, HAS (Hungary); Nemeth, Zoltan, E-mail: hentes@chem.elte.hu; Vertes, Attila [Institute of Chemistry, Eoetvoes Lorand University (Hungary); Nyikos, Lajos [Institute of Nanochemistry and Catalysis, Chemical Research Center, HAS (Hungary)

    2012-03-15

    Langmuir-Blodgett (LB) films of octadecanoyl hydroxamic acid (C18N) complexed with Fe{sup 3 + } ions have been prepared at various subphase pH values. The LB films consisting of different number of layers were investigated by {sup 57}Fe conversion electron Moessbauer spectroscopy (CEM) at room temperature. The CEM detector contained a piece of {alpha}-iron, enriched with {sup 57}Fe, using as an internal standard. The Moessbauer pattern of the C18N/Fe LB films is a doublet with parameters {delta} = 0.35 mm/s and {Delta} = 0.74 mm/s. A gradual increase of the relative occurrence of the doublet compared to the sextet of the internal standard was observed with the increasing number of layers, indicating the nearly uniform distribution of Fe among the LB layers.

  20. Development and validation of a new HPLC-DAD method for quantification of sofosbuvir in human serum and its comparison with LC-MS/MS technique: Application to a bioequivalence study.

    Science.gov (United States)

    Miraghaei, Shahram; Mohammadi, Bahareh; Babaei, Atefeh; Keshavarz, Samira; Bahrami, Gholamreza

    2017-09-15

    Although for many analyses tandem mass spectrometry (LC-MS/MS) systems have significant advantage over the high-performance liquid chromatography with diode array detection (HPLC-DAD) however, the HPLC methods are easier, cheaper and more available to perform. As no published method is available for quantitative HPLC analysis of sofosbuvir (SOF), an orally administered anti-hepatitis drug in human serum, this study was aimed to evaluate applicability of the HPLC technique to quantify sofosbuvir and comparison of the two methods for analytical performance. Following extraction of the drug and an internal standard (Hexobarbital), same chromatographic conditions were used for both the systems. After the chromatographic separation on a reverse phase C18 column using a mobile phase consisting of water (containing formic acid 0.5mL/L) and acetonitrile (57:43; v/v) at a flow rate of 0.8mL/min, the eluate was introduced into a DAD detector set at 261nm, then passed through the mass spectrometry system in single ion monitoring mode (SIM). For UV and mass spectrometry detections the calibration curves were linear over a concentration range of 25-3200 and 10-3200ng/mL, respectively and the linearity was over 0.998 for both the systems. Lower limit of quantification (LLOQ) for mass spectrometry and DAD detections were 10 and 25ng/mL, respectively. In conclusion sensitivity of DAD detection is sufficient enough to determine concentrations down to 0.5% of Cmax which achieved in bioequivalence study of sofosbuvir and meet FDA requirements for these types of studies. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. HPLC: Early and Recent Perspectives.

    Science.gov (United States)

    Karger, Barry L.

    1997-01-01

    Provides a perspective on what it was like in the early days of high-performance liquid chromatography (HPLC) and several of the key developments. Focuses on the advances in HPLC generally, and more specifically for the biological sciences, that were necessary for the method to reach the preeminent stage of today. Contains 20 references. (JRH)

  2. Applications of monolithic columns in liquid chromatography-based clinical chemistry assays.

    Science.gov (United States)

    Bunch, Dustin R; Wang, Sihe

    2011-08-01

    Monolithic columns have slowly been applied to HPLC methods for clinical chemistry testing in the last 10 years. The application areas include therapeutic drug monitoring, drugs of abuse, vitamins, porphyrins, and steroids. In comparison with conventional particulate columns, the monolithic columns may offer shorter chromatography time, more robustness, and better resolution for certain analytes. The potential drawback of large mobile phase consumption may be improved with smaller id columns, which are currently on the market. Methods covered in this review are those searchable in PubMed up to December 2010. This review highlights the emergence of monolithic column technology in HPLC methods used for clinical chemistry testing. The goals of this review are threefold: (i) To identify the areas of clinical chemistry that analytical monolithic columns have been used in HPLC methods. (ii) To demonstrate the application of analytical monolithic columns in HPLC methods using different detection systems. (iii) To discuss the advantages and limitations of the monolithic columns compared with particulate columns in the clinical chemistry applications. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Flavonoids in Different Parts of Lysimachia clethroides Duby Extracted by Ionic Liquid: Analysis by HPLC and Antioxidant Activity Assay

    Directory of Open Access Journals (Sweden)

    Jin-feng Wei

    2017-01-01

    Full Text Available To establish methods for simultaneous determination of isoquercitrin, astragalin in leaves, quercetin, and kaempferol in flowers of Lysimachia clethroides Duby, respectively, the methods were ultrasound-assisted extraction combined with RP-HPLC, and ionic liquid was used as the extraction solvent. Meanwhile, the antioxidant activity of the different extracts of L. clethroides was evaluated. Purospher STAR RP-C18 column (4.6 mm × 250 mm, 5 μm was used for analysis. The flow rate was 0.6 mL·min−1, and the column temperature was 25°C. The detection wavelength was 360 nm. The mobile phases a and b consisted of acetonitrile-0.4% phosphoric acid (18 : 82, v/v, methanol (A, and 0.4% phosphoric acid (B, respectively. Linear ranges were 0.068~1.64, 0.060~1.44, 0.0080~0.19, and 0.0077~0.18 μg for isoquercitrin, astragalin, quercetin, and kaempferol, respectively. The average recoveries of the four constituents were 99.17%, 98.39%, 100.68%, and 98.81%, respectively. The antioxidant activity of the extracts was detected by DPPH, ABTS, and FRAP. Under the optimized conditions, all the test solutions showed a certain antioxidant activity and the ionic liquid extracts were better than that of extract of methanol. Ionic liquid used as the extraction solvent had the potential to extract active ingredients efficiently from L. clethroides, and this method improved the antioxidant activity with accurate and reliable results.

  4. Simultaneous determination of phenolic acids and flavonoids in Chenopodium formosanum Koidz. (djulis) by HPLC-DAD-ESI-MS/MS.

    Science.gov (United States)

    Hsu, B Y; Lin, S W; Inbaraj, B Stephen; Chen, B H

    2017-01-05

    A high performance liquid chromatography-diode array detection-tandem mass spectrometry method (HPLC-DAD-MS/MS) was developed for simultaneous determination of phenolic acids and flavonoids in djulis (Chenopodium formosanum Koidz.), a traditional Chinese herb reported to possess vital biological activities. A high yield of phenolic acids and flavonoids was attained by employing 50% ethanol in water as the extraction solvent and shaking in a 60°C water bath for 3h. A total of 8 phenolic acids and 14 flavonoids were separated and identified within 55min by using a Poroshell 120 EC-C18 column with detection at 280nm, flow rate at 0.8mL/min, column temperature at 35°C, and a gradient solvent system of 0.1% formic acid in water and acetonitrile. Two internal standards caffeic acid and kaempferol-3-O-rutinoside were used for quantitation of phenolic acids and flavonoids in djulis respectively. The amounts of phenolic acids ranged from 11.5±0.8μg/g (caffeoyl-putrescine-derivative (2)) to 1855.3±16.9μg/g (hydroxylphenylacetic acid pentoside), while the flavonoids ranged from 19.93±2.29μg/g (quercetin-3-O-(coumaryl)-rutinoside-pentoside (1)) to 257.3±2.05μg/g (rutin-O-pentoside (2)). A high recovery (89.68-97.20%) and high reproducibility was obtained for both phenolic acids and flavonoids with the relative standard deviation (RSD) for the latter ranging from 0.09-8.22% (intra-day variability) and 0.80-8.48% (inter-day variability). This method may be applied to determination of both phenolic acids and flavonoids in food products and Chinese herbs. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Determination of the Marker Diarylheptanoid Phytoestrogens in Curcuma comosa Rhizomes and Selected Herbal Medicinal Products by HPLC-DAD.

    Science.gov (United States)

    Yingngam, Bancha; Brantner, Adelheid; Jinarat, Damrongsak; Kaewamatawong, Rawiwun; Rungseevijitprapa, Wandee; Suksamrarn, Apichart; Piyachaturawat, Pawinee; Chokchaisiri, Ratchanaporn

    2018-01-01

    A method for quantification of diarylheptanoids in Curcuma comosa rhizomes and selected pharmaceutical preparations was established by using HPLC-diode array detector (DAD). The chromatographic separation of three diarylheptanoids [(3S)-1-(3,4-dihydroxy-phenyl)-7-phenyl-(6E)-6-hepten-3-ol (1), (3R)-1,7-diphenyl-(4E,6E)-4,6-heptadien-3-ol (2), and (3S)-1,7-diphenyl-(6E)-6-hepten-3-ol (3)] was performed on a Luna C 18 analytical column using gradient elution with 0.5% acetic acid in water and acetonitrile with a flow rate of 1 mL/min and a column temperature of 35°C. The calibration curves for the analytes showed good linearity (R 2 >0.999), high precision (relative standard deviation (RSD) <2%) and acceptable recovery (98.35-103.90%, RSD <2%). The limit of detection (LOD) and limit of quantification (LOQ) were 0.06-0.22 and 0.18-0.69 µg/mL, respectively. The results of all validated parameters were within the limits according to the International Conference on Harmonization (ICH) Guidelines. The established method was successfully applied for qualitative and quantitative determination of the three constituents in different samples of C. comosa and some commercial products in capsules. The simplicity, rapidity, and reliability of the method could be useful for the fingerprint analysis and standardization of diarylheptanoids, which are responsible for the estrogenic activity in raw materials and herbal medicinal products of C. comosa.

  6. Evaluation of aminoacids in irradiated beans (Vigna unguiculata (L.) Walp) by high performance liquid chromatography (HPLC)

    Energy Technology Data Exchange (ETDEWEB)

    Lima, Keila S. Cople; Souza, Luciana B.; Coelho, Maysa J.; Lima, Antonio L. Santos; Hernandes, Nilber K. [Instituto Militar de Engenharia (IME), Rio de Janeiro, RJ (Brazil). Secao de Engenharia Nuclear]. E-mail: keila@ime.eb.br; Godoy, Ronoel L.O. [EMBRAPA Agroindustria de Alimentos, Rio de Janeiro, RJ (Brazil)]. E-mail: ronoel@ctaa.embrapa.br

    2007-07-01

    Fradinho-bean (Vigna unguiculata (L.) Walp) is originated from Africa and is known in Brazil as 'caupi', 'corda' or 'macassar'. It is grown in the interior of Northeast Brazil (semi-arid region) and can be found in parts of the North, being one of the most important components of people's diet in those regions. The Northeast area produces around 429,375 ton of fradinho-bean per year. Leguminous plants are very important sources of proteins, vitamins, carbohydrates and minerals. This kind of bean is an excellent source of proteins (around 23- 25% of its nutritional content), being superior to regular beans (Phaseolus vulgaris). The irradiation process is an alternative to avoid post-harvesting losses, without changing the nutritional value of food. This study has the objective to evaluate the effect of different gamma irradiation doses (0.0; 0.5; 1.0; 2.5; 5.0 and 10.0 kGy) on aminoacid content of fradinho-bean by high performance liquid chromatography (HPLC) and the accompanying of the grains during storage time of 6 months. After irradiation, the bean grains went through a milling process in order to make flour for posterior extraction. A liquid chromatographer Waters, model Alliance 2695, with fluorescent detector Waters 2475, having a mobile phase with gradient elution of sodium acetate. acetonitrile and Milli-Q water, was employed. The flux used was 1 mL/min and the injection volume of 10 {mu}L. The column (C 18 150.0 x 3.9 mm) was kept at 36 deg C. The results show that gamma irradiation is a promise process for fradinho bean during conservation storage time of 6 months, until the dose of 10.0 kGy. Even the most radio-sensitive aminoacids like aromatics and basic lateral chains were preserved. (author)

  7. Reverse phase HPLC method for detection and quantification of lupin seed γ-conglutin.

    Science.gov (United States)

    Mane, Sharmilee; Bringans, Scott; Johnson, Stuart; Pareek, Vishnu; Utikar, Ranjeet

    2017-09-15

    A simple, selective and accurate reverse phase HPLC method was developed for detection and quantitation of γ-conglutin from lupin seed extract. A linear gradient of water and acetonitrile containing trifluoroacetic acid (TFA) on a reverse phase column (Agilent Zorbax 300SB C-18), with a flow rate of 0.8ml/min was able to produce a sharp and symmetric peak of γ-conglutin with a retention time at 29.16min. The identity of γ-conglutin in the peak was confirmed by mass spectrometry (MS/MS identification) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The data obtained from MS/MS analysis was matched against the specified database to obtain the exact match for the protein of interest. The proposed method was validated in terms of specificity, linearity, sensitivity, precision, recovery and accuracy. The analytical parameters revealed that the validated method was capable of selectively performing a good chromatographic separation of γ-conglutin from the lupin seed extract with no interference of the matrix. The detection and quantitation limit of γ-conglutin were found to be 2.68μg/ml and 8.12μg/ml respectively. The accuracy (precision and recovery) analysis of the method was conducted under repeatable conditions on different days. Intra-day and inter-day precision values less than 0.5% and recovery greater than 97% indicated high precision and accuracy of the method for analysis of γ-conglutin. The method validation findings were reproducible and can be successfully applied for routine analysis of γ-conglutin from lupin seed extract. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. DEVELOPMENT AND VALIDATION OF HPLC METHOD FOR QUANTIFICATION OF CEFOTAXIME IN PLASMA OF PATANWADI SHEEP

    Directory of Open Access Journals (Sweden)

    Suprita Sinha

    2015-12-01

    Full Text Available Present research work was carried out to study the validation procedure of Cefotaxime using plasma of Patanwadi sheep. The samples were analyzed using RP- HPLC C18 column (250 X 4.6 mm with UV detection (254 nm. The method was developed for extraction of Cefotaxime from plasma of sheep using acetonitrile and was validated. The mobile phase was a mixture of 0.05 M potassium dihydrogen phosphate buffer with pH 6 and acetonitrile (88: 12, v/v. The method was validated with respect to linearity, precision and accuracy. The intra day and inter day precision study of cefotaxime was carried out by estimating the corresponding responses 3 times on the same day and on 3 different days (1st, 2nd and 3rd for 6 different concentration of Cefotaxime (10, 5, 2.5, 0.625, 0.312 and 0.156 µg/ml and the results were reported in terms of relative standard deviation (RSD. At all concentration studied the C.V. (Coefficient of Variance was less than 7%. The recovery of Cefotaxime from plasma varied from 85-89%. The limit of detection (LOD was found to be 0.156 ppm. Calibration graphs showed a linear correlation (r > 0.998 over the concentration ranges of 1.56 – 200 µg mL-1 for plasma. The results obtained indicated good precision of assay. The developed method was found to be simple, sensitive, accurate, precise and reproducible and can be used for analytical studies of Cefotaxime.

  9. Sedimentation assisted preparation of ground particles of silica monolith and their C18 modification resulting in a chromatographic phase of improved separation efficiency.

    Science.gov (United States)

    Ali, Ashraf; Ali, Faiz; Cheong, Won Jo

    2017-11-24

    The sedimentation procedure has been adopted in production of ground silica monolith particles to improve chromatographic separation efficiency of the resultant phase. First, silica monolith particles have been successfully prepared in a large scale by a sol-gel process followed by grinding. The particles after calcination were separated by sedimentation into three zones using an Imhoff sedimentation cone. The particles of the bottom zone were derivatized with a C18 ligand and end-capped. The sedimentation process was found to not only eliminate troublesome minute particles but also narrow down the particle size distribution. The resultant phase was packed in glass lined stainless steel micro-columns. The average number of theoretical plates (N) of the columns for a test mixture was 47,000 and 29,300 for the 300 and 150mm columns (1mm internal diameter), corresponding to 157,000/m and 195,000/m, respectively. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. SENSITIVE METHOD FOR THE DETERMINATION OF VINCRISTINE IN HUMAN SERUM BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AFTER ONLINE COLUMN-EXTRACTION

    NARCIS (Netherlands)

    BLOEMHOF, H; VANDIJK, KN; DEGRAAF, SSN; VENDRIG, DEMM; UGES, DRA

    1991-01-01

    A column-switching high-performance liquid chromatographic method was developed for the determination of vincristine in serum. Sample preparation was carried out by means of on-line column-extraction, using a C18 reversed-phase preconcentration column. This technique is simple (minimizing manual

  11. Selective on-line detection of boronic acids and derivatives in high-performance liquid chromatography eluates by post-column reaction with alizarin

    NARCIS (Netherlands)

    Duval, F.L.; Wardani, P.A.; Zuilhof, H.; Beek, van T.A.

    2015-01-01

    An on-line high-performance liquid chromatography (HPLC) method for the rapid and selective detection of boronic acids in complex mixtures was developed. After optimization experiments at an HPLC flow rate of 0.40 mL/min, the HPLC-separated analytes were mixed post-column with a solution of 75 µM

  12. 13C-18O bonding (Δ47) in deep-sea corals: a calibration study

    Science.gov (United States)

    Kimball, J. B.; Tripati, A.; Dunbar, R. B.; Eagle, R.

    2013-12-01

    Deep-sea corals are a potentially valuable archive of temperature in intermediate and deep waters, regions for which a paucity of temperature data exists. These archives could give valuable insight into the natural variability of areas of the ocean that play an active role in large-scale climate dynamics. Due to significant 'vital effects' (i.e., non-equilibrium mineral compositions) in δ18O, however, deep-sea coral have been challenging to develop as a paleotemperature proxy. Clumped-isotope paleothermometry is a new method that may circumvent some of the known complications with δ18O paleotemperature analysis in deep-sea coral. This geothermometer is based on the ordering of heavy 13C-18O ';clumps' in carbonate minerals. Initial calibration studies have shown that the method is independent from the solution chemistry of the precipitating fluids as well as 'vital effects' in deep-sea corals and other types of carbonates. Some kinetic effects have been observed in tropical corals and speleothems. Here we report new data in order to further develop clumped isotopes as a paleothermometer in deep-sea corals as well as to investigate taxon-specific effects. 13C-18O bond ordering was analyzed in live-collected scleractinian (Enallopsammia sp.) and gorgonian (Isididae and Coralliidae) deep-sea corals. We determined mass 47 anomalies in samples (Δ47), which refers to the parts per thousand excess of 13C-18O-16O in CO2 produced on acid digestion of a sample, relative to the amount predicted to be present if isotopes were randomly distributed amongst all CO2 isotopologues. Measured Δ47 values were compared to in situ temperatures and the relationship between Δ47 and temperature was determined for each group to investigate taxon-specific effects.

  13. SINTESIS SENYAWA C18H26O9 DARI HIPTOLIDA HASIL ISOLASI DAUN HYPTIS PECTINATA

    OpenAIRE

    Meiny Suzery; Ely Kusniawati; Dwi Hudiyanti; Bambang Cahyono

    2012-01-01

    SYNTHESIS OF C18H26O9 COMPOUNDS FROM HYPTOLIDE ISOLATED FROM HYPTIS PECTINATA LEAVES. Isolation of hyptolide has been done from Hyptis pectinata, and alkene group transformation through oxidation reactions using H3B: OEt2 to the isolated compound was also conducted. Product analyses were carried out using TLC, UV spectrometry, IR, and LC-MS. Pure crystal with melting point of 86-87oC was isolated. The yield was 1.75% (w/w). After analysing and compilating of spectroscopic data it was confirme...

  14. Quantitative Determination of α-Arbutin, β-Arbutin, Kojic Acid, Nicotinamide, Hydroquinone, Resorcinol, 4-Methoxyphenol, 4-Ethoxyphenol, and Ascorbic Acid from Skin Whitening Products by HPLC-UV.

    Science.gov (United States)

    Wang, Yan-Hong; Avonto, Cristina; Avula, Bharathi; Wang, Mei; Rua, Diego; Khan, Ikhlas A

    2015-01-01

    An HPLC-UV method was developed for the quantitative analysis of nine skin whitening agents in a single injection. These compounds are α-arbutin, β-arbutin, kojic acid, nicotinamide, resorcinol, ascorbic acid, hydroquinone, 4-methoxyphenol, and 4-ethoxyphenol. The separation was achieved on a reversed-phase C18 column within 30 min. The mobile phase was composed of water and methanol, both containing 0.1% acetic acid (v/v). The stability of the analytes was evaluated at different pH values between 2.3 and 7.6, and the extraction procedure was validated for different types of skin whitening product matrixes, which included two creams, a soap bar, and a capsule. The best solvent system for sample preparation was 20 mM NaH2PO4 containing 10% methanol at pH 2.3. The analytical method was validated for accuracy, precision, LOD, and LOQ. The developed HPLC-UV method was applied for the quantitation of the nine analytes in 59 skin whitening products including creams, lotions, sera, foams, gels, mask sheets, soap bars, tablets, and capsules.

  15. Semi-preparative HPLC purification of δ-tocotrienol (δ-T3) from Elaeis guineensis Jacq. and Bixa orellana L. and evaluation of its in vitro anticancer activity in human A375 melanoma cells.

    Science.gov (United States)

    Beretta, Giangiacomo; Gelmini, Fabrizio; Fontana, Fabrizio; Moretti, Roberta Manuela; Montagnani Marelli, Marina; Limonta, Patrizia

    2017-04-24

    In this work, we report a rapid and convenient HPLC-UV-DAD method for the isolation of δ-T3 on semi-preparative scale from two different vitamin E rich processed, commercially available products obtained from the fruits of Elaeis guineensis Jacq. (oil palm) and from the seeds of Bixa orellana L. (achiote tree). Chromatography was run using reverse phase (RP) C-18 columns and HPLC-grade acetonitrile as mobile phase. The purity of the isolated δ-T3, assessed by GC-MS and 1H NMR was above 98%. The δ-T3 cytotoxic activity found in vitro against the proliferation of human A375 melanoma cells compared to that of the other δ-T3 free tocols mixture suggest its primary role in the experimental anticancer activity observed for palm oil derived products. Taken altogether, the results of this study highlight the importance of the application of suitable purification systems for the preparations of tocotrienols prior to their experimental or clinical testing.

  16. Development and validation of HPLC analytical method for nepafenac in ophthalmic dosage form (suspension).

    Science.gov (United States)

    Usman, Shahnaz; Akram, Muhammad; Aziz, Asif; Ramesh, Venkat; Sarheed, Omar Abdulraheem

    2014-09-01

    The aim of the present study was to develop and validate an analytical method for the estimation of nepafenac as a raw material as well as in dosage form (suspension) by using reverse phase high performance liquid chromatographic (RP-HPLC). The target was to obtain an easy, rapid, reproducible as well as a rugged method. The HPLC system that was used in the proposed study was LC-20AD liquid chromatograph equipped with SPD-20A UV-VIS detector. The separation was performed on C18 column which was attached with loop 20 β l. Elution was done at ambient temperature with a mobile phase consisting of acetonitrile: Water (40: 60v/v) at a flow rate of 1ml/min and at a wavelength of 254 nm. The proposed method was validated as per the ICH guidelines. The retention time for nepafenac was 7.49 minutes (% CV=0.0076). The percentage coefficient variation (CV) of six consecutive peak areas of injections was 0.34% with tailing factor 1.76. The peak area responses were linear within the concentration range of 0.078-20.0 βg/ml (R(2)=0.9993). The sensitivity of the method could be evaluated by limits of detection (LOD) (0.0195 β g/ml) and limits of quantitation (LOQ) (0.039 β g/ml). Nepafenac drug is s in its diluent that could see by intra-day (% CV =0.45-1.96) and inter-day variation (%CV=0.173-1.898%). The accuracy and recovery results of 80%, 100% and 120% were 97.40% to 102.10% with % CV of 0.3201% to 1.3496%. The robustness and ruggedness of the method are significantly broader and is reproducible. It could be used as a more convenient, efficient, easy and time saving method for the analysis of drug in raw material as well as in dosage form (ophthalmic suspension).

  17. Monolithic columns with organic sorbent based on poly-1-vinylimidazole for high performance liquid chromatography

    Science.gov (United States)

    Patrushev, Y. V.; Sidelnikov, V. N.; Yudina, Y. S.

    2017-03-01

    Monolithic chromatographic columns for HPLC with sorbent based on 1-vinylimidazole are prepared. It is shown that changing the 1-vinylimidazole content in the initial solution allows us to change the polarity of columns. An example of aromatic hydrocarbons separation is presented.

  18. Derringer desirability and kinetic plot LC-column comparison approach for MS-compatible lipopeptide analysis

    Directory of Open Access Journals (Sweden)

    Matthias D’Hondt

    2014-06-01

    Full Text Available Lipopeptides are currently re-emerging as an interesting subgroup in the peptide research field, having historical applications as antibacterial and antifungal agents and new potential applications as antiviral, antitumor, immune-modulating and cell-penetrating compounds. However, due to their specific structure, chromatographic analysis often requires special buffer systems or the use of trifluoroacetic acid, limiting mass spectrometry detection. Therefore, we used a traditional aqueous/acetonitrile based gradient system, containing 0.1% (m/v formic acid, to separate four pharmaceutically relevant lipopeptides (polymyxin B1, caspofungin, daptomycin and gramicidin A1, which were selected based upon hierarchical cluster analysis (HCA and principal component analysis (PCA.In total, the performance of four different C18 columns, including one UPLC column, were evaluated using two parallel approaches. First, a Derringer desirability function was used, whereby six single and multiple chromatographic response values were rescaled into one overall D-value per column. Using this approach, the YMC Pack Pro C18 column was ranked as the best column for general MS-compatible lipopeptide separation. Secondly, the kinetic plot approach was used to compare the different columns at different flow rate ranges. As the optimal kinetic column performance is obtained at its maximal pressure, the length elongation factor λ (Pmax/Pexp was used to transform the obtained experimental data (retention times and peak capacities and construct kinetic performance limit (KPL curves, allowing a direct visual and unbiased comparison of the selected columns, whereby the YMC Triart C18 UPLC and ACE C18 columns performed as best. Finally, differences in column performance and the (disadvantages of both approaches are discussed. Keywords: Lipopeptide, Hierarchical cluster analysis (HCA, Principal component analysis (PCA, LC–MS, Kinetic plot, Derringer desirability function

  19. Methacrylate-bonded covalent-organic framework monolithic columns for high performance liquid chromatography.

    Science.gov (United States)

    Liu, Li-Hua; Yang, Cheng-Xiong; Yan, Xiu-Ping

    2017-01-06

    Covalent-organic frameworks (COFs) are a newfangled class of intriguing microporous materials. Considering their unique properties, COFs should be promising as packing materials for high performance liquid chromatography (HPLC). However, the irregular shape and sub-micrometer size of COFs synthesized via the traditional methods render the main obstacles for the application of COFs in HPLC. Herein, we report the preparation of methacrylate-bonded COF monolithic columns for HPLC to overcome the above obstacles. The prepared COF bonded monolithic columns not only show good homogeneity and permeability, but also give high column efficiency, good resolution and precision for HPLC separation of small molecules including polycyclic aromatic hydrocarbons, phenols, anilines, nonsteroidal anti-inflammatory drugs and benzothiophenes. Compared with the bare polymer monolithic column, the COF bonded monolithic columns show enhanced hydrophobic, π-π and hydrogen bond interactions in reverse phase HPLC. The results reveal the great potential of COF bonded monoliths for HPLC and COFs in separation sciences. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. An on-line SPE-HPLC method for effective sample preconcentration and determination of fenoxycarb and cis, trans-permethrin in surface waters.

    Science.gov (United States)

    Šatínský, Dalibor; Naibrtová, Linda; Fernández-Ramos, Carolina; Solich, Petr

    2015-09-01

    A new on-line SPE-HPLC method using fused-core columns for on-line solid phase extraction and large volume sample injection for increasing the sensitivity of detection was developed for the determination of insecticides fenoxycarb and cis-, trans-permethrin in surface waters. The separation was carried out on fused-core column Phenyl-Hexyl (100×4.6 mm), particle size 2.7 µm with mobile phase acetonitrile:water in gradient mode at flow rate 1.0 mL min(-1), column temperature 45°C. Large volume sample injection (1500 µL) to the extraction dimension using short precolumn Ascentis Express RP C-18 (5×4.6 mm); fused-core particle size 2.7 µm allowed effective sample preconcentration and efficient ballast sample matrix removal. The washing mobile phase consisting of a mixture of acetonitrile:water; 30:70, (v/v) was pumped at flow rate of 0.5 mL min(-1) through the extraction precolumn to the waste. Time of the valve switch for transferring the preconcentrated sample zone from the extraction to the separation column was set at 3rd min. Elution of preconcentrated insecticides from the extraction precolumn and separation on the analytical column was performed in gradient mode. Linear gradient elution started from 40% of acetonitrile at time of valve switch from SPE column (3rd min) to 95% of acetonitrile at 7th min. Synthetic dye sudan I was chosen as an internal standard. UV detection at wavelength 225 nm was used and the method reached the limits of detection (LOD) at ng mL(-1) levels for both insecticides. The method showing on-line sample pretreatment and preconcentration with highly sensitive determination of insecticides was applied for monitoring of fenoxycarb and both permethrin isomers in different surface water samples in Czech Republic. The time of whole analysis including on-line extraction, interferences removal, chromatography separation and system equilibration was less than 8 min. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Development and validation of a high-performance liquid chromatography method for determination of ractopamine residue in pork samples by solid phase extraction and pre-column derivatization.

    Science.gov (United States)

    Ding, Guanglong; Li, Deguang; Qin, Jiao; Zhu, Juanli; Wang, Baitao; Geng, Qianqian; Guo, Mingcheng; Punyapitak, Darunee; Cao, Yongsong

    2015-08-01

    Ractopamine (RAC) has been approved as a feed additive for swine, cattle or turkey, and is likely to have residue in edible animal products and may pose a potential risk for consumer health. Therefore, it is essential to establish a method to detect the residue of RAC in animal products. This work presents a rapid and sensitive HPLC method for the determination of RAC in pork samples with pre-column derivatization. The RAC derivative was separated on a kromasil C18 column and detected at 284nm with a UV detector. The detection capability (CCβ) was 0.078μgg(-1) and the linearity was established over the concentration range of 0.15-100.0μgg(-1). The overall mean recovery in spike range of 0.2μgg(-1) to 100μgg(-1) was 89.9% with the overall mean relative standard deviation of 4.1%. This method can be used for the quantification of RAC in pork samples and help to establish adequate monitoring of the residue of RAC. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Study of clean up procedures using Charcoal-Alumina-Celite column, immunoaffinity column and strata x column to determine deoxynivalenol by high performance liquid chromatography in wheat

    Directory of Open Access Journals (Sweden)

    Jacqueline Cea

    2011-04-01

    Full Text Available Fusarium graminearum is the most common toxic fungal species affecting grains in Uruguay. Since 1977, due to favorable climate conditions, there have been harvests with prominent Fusarium Head Blight in wheat. This were in 1984, 1990, 1993, 1996 and 2001.Natural Toxin Department of Technological Laboratory of Uruguay as National Reference Laboratory, is continuously improving the analytical methods in order to have a good response to the industry requirements and to the monitoring programs for import and exports commodities. The objective of this work was to compare different clean up methods in order to select the best one for routine determination of deoxynivalenol (DON in wheat (grain and flour.  Charcoal-alumina-celite (7+5+3,  immunoaffinity columns DONPREP R-Biopharm Rhone and Strata X 33 m polymeric sorbent  Phenomenex columns were used to perform the study. Considering as reference analytical method the internal protocol  PEC.TOX.063 accredited by United Kingdom Accreditation Service (UKAS  following the ISO 17025 requirements, and  based on AOAC method 986.17( chapter 49, 2002 for extraction and clean-up  and on J.AOAC 70(3, 1987:479-483 for the high performance liquid chromatography (HPLC detection, two more clean up methods were evaluated. In all of them PEC.TOX.063 detection procedure was carried out. PEC.TOX.063 used for the clean up an in house column chromatography prepared with charcoal-alumina-celite (7+5+3 . Extraction was performed using acetonitrile- water (84+16 and an aliquot of the extract was passed through the column. Extract was dryed under vacuum and DON detected by HPLC using photodiode array detector. For the method that used immunoaffinity columns, water was the extraction solvent, and manufacture protocol was followed up. An aliquot of the extract was passed through the column. Column was washed using water and DON eluated using methanol 100%. For the method that used Strata X column for the clean up, the

  3. A new method for rapid determination of indole-3-carbinol and its condensation products in nutraceuticals using core-shell column chromatography method.

    Science.gov (United States)

    Fibigr, Jakub; Šatínský, Dalibor; Havlíková, Lucie; Solich, Petr

    2016-02-20

    Indole-3-carbinol is a natural glucosinolate known for prevention of human breast, prostate and other types of cancer and it started to be used in commercial preparations, as food supplements. However no analytical method has been proposed for quality control of nutraceuticals with this substance yet. In this paper a new high-performance liquid chromatography (HPLC) method using core-shell column for separation of indole-3-carbinol and its condensation/degradation products was developed and used for the quantitative determination of indole-3-carbinol in nutraceuticals. Separation of indole-3-carbinol, its condensation/degradation products and internal standard ethylparaben was performed on the core-shell column Kinetex 5μ XB-C18 100A (100×4.6mm), particle size 5.0μm, with mobile phase acetonitrile/water according to the gradient program at a flow rate of 1.25mLmin(-1) and at temperature 50°C. The detection wavelength was set at 270nm. Under the optimal chromatographic conditions good linearity of determination was achieved. Available commercial samples of nutraceuticals were extracted with 100% methanol using ultrasound bath. A 5-μL sample volume of the supernatant was directly injected into the HPLC system. The developed method provided rapid and accurate tool for quality control of nutraceuticals based on cruciferous vegetable extracts with indole-3-carbinol content. The presented study showed that the declared content of indole-3-carbinol significantly varied in the different nutraceuticals available on the market. Two analyzed preparations showed the presence of condensation/degradation products of indole-3-carbinol which were not officially declared by the manufacturer. Moreover, further two analyzed nutraceutical preparations showed absolutely no content of declared amount of indole-3-carbinol. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Electrospray[+] tandem quadrupole mass spectrometry in the elucidation of ergot alkaloids chromatographed by HPLC: screening of grass or forage samples for novel toxic compounds.

    Science.gov (United States)

    Lehner, Andreas F; Craig, Morrie; Fannin, Neil; Bush, Lowell; Tobin, Tom

    2005-11-01

    Ergot alkaloids are mycotoxins generated by grass and grain pathogens such as Claviceps, for example. Ergot alkaloid-poisoning syndromes, such as tall fescue toxicosis from endophyte-infected tall fescue grass, are important veterinary problems for cattle, horses, sheep, pigs and chickens, with consequent impact on food, meat and dairy industries. Damage to livestock is of the order of a billion dollars a year in the United States alone. HPLC with UV and fluorescence detection are the predominant means of ergot alkaloid determination, with focus on quantitation of the marker compound ergovaline, although ELISA methods are undergoing investigation. These techniques are excellent for rapid detection, but of poor specificity in defining new or poorly characterized ergot alkaloids and related compounds. This paper demonstrates the facility of using electrospray(+) mass spectrometry with multiple reaction monitoring (MRM) detection during chromatographic examination of ergot alkaloid standards of lysergic acid, lysergol, ergonovine, ergovaline, ergotamine, ergocornine, ergocryptine and ergocrystine by HPLC. Ergoline-8 position epimers could be separated on the gradient HPLC system for ergocornine, ergocrystine and ergonovine and appeared as shoulders for ergotamine and ergovaline; epimers generally showed different patterns of relative intensity for specific MRM transitions. There was reasonable correspondence between retention of standards on the 2-mm ESI(+)MS phenyl-hexyl-based reverse phase column and those on the 4-mm C18-based column. Since up to 10% of clinical cases involving toxin exposure display unidentified chromatographic peaks, 11 samples of feed components associated with such cases were studied with developed MRM methods to attempt elucidation of crucial components if possible. Ergotamine appeared in all, ergovaline appeared in five and ergocornine appeared in six; ergonovine, ergocryptine, ergocrystine and lysergol also appeared in several. In addition

  5. Investigation of a new core-shell particle column for ion-pair reversed-phase liquid chromatography analysis of oligonucleotides.

    Science.gov (United States)

    Biba, Mirlinda; Welch, Christopher J; Foley, Joe P

    2014-08-05

    A new core-shell particle column showed excellent performance and durability for separation of short (∼21-mer) ribonucleic acid (RNA) oligonucleotides by ion-pair reversed-phase liquid chromatography (IP-RPLC). Previously investigated core-shell C18 columns showed excellent peak shapes and separations of closely eluting impurities by IP-RPLC. However, these columns showed only modest long-term stability at the neutral pH and elevated column temperatures of ≥60°C, typically used for IP-RPLC analysis of oligonucleotides. The newly introduced SunShell C18 column provided separations comparable to the previously evaluated core-shell columns, but with significantly improved long-term column stability when operated at neutral pH and elevated column temperature. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Modeling Stone Columns

    OpenAIRE

    Castro Gonzalez, Jorge

    2017-01-01

    This paper reviews the main modeling techniques for stone columns, both ordinary stone columns and geosynthetic-encased stone columns. The paper tries to encompass the more recent advances and recommendations in the topic. Regarding the geometrical model, the main options are the "unit cell", longitudinal gravel trenches in plane strain conditions, cylindrical rings of gravel in axial symmetry conditions, equivalent homogeneous soil with improved properties and three-dimensional models, eith...

  7. Slender CRC Columns

    DEFF Research Database (Denmark)

    Aarup, Bendt; Jensen, Lars Rom; Ellegaard, Peter

    2005-01-01

    CRC is a high-performance steel fibre reinforced concrete with a typical compressive strength of 150 MPa. Design methods for a number of structural elements have been developed since CRC was invented in 1986, but the current project set out to further investigate the range of columns for which...... current design guides can be used. The columns tested had a slenderness varying from 1.11 to 12.76 and a reinforcement ratio (area of rebar to area of concrete) ranging from 0 to 8.8 %. A total of 77 tests were carried out - 61 columns were tested in ambient conditions and 16 columns were tested...

  8. Current HPLC Methods for Assay of Nano Drug Delivery Systems.

    Science.gov (United States)

    Tekkeli, Serife Evrim Kepekci; Kiziltas, Mustafa Volkan

    2017-01-01

    In nano drug formulations the mechanism of release is a critical process to recognize controlled and targeted drug delivery systems. In order to gain high bioavailability and specificity from the drug to reach its therapeutic goal, the active substance must be loaded into the nanoparticles efficiently. Therefore, the amount in biological fluids or tissues and the remaining amount in nano carriers are very important parameters to understand the potential of the nano drug delivery systems. For this aim, suitable and validated quantitation methods are required to determine released drug concentrations from nano pharmaceutical formulations. HPLC (High Performance Liquid Chromatography) is one of the most common techniques used for determination of released drug content out of nano drug formulations, in different physical conditions, over different periods of time. Since there are many types of HPLC methods depending on detector and column types, it is a challenge for the researchers to choose a suitable method that is simple, fast and validated HPLC techniques for their nano drug delivery systems. This review's goal is to compare HPLC methods that are currently used in different nano drug delivery systems in order to provide detailed and useful information for researchers. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  9. HPLC-Chip/MS technology in proteomic profiling.

    Science.gov (United States)

    Vollmer, Martin; van de Goor, Tom

    2009-01-01

    HPLC-chip/MS is a novel nanoflow analytical technology conducted on a microfabricated chip that allows for highly efficient HPLC separation and superior sensitive MS detection of complex proteomic mixtures. This is possible through on-chip preconcentration and separation with fluidic connection made automatically in a leak-tight fashion. Minimum precolumn and postcolumn peak dispersion and uncompromised ease of use result in compounds eluting in bands of only a few nanoliters. The chip is fabricated out of bio-inert polyimide-containing channels and integrated chip structures, such as an electrospray emitter, columns, and frits manufactured by laser ablation technology. Meanwhile, a variety of HPLC-chips differing in design and stationary phase are commercially available, which provide a comprehensive solution for applications in proteomics, glycomics, biomarker, and pharmaceutical discovery. The HPLC-chip can also be easily integrated into a multidimensional separation workflow where different orthogonal separation techniques are combined to solve a highly complex separation problems. In this chapter, we describe in detail the methodological chip usage and functionality and its application in the elucidation of the protein profile of human nucleoli.

  10. [Simultaneous separation and detection of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate by RP-HPLC and structure confirmation].

    Science.gov (United States)

    Zhao, Yan-Yan; Liu, Li-Yan; Han, Yuan-Yuan; Li, Yue-Qiu; Wang, Yan; Shi, Min-Jian

    2013-08-01

    A simple, fast and sensitive analytical method for the simultaneous separation and detection of 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B by RP-HPLC and drug quality standard was established. The structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate have been confirmed. Reference European Pharmacopoeia EP7.0 version, British Pharmacopoeia 2012 version, National Drug Standards of China (WS 1-XG-2002), domestic and international interrelated literature were referred to select the composition of mobile phase. The experimental parameters including salt concentration, pH, addition quantities of organic solvent, column temperature and flow rate were optimized. Finally, the assay was conducted on a Durashell-C18 column (250 mm x 4.6 mm, 5 microm) with 0.01 mol x mL(-1) ammonium perchlorate (add ammonia to adjust the pH value to 8.2) -methanol (48 : 52) as mobile phase at the flow rate of 0.8 mL x min(-1), and the detection wavelength was set at 254 nm. The column temperature was 50 degrees C and the injection volume was 10 microL. The MS, NMR, UV and RP-HPLC were used to confirm the structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate. Under the optimized separation conditions, the calibration curves of 18 alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B showed good linearity within the concentration of 0.50-100 microg x mL(-1) (r = 0.999 9). The detection limits for 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B were 0.15, 0.10, 0.10, 0.15 microg x mL(-1) respectively. The method is sensitive, reproducible and the results are accurate and reliable. It can be used for chiral resolution of 18alpha-glycyrrhizinic acid, 18Pbeta-glycyrrhizinic acid, and detection content of principal component and

  11. HPLC-MS/MS analysis of peramivir in rat plasma: Elimination of matrix effect using the phospholipid-removal solid-phase extraction method.

    Science.gov (United States)

    Lei, Mingdao; Gan, Wei; Sun, Yongbing

    2018-03-01

    A simple HPLC-MS/MS method has been developed for the determination of peramivir in rat plasma in the present study. The analytes were separated on a C18 column (50 × 2.1 mm, 1.7 μm) and a triple-quadrupole mass spectrometer equipped with an electrospray ionization source was applied for the detection. A phospholipid-free cartridge solid-phase extraction was used to pretreat the plasma and eliminate the endogenous phospholipid. The in-source collision-induced dissociation approach showed that this pretreatment could result in negligible ion suppression from the extracted sample and could produce cleaner samples when compared with the protein precipitation. The method was linear over the concentration range of 0.12-1200.0 ng/mL for peramivir. The method was validated and successfully applied to a pharmacokinetic study after peramivir was orally and intravenously administered to Sprague-Dawley rats. Copyright © 2017 John Wiley & Sons, Ltd.

  12. Synchronous determination with double-wavelength by RP-HPLC-UV and optimization of ultrasound-assisted extraction of phenolic acids from Caragana species using response surface methodology.

    Science.gov (United States)

    Zeng, Zhi; Ji, Zhongyin; Hu, Na; Chen, Shasha; Bai, Bo; Wang, Honglun; Suo, Yourui

    2017-06-05

    The utilization of Caragana korshinskii Kom. (CK) is currently concentrated on its ecological and fuel functions. Little attention has been devoted to the analysis of their phenolic acid (PA) components. To obtain more data for further utilization of CK, a new analysis protocol was tested to determine PAs synchronously by RP-HPLC-UV with double-wavelength (280nm and 320nm) detection. Specifically, separation of PA components was performed on a Hypersil Gold C18 reverse phase column with gradient elution. A four-factor-three-level Box-Behnken design was implemented for optimization of PA extraction. The results demonstrated that CK were rich primarily in chlorogenic acid, vanillic acid, caffeic acid and rosmarinic acid. The total content of PAs in CK leaves was the highest compared with its other parts. The distribution of total flavonoid content of CK was leaves>flowers>bark, while that of the total phenolic content of CK was flowers>leaves>bark. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Development and Validation of a Reversed-Phase HPLC Method for the Quantitative Determination of Ten Polyphenols Extracted from Apple Peel.

    Science.gov (United States)

    Ran, Junjian; Sun, Huadi; Zhu, Mingming; Chen, Juan; Zhao, Ruixiang

    2016-01-01

    A method based on a reversed-phase HPLC method was established, optimized, and validated for the separation and quantitation of 10 polyphenols extracted from the peel of apple species. A bidentate reversed-phase C18 column was used as stationary phase, and an acidified water buffer and methanol were used as mobile phase. The polyphenols were well separated and detected using UV at 280 and 320 nm. Validation parameters, such as linearity, LOD, LOQ, accuracy, and precision, were acceptable for all 10 polyphenols. The proposed method has enough linearity with correlation coefficient >0.99 within the investigated range for all tested polyphenols. The LOD was 0.24 μg/mL for ellagic acid and polyphenols. The LOQ was 9.39 × 10(-2) μg/mL for chlorogenic acid, and ellagic acid, 2.82 × 10(-2) μg/mL for caffeic acid and >0.1 μg/mL for all other polyphenols. Recovery was within the acceptable range from 98.38 to 100.39% for all polyphenols standards. Satisfactory precision was achieved for both intra- and interday assay, with RSD polyphenols from apple peel.

  14. Simultaneous determination of four phenolic compounds in extracts of aerial parts of Ipomoea pes-caprae (L. R. Br. (Convolvulaceae by HPLC-UV

    Directory of Open Access Journals (Sweden)

    Daniela Maes Dutra

    2014-01-01

    Full Text Available Among other applications, Ipomoea pes-caprae is popularly used to treat jellyfish stings, supporting the development of a product for dermatological use. Hydroethanolic spray-dried extract was chosen for the further development of phytomedicines, and a stability-indicative HPLC-UV method was developed and validated for the determination of isoquercitrin and isochlorogenic acids A, B and C. The method was developed using a C18 column (250 x 4.6 mm, 5 µm with an acetonitrile:water mobile phase at pH 3.0 in a gradient run. The four constituents and other unidentified components of the extract were appropriately resolved without interference of degradation products after stress tests (acid, alkali, neutral, oxidant, photolysis. The method showed linearity in the isoquercitrin concentration range from 5.0-50.0 µg mL-1, with adequate precision (RSD% < 2.5% for the intra- and inter-day studies, accuracy (recovery of 100.0 ± 2.0%, and robustness. Both the herbal drug and spray-dried extract of I. pes-caprae were subjected to stability studies in accelerated and long-term conditions over four months. The samples maintained their characteristics and marker contents (< 10% of variation.

  15. An HPLC-ESI-MS method for analysis of loureirin A and B in dragon's blood and application in pharmacokinetics and tissue distribution in rats.

    Science.gov (United States)

    Zhao, Haiyan; Chen, Zilin

    2013-04-01

    A sensitive HPLC-ESI-MS method for the simultaneous determination of loureirin A (LA) and loureirin B (LB) in rat plasma and tissues was developed, and the pharmacokinetic and tissue distribution characteristics of LA and LB were investigated after gavage administration. The samples were pretreated by protein precipitation with ethyl acetate and then separated on a Welch Ultimate XB-C18 column with water-acetonitrile (42:58, v/v) containing 0.1% glacial acetic acid as the mobile phase. The analytes were detected with no interference in multiple reaction monitoring (MRM) mode on an electrospray ionization ion trap mass spectrometer. The analytes showed good linearity over a wide concentration range and the lowest limit of quantification (LLOQ) was 5 ng/mL for LA and 2ng/mL for LB in matrices. The pharmacokinetic curves of both analytes were best fitted to one-compartment model. It suggested that the analytes absorbed and distributed very quickly in rats. Tissue distribution results showed that the analytes had a wide distribution in tissues and the highest levels for LA and LB were observed in liver followed by kidney, lung, spleen, heart and cerebrum. This work provided the pharmacokinetics and tissue distribution characteristics of LA and LB, which would be instructive for their clinical regiment design. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Development and validation of a stability-indicating high performance liquid chromatographic (HPLC) method for the determination of related substances of micafungin sodium in drug substances.

    Science.gov (United States)

    Zhu, Shengsheng; Meng, Xiang; Su, Xin; Luo, Yongwei; Sun, Zuyue

    2013-10-24

    An isocratic, sensitive and stability-indicating high performance liquid chromatographic (HPLC) method for separation and determination of the related substances of micafungin sodium was developed. The chromatographic separation was achieved on Agilent Zorbax SB-C18 column (250 × 4.6 mm, 5 μm). Forced degradation study confirmed that the newly developed method was specific and selective to the degradation products. The performance of the method was validated according to the present ICH guidelines for specificity, linearity, accuracy, precision and robustness. Regression analysis showed correlation coefficient value greater than 0.999 for micafungin sodium and its six impurities. Limit of detection of impurities was in the range of 0.006%-0.013% indicating the high sensitivity of the newly developed method. Accuracy of the method was established based on the recovery obtained between 98.2% and 102.0% for all impurities. RSD obtained for the repeatability and intermediate precision experiments, was less than 1.0%. The method was successfully applied to quantify related substances of micafungin sodium in bulk drugs.

  17. Quantitative and qualitative analysis of the novel antitumor 1,3,4-oxadiazole derivative (GLB) and its metabolites using HPLC-UV and UPLC-QTOF-MS

    Science.gov (United States)

    Li, Pu; Wang, Xin; Li, Jian; Meng, Zhi-Yun; Li, Shu-Chun; Li, Zhong-Jun; Lu, Ying-Yuan; Ren, Hong; Lou, Ya-Qing; Lu, Chuang; Dou, Gui-Fang; Zhang, Guo-Liang

    2015-01-01

    Fructose-based 3-acetyl-2,3-dihydro-1,3,4-oxadiazole (GLB) is a novel antitumor agent and belongs to glycosylated spiro-heterocyclic oxadiazole scaffold derivative. This research first reported a simple, specific, sensitive and stable high performance liquid chromatography -ultraviolet detector (HPLC-UV) method for the quantitative determination of GLB in plasma. In this method, the chromatographic separation was achieved with a reversed phase C18 column. The calibration curve for GLB was linear at 300 nm. The lower limit of quantification was 10 ng/mL. The precision, accuracy and stability of the method were validated adequately. This method was successfully applied to the pharmacokinetic study in rats for detection of GLB after oral administration. Moreover, the structures of parent compound GLB and its two major metabolites M1 and M2 were identified in plasma using an ultra performance liquid chromatography- electrospray ionization-quadrupole-time of flight- mass spectrometry (UPLC-ESI-QTOF-MS) method. Our results indicated that the di-hydroxylation (M1) and hydroxylation (M2) of GLB are the major metabolites. In conclusion, the present study provided valuable information on an analytical method for the determination of GLB and its metabolites in rats, can be used to support further developing of this antitumor agent. PMID:26148672

  18. Simultaneous Determination and Pharmacokinetic Study of Six Components in Rat Plasma by HPLC-MS/MS after Oral Administration of Acanthopanax sessiliflorus Fruit Extract

    Directory of Open Access Journals (Sweden)

    Peng Du

    2016-12-01

    Full Text Available A specific and reliable HPLC-MS/MS method was developed and validated for the simultaneous determination of protocatechuic acid (PCA, scopolin, (−-pinoresinol-4,4′-di-O-β-d-glucopyranoside (PDG, acanthoside D, acanthoside B and hyperin in rat plasma for the first time. The analytes were separated on a C18 column (50 × 2.1 mm, 1.8 µm and a triple-quadrupole mass spectrometer equipped with an electrospray ionization (ESI source was used for detection. The rat plasma sample was prepared using the protein precipitation procedure. The calibration curves were linear over a concentration range of 1.2–1200.0 ng/mL for PCA, 0.96–960.0 ng/mL for scopolin, 1.12–1120.0 ng/mL for PDG, 1.32–1320.0 ng/mL for acanthoside D, 0.99–990.0 ng/mL for acanthoside B and 1.01–1010.0 ng/mL for hyperin. The intra-day and inter-day precision was less than 11.4% and the relative error (RE was all within ±15%. The validated method was successfully applied to assess the pharmacokinetics characteristics after the extracts of Acanthopanax sessiliflorus fruits were orally administered to the Sprague-Dawley rat.

  19. Development and Validation of a RP-HPLC Method for Estimation of Montelukast Sodium in Bulk and in Tablet Dosage Form.

    Science.gov (United States)

    Singh, R M; Saini, P K; Mathur, S C; Singh, G N; Lal, B

    2010-03-01

    The present work describes a simple, precise and accurate HPLC method for estimation of montelukast sodium in bulk and in tablet dosage form. The separation was achieved by using octadecylsilane column (C18) and acetonitrile:1 mM sodium acetate adjusted to pH 6.3 with acetic acid in proportion of 90:10 v/v as mobile phase, at a flow rate of 1.5 ml/min. Detection was carried out at 285 nm. The retention time of montelukast sodium was found to be 3.4 min. The limit of detection was found 1.31 µg/ml and limit of quantification 3.97 µg/ml. The accuracy and reliability of the proposed method was ascertained by evaluating various validation parameters like linearity (1-100 µg/ml), precision, accuracy and specificity according to ICH guidelines. The proposed method provides an accurate and precise quality control tool for routine analysis of montelukast sodium in bulk and in tablet dosage form.

  20. A stability indicating HPLC method for determination of mebeverine in the presence of its degradation products and kinetic study of its degradation in oxidative condition.

    Science.gov (United States)

    Souri, E; Aghdami, A Negahban; Adib, N

    2014-01-01

    An HPLC method for determination of mebeverine hydrochloride (MH) in the presence of its degradation products was developed. The degradation of MH was studied under hydrolysis, oxidative and photolysis stress conditions. Under alkaline, acidic and oxidative conditions, degradation of MH was observed. The separation was performed using a Symmetry C18 column and a mixture of 50 mM KH2PO4, acetonitrile and tetrahydrfuran (THF) (63:35:2; v/v/v) as the mobile phase. No interference peaks from degradation products in acidic, alkaline and oxidative conditions were observed. The linearity, accuracy and precision of the method were studied. The method was linear over the range of 1-100 μg/ml MH (r(2)>0.999) and the CV values for intra-day and inter-day variations were in the range of 1.0-1.8%. The limit of quantification (LOQ) and the limit of detection (LOD) of the method were 1.0 and 0.2 μg/ml, respectively. Determination of MH in pharmaceutical dosage forms was performed using the developed method. Furthermore the kinetics of the degradation of MH in the presence of hydrogen peroxide was investigated. The proposed method could be a suitable method for routine quality control studies of mebeverine dosage forms.

  1. A simple reversed phase high-performance liquid chromatography (RP-HPLC method for determination of curcumin in aqueous humor of rabbit

    Directory of Open Access Journals (Sweden)

    Akhilesh Mishra

    2014-01-01

    Full Text Available This article describes a simple and rapid method for determination of curcumin (diferuloylmethane in aqueous humor of rabbit using high-performance liquid chromatography (HPLC. Analysis was performed using a C-18 column (250 × 4.6 mm, 5 μ luna by isocratic elution with a mobile phase containing 25 mM potassium dihydrogen orthophosphate (pH 3.5: Acetonitrile (40:60 and detection at 424 nm using a photodiode array (PDA detector for curcumin. The regression data for curcumin showed a good linear relationship with r 2 > 0.998 over the concentration range of 0.1-10 μg ml−1 . Relative standard deviations (RSD for the intraday and interday coefficient of variations for the assay were less than 5.0 and 8.5, respectively. The recovery of the method was between 79.8-83.6%. The quantification limit of the method for curcumin was 0.01 μg ml−1 . This method has good accuracy, precision, and quantitation limit. It is also concluded that the method is useful for measuring very low curcumin concentrations in aqueous humor.

  2. Development and Validation of a Stability-Indicating RP-HPLC Method by a Statistical Optimization Process for the Quantification of Asenapine Maleate in Lipidic Nanoformulations.

    Science.gov (United States)

    Managuli, Renuka S; Kumar, Lalit; Chonkar, Ankita D; Shirodkar, Rupesh K; Lewis, Shaila; Koteshwara, Kunnatur B; Reddy, Meka Sreenivasa; Mutalik, Srinivas

    2016-09-01

    A stability-indicating RP-HPLC method was developed for quantification of asenapine maleate (ASPM) in lipid nanoformulations. The proposed method was used to assess intrinsic stability of ASPM by conducting force degradation study. The results indicated no considerable degradation of ASPM on subjecting it to hydrolytic, oxidative, thermal and photolytic stresses. The method was validated according to ICH Q2(R1) guidelines by employing Full factorial design using Design-Expert(®) software. ASPM was precisely and accurately quantified in nanoparticles by separating it on Hyperclone BDS C18 using 80-20% v/v mixture of potassium phosphate solution containing 0.1% v/v triethylamine and acetonitrile. The effect of flow rate, pH, acetonitrile content and column temperature was assessed on method responses. The current method was linear in the range of 0.1-20 µg/mL with limit of detection (LOD) and limit of quantification (LOQ) of 29 and 89 ng/mL, respectively. The method was precise and accurate in the determination of ASPM with peak area RSD and recovery of ASPM content in bulk and lipid nanoformulations. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. Validation of a SPE HPLC-UV method for the quantification of a new ER-specific photosensitizer OR-141 in blood serum using total error concept.

    Science.gov (United States)

    Bony, Emilie; Mace, Yohan; Pinto, Adán; Delvaux, David; Kiss, Robert; Riant, Olivier; Feron, Olivier; Quetin-Leclercq, Joëlle

    2017-07-15

    The aim of this work is to develop the first validated HPLC-UV method quantification in blood serum for a new endoplasmic reticulum (ER)-specific benzophenazine photosensitizer (OR-141). A fast solid phase extraction (SPE) cleaning sample procedure was achieved on C18 encapped (ec) SPE cartridges and the separation was performed on a RP-18e column (5μM) using an isocratic elution with methanol. The method has been fully validated according to accuracy profiles based on total error and tolerance intervals. Calibration was performed in the matrix and trueness (<4.25% relative bias), repeatability (<4.75% relative standard deviation (RSD)), intermediate precision (<5.37% RSD), selectivity, response function, linearity, and dilution effect were evaluated for both OR-141 regio-isomers. Afterwards the developed method was successfully applied to perform the quantitative determination of OR-141 in mouse blood serum samples in a preliminary pharmacokinetic study. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. HPLC-DAD DETERMINATION OF BERBERINE, CHELERYTHRINE AND SANGUINARINE IN THE MEXICAN PRICKLY POPPY (Argemone mexicana L. PAPAVERACEAE, A MEDICINAL PLANT

    Directory of Open Access Journals (Sweden)

    Jorge F. Xool-Tamayo

    Full Text Available A sensitive, simple, rapid and reliable HPLC-DAD method for the analysis of the benzylisoquinoline alkaloids (BIA's content in Argemone mexicana (Papaveraceae is presented. This method allows the simultaneous separation and quantitation of berberine (Bn, chelerythrine (C and sanguinarine (S in extracts from A. mexicana tissues, reducing time of analysis in comparison to previous reports. Alkaloids were separated on a C18 Hypersil Gold column using an acetonitrile gradient (20 to 70% in 1% acetic acid in water. Alkaloids were identified based on retention times and UV spectra and quantified at 254 nm. Linearity between 0.5-20 µg mL-1 was observed for Bn, C and S, with limits of detection (LOD and quantitation (LOQ of 0.11 and 0.33 for Bn, 0.10 and 0.30 for C and 0.05 and 0.15 for S, respectively. Maximal intra- and inter-day variation values were < 0.49% in all cases, with alkaloids' recoveries higher than 95%. System suitability tests (SST, including resolution (Rs, retention factor (K', selectivity (α, tailing factor and number of theoretical plates were performed according to the United States Pharmacopeia (USP, fulfilling recommended values. The method proved to be efficient and reproducible when analyzing different tissues of field-collected A. mexicana plants.

  5. Development of a simple and sensitive HPLC-UV method for the simultaneous determination of cannabidiol and Δ(9)-tetrahydrocannabinol in rat plasma.

    Science.gov (United States)

    Zgair, Atheer; Wong, Jonathan C M; Sabri, Akmal; Fischer, Peter M; Barrett, David A; Constantinescu, Cris S; Gershkovich, Pavel

    2015-10-10

    There has been increased interest in the medical use of cannabinoids in recent years, particularly in the predominant natural cannabinoids, cannabidiol (CBD) and Δ(9)-tetrahydrocannabinol (THC). The aim of the current study was to develop a sensitive and reliable method for the quantification of CBD and THC in rat plasma. A combination of protein precipitation using cold acetonitrile and liquid-liquid extraction using n-hexane was utilised to extract CBD and THC from rat plasma. Samples were then evaporated and reconstituted in acetonitrile and 30 μL was injected into an HPLC system. Separation was achieved using an ACE C18-PFP 150 mm × 4.6 mm, 3 μm column at 55 °C with isocratic elution using a mobile phase consisting of acetonitrile-water (62:38, v/v) at 1 mL/min for 20 min. Both cannabinoids, as well as the internal standard (4,4-dichlorodiphenyltrichloroethane, DDT) were detected at 220 nm. Our new method showed linearity in the range of 10-10,000 ng/mL and a lower limit of quantification (LLOQ) of 10 ng/mL for both cannabinoids, which is comparable to previously reported LC-MS/MS methods. Inter- and intra-day precision and accuracy were below 15% RSD and RE, respectively. To demonstrate the suitability of the method for in vivo studies in rats, the assay was applied to a preliminary pharmacokinetic study following IV bolus administration of 5 mg/kg CBD or THC. In conclusion, a simple, sensitive, and cost-efficient HPLC-UV method for the simultaneous determination of CBD and THC has been successfully developed, validated and applied to a pharmacokinetic study in rats. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. HPLC Determination of α-keto Acids in Human Serum and Urine after Derivatization with 4-Nitro-1,2-phenylenediamine

    Directory of Open Access Journals (Sweden)

    Shamroz Bano Sahito

    2013-06-01

    Full Text Available The determination of α-keto acids has clinical importance, because these are intermediates in a number of biochemical processes. This work reports the development of an HPLC procedure for the analysis α-keto acids in blood and urine samples after derivatization with 4-nitro-1,2-phenylenediamine (NPD. Nine α-keto acids: glyoxylic acid (GA, pyruvic acid (PYR, 2-oxobutyric acid (KB, 3-methyl-2-oxobutyric acid (MKBA, 3-methyl-2-oxovaleric acid (K3MVA, 2-oxoglutaric acid (KG, 4-methyl-2-oxovaleric acid (K4MVA, 2-oxohexanoic acid (KHA and phenylpyruvic acid (PPY were derivatized with (NPD at pH 3 and separated on a Zorbax 300 SB-C18 HPLC column (4.6x150mm id and photodiode array detection at 255 nm. The isocratic elution was performed with methanol: water: acetonitrile (42: 56:2, v/ v/ v with a flow rate 0.9 mL/min. The keto acids separated within 14 min. The method was repeatable with a relative standard deviation (RSD of 0.1-2.9% for each of the α-keto acids. The limits of detection and quantitation were obtained within the range 0.05-0.26 µg/ mL and 0.15-0.8 µg/ mL respectively. The method was applied for determination of α-keto acids from a pharmaceutical preparation, human serum and urine samples of healthy volunteers and diabetic patients. The results were further confirmed by standard addition technique. The method is rapid and simple and is suitable for the separation and determination of α-keto acids from clinical samples.

  7. Development of HPLC and NACE methods for the simultaneous determination of benzoic and sorbic acids in sour snap beans containing oil.

    Science.gov (United States)

    Han, Po; Jia, Zhenmin; Liu, Mian; Li, Yongbo; Liu, Hongxia; Yang, Hui; Wang, Xie; Ban, Fuguo; Zhang, Shusheng

    2007-11-01

    The practical methods were developed for the simultaneous determination of benzoic acid (BA) and sorbic acid (SA) in sour snap bean samples containing oil. BA and SA in the samples were extracted by ultrasonication with water, followed by cleanup procedures with precipitation for removing the potential proteins and with petroleum ether liquid-liquid extraction for removing the edible oil contained in the samples. The HPLC method was developed using Supelco C18 (250 mm x 4.6 mm id, 5 microm) as column, MeOH-20 mM NH(4)Ac (25:75 v/v) at 1.0 mL/min as the mobile phase and 230 nm as the detection wavelength. The optimal NACE method was established with a running buffer of 20.0 mM NH(4)Ac in 95% MeOH (pH* 10.6), and an applied voltage of -30 kV over a capillary of 50 microm id x 48.5 cm (40 cm to the detector window), which gave a baseline separation of BA and SA, and as well as of the blank matrix within ca. 10 min. Both HPLC and NACE methods gave the relatively lower limits of quantification at about 0.01-0.02 and 0.04-0.05 mg/kg, respectively, whereas the overall recoveries were larger than 85.0%. The proposed methods have been successfully applied to measure 15 real sour bean samples and the content profile of BA and SA in sour bean samples was obtained and evaluated.

  8. Associations between major fatty acids in plant oils fed to dairy goats and C18 isomers in milk fat.

    Science.gov (United States)

    Martínez Marín, Andrés L; Gómez-Cortés, Pilar; Núñez Sánchez, Nieves; Juárez, Manuela; Garzón Sigler, Ana I; Peña Blanco, Francisco; de la Fuente, Miguel Angel

    2015-05-01

    Relationships between fatty acids (FAs) in plant oils included in goat diets and milk fat C18 isomers were determined by Principal Factor Analysis (PFA). The three first principal factors (PF1, PF2 and PF3) accounted for 64.5% of the total variation in milk FAs contents. Fatty acids with a double bond at carbons 13, 14, 15 or 16 had high (>0.6) and positive loadings for PF1, trans-4 to trans-8 C18:1 for PF2, whereas trans-10 C18:1, trans-11 C18:1 and cis-9 trans-11 C18:2 showed high and positive loadings for PF3. Pearson's correlations supported that PF1, PF2 and PF3 were related to α-linolenic, oleic and linoleic acid intakes, respectively. Our results show that the quantitatively main FAs in plant lipids supplemented to dairy ruminants are often the main cause of the observed changes in milk C18 isomer contents. However, sometimes the observed changes are caused, or at least are influenced, by other FAs present in lower quantities in the plant lipids. Thus, using mixtures of plant oils with differently unsaturated main FAs could be a way of tailoring milk fat composition to a pre-designed pattern.

  9. Inflatable Column Structure

    Science.gov (United States)

    Hedgepeth, J. M.

    1985-01-01

    Lightweight structural member easy to store. Billowing between circumferential loops of fiber inflated column becomes series of cells. Each fiber subjected to same tension along entire length (though tension is different in different fibers). Member is called "isotensoid" column. Serves as jack for automobiles or structures during repairs. Also used as support for temporary bleachers or swimming pools.

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

    African Journals Online (AJOL)

    Purpose: To develop a stability indicating RP-HPLC method for a combination drug product containing a high dose of paracetamol (PR) and low doses of domperidone (DM) and tramadol HCL (TR). Methods: The analytes are well separated by a reverse phase column and an isocratic mobile phase consisting of 0.1 %v/v ...

  11. Development and Validation of a RP-HPLC Method for Assay of ...

    African Journals Online (AJOL)

    1Polymer Research Lab, Department of Chemistry, Faculty of Science, 2Department of Pharmaceutics, Faculty of Pharmacy,. King Abdulaziz ... Purpose: To develop and validate a novel reverse phase high performance liquid chromatographic. (RP-HPLC) .... instrument with Eclipse XDB® column 5 µm (4.6 x. 150 mm).

  12. Determination of organic acids evolution during apple cider fermentation using an improved HPLC analysis method

    NARCIS (Netherlands)

    Zhang, H.; Zhou, F.; Ji, B.; Nout, M.J.R.; Fang, Q.; Zhang, Z.

    2008-01-01

    An efficient method for analyzing ten organic acids in food, namely citric, pyruvic, malic, lactic, succinic, formic, acetic, adipic, propionic and butyric acids, using HPLC was developed. Boric acid was added into the mobile phase to separate lactic and succinic acids, and a post-column buffer

  13. Application of HPLC capacity coefficients to characterize the sorption of polycyclic aromatic compounds to humic acid

    DEFF Research Database (Denmark)

    Nielsen, T.; Helweg, C.; Siigur, K.

    1997-01-01

    The sorption coefficients to humic acid of 46 PAC having a wide range in polarity were compared with the capacity coefficients of the PAC to a non-polar HPLC column material (ODS) and a polar one (Diol). It is shown that polar interactions contribute to the sorption of polar PAC in addition...

  14. Analysis of several irdoid and indole precursors of terpenoid indole alkaloids with a single HPLC run

    DEFF Research Database (Denmark)

    Dagnino, Denise; Schripsema, Jan; Verpoorte, Robert

    1996-01-01

    An isocratic HPLC system is described which allows the separation of the iridoid and indole precursors of terpenoid indole alkaloids, which are present in a single crude extract. The system consists of a column of LiChrospher 60 RP select B 5 my, 250x4 mm (Merck) with an eluent of 1 % formic acid...

  15. Isolation and characterisation of selected germander diterpenoids from authenticated Teucrium chamaedrys and T. canadense by HPLC, HPLC-mS and NMR.

    Science.gov (United States)

    Sundaresan, P Ramnathan; Slavoff, Sarah A; Grundel, Erich; White, Kevin D; Mazzola, Eugene; Koblenz, Daniel; Rader, Jeanne I

    2006-01-01

    Teucrium species, such as germander, are rich in neo-clerodane diterpenoids and have been used in traditional folk medicine for their stimulant, diuretic, antipyretic and antiseptic properties. However, the furano neo-clerodane diterpenoids present in germander have been implicated in the in vivo hepatotoxicity of this botanical. In this study, authenticated germander (Teucrium chamaedrys L. and Teucrium canadense L.) was used as the source material. Methanol extracts of powdered plant mate rial were prepared and analysed by HPLC using Synergi Max-RP columns with monitoring at 220 nm. Limited amounts of teucrin A and other diterpenoid standards were analysed on a Synergi Max-RP column in order to determine their retention times and to generate calibration curves. The same standards were subjected to concurrent mass spectral analysis. Teucrin A and diterpenoids such as dihydroteugin, teuflin, teuflidin and teucvidin were tentatively identified in the plant extracts by HPLC-MS and 1H-NMR experiments. For the isolation of teucrium diterpenoids on a semipreparative scale, a solid-phase extraction method was developed for the first time using styrene divinylbenzene and strata-X sorbents for teucrin A and teuflin, respectively. Semi-preparative HPLC of the methanol extract of the powdered aerial parts of Teucrium plants was carried out on a semipreparative Synergi Max-RP column with photodiode array detection in order to confirm the identities of some diterpenoids by HPLC-MS and NMR.

  16. JCE Feature Columns

    Science.gov (United States)

    Holmes, Jon L.

    1999-05-01

    The Features area of JCE Online is now readily accessible through a single click from our home page. In the Features area each column is linked to its own home page. These column home pages also have links to them from the online Journal Table of Contents pages or from any article published as part of that feature column. Using these links you can easily find abstracts of additional articles that are related by topic. Of course, JCE Online+ subscribers are then just one click away from the entire article. Finding related articles is easy because each feature column "site" contains links to the online abstracts of all the articles that have appeared in the column. In addition, you can find the mission statement for the column and the email link to the column editor that I mentioned above. At the discretion of its editor, a feature column site may contain additional resources. As an example, the Chemical Information Instructor column edited by Arleen Somerville will have a periodically updated bibliography of resources for teaching and using chemical information. Due to the increase in the number of these resources available on the WWW, it only makes sense to publish this information online so that you can get to these resources with a simple click of the mouse. We expect that there will soon be additional information and resources at several other feature column sites. Following in the footsteps of the Chemical Information Instructor, up-to-date bibliographies and links to related online resources can be made available. We hope to extend the online component of our feature columns with moderated online discussion forums. If you have a suggestion for an online resource you would like to see included, let the feature editor or JCE Online (jceonline@chem.wisc.edu) know about it. JCE Internet Features JCE Internet also has several feature columns: Chemical Education Resource Shelf, Conceptual Questions and Challenge Problems, Equipment Buyers Guide, Hal's Picks, Mathcad

  17. Characterization of cis- and trans-octadecenoic acid positional isomers in edible fat and oil using gas chromatography-flame ionisation detector equipped with highly polar ionic liquid capillary column.

    Science.gov (United States)

    Yoshinaga, Kazuaki; Asanuma, Masaharu; Mizobe, Hoyo; Kojima, Koichi; Nagai, Toshiharu; Beppu, Fumiaki; Gotoh, Naohiro

    2014-10-01

    In this study, the characterisation of all cis- and trans-octadecenoic acid (C18:1) positional isomers in partially hydrogenated vegetable oil (PHVO) and milk fat, which contain several cis- and trans-C18:1 positional isomers, was achieved by gas chromatography-flame ionisation detector equipped with a highly polar ionic liquid capillary column (SLB-IL111). Prior to analysis, the cis- and trans-C18:1 fractions in PHVO and milk fat were separated using a silver-ion cartridge. The resolution of all cis-C18:1 positional isomers was successfully accomplished at the optimal isothermal column temperature of 120 °C. Similarly, the positional isomers of trans-C18:1, except for trans-6-C18:1 and trans-7-C18:1, were separated at 120 °C. The resolution of trans-6-C18:1 and trans-7-C18:1 isomers was made possible by increasing the column temperature to 160 °C. This analytical method is suitable for determining the cis- and trans-C18:1 positional isomers in edible fats and oils. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Análise, por CLAE, de carotenóides de cinco linhagens de Rhodotorula HPLC analysis of carotenoids from five Rhodotorula strains

    Directory of Open Access Journals (Sweden)

    Fabio M. Squina

    2003-09-01

    Full Text Available Um método por cromatografia líquida de alta eficiência (CLAE foi otimizado para a análise da composição de carotenóides de cinco linhagens de Rhodotorula.A extração com ruptura mecânica da parede celular da levedura com areia tratada mostrou ser mais eficiente que a ruptura química com dimetilsulfóxido. Os carotenóides foram separados e quantificados por CLAE em coluna de C18 utilizando como fase móvel acetonitrila/metanol (0,1% trietilamina/acetato de etila (75:15:10 e 100% metanol (0,1% trietilamina entre as injeções, com vazão de 1 mL/min. Em todas as linhagens, os carotenóides majoritários encontrados foram torularrodina, toruleno, ³-caroteno e ²-caroteno. Os teores totais de carotenóides, em µg/g, foram de 251,7 em R. glutinis,123,5 em R. rubra,113,2 em R. araucariae,105,8 em R. lactosa ede 103,7 em R. minuta.A method for extraction and HPLC separation of carotenoids from fiveRhodotorula strains was optimized. The extraction by mechanical disruption of the yeast cell wall with fine treated sand was shown to be more efficient than chemical disruption with dimethylsulfoxide. The carotenoids were separated and quantified by HPLC on a C18 column using as mobile phase acetonitrile/methanol (0.1% triethylamina/ethyl acetate (75:15:10 with 100% methanol (0.1% triethylamine between the injections, at a flow rate of 1.0 mL/min. In all strains, the major carotenoids found were torularhodin, torulene, ³-carotene and ²-carotene. The total carotenoid contents, in µg/g, obtained were 251.7 for R. glutinis,123.5 for R. rubra,113.2 for R. araucariae,105.8 for R. lactosaand 103.7 for R. minuta.

  19. Nuclear reactor control column

    Science.gov (United States)

    Bachovchin, Dennis M.

    1982-01-01

    The nuclear reactor control column comprises a column disposed within the nuclear reactor core having a variable cross-section hollow channel and containing balls whose vertical location is determined by the flow of the reactor coolant through the column. The control column is divided into three basic sections wherein each of the sections has a different cross-sectional area. The uppermost section of the control column has the greatest cross-sectional area, the intermediate section of the control column has the smallest cross-sectional area, and the lowermost section of the control column has the intermediate cross-sectional area. In this manner, the area of the uppermost section can be established such that when the reactor coolant is flowing under normal conditions therethrough, the absorber balls will be lifted and suspended in a fluidized bed manner in the upper section. However, when the reactor coolant flow falls below a predetermined value, the absorber balls will fall through the intermediate section and into the lowermost section, thereby reducing the reactivity of the reactor core and shutting down the reactor.

  20. Two-dimensional chromatography based on on-line HPLC-DPPH bioactivity-guided assay for the preparative isolation of analogue antioxidant compound from Arenaria kansuensis.

    Science.gov (United States)

    Cui, Yulei; Shen, Na; Yuan, Xiang; Dang, Jun; Shao, Yun; Mei, Lijuan; Tao, Yanduo; Wang, Qilan; Liu, Zenggen

    2017-03-01

    Traditional Tibetan medicine is important for discovery of drug precursors. However, information about the chemical composition of traditional Tibetan medicine is very limited due to the lack of appropriate chromatographic purification methods. In the present work, A. kansuensis was taken as an example and a novel two-dimensional reversed-phase/hydrophilic interaction liquid chromatography(HILIC) method based on on-line HPLC-DPPH bioactivity-guided assay was developed for the purification of analogue antioxidant compounds with high purity from the extract of A. kansuensis. Based on the separation results of many different chromatographic stationary phases, the first-dimensional (1D) preparation was carried on a RP-C18HCE prep column, and 2 antioxidant fractions were obtained from the 800mg crude sample with a recovery of 56.7%. A HILIC-XAmide prep column was selected for the second-dimensional (2D) preparation. Finally, a novel antioxidant β-carboline Alkaloids (Glusodichotomine AK) and 4 known compounds (Tricin, Homoeriodictyol, Luteolin, Glucodichotomine B) were purified from A. kansuensis. The purity of the compounds isolated from the crude extract was >98%, which indicated that the method built in this work was efficient to manufacture single analogue antioxidant compounds of high purity from the extract of A. kansuensis. Additionally, this method showed great potential in the preparation of analogue structure antioxidant compounds and can serve as a good example for the purification of analogue structure antioxidant carboline alkaloids and flavonoids from other plant materials. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. A validated stability-indicating HPLC method for determination of brimonidine tartrate in BRI/PHEMA drug delivery systems.

    Science.gov (United States)

    Sun, Jianguo; Zhang, Xiuwen; Huang, Taomin

    2017-07-11

    A simple, rapid and accurate stability-indicating reverse phase high performance liquid chromatography (RP-HPLC) was developed and validated for the determination of brimonidine tartrate in brimonidine tartrate/poly(2-hydroxyethyl methacrylate) (BRI/PHEMA) drug delivery contact lenses and pharmaceutical formulations. Optimum chromatographic conditions for separating brimonidine tartrate from other impurities in the leaching liquor of BRI/PHEMA drug delivery contact lenses or pharmaceutical formulations have been achieved by using a Diamonsil C18 column (150 mm × 4.6 mm, 5 μm) as a stationary phase and a mixture solution of phosphate buffer (10 mM, pH3.5) containing 0.5% triethlamine and methanol (85:15, v/v) as a mobile phase at a flow rate of 1 mL/min. The theoretical plates for the brimonidine tartrate measurement were calculated to be 8360 when detection was performed at 246 nm using a diode array detector. The proposed method was validated in accordance with ICH guidelines with respect to linearity, accuracy, precision, robustness, specificity, limit of detection and quantitation. Regression analysis showed a good correlation (R2 > 0.999) for brimonidine tartrate in the concentration range of 0.01-50 μg/mL. The peak purity factor is ≥980 for the analyte after all types of stress tests, indicating an excellent separation of brimonidine tartrate peak from other impurities. The measurement course could be completed within 10 min, which was very quick, effective and convenient. Overall, the proposed stability-indicating method was suitable for routine quality control and drug analysis of brimonidine tartrate in BRI/PHEMA drug delivery contact lenses and other pharmaceutical formulations.

  2. Simultaneous HPLC assay for pretomanid (PA-824), moxifloxacin and pyrazinamide in an inhaler formulation for drug-resistant tuberculosis.

    Science.gov (United States)

    Momin, Mohammad A M; Thien, Sim J; Krittaphol, Woravimol; Das, Shyamal C

    2017-02-20

    A simple and sensitive reversed phase HPLC method has been developed for the simultaneous quantitation of pretomanid (PA-824), moxifloxacin and pyrazinamide in a combination spray-dried powder formulation for inhalation, without any use of an internal standard. Good resolution of the analytes was achieved on a Luna C18 (2), 150×4.6mm, 5μm, 100Å column using gradient elution with a mobile phase containing methanol and triethylamine phosphate buffer (pH 2.5) at a flow rate of 1.0mL/min in a total run time of 25min. Pyrazinamide, moxifloxacin and pretomanid (PA-824) were detected at wavelengths (retention times) of 269nm (3.80min), 296nm (7.94min) and 330nm (17.46min), respectively. The assay was linear for all analytes in the concentration range 2.5-100μg/mL (correlation coefficients >0.999) with LODs and LLOQs (μg/mL) of pretomanid (PA-824) 0.51 and 1.56, moxifloxacin 0.06 and 0.19 and pyrazinamide 0.35 and 1.06, respectively. Recoveries of the three drugs were 99.6-106.8% with intra- and inter-day precisions (as relative standard deviation) of <7%. The method was successfully applied to an evaluation of content uniformity and freedom from interference by l-leucine of a spray-dried combination powder for inhalation. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Development and Validation of an RP-HPLC Method for CB13 Evaluation in Several PLGA Nanoparticle Systems

    Directory of Open Access Journals (Sweden)

    J. Álvarez-Fuentes

    2012-01-01

    Full Text Available A simple, fast, and reversed-phase high-performance liquid chromatographic (RP-HPLC method has been developed and validated for determining of a cannabinoid derivate, which displays potent antihyperalgesic activity, 1-naphthalenyl[4-(pentyloxy-1-naphthalenyl]methanone (CB13 into PLGA nanoparticles. Separation was achieved in a C18 column using a mobile phase consisting of two solvents: solvent A, consisting of acetonitrile : water : acetic acid (75 : 23.7 : 1.3 v/v, and solvent B, consisting of acetonitrile. An isocratic method (70 : 30 v/v, with a flow rate of 1.000 mL/min, and a diode array detector were used. The developed method was precise, accurate, and linear over the concentration range of analysis with a limit of detection and a limit of quantification of 0.5 and 1.25 μg/mL, respectively. The developed method was applied to the analysis of CB13 in nanoparticles samples obtained by three different procedures (SEV, FF, and NPP in terms of encapsulation efficiency and drug release. Nanoparticles size and size distribution were also evaluated founding that NPP method presented the most lowest particle sizes with narrow-size distribution (≈320 nm and slightly negative zeta potential (≈−25 mV which presumes a suitable procedure for the synthesis of PLGA-CB13 nanoparticles for oral administration.

  4. A rapid and validated HPLC method to quantify racecadotril metabolite, thiorphan, in human plasma using solid-phase extraction.

    Science.gov (United States)

    Xu, Fan; Yang, Lingli; Xu, Guili

    2008-01-01

    A HPLC method with UV detection was developed and validated for the determination of thiorphan in human plasma. Nevirapine was used as the internal standard. Separation was performed by a Waters sunfire C18 reversed-phase column maintained at 35 degrees C. The mobile phase was a mixture of 0.05 M phosphate buffer with the pH adjusted to 2.6 and acetonitrile (74:26, v/v) at a flow rate of 1.0 mL/min. The UV detector was set at 210 nm. An original pre-treatment of plasma samples was developed, based on solid-phase extraction (SPE) with solid-phase extraction cartridges (Oasis HLB 3 mL, 60 mg). The extraction recovery for plasma samples of thiorphan at 0.1, 0.4 and 2.0 microg/mL was 93.5%, 98.2% and 97.8%, respectively. The calibration curve was linear with the correlation coefficient (r) above 0.9998. Linearity was verified over the range of 0.05-4 microg/mL thiorphan in plasma. The limit of quantification (LOQ) is 0.05 microg/mL. The mean accuracy was 92.7-99.6%. The coefficient of variation (precision) in the within- and between-batch was 2.2-8.4% and 4.1-8.1%, respectively. This method is simple, economical and specific, and has been used successfully in a pharmacokinetic study of thiorphan.

  5. Development and Validation of a HPLC Method to Determine Griseofulvin in Rat Plasma: Application to Pharmacokinetic Studies

    Directory of Open Access Journals (Sweden)

    Bo Wei

    2008-01-01

    Full Text Available A simple, specific, sensitive, and rapid high performance liquid chromatography (HPLC method for the determination of griseofulvin in small volumes of rat plasma was developed and validated using warfarin as an internal standard. Biological sample preparation involved simple extraction with acetonitrile, followed by dilution with aqueous mobile phase buffer (20 mM sodium dihydrogen phosphate, pH 3.5 to eliminate any chromatographic solvent effects. Griseofulvin and warfarin were baseline separated and quantitated on a C18 reversed phase column (4.6 × 150 mm, 3.5 µ m, using a mobile phase composed of a 20 mM aqueous solution of sodium dihydrogen phosphate-acetonitrile (55:45, v/v, pH 3.5 delivered at a flow rate of 1.0 mL/min, and with fluorescence detection (λexcitation = 300 nm, λemission = 418 nm. The method was proven to be linear over a plasma griseofulvin concentration range of 10 to 2500 ng/mL with a mean correlation coefficient of 0.9996. The intra-day and inter-day accuracy (relative error were in the range of 0.89% to 9.26% and 0.71% to 7.68%, respectively. The within-day precision (coefficient of variation was less than 3.0% and the between-day precision was less than 7.5%. The mean recovery of griseofulvin from rat plasma was found to be 99.2%. The limit of detection (LOD and the limit of quantification (LOQ of griseofulvin were determined to be 1 ng/mL and 10 ng/mL, respectively. The developed method was successfully applied to quantitatively assess the pharmacokinetics of griseofulvin in rats following a single 50 mg/kg oral dose of the drug.

  6. Simultaneous Determination of Four Triterpenoid Saponins in Aralia elata Leaves by HPLC-ELSD Combined with Hierarchical Clustering Analysis.

    Science.gov (United States)

    Sun, Yichun; Li, Baimei; Lin, Xiaoting; Xue, Juan; Wang, Zhibin; Zhang, Hongwei; Jiang, Hai; Wang, Qiuhong; Kuang, Haixue

    2017-05-01

    Aralia elata leaves are known to have several biological activities, including anti-arrythmia, antitumor, anti-inflammatory, anti-fatigue, antimicrobial and antiviral effects. Our previous study found that triterpenoid saponins from the leaves of A. elata had antitumor effects. Quantification of the triterpenoids is important for the quality control of A. elata leaves. To establish high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) for the simultaneous determination of four major triterpenoid saponins, including Aralia-saponin IV, Aralia-saponin VI, 3-O-β-d- glucopyranosyl-(1 → 3)-β-d-glucopyranosyl-(1 → 3)-β-d-glucopyranosyl oleanolic acid 28-O-β-d-glucopyranoside (Aralia-saponin TTP)and Aralia-saponin V. The separation was carried out on a Dikma Diamonsil C 18 column (4.6 mm × 250 mm, 5 μm) efficiently with gradient elution consisting of acetonitrile and water. All calibration curves showed good linear regression (R 2  > 0.9996) within the ranges of tested concentrations. This validated method was applied to determine the contents of the four major triterpenoid saponins in 53 samples from different regions of northeast China. Hierarchical clustering analysis was first used to classify and differentiate Aralia elata leaves. The method developed was successfully applied to analyse four major triterpenoid saponins in Aralia elata leaves which is helpful for quality control of the herb. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  7. Assessment by HPLC of the degradation behavior of acitretin under hydrolytic, oxidative, photolytic and thermal stress conditions

    Directory of Open Access Journals (Sweden)

    Pawan K. Porwal

    2014-12-01

    Full Text Available Acitretin is a photosensitive oral retinoid with very limited data available on its degradation. The official HPLC method for acitretin determination was insufficient to resolve the degradation products generated during stability studies. Therefore, an isocratic RP-HPLC–UV method was developed for the determination of acitretin in the presence of its related impurities and degradation products. Efficient chromatographic separation was achieved on a Thermo beta-basic column C18 (100 mm×4.6 mm, 5 μm with mobile phase containing 0.3% (v/v glacial acetic acid with acetonitrile (ACN and isopropyl alcohol (IPA in an isocratic ratio of 70:30 at a flow rate of 1.0 mL/min with the eluent monitored at 360 nm. The method was validated for specificity, linearity, precision, accuracy and robustness. The calibration plot was linear over the concentration range of 50–150 μg/mL with a correlation coefficient (r2 of 0.999. The proposed method was used to investigate the degradation kinetics of acitretin under the different degradative conditions. The degradation rate constant (K, half-life (t1/2, and t90 were calculated. Degradation of acitretin followed pseudo-first-order kinetics. The drug was found to be less stable under acidic and photolytic degradation conditions: the photolytic degradation constants for acitretin in sunlight and UV light were 0.002698% and 0.0008402% min−1, respectively. The LOD for acitretin and the known impurities were at a level below 0.02%. The method shows consistent recoveries for ACTR (99.8%–101.2% and also for its known impurities (97.2–101.3%. The method was found to be accurate, precise, linear, specific, sensitive, rugged, robust, and useful for characterizing the stability of this chemical.

  8. Simultaneous stability-indicating HPLC method for the determination of cisapride, methylparaben and propylparaben in oral suspension

    Directory of Open Access Journals (Sweden)

    Jutima Boonleang

    2010-08-01

    Full Text Available A simultaneous stability-indicating HPLC method for the determination of cisapride, methylparaben and propylparabenin oral suspensions has been developed and validated. Baseline separation was achieved on a C18 column at room temperature(25°C by gradient elution with mobile phase consisting of solvent A: 10% v/v acetonitrile in 0.13% w/v sodium-1-pentanesulfonate pH 8 and solvent B: acetonitrile. The gradient program was as follows: 0-5 min: 20 to 56% solvent B; 5-7min: 56 to 85% solvent B; 7-10 min: 85% solvent B. The flow rate of mobile phase was 1.2 mL/min. The injection volume was20 L. Detection and peak purity assessments were performed by photo-diode array detector set at 275 nm with scan modein the range of 190-400 nm. The method was selective, accurate and precise. It provided chromatograms with good peak shapeand acceptable resolutions of greater than 4.4 for all analytes including the degradation products formed in oral suspensionsin about 8.5 min. All analyte peaks were pure. The accuracy of all analytes was in the range of 99.20-100.6%. The within-runand between-run relative standard deviations were less than 1.50%. The calibration curves for cisapride, methylparaben, andpropylparaben were linear over the concentration range of 10.0-75.0 g/mL, 8.0-100.0 g/mL, and 0.8-10.0 g/mL, respectivelywith r2 greater than 0.999. This developed method was successfully applied to the stability study of cisapride, methylparabenand propylparaben in oral suspension formulations.

  9. Human low density lipoprotein (LDL) and human serum albumin (HSA) co-adsorption onto the C18-silica gradient surface

    Energy Technology Data Exchange (ETDEWEB)

    Hlady, V. [Utah Univ., Salt Lake City, UT (United States). Dept. of Bioengineering; Ho, C.H.

    2001-02-01

    Co-adsorption kinetics of human low density lipoprotein (LDL) and serum albumin (HSA) on hydrophilic/hydrophobic gradient silica surface were studied using Total Internal Reflection Fluorescence (TIRF) and autoradiography. Two experimental systems were examined: (1) fluorescein-labeled LDL (FITC-LDL) adsorption from a FITC-LDL+HSA solution mixture onto the octadecyldimethylsilyl (C18)-silica gradient surface, and (2) the FITC-LDL adsorption onto the HSA pre-adsorbed on the C18-silica gradient surface. Experiments with fluorescein-labeled albumin (FITC-HSA) and unlabeled LDL have been performed in parallel. The adsorption kinetics of FITC-LDL onto the hydrophilic silica was found to be transport-limited and not affected by co-adsorption of HSA. A slower adsorption kinetics of lipoprotein onto the silica with pre-adsorbed HSA layer resulted from a slow appearance of LDL binding sites exposed by the process of HSA desorption. In the region of increasing surface density of C18 groups, the FITC-LDL adsorption rate fell below the transport-limited adsorption rate, except in the very early adsorption times. Pre-adsorption of HSA onto the C18-silica gradient region resulted in a significant decrease of both the FITC-LDL adsorption rate and adsorbed amount. The lowest FITC-LDL adsorption was found in the region of C18 self-assembled monolayer, where the pre-adsorbed HSA layer almost completely eliminated lipoprotein binding. (orig.)

  10. Literatuuronderzoek HPLC-methoden voor vitamine E

    NARCIS (Netherlands)

    Altena, A.; Hollman, P.C.H.

    1985-01-01

    Doel van dit onderzoek is: het inventariseren van HPLC-methoden voor vitamine E, eventueel in combinatie met vitamine A, in levensmiddelen. Een overzicht van de in de literatuur beschreven HPLC-methoden vanaf ca. 1977 wordt gegeven.

  11. Mass transfer kinetics, band broadening and column efficiency.

    Science.gov (United States)

    Gritti, Fabrice; Guiochon, Georges

    2012-01-20

    Important progress was recently made in our understanding of the physico-chemical aspects of mass transfer kinetics in chromatographic columns, in methods used for accurate determination of the different contributions to the height equivalent to a theoretical plate (HETP), and in the application of these advances to the elucidation of mass transfer mechanisms in columns packed with recent chromatographic supports (sub-2 μm fully porous particles, sub-3 μm core-shell particles, and monoliths). The independent contributions to the HETP are longitudinal diffusion, eddy dispersion, liquid-solid mass transfer (including trans-particle or trans-skeleton mass transfer and external film mass transfer), and the contributions caused by the thermal heterogeneity of the column. The origin and importance of these contributions are investigated in depth. This work underlines the areas in which improvements are needed, an understanding of the contribution of the external film mass transfer term, a better design of HPLC instruments providing a decrease of the extra-column band broadening contributions to the apparent HETP, the development of better packing procedures giving more radially homogeneous column beds, and new packing materials having a higher thermal conductivity to eliminate the nefarious impact of heat effects in very high pressure liquid chromatography (vHPLC) and supercritical fluid chromatography (SFC). Copyright © 2011 Elsevier B.V. All rights reserved.

  12. Microminiature gas chromatographic column

    Science.gov (United States)

    Donaldson, R. W., Jr.

    1972-01-01

    Techniques commonly used for fabrication of integrated circuits are utilized to produce long capillary tubes for microminiature chromatographs. Method involves bonding of flat silicon plate to top of spirally grooved silicon chip to close groove and form capillary column.

  13. Towards Atomic Column-by-Column Spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Pennycook, S.J.; Rafferty, B.

    1998-09-06

    The optical arrangement of the scanning transmission electron microscope (STEM) is ideally suited for performing analysis of individual atomic columns in materials. Using the incoherent Z-contrast image as a reference, and arranging incoherent conditions also for the spectroscopy, a precise correspondence is ensured between features in the inelastic image and elastic signals. In this way the exact probe position needed to maximise the inelastic signal from a selected column can be located and monitored during the analysis using the much higher intensity elastic signal. Although object functions for EELS are typically less than 1 {Angstrom} full width at half maximum, this is still an order of magnitude larger than the corresponding object functions for elastic (or diffuse) scattering used to form the Z-contrast image. Therefore the analysis is performed with an effective probe that is significantly broader than that used for the reference Z-contrast image. For a 2.2 {Angstrom} probe the effective probe is of the order of 2.5 {Angstrom}, while for a 1.3 {Angstrom} probe the effective probe is 1.6 {Angstrom}. Such increases in effective probe size can significantly reduce or even eliminate contrast between atomic columns that are visible in the image. However, this is only true if we consider circular collector apertures. Calculations based upon the theory of Maslen and Rossouw (Maslen and Rossouw 1984; Rossouw and Maslen 1984) show that employing an annular aperture can reduce the FWHM of the inelastic object function down to values close 0.1 {Angstrom}. With practical aperture sizes it should be possible to achieve this increased spatial resolution without loosing too much signal.

  14. Are Extracted Materials Truly Representative of Original Samples? Impact of C18 Extraction on CDOM Optical and Chemical Properties.

    Science.gov (United States)

    Andrew, Andrea A; Del Vecchio, Rossana; Zhang, Yi; Subramaniam, Ajit; Blough, Neil V

    2016-01-01

    Some properties of dissolved organic matter (DOM) and chromophoric dissolved organic matter (CDOM) can be easily measured directly on whole waters, while others require sample concentration and removal of natural salts. To increase CDOM content and eliminate salts, solid phase extraction (SPE) is often employed. Biases following extraction and elution are inevitable, thus raising the question of how truly representative the extracted material is of the original. In this context, we investigated the wavelength dependence of extraction efficiency for C18 cartridges with respect to CDOM optical properties using samples obtained from the Middle Atlantic Bight (MAB) and the Equatorial Atlantic Ocean (EAO). Further, we compared the optical changes of C18 extracts and the corresponding whole water following chemical reduction with sodium borohydride (NaBH4). C18 cartridges preferentially extracted long-wavelength absorbing/emitting material for samples impacted by riverine input. Extraction efficiency overall decreased with offshore distance away from riverine input. Spectral slopes of C18-OM samples were also almost always lower than those of their corresponding CDOM samples supporting the preferential extraction of higher molecular weight absorbing material. The wavelength dependence of the optical properties (absorption, fluorescence emission, and quantum yield) of the original water samples and their corresponding extracted material were very similar. C18 extracts and corresponding water samples further exhibited comparable optical changes following NaBH4 reduction, thus suggesting a similarity in nature (structure) of the optically active extracted material, independent of geographical locale. Altogether, these data suggested a strong similarity between C18 extracts and corresponding whole waters, thus indicating that extracts are representative of the CDOM content of original waters.

  15. Are extracted materials truly representative of original samples? Impact of C18 extraction on CDOM optical and chemical properties

    Directory of Open Access Journals (Sweden)

    Andrea A Andrew

    2016-02-01

    Full Text Available Some properties of dissolved organic matter (DOM and chromophoric dissolved organic matter (CDOM can be easily measured directly on whole waters, while others require sample concentration and removal of natural salts. To increase CDOM content and eliminate salts, solid phase extraction is often employed. Biases following extraction and elution are inevitable, thus raising the question of how truly representative the extracted material is of the original. In this context, we investigated the wavelength dependence of extraction efficiency for C18 cartridges with respect to CDOM optical properties using samples obtained from the Middle Atlantic Bight (MAB and the Equatorial Atlantic Ocean (EAO. Further, we compared the optical changes of C18 extracts and the corresponding whole water following chemical reduction with sodium borohydride (NaBH4.C18 cartridges preferentially extracted long-wavelength absorbing/emitting material for samples impacted by riverine input. Extraction efficiency overall decreased with offshore distance away from riverine input. Spectral slopes of C18-OM samples were also almost always lower than those of their corresponding CDOM samples supporting the preferential extraction of higher molecular weight absorbing material. The wavelength dependence of the optical properties (absorption, fluorescence emission and quantum yield of the original water samples and their corresponding extracted material were very similar. C18 extracts and corresponding water samples further exhibited comparable optical changes following NaBH4 reduction, thus suggesting a similarity in nature (structure of the optically active extracted material, independent of geographical locale. Altogether, these data suggested a strong similarity between C18 extracts and corresponding whole waters, thus indicating that extracts are representative of the CDOM content of original waters.

  16. Comparison of Separation of Seed Oil Triglycerides Containing Isomeric Conjugated Octadecatrienoic Acid Moieties by Reversed-Phase HPLC

    OpenAIRE

    Anh Van Nguyen; Victor Deineka; Lumila Deineka; Anh Vu Thi Ngoc

    2017-01-01

    Relative retention analysis and increment approach were applied for the comparison of triglycerides (TGs) retention of a broad set of plant seed oils with isomeric conjugated octadecatrienoic acids (CLnA) by reversed-phase HPLC for “propanol-2-acetonitrile” mobile phases and Kromasil 100-5C18 stationary phase with diode array detection (DAD) and mass spectrometric (MS) detection. The subjects of investigation were TGs of seed oils: Calendula officinalis, Catalpa ovata, Jacaranda mimosifolia, ...

  17. DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF IVERMECTIN AND CLORSULON IN IVERCAM INJECTION

    OpenAIRE

    Vegad Kunjal L*, Paranjape Dipty B., Shah Dhwani A., Patel Ekta D., Patel Yogesh K., Patel Kaushik R.

    2017-01-01

    A precise, simple, accurate and selective method was developed and validate for estimation of Ivermectin and Clorsulon in Ivercam injection, Reversed phase high performance liquid chromatographic (RP-HPLC) method was developed for routine quantification of Ivermectin and Clorsulon in laboratory prepared mixtures as well as in combined dosage form. Chromatographic separation was achieved on a BDS hypersil C18 (5μ, 250 x 4.6 mm) utilizing mobile phase of filtered and degassed mixture of 60 phos...

  18. 26 CFR 1.381(c)(18)-1 - Depletion on extraction of ores or minerals from the waste or residue of prior mining.

    Science.gov (United States)

    2010-04-01

    ... the waste or residue of prior mining. 1.381(c)(18)-1 Section 1.381(c)(18)-1 Internal Revenue INTERNAL... mining. (a) Carryover requirement. Section 381(c)(18) provides that the acquiring corporation in a... corporation after the date of distribution or transfer for the purpose of determining the applicability of...

  19. High-resolution study of oscillator strengths and predissociation rates for 13C18O . W-X bands and Rydberg complexes between 92.9 and 93.5 nm

    Science.gov (United States)

    Eidelsberg, M.; Lemaire, J. L.; Federman, S. R.; Heays, A. N.; Stark, G.; Lyons, J. R.; Gavilan, L.; de Oliveira, N.

    2017-06-01

    We carried out experiments at the SOLEIL synchrotron facility to acquire data for modelling CO photochemistry in the vacuum ultraviolet. We report oscillator strengths and predissociation rates for four vibrational bands associated with transitions from the v = 0 level of the X1Σ+ ground state to the v = 0-3 vibrational levels of the core excited W1Π Rydberg state, and for three overlapping bands associated with the 4pπ, 5pπ, and 5pσ Rydberg states between 92.9 and 93.4 nm in 13C18O. These results complete those obtained in the same conditions for 12C16O, 13C16O, and 12C18O recently published by us, and extend the development of a comprehensive database of line positions, oscillator strengths, and linewidths of photodissociating transitions for CO isotopologues. Absorption spectra were recorded using the Vacuum UltraViolet Fourier Transform Spectrometer (VUV-FTS) installed on the Dichroïsme Et Spectroscopie par Interaction avec le Rayonnement Synchrotron (DESIRS) beamline at SOLEIL. The resolving power of the measurements, R = 300 000 to 400 000, allows the analysis of individual line strengths and widths within the bands. Gas column densities in the differentially pumped system were calibrated using the B-X (0-0) band at 115.1 nm in 13C18O.

  20. Concurrent estimation of amlodipine besylate, hydrochlorothiazide and valsartan by RP-HPLC, HPTLC and UV-spectrophotometry.

    Science.gov (United States)

    Sharma, Manish; Kothari, Charmy; Sherikar, Omkar; Mehta, Priti

    2014-01-01

    Accurate, sensitive and reproducible reversed-phase high-performance liquid chromatography (RP-HPLC), high-performance thin-layer chromatography (HPTLC) and ultraviolet (UV) spectrophopometric methods were developed for the concurrent estimation of amlodipine besylate (AMLO), hydrochlorothiazide (HCTZ) and valsartan (VALS) in bulk and combined tablet dosage forms. For the RP-HPLC method, separation was achieved on a C18 column using potassium dihydrogen orthophosphate buffer (50 mM, pH 3.7) with 0.2% triethylamine as the modifier and acetonitrile in the ratio of 56:44 (v/v) as the mobile phase. Quantification was achieved using a photodiode array detector at 232 nm over a concentration range of 2-25 µg/mL for AMLO, 5-45 µg/mL for HCTZ and 20-150 µg/mL for VALS. For the HPTLC method, the drugs were separated by using ethyl acetate-methanol-toluene-ammonia (7.5:3:2:0.8, v/v/v/v) as the mobile phase. Quantification was achieved using UV detection at 242 nm over a concentration range of 100-600 ng/spot for AMLO, 150-900 ng/spot for HCTZ and 1,200-3,200 ng/spot for VALS. The UV-spectrophotometric simultaneous equation method was based on the measurement of absorbance at three wavelengths; i.e., at 237.6 nm (λmax of AMLO), 270.2 nm (λmax of HCTZ) and 249.2 nm (λmax of VALS) in methanol. Quantification was achieved over the concentration range of 2-20 µg/mL for AMLO, 5-25 µg/mL HCTZ and 10-50 µg/mL for VALS. All methods were validated according to International Conference on Harmonization guidelines and successfully applied to marketed pharmaceutical formulations. Additionally, the three methods were compared statistically by an analysis of variance test, which revealed no significant difference between the proposed methods with respect to accuracy and precision.

  1. A rapid and validated HPLC method to quantify sphingosine 1-phosphate in human plasma using solid-phase extraction followed by derivatization with fluorescence detection

    NARCIS (Netherlands)

    Butter, Jan J.; Koopmans, Richard P.; Michel, Martin C.

    2005-01-01

    We describe the development and validation of analytical methodology for the determination of sphingosine 1-phosphate (S1P) in plasma. It uses solid-phase extraction (SPE) followed by an automated reversed-phase gradient HPLC column-switching system with a pre-column derivatization with

  2. SINTESIS SENYAWA C18H26O9 DARI HIPTOLIDA HASIL ISOLASI DAUN HYPTIS PECTINATA

    Directory of Open Access Journals (Sweden)

    Meiny Suzery

    2012-05-01

    Full Text Available SYNTHESIS OF C18H26O9 COMPOUNDS FROM HYPTOLIDE ISOLATED FROM HYPTIS PECTINATA LEAVES. Isolation of hyptolide has been done from Hyptis pectinata, and alkene group transformation through oxidation reactions using H3B: OEt2 to the isolated compound was also conducted. Product analyses were carried out using TLC, UV spectrometry, IR, and LC-MS. Pure crystal with melting point of 86-87oC was isolated. The yield was 1.75% (w/w. After analysing and compilating of spectroscopic data it was confirmed as hyptolide compound. Hydroboration of this compound (followed by hydrolysis using H2O2 under alkaline conditions produce its alcohol derivatives, with 28.9% the percentage of transformation, it was demonstrated by LCMS data. IR spectrum at 3600cm-1, confirming the replacement of hydroxyl bond by alkene. Regioselectivity of addition reaction is proposed through simulation with Chem Office. The reaction product was suspected as 6-hydroxy-7-(6-oxo-3,6-dihydro-2H-pyran-2-yl heptane-2,3,5-tryil triacetate. Extension of reaction time to 24 hours, has increase hydroboration product to 78.3%. This research has opened other studies of natural materials in accordance to the roadmap set.  Telah dilakukan isolasi hiptolida dari bahan alam Hyptis pectinata, dan transformasinya melalui reaksi oksidasi menggunakan H3B:OEt2 terhadap gugus alkena pada senyawa hasil isolasi. Analisis produk dilakukan menggunakan KLT, spektrometri UV, IR, dan LC-MS. Kristal murni dengan titik leleh 86-87oC berhasil diisolasi dengan rendemen 1,75 % (b/b, dirujuk sebagai senyawa hiptolida setelah melalui analisis dan kompilasi data-data spektroskopi. Hidroborasi terhadap senyawa hiptolida (yang diikuti hidrolisis menggunakan H2O2 dalam suasana basa menghasilkan senyawa alkohol turunannya, dengan persentase transformasi sebesar 28,9%, dapat ditunjukkan melalui data LCMS. Data spectrum IR menunjukkan adanya puncak pada 3600cm-1, memperkuat dugaan  adanya ikatan hidroksil menggantikan gugus

  3. Comparison of Separation of Seed Oil Triglycerides Containing Isomeric Conjugated Octadecatrienoic Acid Moieties by Reversed-Phase HPLC

    Directory of Open Access Journals (Sweden)

    Anh Van Nguyen

    2017-12-01

    Full Text Available Relative retention analysis and increment approach were applied for the comparison of triglycerides (TGs retention of a broad set of plant seed oils with isomeric conjugated octadecatrienoic acids (CLnA by reversed-phase HPLC for “propanol-2-acetonitrile” mobile phases and Kromasil 100-5C18 stationary phase with diode array detection (DAD and mass spectrometric (MS detection. The subjects of investigation were TGs of seed oils: Calendula officinalis, Catalpa ovata, Jacaranda mimosifolia, Centranthus ruber, Momordica charantia, Trichosanthes anguina, Punica granatum, Thladiantha dubia, Valeriana officinalis, and Vernicia montana. It was found that a sequence of elution of TGs of the same types is the same without any inversions for full range of mobile phase compositions: punicic (C18:39Z11E13Z < jacaric (C18:38Z10E12Z < catalpic (C18:39E11E13Z < α-eleostearic (C18:39Z11E13E < calendic (C18:38E10E12Z < β-eleostearic (C18:39E11E13E < all-E calendic (C18:38E10E12E acids. TGs and fatty acid compositions were calculated for all oil samples. Regularities of solute retentions as a function of isomeric conjugated octadecatrienoic acid moiety structure are discussed. Thus, it was proven that it is possible to differentiate TGs of complex composition with moieties of all natural CLnA by retention control accomplished by electronic spectra comparison, even though there are only three types of electronic-vibration spectra for seven isomeric CLnA.

  4. Quality control of PET radiopharmaceuticals using HPLC with electrochemical detection.

    Science.gov (United States)

    Nakao, Ryuji; Furutuka, Kenji; Yamaguchi, Masatoshi; Suzuki, Kazutoshi

    2006-04-01

    The usefulness of high-performance liquid chromatography with electrochemical detection (HPLC/ECD) in the quality control of positron emission tomography (PET) radiopharmaceuticals was evaluated for a number of substances. Chromatographic separation was performed using a reversed phase column and acetonitrile or 20 mM sodium phosphate buffer as the mobile phase. The effluent from the column was introduced into an electrochemical detector equipped with a glassy carbon electrode versus Ag/AgCl electrode operated in the direct current mode. In 19 of 21 PET radiopharmaceuticals studied, the compounds and corresponding precursors used in the synthesis of the radiopharmaceuticals could be successfully detected by the HPLC/ECD method. For 17 compounds with electroactive functional groups, such as aliphatic amines, phenols and aromatic amines, the detection limits were ppb levels for a 20-mul injection volume; this was significantly better compared with ultraviolet (UV) detection. This method could be applied to the analysis of [11C]MP4A, useful PET radiopharmaceutical for measuring acetylcholinesterase activity in the brain with no available UV absorbance.

  5. Variable-gradient generator for micro- and nano-HPLC.

    Science.gov (United States)

    Cappiello, Achille; Famiglini, Giorgio; Fiorucci, Chiara; Mangani, Filippo; Palma, Pierangela; Siviero, Antonella

    2003-03-01

    A new, simple device generates accurate nano- and microflow rate gradients from any conventional HPLC system. The core of the new device is represented by an electric-actuated, computer-controlled, multiposition HPLC valve. The valve hosts six reservoirs for as many different mobile-phase compositions of increasing strength. A low flow rate stream pushes the weakest solvent through the column as long as required and at the desired flow rate, until the chromatographic run is started. From this time on, the electric actuation allows one to select which reservoir will be on-line with the column and for how long, thus generating a specific solvent gradient, through a sequence of controlled segments of precise mobile-phase composition. This permits one not only to exactly reproduce the programmed slope but also to achieve different gradient shapes (i.e., linear, convex, concave) for different separation needs. The new device has proven to be reliable and reproducible even at the lowest flow rate tested (250 nL x min(-1)) and in different chromatographic conditions.

  6. Trace anti-inflammatory β-carboline alkaloid identified in Arenaria kansuensis by two-dimensional chromatography coupled with UniElut C18AEX based solid-phase extraction re-enrichment technology.

    Science.gov (United States)

    Cui, Yulei; Shen, Na; Wang, Shuo; Mei, Lijuan; Liu, Zenggen; Dang, Jun; Tao, Yanduo

    2017-11-15

    Traditional Chinese medicine is important for discovery of drug precursors. However, information about trace chemical composition of them is very limited due to the lack of appropriate enrichment and chromatographic purification methods In our work, A. kansuensis was taken as an example, a novel two-dimensional reversed-phase/hydrophilic interaction liquid chromatography coupled with UniElut C18AEX solid-phase extraction re-enrichment method based on anti-inflammatory bioactivity-guided assay was developed for gathering and purifying trace β-carboline alkaloids with high purity from the ethyl acetate extract of A. kansuensis. Extraction with ethyl acetate as the first enrichment method, then, a UniElut C18AEX column was employed to re-enrich trace fraction which was hardly detected by diode array detector during high performance liquid chromatography analysis, eighteen grams of UniElut C18AEX was used as sorbent material to pack a 60mL pipette tip for the extraction of β-carboline alkaloids from 100mL of ethyl acetate sample. The whole extraction process was finished in 10min, and the volume of eluent used was only 120mL. The enriching fraction (100mg) was used for the following two-dimensional purification. First-dimensional preparation was carried on a RP-Megress-C18 prep column, and four anti-inflammatory fractions were obtained from the 100mg re-enriching sample with a recovery of 66.9%. A HILIC-XAmide prep column was selected for the second dimensional preparation. Finally, two pair of analogue β-carboline alkaloids and one other β-carboline alkaloid were purified from A. kansuensis. The purity of the isolated compounds was ≫>98%, which indicated that the method was efficient to re-enrich and manufacture single trace β-carboline alkaloids with high purity from A. kansuensis. Additionally, this method showed great potential to serve as a good example for the purification and enrichment of analogue structure anti-inflammation carboline alkaloids from

  7. A simple HPLC method for determination of permethrin residues in wine.

    Science.gov (United States)

    Shishovska, Maja A; Trajkovska, Vera P; Stefova, Marina T

    2010-10-01

    An isocratic High Performance Liquid Chromatographic (HPLC) method was optimized for 3-phenoxybenzyl (1RS)-cis-trans-3-(2,2-dichlorovinyl)-2,2-dimethyl-cyclopropanecarboxylate (permethrin) residues identification and quantification in wine matrix. Analytical reverse phase (RP) C-18 column was used (25 cm × 4 mm i.d., 5 μ m) with mobile phase consisting of acetonitrile and water in ratio 70 %/30 % (v v(-1)), flow-rate 2.0 mL min(-1), UV-detection at 215 nm and controlled oven temperature at 25°C. The peaks of isomers were identified with the retention times as compared to standard cis-/trans- mixture and confirmed with characteristic spectra using photodiode array detector. Under these conditions, permethrin isomers were well separated with resolution 2.8 and no interference with the naturally present wine compounds was observed. The method was validated for linearity, precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ). Linear regression analysis data proved a good linear relationship (correlation coefficients, r(2), for cis- and trans-isomer are: 0.9995 and 0.9997, respectively) between response of the detector and concentration of permethrin isomers over a wide concentration range for both isomers (0.55 mg L(-1) -4.40 mg L(-1)). Experimental data showed mean recoveries between 93.95% and 96.58% with RSD values in range: 0.89% -3.69%. The effect of ethanol content in the solvent on permethrin isomers peak areas was also studied and 60% v v(-1) ethanol was found to be optimal for sample preparation. The method was successfully tested on 20 commercial wine samples from the market in which no permethrin was detected. Thus, it was proved that it is suitable for routine permethrin residues analysis. The proposed method is suitable for routine analysis because of the simple sample preparation, acceptable run-time, low cost and its applicability with conventional instruments.

  8. Analysis of trifluoroacetic acid and other short-chain perfluorinated acids (C2-C4) in precipitation by liquid chromatography-tandem mass spectrometry: comparison to patterns of long-chain perfluorinated acids (C5-C18).

    Science.gov (United States)

    Taniyasu, Sachi; Kannan, Kurunthachalam; Yeung, Leo W Y; Kwok, Karen Y; Lam, Paul K S; Yamashita, Nobuyoshi

    2008-07-07

    A method has been developed to measure 29 perfluorinated acids (PFAs) including short-chain perfluorocarboxylates (PFCAs) such as trifluoroacetic acid (TFA; C2) and long-chain PFCAs, perfluoroalkylsulfonates, fluorotelomer acids, and two perfluorooctylsulfonamides in water matrices. The method involves solid phase extraction (SPE) using a weak anion-exchange (WAX) cartridge, an ion-exchange high-performance liquid chromatography (HPLC) column separation, and tandem mass spectrometry (MS/MS) detection. To our knowledge, this is the first HPLC-MS/MS method to determine TFA in water at sub-ng L(-1) concentrations. The method is selective, simple, and robust, capable of measuring 29 PFAs in a single analysis, with overall recoveries of the target analytes ranging from 75% to 132%. The method was applied to the analysis of rainwater samples collected from two cities in Japan. TFA and several short-chain PFAs were the major compounds found in rainwater.

  9. Development of a new extraction technique and HPLC method for the analysis of non-psychoactive cannabinoids in fibre-type Cannabis sativa L. (hemp).

    Science.gov (United States)

    Brighenti, Virginia; Pellati, Federica; Steinbach, Marleen; Maran, Davide; Benvenuti, Stefania

    2017-09-05

    The present work was aimed at the development and validation of a new, efficient and reliable technique for the analysis of the main non-psychoactive cannabinoids in fibre-type Cannabis sativa L. (hemp) inflorescences belonging to different varieties. This study was designed to identify samples with a high content of bioactive compounds, with a view to underscoring the importance of quality control in derived products as well. Different extraction methods, including dynamic maceration (DM), ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE) and supercritical-fluid extraction (SFE) were applied and compared in order to obtain a high yield of the target analytes from hemp. Dynamic maceration for 45min with ethanol (EtOH) at room temperature proved to be the most suitable technique for the extraction of cannabinoids in hemp samples. The analysis of the target analytes in hemp extracts was carried out by developing a new reversed-phase high-performance liquid chromatography (HPLC) method coupled with diode array (UV/DAD) and electrospray ionization-mass spectrometry (ESI-MS) detection, by using an ion trap mass analyser. An Ascentis Express C18 column (150mm×3.0mm I.D., 2.7μm) was selected for the HPLC analysis, with a mobile phase composed of 0.1% formic acid in both water and acetonitrile, under gradient elution. The application of the fused-core technology allowed us to obtain a significant improvement of the HPLC performance compared with that of conventional particulate stationary phases, with a shorter analysis time and a remarkable reduction of solvent usage. The analytical method optimized in this study was fully validated to show compliance with international requirements. Furthermore, it was applied to the characterization of nine hemp samples and six hemp-based pharmaceutical products. As such, it was demonstrated to be a very useful tool for the analysis of cannabinoids in both the plant material and its derivatives for

  10. Using a simple HPLC approach to identify the enzymatic products of UTL-5g, a small molecule TNF-α inhibitor, from porcine esterase and from rabbit esterase.

    Science.gov (United States)

    Swartz, Kenneth; Zhang, Yiguan; Valeriote, Frederick; Chen, Ben; Shaw, Jiajiu

    2013-12-01

    UTL-5g is a novel small-molecule chemoprotector that lowers hepatotoxicity, nephrotoxicity, and myelotoxicity induced by cisplatin through TNF-α inhibition among other factors. As a prelude to investigating the metabolites of UTL-5g, we set out to identify the enzymatic products of UTL-5g under the treatment of both porcine liver esterase (PLE) and rabbit liver esterase (RLE). First, a number of mixtures made by UTL-5g and PLE were incubated at 25°C. At predetermined time points, individual samples were quenched by acetonitrile, vortexed, and centrifuged. The supernatants were then analyzed by reversed-phase HPLC (using a C18 column). The retention times and UV/vis spectra of individual peaks were compared to those of UTL-5g and its two postulated enzymatic products; thus the enzymatic products of UTL-5g were tentatively identified. Secondly, a different HPLC method (providing different retentions times) was used to cross-check and to confirm the identities of the two enzymatic products. Based on the observations, it was concluded that under the treatment of PLE, the major enzymatic products of UTL-5g were 5-methyliosxazole-3-carboxylic acid (ISOX) and 2,4-dichloroaniline (DCA). Treatment of UTL-5g by RLE also provided the same enzymatic products of UTL-5g from esterase. These results indicate that the peptide bond in UTL-5g was cleaved by PLE/RLE. Michaelis-Menten kinetics showed that the Km values of UTL-5g were 2.07mM with PLE and 0.37mM with RLE indicating that UTL-5g had a higher affinity with RLE. In summary, by a simple HPLC approach, we have concluded that the peptide bond in UTL-5g was cleaved by esterase from either porcine liver or rabbit liver in vitro and afforded DCA (at a mole ratio of 1:1) and ISOX. However, further studies are needed in order to determine whether UTL-5g is metabolized by microsomal enzymes to produce ISOX and DCA. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. 40 CFR Appendix 8 to Subpart A of... - Reference C16-C18 Internal Olefin Drilling Fluid Formulation

    Science.gov (United States)

    2010-07-01

    ... Drilling Fluid Formulation 8 Appendix 8 to Subpart A of Part 435 Protection of Environment ENVIRONMENTAL... Internal Olefin Drilling Fluid Formulation The reference C16-C18 internal olefin drilling fluid used to determine the drilling fluid sediment toxicity ratio and compliance with the BAT sediment toxicity discharge...

  12. Comparative evaluation of manganese peroxidase- and Mn(III)-initiated peroxidation of C18 unsaturated fatty acids by different methods

    Science.gov (United States)

    Alexander N. Kapich; Tatyana V. Korneichik; Kenneth E. Hammel; Annele Hatakka

    2011-01-01

    The peroxidation of C18 unsaturated fatty acids by fungal manganese peroxidase (MnP)/Mn(II) and by chelated Mn(III) was studied with application of three different methods: by monitoring oxygen consumption, by measuring conjugated dienes and by thiobarbituric acid-reactive substances (TBARS) formation. All tested polyunsaturated fatty acids (PUFAs) were oxidized by MnP...

  13. Enantioselective synthesis of the C18-C25 segment of lasonolide A by an oxonia-cope prins cascade.

    Science.gov (United States)

    Dalgard, Jackline E; Rychnovsky, Scott D

    2005-04-14

    [reaction: see text] A 2-oxonia-Cope Prins cascade was developed that led to a facile and stereoselective synthesis of the C18-C25 segment of lasonolide A. The strategy nicely handles the introduction of the quaternary center in the tetrahydropyran ring, and all of the stereogenic centers in the product arise from a single stereocenter introduced in a catalytic enantioselective reaction.

  14. The kinetics of the ordering of 13C-18O bonds in calcite and apatite

    Science.gov (United States)

    Stolper, D. A.; Halevy, I.; Eiler, J. M.

    2011-12-01

    Eiler and Schauble (2004) showed that the isotopes of C and O are not randomly distributed within single phases such as CO2 gas and carbonates, and in particular, that heavy isotopes of C and O tend to bond preferentially (clump) at lower temperatures. Consequently, the measurement of the deviation from a random distribution of C and O isotope distributions in a single phase can be used as a thermometer. As with other geothermometers based on homogeneous or heterogeneous equilibria, the clumped-isotope thermometer is susceptible to resetting (e.g., if the phase is reheated or experiences slow cooling). Thus, clumped-isotope "temperatures" of phases that have experienced complex thermal histories may, in fact, be closure temperatures, the interpretation of which requires quantification of the kinetics of redistribution of C and O isotopes as a function of temperature. These kinetics have received increasing attention (Dennis and Schrag, 2010; Passey 2010), and are likely to be critical for understanding clumped-isotope temperatures of samples that have been buried for long periods of time. To better constrain these kinetics we performed experiments on natural optical calcite from Mexico and carbonate-bearing apatite from the Siilinjarvi carbonatite (Finland). For each experiment, multiple single crystal grains (~2 mm in diameter) of calcite or apatite were loaded in open Pt capsules, pressurized with Ar gas, and held at 400-700 °C, 550 bars using a rapid quench TZM apparatus for 5 min to 520 hrs. After quenching, 13C-18O clumping was measured in the samples; the change from the initial Δ47 with time for each phase at each temperature was fit to simple mechanistic models of isotope exchange between sites in these phases. One conclusion of the experimental study is that resetting the internal ordering of carbonate groups proceeds more rapidly in calcites than in apatites. For example, heating apatite at 400 °C results in no change in clumping over a 24 hr period

  15. Estimation of Lamotrigine by RP-HPLC Method

    Directory of Open Access Journals (Sweden)

    D. Anantha Kumar

    2010-01-01

    Full Text Available A rapid and reproducible reverse phase high performance liquid chromatographic method has been developed for the estimation of lamotrigine in its pure form as well as in pharmaceutical dosage forms. Chromatography was carried out on a Luna C18 column using a mixture of potassium dihydrogen phosphate buffer (pH 7.3 and methanol in a ratio of 60:40 v/v as the mobile phase at a flow rate of 1.0 mL/min. The detection was done at 305 nm. The retention time of the drug was 6.1 min. The method produced linear responses in the concentration range of 10 to 70 μg/mL of lamotrigine. The method was found to be reproducible for analysis of the drug in tablet dosage forms.

  16. Purification and characterization of a novel antithrombotic peptide from Scolopendra subspinipes mutilans

    National Research Council Canada - National Science Library

    Kong, Yi; Huang, Shi-Long; Shao, Yu; Li, Shuai; Wei, Ji-Fu

    2013-01-01

    .... A novel antithrombotic peptide was isolated from Scolopendra subspinipes mutilans using a combination of ultrafiltration, Sephadex G-50 column, Source 15Q anion exchange column and RP-HPLC C18 column...

  17. Ideal versus real automated twin column recycling chromatography process.

    Science.gov (United States)

    Gritti, Fabrice; Leal, Mike; McDonald, Thomas; Gilar, Martin

    2017-07-28

    The full baseline separation of two compounds (selectivity factors αchromatography is used to confirm that the speed-resolution performance of the TCRSP is intrinsically superior to that of the single-column process. This advantage is illustrated in this work by developing an automated TCRSP for the challenging separation of two polycyclic aromatic hydrocarbon (PAH) isomers (benzo[a]anthracene and chrysene) in the reversed-phase retention mode at pressure smaller than 5000psi. The columns used are the 3.0mm×150mm column packed with 3.5μm XBridge BEH-C18 material (α=1.010) and the 3.0mm or 4.6mm×150mm columns packed with the same 3.5μm XSelect HSST3 material (α=1.025). The isocratic mobile phase is an acetonitrile-water mixture (80/20, v/v). Remarkably, significant differences are observed between the predicted retention times and efficiencies of the ideal TCRSP (given by the number of cycles multiplied by the retention time and efficiency of one column) and those of the real TCRSP. The fundamental explanation lies in the pressure-dependent retention of these PAHs or in the change of their partial molar volume as they are transferred from the mobile to the stationary phase. A revisited retention and efficiency model is then built to predict the actual performance of real TCRSPs. The experimental and calculated resolution data are found in very good agreement for a change, Δvm=-10cm3/mol, of the partial molar volume of the two PAH isomers upon transfer from the acetonitrile-water eluent mixture to the silica-C18 stationary phase. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Solid-Phase Extraction and Reverse-Phase HPLC: Application to Study the Urinary Excretion Pattern of Benzophenone-3 and its Metabolite 2,4-Dihydroxybenzophenone in Human Urine

    Directory of Open Access Journals (Sweden)

    Anne Farbrot

    2008-01-01

    Full Text Available Background: Benzophenone-3 (BZ-3 is a common ultraviolet (UV absorbing compound in sunscreens. It is the most bioavailable species of all UV-absorbing compounds after topical application and can be found in plasma and urine.Objectives: The aim of this study was to develop a reverse-phase high performance liquid chromatography (HPLC method for determining the amounts BZ-3 and its metabolite 2,4-dihydroxybenzophenone (DHB in human urine. The method had to be suitable for handling a large number of samples. It also had to be rapid and simple, but still sensitive, accurate and reproducible. The assay was applied to study the urinary excretion pattern after repeated whole-body applications of a commercial sunscreen, containing 4% BZ-3, to 25 healthy volunteers.Methods: Each sample was analyzed with regard to both conjugated/non-conjugated BZ-3 and conjugated/non-conjugated DHB, since both BZ-3 and DHB are extensively conjugated in the body. Solid-phase extraction (SPE with C8 columns was followed by reverse-phase HPLC. For separation a Genesis C18 column was used with an acethonitrile-water mobile phase and the UV-detector was set at 287 nm.Results: The assay was linear r2 > 0.99, with detection limits for BZ-3 and DHB of 0.01 μmol L−1 and 0.16 μmol L−1 respectively. Relative standard deviation (RSD was less than 10% for BZ-3 and less than 13% for DHB.The excretion pattern varied among the human volunteers; we discerned different patterns among the individuals.Conclusions: The reverse-phase HPLC assay and extraction procedures developed are suitable for use when a large number of samples need to be analyzed and the method fulfilled our objectives. The differences in excretion pattern may be due to differences in enzyme activity but further studies, especially about genetic polymorphism, need to be performed to verify this finding.

  19. Sensitive mercury speciation by reversed-phase column high-performance liquid chromatography with UV-visible detection after solid-phase extraction using 6-mercaptopurine and dithizone.

    Science.gov (United States)

    Hashemi-Moghaddam, Hamid; Saber-Tehrani, Mohamad

    2008-01-01

    A highly selective and sensitive method was developed for preconcentration of inorganic and organic mercury compounds followed by reversed-phase column high-performance liquid chromatography (RP-HPLC) with UV-visible detection. The method was based on the reaction of mercury with 6-mercaptopurine and solid-phase extraction (SPE) of the complex on an octadecylsilane (C18) cartridge. The complex was then treated with ammoniacal dithizone solution, and the complexes of inorganic and organic mercury with dithizone were eluted by methanol. The speciation analysis of methylmercury (MeHg), phenylmercury (PhHg), and inorganic Hg (II) was carried out by RP-HPLC. Some experimental variables that influence the SPE and derivatization, such as pH, chelating and derivatizing agent concentration, and surfactant addition, were investigated. The calibration graphs of MeHg, PhHg, and Hg (II) were linear [correlation coefficient (r) > 0.999] from the detection limits (0.12, 0.16, and 0.14 ng) to 8.5, 6.0, and 6.7 ng Hg, respectively. By applying the SPE procedure, a 100-fold concentration of the sample was obtained. The procedure was applied to sea water and tuna fish samples. The method's accuracy was investigated by using tuna fish certified reference material BCR 464 and by spiking the samples with different amounts of MeHg, PhHg, and Hg (II). The average recoveries of MeHg, PhHg, and Hg (II) from spiked samples (0.1-2.0 microg/L Hg) were 96 +/- 4, 98 +/- 3, and 104 +/- 4%, respectively.

  20. Columns in Clay

    Science.gov (United States)

    Leenhouts, Robin

    2010-01-01

    This article describes a clay project for students studying Greece and Rome. It provides a wonderful way to learn slab construction techniques by making small clay column capitols. With this lesson, students learn architectural vocabulary and history, understand the importance of classical architectural forms and their influence on today's…

  1. Practical column design guide

    CERN Document Server

    Nitsche, M

    2017-01-01

    This book highlights the aspects that need to be considered when designing distillation columns in practice. It discusses the influencing parameters as well as the equations governing them, and presents several numerical examples. The book is intended both for experienced designers and for those who are new to the subject.

  2. Improvement in HPLC separation of acetic acid and levulinic acid in the profiling of biomass hydrolysate.

    Science.gov (United States)

    Xie, Rui; Tu, Maobing; Wu, Yonnie; Adhikari, Sushil

    2011-04-01

    5-Hydroxymethylfurfural (HMF) and furfural could be separated by the Aminex HPX-87H column chromatography, however, the separation and quantification of acetic acid and levulinic acid in biomass hydrolysate have been difficult with this method. In present study, the HPLC separation of acetic acid and levulinic acid on Aminex HPX-87H column has been investigated by varying column temperature, flow rate, and sulfuric acid content in the mobile phase. The column temperature was found critical in resolving acetic acid and levulinic acid. The resolution for two acids increased dramatically from 0.42 to 1.86 when the column temperature was lowered from 60 to 30 °C. So did the capacity factors for levulinic acid that was increased from 1.20 to 1.44 as the column temperature dropped. The optimum column temperature for the separation was found at 45 °C. Variation in flow rate and sulfuric acid concentration improved not as much as the column temperature did. Published by Elsevier Ltd.

  3. The quantitative impact of the mesopore size on the mass transfer mechanism of the new 1.9μm fully porous Titan-C18 particles. I: analysis of small molecules.

    Science.gov (United States)

    Gritti, Fabrice; Guiochon, Georges

    2015-03-06

    Previous data have shown that could deliver a minimum reduced plate height as small as 1.7. Additionally, the reduction of the mesopore size after C18 derivatization and the subsequent restriction for sample diffusivity across the Titan-C18 particles were found responsible for the unusually small value of the experimental optimum reduced velocity (5 versus 10 for conventional particles) and for the large values of the average reduced solid-liquid mass transfer resistance coefficients (0.032 versus 0.016) measured for a series of seven n-alkanophenones. The improvements in column efficiency made by increasing the average mesopore size of the Titan silica from 80 to 120Å are investigated from a quantitative viewpoint based on the accurate measurements of the reduced coefficients (longitudinal diffusion, trans-particle mass transfer resistance, and eddy diffusion) and of the intra-particle diffusivity, pore, and surface diffusion for the same series of n-alkanophenone compounds. The experimental results reveal an increase (from 0% to 30%) of the longitudinal diffusion coefficients for the same sample concentration distribution (from 0.25 to 4) between the particle volume and the external volume of the column, a 40% increase of the intra-particle diffusivity for the same sample distribution (from 1 to 7) between the particle skeleton volume and the bulk phase, and a 15-30% decrease of the solid-liquid mass transfer coefficient for the n-alkanophenone compounds. Pore and surface diffusion are increased by 60% and 20%, respectively. The eddy dispersion term and the maximum column efficiency (295000plates/m) remain virtually unchanged. The rate of increase of the total plate height with increasing the chromatographic speed is reduced by 20% and it is mostly controlled (75% and 70% for 80 and 120Å pore size) by the flow rate dependence of the eddy dispersion term. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. HPLC method development for the simultaneous analysis of amlodipine and valsartan in combined dosage forms and in vitro dissolution studies

    Directory of Open Access Journals (Sweden)

    Mustafa Çelebier

    2010-12-01

    Full Text Available A simple, rapid and reproducible HPLC method was developed for the simultaneous determination of amlodipine and valsartan in their combined dosage forms, and for drug dissolution studies. A C18 column (ODS 2, 10 μm, 200 x 4.6 mm and a mobile phase of phosphate buffer (pH 3.6 , 0.01 mol L-1:acetonitrile: methanol (46:44:10 v/v/v mixture were used for separation and quantification. Analyses were run at a flow-rate of 1 mL min-1 and at ambient temperature. The injection volume was 20 μL and the ultraviolet detector was set at 240 nm. Under these conditions, amlodipine and valsartan were eluted at 7.1 min and 3.4 min, respectively. Total run time was shorter than 9 min. The developed method was validated according to the literature and found to be linear within the range 0.1 - 50 μg mL-1 for amlodipine, and 0.05 - 50 μg mL-1 for valsartan. The developed method was applied successfully for quality control assay of amlodipine and valsartan in their combination drug product and in vitro dissolution studies.Desenvolveu-se método de HPLC rápido e reprodutível para a determinação simultânea de anlodipino e valsartana em suas formas de associação e para os estudos de dissolução dos fármacos. Utilizaram-se coluna C18 (ODS 2, 10 μm, 200 x 4,6 mm e fase móvel tampão fosfato (pH 3,6, 0,01 mol L-1:acetonitrila: metanol para a separação e a quantificação. As análises foram efetuadas com velocidade de fluxo de 1 mL min-1 e à temparatura ambiente O volume de injeção foi de 20 μL e utilizou-se detector de ultravioleta a 240 nm. Sob essas condições, anlodipino e valsartana foram eluídas a 7,1 min e 3,4 min, respectivamente. O tempo total de corrida foi menor que 9 min. O método desenvolvido foi validado de acordo com a literatura e se mostrou linear na faixa de 0,1-50 μg mL-1 para anlodipino e de 0,05-50 μg mL-1 para valsartana. O método desenvolvido foi aplicado com sucesso para ensaios de controle de qualidade de associações de

  5. Validation of a rapid and sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay for the simultaneous determination of existing and new antiretroviral compounds.

    Science.gov (United States)

    Else, Laura; Watson, Victoria; Tjia, John; Hughes, Andrew; Siccardi, Marco; Khoo, Saye; Back, David

    2010-06-01

    Clinical pharmacokinetic studies of antiretrovirals require accurate and precise measurement of plasma drug concentrations. Here we describe a simple, fast and sensitive HPLC-MS/MS method for determination of the commonly used protease inhibitors (PI) amprenavir, atazanavir, darunavir, lopinavir, ritonavir, saquinavir and the non-nucleoside reverse transcriptase inhibitor (NNRTI) nevirapine, as well as the more recent antiretrovirals, the CCR5 antagonist maraviroc and the "second generation" NNRTI etravirine and rilpivirine. An internal standard (quinoxalone; QX) was added to plasma aliquots (100 microl) prior to protein precipitation with acetonitrile (500 microl) followed by centrifugation and addition of 0.05% formic acid (200 microl) to the supernatant. Chromatographic separation was achieved using a gradient (acetonitrile and 0.05% formic acid) mobile phase on a reverse-phase C18 column. Detection was via selective reaction monitoring (SRM) operating in positive ionization mode on a triple-quadrupole mass spectrometer. All compounds eluted within a 5 min run time. Calibration curves were validated over concentration ranges reflecting therapeutic concentrations observed in HIV-infected patients from pharmacokinetic data reported in the literature. Correlation coefficients (r2) exceeded 0.998. Inter- and intra-assay variation ranged between 1% and 10% and % recovery exceeded 90% for all analytes. The method described is being successfully applied to measure plasma antiretroviral concentrations from samples obtained from clinical pharmacokinetic studies. Copyright 2010 Elsevier B.V. All rights reserved.

  6. Preparation of bi-layer-polymer coated silica using atom transfer radical polymerization and its application as restricted access phase for HPLC separation of hydrophobic molecules in biological fluids.

    Science.gov (United States)

    Xu, Dan; Dong, Xiangchao; Zhang, Haiyan; Wang, Huaisong; Jiang, Ping; Zhang, Min

    2012-07-01

    A novel restricted access material was prepared by surface initiated atom transfer radical polymerization. The bi-layer-polymer structures were created on the surface of silica layer-by-layer. The inner layer was composed of poly(styrene-co-divinylbenzene), which was grafted first for binding small molecules based on hydrophobic and π-π interactions. The poly(styrene-co-divinylbenzene) bonded silica has good selectivity for aromatic hydrocarbons. It also has hydrophobicity and column efficiency similar to a C(18) bonded silica. The material has shown good ability of protein exclusion after grafting hydrophilic polymer on the external surface while its hydrophobicity and selectivity do not have obvious change. It demonstrated that the material is still qualified for hydrophobic extraction. In the study, the relations between the polymer structures and chromatographic properties of the materials were investigated. The synthetic conditions were optimized. The results have shown that the material prepared in the study has application potential in the HPLC analysis of hydrophobic molecules from biological samples by direct injection. It demonstrated that atom transfer radical polymerization can be used as a method in the preparation of restricted access material. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Development and Validation of a New HPLC Method for the Simultaneous Determination of Paracetamol, Ascorbic Acid, and Pseudoephedrine HCl in their Co-formulated Tablets. Application to in vitro Dissolution Testing.

    Science.gov (United States)

    Ibrahim, Fawzia; El-Enany, Nahed; El-Shaheny, Rania N; Mikhail, Ibraam E

    2015-01-01

    The first HPLC method was developed for the simultaneous determination of paracetamol (PC), ascorbic acid (AA), and pseudoephedrine HCl (PE) in their co-formulated tablets. Separation was achieved on a C18 column in 5 min using a mobile phase composed of methanol-0.05 M phosphate buffer (35:65, v/v) at pH 2.5 with UV detection at 220 nm. Linear calibration curves were constructed over concentration ranges of 1.0 - 50.0, 3.0 - 60.0 and 3.0 - 80.0 μg mL(-1) for PC, AA, and PE, respectively. The method was validated and applied for the simultaneous determination of these drugs in their tablets with average % recoveries of 101.17 ± 0.67, 98.34 ± 0.77, and 98.95 ± 1.11%, for PC, AA, and PE, respectively. The proposed method was also used to construct in vitro dissolution profiles of the co-formulated tablets containing the three drugs.

  8. Simultaneous investigation of sesquiterpenes, pyrrolizidine alkaloids and N-oxides in Butterbur (Petasites hybridus) with an offline 2D-combination of HPLC-UV and LC-MMI-ToF-MS.

    Science.gov (United States)

    Aydın, Ahmet Alper; Letzel, Thomas

    2013-11-01

    With the worldwide rapid increasing interest in the use of natural products as dietary supplements, medical remedies and functional foods, it has been accepted that omitting the plant constituents with potential adverse effects was a huge fault of the past. Several countries developed regulations to limit the consumption of such products in the markets. Among these natural products, butterbur (Petasites) has been used for years as herbal supplement for its anti-spasmodic and anti-inflammatory effects. However, its hepatotoxic alkaloid content limits the direct usage. In this study, investigation of sesquiterpenes and pyrrolizidine alkaloids (PAs) together with their N-oxide forms has been conducted with an offline 2D-combination using high-performance liquid chromatography with UV detection (HPLC-UV) and liquid chromatography - multi mode ionization - time-of-flight mass spectrometry (LC-MMI-ToF-MS) for plant screening. The content has been qualitatively investigated to provide information on the constituents of the plant rhizomes extracted using ethanol. Besides the reported hepatotoxic and medically bio-active plant constituents, a strategy has been suggested for estimating the retention order and retention times with respect to calculated logD (distribution coefficient) and hydrophobicity distributions on C18 reversed-phase column when all standard compounds are not available in laboratory. In this sense, the influence of calculated logD and hydrophobicity distributions on retention time has been clarified via available PA and PA-N-oxide standards. The ethanolic extract of Petasites hybridus has been used for examination of the strategy in a real-sample model. Additionally, the advantages of the developed HPLC-UV and LC-MMI-ToF-MS combination have been discussed with respect to the presented results. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Determination of theophylline by HPLC and GC-IDMS, the effect of chemically similar xanthine derivatives on the specificity of the method and the possibility of paracetamol as interfering substance.

    Science.gov (United States)

    Kress, Michael; Meissner, Dieane; Kaiser, Patricia; Hanke, Rainer; Wood, William Graham

    2002-01-01

    The aim of this study was to develop and compare high-performance liquid chromatography (HPLC) and gas chromatography coupled with isotope dilution-mass spectrometry (GC-IDMS) methods with a common extraction procedure for the determination of substituted xanthines in biological matrices such as serum and urine. For HPLC both isocratic and gradient methods were evaluated. Difficulties occurred in separation of all 6 xanthines of interest - uric acid, theobromine, theophylline, paraxanthine, caffeine and 1,3-dimethyl-7-(2-hydroxyethyl) xanthine as internal standard. In addition, paracetamol was seen to interfere at higher concentrations, which meant that a system had to be developed to separate all 7 components of interest. The final solution chosen consisted of precipitation of serum samples with 6 mol/l trichloroacetic acid followed by neutralisation with 3 mol/l KOH and chromatography on a 150 x 4.6 mm Nautilus C-18 column (Macherey & Nagel) using an isocratic elution consisting of 0.02 mol/l acetate-phosphate buffer, pH 3.0 containing 9.6% v/v acetonitrile and monitoring at 273 +/- 7 nm. Comparisons with GC-IDMS and FPIA were acceptable. Run times of 10 minutes were possible. An additional "safe time" of 5 minutes was allowed to elute any substances with similar absorption maxima which were sometimes present in commercial control sera. Precision of the method was 1.64% (intra-assay) and 2.87% (inter-assay) at 4.1 mg/l and 1.51% respectively 2.15% at 25 mg/l including extraction and measurement steps. Recovery was between 86 and 101% between 1.25 and 100 mg/l and peak time deviations for all 7 components between 0.07% and 0.34% (coefficient of variation) in 7 consecutive measurements.

  10. HPLC determination of certain flavonoids and terpene lactones in selected Ginkgo biloba L. phytopharmaceuticals.

    Science.gov (United States)

    Mesbah, Mostafa K; Khalifa, Sherief I; El-Gindy, Alaa; Tawfik, Kamilia A

    2005-01-01

    The biologically active secondary metabolites of Ginkgo biloba extract EGb 761 in phytopharmaceuticals were analyzed using two simple, rapid, accurate and sensitive HPLC methods. The proposed methods were successfully applied in the determination of terpenes and flavonoids in four phytopharmaceutical preparations selected from the Egyptian market. The terpenes; ginkgolide A, ginkgolide B, and bilobalide were analyzed using RP 18 column with a mobile phase consisting of water/methanol/isopropanol (72.5:17.5:10, v/v) at a flow rate of 1 ml min-1 and UV detection at 220 nm. The flavonoids; quercetin and kaempferol were analyzed using RP 18 column in a step gradient elution with acetonitrile and water at pH 3.3 and flow rate of 1.5 ml min-1 with UV detection at 370 nm. The two HPLC methods were completely validated.

  11. Nine Words - Nine Columns

    DEFF Research Database (Denmark)

    Trempe Jr., Robert B.; Buthke, Jan

    2016-01-01

    This book records the efforts of a one-week joint workshop between Master students from Studio 2B of Arkitektskolen Aarhus and Master students from the Harbin Institute of Technology in Harbin, China. The workshop employed nine action words to instigate team-based investigation into the effects o...... as formwork for the shaping of wood veneer. The resulting columns ‘wear’ every aspect of this design pipeline process and display the power of process towards an architectural resolution....

  12. Brief on -Hyphenated Methods of HPLC for Determining the Presence of Solutes.

    Directory of Open Access Journals (Sweden)

    Mohamad Taleuzzaman

    2017-02-01

    Full Text Available HPLC is the tool in liquid chromatography is unique because of particle size, smaller particle in the stationary phase, increase efficiency of a separation. However, if the particles are made smaller, capillary action increases and it becomes more difficult to drain the column under gravity. For quantitative analysis different types of detector is used in conjunction with HPLC which give precise and accurate result and it is apply according to the nature of the substance. Various types of detectors used in HPLC are mass spectrometry, infrared spectroscopy, visible spectroscopy, ultraviolet spectroscopy, fluorescence spectroscopy, nuclear magnetic resonance, conductivity measurement, and refractive index measurement. Each detector has its assets, limitations and sample types for which it is most effective. The recent development of the so-called hyphenated techniques has improved the ability to separate and identify multiple entities within a mixture.

  13. [Separation and preparation of indole alkaloids in Lycorma delicatula White. by HPLC].

    Science.gov (United States)

    Xue, G; Yuan, S

    1996-09-01

    A HPLC method for separating and preparing indole alkaloids is described. HPLC conditions for analysis: BIO-RAD series 700 HPLC, model 700 data station, UV: model 1749 UV-VIS monitor, column: BIO-RAD Hi-pore RP318, 250 mm x 10 mm, mobile phase: 80% methanol-H2O(gradient), flow rate: 1.5 ml/min, detection wavelength: 254 nm. On the basis of spectral (1HNMR, 13CNMR, H-H COSY, MS, DEPT) and chemical evidence, the structures of two compounds were elucidated as beta-yohimbine (yohimban-16-carboxylic acid-17-hydroxy methylester (3 alpha, 16 alpha, 17 beta)) and ajmalicine (oxayohimban-16-carboxylic acid-16,17-didehydro-19-ethyl methyl ester (19 alpha)).

  14. Validated Reverse Phase HPLC Method for the Determination of Impurities in Etoricoxib

    Directory of Open Access Journals (Sweden)

    S. Venugopal

    2011-01-01

    Full Text Available This paper describes the development of reverse phase HPLC method for etoricoxib in the presence of impurities and degradation products generated from the forced degradation studies. The drug substance was subjected to stress conditions of hydrolysis, oxidation, photolysis and thermal degradation. The degradation of etoricoxib was observed under base and oxidation environment. The drug was found stable in other stress conditions studied. Successful separation of the drug from the process related impurities and degradation products were achieved on zorbax SB CN (250 x 4.6 mm 5 μm particle size column using reverse phase HPLC method. The isocratic method employed with a mixture of buffer and acetonitrile in a ratio of 60:40 respectively. Disodium hydrogen orthophosphate (0.02 M is used as buffer and pH adjusted to 7.20 with 1 N sodium hydroxide solution. The HPLC method was developed and validated with respect to linearity, accuracy, precision, specificity and ruggedness.

  15. Impact of reversed phase column pairs in comprehensive two-dimensional liquid chromatography.

    Science.gov (United States)

    Allen, Robert C; Barnes, Brian B; Haidar Ahmad, Imad A; Filgueira, Marcelo R; Carr, Peter W

    2014-09-26

    A major issue in optimizing the resolving power of two-dimensional chromatographic separations is the choice of the two phases so as to maximize the distribution of the analytes over the separation space. In this work, we studied the choice of appropriate reversed phases to use in on-line comprehensive two-dimensional liquid chromatography (LC×LC). A set of four chemically different conventional bonded reversed phases was used in the first dimension. The second dimension column was either a conventional bonded C18 phase or a carbon-clad phase (CCP). The LC×LC chromatograms and contour plots were all rather similar indicating that the selectivities of the two phases were also similar regardless of the reverse phase column used in the first dimension. Further, the spatial coverage seen with all four first dimension stationary phases when paired with a second dimension C18 phase were low and the retention times were strongly correlated. However, when the C18 column was replaced with the CCP column much improved separations were observed with higher spatial coverages, greater orthogonalities and significant increases in the number of observed peaks. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. High Performance Liquid Chromatographic Analysis of Phytoplankton Pigments Using a C16-Amide Column

    Science.gov (United States)

    A reverse-phase high performance liquid chromatographic (RP-HPLC) method was developed to analyze in a single run, most polar and non-polar chlorophylls and carotenoids from marine phytoplankton. The method is based on a RP-C16-Amide column and a ternary gradient system consistin...

  17. Behavior of Columns During Earthquakes

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The behavior of columns during earthquakes is very important since column failures may lead to additional structural failures and result in total building collapses....

  18. An improved HPLC method for determination of colocynthin in colocynth

    Directory of Open Access Journals (Sweden)

    M. Shekarchi

    2015-10-01

    Full Text Available Background and objectives: Colocynthin is the major active secondary metabolite of colocynth, Citrullus colocynthis (L. Schrad, which has been used in traditional and ethno medicine of many countries.  It could be considered as an active marker for quality control of colocynth and its herbal products. Analysis and standardization of colocynth and its herbal preparations are a critical issue for their safe applications in phytotherapy and traditional medicine. In the present work, a simple and efficient sample preparation was developed and optimized through combination of matrix solid phase dispersion and ultrasonic assisted extraction. In addition, analytical reversed-phase HPLC method was optimized for analyzing the concentration of colocynthin in colocynth pulp. Methods: Powdered colocynth pulp was grinded with diatomaceous earth to obtain a homogenous mixture. The blend was mixed with methanol and extracted by sonication, followed by centrifugation and filtration. The analytical chromatographic separation was carried out using Luna C18 in isocratic elution with methanol: isopropanol: water: triflouroacetic acid (30:10:60:0.1 v/v. The method was validated as well.  Results: The validation parameters were determines as follows, linear range (r2 = 0.999, 75-500 μg/mL, precision (intra-day < 2.7%, inter-day = 4.4% and accuracy measured via determination of recovery (90-107%. The limit of detection and quantization were calculated 8.5 and 25.7 μg/mL, respectively. Conclusion: Regarding the relatively high content of colocynthin in colocynth pulp, the validated HPLC method could be applied for quality control of colocynth pulp used in Traditional Persian Medicine.

  19. Simple Determination of Plasma Ponatinib Concentration Using HPLC.

    Science.gov (United States)

    Yasu, Takeo; Momo, Kenji; Kobayashi, Shunsuke; Kuroda, Seiichirou; Tojo, Arinobu

    2018-02-01

    Ponatinib, a novel tyrosine kinase inhibitor marketed in 2016, is a key drug used for treating chronic myeloid leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia. This study aimed to develop a simple method for determining plasma ponatinib concentration. The analysis required extraction of a 400-µL sample of plasma and precipitation of proteins using an Oasis HLB cartridge. Ponatinib and bosutinib, which is used as an internal standard, were separated by HPLC using a mobile phase of acetonitrile : 0.037 mol/L KH2PO4 (pH 4.5) (39 : 61, v/v) on a Capcell Pack C18 MG II (25×4.6 mm) monitored at 250 nm, with a flow rate of 1.0 mL/min. This assay method was then used for determining plasma ponatinib concentration in a 42-year-old man treated with ponatinib at 15 mg/d. The calibration curve was found to be linear for the plasma concentration range of 5-250 ng/mL with a regression coefficient (r2) of 0.9999. The coefficients of intra-day and inter-day validation under these concentrations were 2.1-6.0 and 4.5-8.0%, respectively. The assay accuracy was -1.5-9.0%, and the recovery was greater than 86%. The plasma concentration of the patient at 2.5 and 3 h after 15 mg ponatinib administration was 43.6 and 49.3 ng/mL, respectively. This method of HPLC equipped with UV detection for determining plasma ponatinib concentration has several advantages, such as simplicity and applicability to routine therapeutic drug monitoring at hospital laboratories.

  20. Acquisition of HPLC-Mass Spectrometer

    Science.gov (United States)

    2015-08-18

    31-Jan-2015 Approved for Public Release; Distribution Unlimited Final Report: Acquisition of HPLC -Mass Spectrometer The views, opinions and/or findings...published in peer-reviewed journals: Final Report: Acquisition of HPLC -Mass Spectrometer Report Title The acquisition of the mass spectrometer has been a

  1. Core-shell column Tanaka characterization and additional tests using active pharmaceutical ingredients.

    Science.gov (United States)

    Ludvigsson, Jufang Wu; Karlsson, Anders; Kjellberg, Viktor

    2016-12-01

    In the last decade, core-shell particles have gained more and more attention in fast liquid chromatography separations due to their comparable performance with fully porous sub-2 μm particles and their significantly lower back pressure. Core-shell particles are made of a solid core surrounded by a shell of classic fully porous material. To embrace the developed core-shell column market and use these columns in pharmaceutical analytical applications, 17 core-shell C18 columns purchased from various vendors with various dimensions (50 mm × 2.1 mm to 100 mm × 3 mm) and particle sizes (1.6-2.7 μm) were characterized using Tanaka test protocols. Furthermore, four selected active pharmaceutical ingredients were chosen as test probes to investigate the batch to batch reproducibility for core-shell columns of particle size 2.6-2.7 μm, with dimension of 100 × 3 mm and columns of particle size 1.6 μm, with dimension 100 × 2.1 mm under isocratic elution. Columns of particle size 2.6-2.7 μm were also tested under gradient elution conditions. To confirm the claimed comparable efficiency of 2.6 μm core-shell particles as sub-2 μm fully porous particles, column performances of the selected core-shell columns were compared with BEH C18 , 1.7 μm, a fully porous column material as well. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Column: Every Last Byte

    Directory of Open Access Journals (Sweden)

    Simson Garfinkel

    2011-06-01

    Full Text Available Inheritance powder is the name that was given to poisons, especially arsenic, that were commonly used in the 17th and early 18th centuries to hasten the death of the elderly. For most of the 17th century, arsenic was deadly but undetectable, making it nearly impossible to prove that someone had been poisoned. The first arsenic test produced a gas—hardly something that a scientist could show to a judge. Faced with a growing epidemic of poisonings, doctors and chemists spent decades searching for something better.(see PDF for full column

  3. TLC-Densitometric and RP-HPLC Methods for Simultaneous Determination of Dexamethasone and Chlorpheniramine Maleate in the Presence of Methylparaben and Propylparaben.

    Science.gov (United States)

    Farid, Nehal F; Naguib, Ibrahim A; Moatamed, Radwa S; El Ghobashy, Mohamed R

    2017-01-01

    Validated simple, sensitive, and highly selective methods are applied for the quantitative determination of dexamethasone and chlorpheniramine maleate in the presence of their reported preservatives (methylparaben and propylparaben), whether in pure forms or in pharmaceutical formulation. TLC is the first method, in which dexamethasone, chlorpheniramine maleate, methylparaben, and propylparaben are separated on silica gel TLC F254 plates using hexane-acetone-ammonia (5.5 + 4.5 + 0.5, v/v/v) as the developing phase. Separated bands are scanned at 254 nm over a concentration range of 0.1-1.7 and 0.4-2.8 μg/band, with mean ± SD recoveries of 99.12 ± 0.964 and 100.14 ± 0.962%, for dexamethasone and chlorpheniramine maleate, respectively. Reversed-phase HPLC is the second method, in which a mixture of dexamethasone and chlorpheniramine maleate, methylparaben, and propylparaben is separated on a reversed-phase silica C18 (5 μm particle size, 250 mm, 4.6 mm id) column using 0.1 M ammonium acetate buffer-acetonitrile (60 + 40, v/v, pH 3) as the mobile phase. The drugs were detected at 220 nm over a concentration range of 5-50 μg/mL, 2-90 μg/mL, 4-100 μg/mL, and 7-50 μg/mL, with mean ± SD recoveries of 100.85 ± 0.905, 99.67 ± 1.281, 100.20 ± 0.906, and 99.81 ± 0.954%, for dexamethasone, chlorpheniramine maleate, methylparaben paraben, and propylparaben, respectively. The advantages of the suggested methods over previously reported methods are the ability to detect lower concentrations of the main drugs and to show better resolution of interfering preservatives; hence, these methods could be more reliable for routine QC analyses.

  4. A new quantitation method of protodioscin by HPLC-ESI-MS/MS in rat plasma and its application to the pharmacokinetic study.

    Science.gov (United States)

    Zhang, Xinxin; Guo, Zengjun; Li, Jing; Ito, Yoichiro; Sun, Wenji

    2016-02-01

    A specific high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS method) was established for determining the concentration of protodioscin (PG) in rat plasma after intragastric administration of its standard form. Ginsenoside Rb1 was selected as the internal standard (IS). The plasma sample was prepared using one-step deproteinization procedure by adding three parts of acetonitrile to precipitate proteins. The chromatographic separation was accomplished on an Inersil ODS-3 C18 column (250 × 4.6 mm, 5 μm) with a mobile phase composed with acetonitrile and water containing 0.1% formic acid under a gradient elution mode at a flow rate of 1 mL min(-1). A 3:1 portion of the eluent after a microsplit was detected on a triple quadrupole tandem mass spectrometer coupled with electrospray ionization (ESI) in positive ion and multiple reaction monitoring (MRM) scanning modes. The mass transitions were selected as 888.1 → 1050.2 for PG and 948.2 → 1110.3 for IS, respectively. After careful validation, the plasma samples were always stable under different storage conditions. These analytical results rendered sensitive, selective, and reliable values by this established method which displayed high accuracy, adequate extracted recoveries, and almost negligible matrix effects. This method was applied to the pharmacokinetic studies on PG level in the rat plasma and its pharmacokinetic effect. The results of our studies suggest that the present method may be a useful tool for further clinical study of PG. Copyright © 2015. Published by Elsevier Inc.

  5. Novel acyclonucleoside analog bearing a 1,2,4-triazole–Schiff base: Synthesis, characterization and analytical studies using square wave-adsorptive stripping voltammetry and HPLC

    Directory of Open Access Journals (Sweden)

    Ali F. Alghamdi

    2017-09-01

    Full Text Available New acyclonucleoside analogs tethered by a 1,2,4-triazole scaffold were synthesized through the condensation of 4-amino-5-(2-phenyleth-1-yl-2,4-dihydro-3H-1,2,4-triazole-3-thione (2 with benzaldehyde followed by the alkylation of the resulting Schiff base (3with 2-bromoethanol, 3-chloropropanol and/or 3-chloropropan-1,2-diol. Voltammetric studies were carried out for the analysis of 1 × 10−6 mol L−1 of the newly synthesized acyclonucleoside analogs (4–6 using square wave-adsorptive stripping voltammetry (SW-AdSV. The sharp voltammetric peak and high reduction current were recorded using a Britton–Robinson B–R pH 10 buffer at Ep = −1250 mV on the hanging mercury drop surface (HMDE and Ag/AgCl reference electrode. Several experimental conditions were studied, such as the supporting electrolytes, the pH, and the accumulation time, as well as the potential, the scan rate, the frequency and the step potential for 4-benzylideneamino-5-(2-phenyleth-1-yl-3-[(2,3-dihydroxyprop-1-ylthio]-1,2,4-triazole (6. The analytical performance of the voltammetric technique was investigated through the analysis of the calibration curve, the detection limit, the recovery and the stability. The voltammetric analytical applications were evaluated by the recovery of compound (6 in the urine and plasma samples. The HPLC technique was also applied for the separation of compound (6 from interference using a C-18 (5 μm column with UV detection at 254 nm.

  6. Fourth-derivative synchronous spectrofluorimetry and HPLC with fluorescence detection as two analytical techniques for the simultaneous determination of itopride and domperidone.

    Science.gov (United States)

    Ibrahim, Fawzia; Nasr, Jenny Jeehan

    2016-02-01

    Two simple, rapid and sensitive methods, namely, fourth-derivative synchronous spectrofluorimetry (method I) and HPLC with fluorescence detection (method II) were developed for the simultaneous analysis of a binary mixture of itopride HCl (ITP) and domperidone (DOM) without prior separation. The first method was based on measuring the fourth derivative of the synchronous fluorescence spectra of the two drugs at Δλ = 40 nm in methanol. The different experimental parameters affecting the synchronous fluorescence of the studied drugs were carefully optimized. Chromatographic separation was performed in < 6.0 min using a RP C18 column (250 mm × 4.6 mm i.d., 5 µm particle size) with fluorescence detection at 344 nm after excitation at 285 nm. A mobile phase composed of a mixture of 0.02 M phosphate buffer with acetonitrile in a ratio of 55 : 45, pH 4.5, was used at a flow rate of 1 mL/min. Linearity ranges were found to be 0.1-2 µg/mL for ITP in both methods, whereas those for DOM were found to be 0.08-2 and 0.05-1.5 µg/mL in methods I and II, respectively. The proposed methods were successfully applied for the determination of the studied drugs in synthetic mixtures and laboratory-prepared tablets. Copyright © 2015 John Wiley & Sons, Ltd.

  7. A validated stability-indicating HPLC with photodiode array detector (PDA) method for the stress tests of Monascus purpureus-fermented rice, red yeast rice.

    Science.gov (United States)

    Li, Yong-Guo; Liu, Hong; Wang, Zheng-Tao

    2005-09-01

    A stability-indicating reversed phase high-performance liquid chromatography (RP-HPLC) with photodiode array (PDA) detection method was developed and validated for the assay of monacolin series compounds including monacolin K, L, J and their hydroxyl acid forms as well as dehydroxymonacolin K simultaneously in Monascus purpureus-fermented rice, red yeast rice. Well-resolved peaks of seven main compounds of monacolin family were profiled on a C(18) reverse-phase column using a linear gradient of 0.1% trifluoroacetic acid and acetonitrile as the mobile phase, and the detection wavelength was set at 237nm. The method was validated with respect to specificity, chromatographic parameters, linearity, precision, accuracy, limits of detection and quantitation. The stability stress testing for fermented red yeast rice powder was carried out to show the effects of high temperature (80 degrees C), high humidity at room temperature (92.5% RH, 25 degrees C), high humidity at high temperature (75% RH, 60 degrees C) and light (sunlight) in solid state. The results exhibited that monacolins decreased significantly under the conditions of high humidity at high temperature (75% RH, 60 degrees C) and sunlight. Monacolin K and its hydroxyl acid form would be dehydrolyzed and turned to dehydromonacolin K at high temperature (80 degrees C) while the monacolin K, J and L would be transformed into their corresponding hydroxyl acid forms under the condition of high humidity (92.5% RH, 25 degrees C). The indication is that monacolins in red yeast rice powder are light-sensitive and thermal-sensitive. Therefore, it has been suggested that the preparations containing monacolins be stored in the place of cool and lightproof. The proposed degradation pathways were discussed as well. The multi-components assay for stability of botanical products could provide much more information than the normal marker-orientation method.

  8. A Sensitive and Robust Ultra HPLC Assay with Tandem Mass Spectrometric Detection for the Quantitation of the PARP Inhibitor Olaparib (AZD2281 in Human Plasma for Pharmacokinetic Application

    Directory of Open Access Journals (Sweden)

    Jeffrey Roth

    2014-06-01

    Full Text Available Olaparib (AZD2281 is an orally active PARP-1 inhibitor, primarily effective against cancers with BRCA1/2 mutations. It is currently in Phase III development and has previously been investigated in numerous clinical trials, both as a single agent and in combination with chemotherapy. Despite this widespread testing, there is only one published method that provides assay details and stability studies for olaparib alone. A more sensitive uHPLC-MS/MS method for the quantification of olaparib in human plasma was developed, increasing the range of quantification at both ends (0.5–50,000 ng/mL compared to previously published methods (10–5,000 ng/mL. The wider range encompasses CMAX levels produced by typical olaparib doses and permits better pharmacokinetic modeling of olaparib elimination. This assay also utilizes a shorter analytical runtime, allowing for more rapid quantification and reduced use of reagents. A liquid-liquid extraction was followed by chromatographic separation on a Waters UPLC® BEH C18 column (2.1 × 50 mm, 1.7 µm and mass spectrometric detection. The mass transitions m/z 435.4→281.1 and m/z 443.2→281.1 were used for olaparib and the internal standard [2H8]-olaparib, respectively. The assay proved to be accurate (<9% deviation and precise (CV < 11%. Stability studies showed that olaparib is stable at room temperature for 24 h. in whole blood, at 4 °C for 24 h post-extraction, at −80 °C in plasma for at least 19 months, and through three freeze-thaw cycles. This method proved to be robust for measuring olaparib levels in clinical samples from a Phase I trial.

  9. Feasibility and application of an HPLC/UVD to determine dinotefuran and its shorter wavelength metabolites residues in melon with tandem mass confirmation.

    Science.gov (United States)

    Rahman, Md Musfiqur; Park, Jong-Hyouk; Abd El-Aty, A M; Choi, Jeong-Heui; Yang, Angel; Park, Ki Hun; Nashir Uddin Al Mahmud, Md; Im, Geon-Jae; Shim, Jae-Han

    2013-01-15

    A new analytical method was developed for dinotefuran and its metabolites, MNG, UF, and DN, in melon using high-performance liquid chromatography (HPLC) coupled with an ultraviolet detector (UVD). Due to shorter wavelength, lower sensitivity to UV detection, and high water miscibility of some metabolites, QuEChERs acetate-buffered version was modified for extraction and purification. Mobile phases with different ion pairing or ionisation agents were tested in different reverse phase columns, and ammonium bicarbonate buffer was found as the best choice to increase the sensitivity of target analytes to the UV detector. After failure of dispersive SPE clean-up with primary secondary amine, different solid phase extraction cartridges (SPE) were used to check the protecting capability of analytes against matrix interference. Finally, samples were extracted with a simple and rapid method using acetonitrile and salts, and purified through C(18)SPE. The method was validated at two spiking levels (three replicates for each) in the matrix. Good recoveries were observed for all of the analytes and ranged between 70.6% and 93.5%, with relative standard deviations of less than 10%. Calibration curves were linear over the calibration ranges for all the analytes with r(2)≥ 0.998. Limits of detection ranged from 0.02 to 0.05 mg kg(-1), whereas limits of quantitation ranged from 0.06 to 0.16 mg kg(-1) for dinotefuran and its metabolites. The method was successfully applied to real samples, where dinotefuran and UF residues were found in the field-incurred melon samples. Residues were confirmed via LC-tandem mass spectrometry (LC-MS/MS) in positive-ion electrospray ionisation (ESI(+)) mode. Copyright © 2012 Elsevier Ltd. All rights reserved.

  10. Storage stability study of porcine hepatic and intestinal cytochrome P450 isoenzymes by use of a newly developed and fully validated highly sensitive HPLC-MS/MS method.

    Science.gov (United States)

    Schelstraete, Wim; Devreese, Mathias; Croubels, Siska

    2018-01-11

    Microsomes are an ideal medium to investigate cytochrome P450 (CYP450) enzyme-mediated drug metabolism. However, before microsomes are prepared, tissues can be stored for a long time. Studies about the stability of these enzymes in porcine hepatic and intestinal tissues upon storage are lacking. To be able to investigate CYP450 stability in microsomes prepared from these tissues, a highly sensitive and rapid HPLC-MS/MS method for the simultaneous determination of six CYP450 metabolites in incubation medium was developed and validated. The metabolites, paracetamol (CYP1A), 7-hydroxy-coumarin (CYP2A), 1-hydroxy-midazolam (CYP3A), 4-hydroxy-tolbutamide (CYP2C), dextrorphan (CYP2D), and 6-hydroxy-chlorzoxazone (CYP2E) were extracted with ethyl acetate at pH 1.0, followed by evaporation and separation on an Agilent Zorbax Eclipse Plus C18 column. The method was fully validated in a GLP-compliant laboratory according to European guidelines and was highly sensitive (LOQ = 0.25-2.5 ng/mL), selective, had good precision (RSD-within, 1.0-9.1%; RSD-between, 1.0-18.4%) and accuracy (within-run, 83.3-102%; between-run, 78.5-102%), and showed no relative signal suppression and enhancement. Consequently, this method was applied to study the stability of porcine hepatic and intestinal CYP450 isoenzymes when tissues were stored at - 80 °C. The results indicate that porcine CYP450 isoenzymes are stable in tissues at least up to 4 months when snap frozen and stored at - 80 °C. Moreover, the results indicate differences in porcine CYP450 stability compared to rat, rabbit, and fish CYP450, as observed by other research groups, hence stressing the importance to investigate the CYP450 stability of a specific species.

  11. Separation of quercetin, sexangularetin, kaempferol and isorhamnetin for simultaneous HPLC determination of flavonoid aglycones in inflorescences, leaves and fruits of three Sorbus species.

    Science.gov (United States)

    Olszewska, Monika

    2008-11-04

    A reversed-phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated for the simultaneous determination of four flavonol aglycones (quercetin, QU; sexangularetin, SX; kaempferol, KA; isorhamnetin, IS) in hydrolyzed extracts from different plant parts of Sorbus aucuparia L., Sorbus aria (L.) Crantz. and Sorbus intermedia (Ehrh.) Pers. Separation of the four compounds was accomplished on a C18 Lichrosphere 100 column (5 microm, 250 mm x 4.6mm, i.d.) with a methanol gradient elution and recorded at 370 nm. The high resolution of critical bands - SX, KA and IS - was achieved with retention of the last peak (IS) in 19.5 min. The equilibration of the standard mixture by addition of HCl to an acid concentration equal that of hydrolyzed extracts injected was found to be necessary when minimizing calibration error. The correlation coefficients of all the calibration curves showed good linearity (r>0.9991) over the test range. The relative standard deviation of the method was less than 2.8% for intra- and inter-day assays, and the average recoveries were between 95.5 and 102.5%. High sensitivity was demonstrated with detection limits between 0.050 and 0.085 microg/ml. The level of total aglycones was found to be in the range of 687-1,515 mg/100g of dry weight in the inflorescences, 424-1,078 mg/100g in the leaves and 20-60 mg/100g in the fruits depending on the Sorbus species.

  12. Analysis of coal extracts by HPLC-ESI-MS

    Energy Technology Data Exchange (ETDEWEB)

    Jie Feng; Cui-Ping Ye; Wen-Ying Li; Ke-Chang Xie [Taiyuan University of Technology, Taiyuan (China). Key Laboratory of Coal Science and Technology

    2003-07-01

    In order to get the structure information of the coal pyridine extracts, the methods based on reverse-phase high-performance liquid chromatography (RP-HPLC) with photodiode array detector (PDA) and electro-spray ionization mass spectrometer (ESI-MS) has been developed. For the purpose of classifying the mixture according to the polarity of the compound before HPLC/MS analysis, column chromatography packed with silica gel was applied to separate coal pyridine extracts into three fractions: acetonitrile (lower), chloroform (middle), pyridine (higher). The fraction of chloroform (CHCl{sub 3}) was analyzed in this article. In the mass range of 150-1500amu, the components observed include 314.3/369.1, and 301.3/369.1, which should be the plasticizer in the solvent. Other peaks included a series of molecular ions from the beginning of 541.3amu with m/z difference 74amu (C{sub 3}H{sub 6}O{sub 2} or C{sub 4}H{sub 10}O). 541.3amu (ESI+) is the elemental structure in the series; this implied that the higher molecular mass parts in coal might consist of some basic units. And two valuable compounds containing nitrogen atom were acquired at m/z 393.5amu (C{sub 27}H{sub 39}NO) and 334.5amu (C{sub 20}H{sub 38}N{sub 4}). 11 refs., 5 figs., 1 tab.

  13. Core-Shell Columns in High-Performance Liquid Chromatography: Food Analysis Applications

    Directory of Open Access Journals (Sweden)

    Raffaella Preti

    2016-01-01

    Full Text Available The increased separation efficiency provided by the new technology of column packed with core-shell particles in high-performance liquid chromatography (HPLC has resulted in their widespread diffusion in several analytical fields: from pharmaceutical, biological, environmental, and toxicological. The present paper presents their most recent applications in food analysis. Their use has proved to be particularly advantageous for the determination of compounds at trace levels or when a large amount of samples must be analyzed fast using reliable and solvent-saving apparatus. The literature hereby described shows how the outstanding performances provided by core-shell particles column on a traditional HPLC instruments are comparable to those obtained with a costly UHPLC instrumentation, making this novel column a promising key tool in food analysis.

  14. of reinforced concrete columns

    Directory of Open Access Journals (Sweden)

    Anna Szcześniak

    2015-09-01

    Full Text Available The paper presents the modification of the dynamic relaxation method in order to increase its effectiveness in the range of the post-critical analysis. For this purpose, the arc-length parameter on the equilibrium path was introduced into the computational procedure. The additional constraints equation that combines increment of load parameter and the vector of displacement increments with the arc-length increment on the solution path was incorporated to analysis of the equations of motion. Solution of nonlinear equilibrium equations is obtained recursively in subsequent pseudo-time instants. The proposed method allows for tracking the global softening phenomenon of the structural element in the post-critical range, which leads to failure. We ran numerical experiments for the reinforced concrete eccentrically loaded column. Our comparative analysis with previously published numerical results demonstrated that the proposed computational method is effective.[b]Keywords[/b]: reinforced concrete columns, dynamic relaxation method, arc-length method, load-carrying capacity

  15. Effect of silage type and concentrate level on conjugated linoleic acids, trans-C18 : 1 isomers and fat content in milk from dairy cows

    DEFF Research Database (Denmark)

    Nielsen, Torben Skov; Straarup, Ellen Marie; Jungersen, Mogens Vestergaard

    2006-01-01

    :1 and reduced cis9, trans11-CLA and trans11-C18:1 when maize but not grass silage was provided. The results suggest that high levels of concentrate (grain) do not significantly alter the pattern of PUFA biohydrogenation in the rumen, the concentration of CLA and trans-C18:1 isomers in milk or cause milk fat...

  16. Monolitní stacionární fáze pro HPLC stanovení biologicky aktivních látek I.

    OpenAIRE

    Krčmová, Lenka

    2006-01-01

    DEVELOPMENT AND VALIDATION OF HPLC METHOD FOR DETERMINATION OF RETINYL ESTERS IN HUMAN SERUM USING MONOLITHIC COLUMN Absorption test of vitamin A is used for monitoring of intestinal permeability and may represent a sensitive indicator of intestinal damage. In this study, a simple and rapid reversed- phase high-performance liquid chromatography (RP-HPLC) procedure for selective and sensitive determination of retinol, -tocopherol, retinyl-palmitate and retinyl-stearate in blood serum has been...

  17. Expermental Studies of quantitative evaluation using HPLC

    Directory of Open Access Journals (Sweden)

    Ki Rok Kwon

    2005-06-01

    Full Text Available Methods : This study was conducted to carry out quantitative evaluation using HPLC Content analysis was done using HPLC Results : According to HPLC analysis, each BVA-1 contained approximately 0.36㎍ melittin, and BVA-2 contained approximately 0.54㎍ melittin. But the volume of coating was so minute, slight difference exists between each needle. Conclusion : Above results indicate that the bee venom acupuncture can complement shortcomings of syringe usage as a part of Oriental medicine treatment, but extensive researches should be done for further verification.

  18. HPLC for quality control of polyimides

    Science.gov (United States)

    Young, P. R.; Sykes, G. F.

    1979-01-01

    High Pressure Liquid Chromatography (HPLC) as a quality control tool for polyimide resins and prepregs are presented. A data base to help establish accept/reject criteria for these materials was developed. This work is intended to supplement, not replace, standard quality control tests normally conducted on incoming resins and prepregs. To help achieve these objectives, the HPLC separation of LARC-160 polyimide precursor resin was characterized. Room temperature resin aging effects were studied. Graphite reinforced composites made from fresh and aged resin were fabricated and tested to determine if changes observed by HPLC were significant.

  19. Efficient control of the rheological and surface properties of surfactant solutions containing C8-C18 fatty acids as cosurfactants.

    Science.gov (United States)

    Mitrinova, Z; Tcholakova, S; Popova, Z; Denkov, N; Dasgupta, Bivash R; Ananthapadmanabhan, K P

    2013-07-02

    Systematic experimental study is performed about the effects of chain length (varied between C8 and C18) and concentration of fatty acids (FAc), used as cosurfactants to the mixture of the anionic surfactant SLES and the zwitterionic surfactant CAPB. The following properties are studied: bulk viscosity of the concentrated solutions (10 wt % surfactants), dynamic and equilibrium surface tensions, surface modulus, and foam rheological properties for the diluted foaming solutions (0.5 wt % surfactants). The obtained results show that C8-C10 FAc induce formation of wormlike micelles in the concentrated surfactant solutions, which leads to transformation of these solutions into viscoelastic fluids with very high apparent viscosity. The same FAc shorten the characteristic adsorption time of the diluted solutions by more than 10 times. In contrast, C14-C18 FAc have small effect on the viscosity of the concentrated solutions but increase the surface modulus above 350 mN/m, which leads to higher friction inside sheared foams and to much smaller bubbles in the formed foams. The intermediate chain C12 FAc combines some of the properties seen with C10 FAc and other properties seen with C14 FAc. These results clearly demonstrate how appropriate cosurfactants can be used for efficient control of the rheological properties of concentrated surfactant solutions and of some important foam attributes, such as bubble size and foam rheology.

  20. Preparation of Magnetic Sorbent with Surface Modified by C18for Removal of Selected Organic Pollutants from Aqueous Samples

    Science.gov (United States)

    Kuráň, Pavel; Pilnaj, Dominik; Ciencialová, Lucie; Pšenička, Martin

    2017-12-01

    Magnetic sorbents have great potential in environmental applications due to their simple synthesis and separation in magnetic field, usability in heterogeneous systems and low toxicity. Possible syntheses, surface modifications and characteristics were described by Li et al 2013. This type of solid-phase extraction is being successfully used in various fields as health care, microbiology, biotechnologies or sample preconcentration in analytical chemistry. In this preliminary study we report on the preparation and application of magnetically separable sorbent with surface modified by C18 alkyl chain for purification of water contaminated by environmentally hazardous organic compounds. Magnetic cores were co-precipitated from Fe2+ and Fe3+ chlorides in alkalic aqueous solution. Surface of synthetized Fe3O4 was modified with SiO2 by tetraethylorthosilicate to assure physico-chemical stability. Furthermore, Fe3O4/SiO2 complex has been treated by C18 functional group, which provides good affinity towards hydrophobic substances in water. Efficiency of sorption under various conditions has been examined on benzene, toluene, ethylbenzene and xylenes (BTEX), compounds found in petroleum products which contaminate air, soil and groundwater near of store tanks. Sorption kinetics was followed by gas chromatography with mass spectrometry. The preliminary sorption kinetics data and efficiency of BTEX removal point at the possible application of prepared magnetic sorbent for BTEX removal, especially for ethylbenzene and xylenes.

  1. Determination of cuticular and internal fatty acids of Chorthippus brunneus males and females using HPLC-LLSD and GC-MS.

    Science.gov (United States)

    Gołębiowski, Marek; Cerkowniak, Magdalena; Ostachowska, Aleksandra; Boguś, Mieczysława I; Stepnowski, Piotr

    2016-08-01

    Insects are of growing significance in veterinary medicine and human healthcare; therefore, an understanding of their biology is very important. The cuticular and internal fatty acid compositions of Chorthippus brunneus males and females have been studied for the first time. The lipids of males and females were separated into classes of compounds using high-performance liquid chromatography with a laser light scattering detector. The free fatty acid (FFA) fractions obtained by HPLC were silylated and then analyzed by GC-MS. The cuticular lipids of males contained 15 saturated, four unsaturated with even-numbered and two unsaturated with odd-numbered carbon chains, FFAs ranging from C8 to C25. The major free fatty acids in males were C16 (20.8%), C18:2 (8.5%), C18:1 (32.9%) and C18 (24.4%). The cuticular lipids of females contained 17 saturated, four monounsaturated and two diunsaturated free fatty acids ranging from C8 to C30. The major cuticular fatty acids in females were C16 (25.1%), C18:2 (6.2%), C18:1 (23.7%) and C18:0 (33.2%). The internal FFAs of males consisted of 20 compounds ranging from C8 to C26. Four of these compounds were detected as major compounds: C16 (14.1%), C18:2 (21.6%), C18:1 (38.0%) and C18 (22.5%). Among 18 internal free fatty acids of females, C16 (22.3%), C18:2 (10.9%), C18:1 (40.2%) and C18 (20.5%) were the most abundant compounds. The following cuticular fatty acids present in the lipids of females were absent in the lipids of males: C26, C27 and C30. On the other hand, only C24 was absent from the cuticular lipids of females. Only C10 and C24 internal fatty acids present in the lipids of males were absent in the lipids of females. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  2. SPIRAL CONTACTOR FOR SOLVENT EXTRACTION COLUMN

    Science.gov (United States)

    Cooley, C.R.

    1961-06-13

    The patented extraction apparatus includes a column, perforated plates extending across the column, liquid pulse means connected to the column, and an imperforate spiral ribbon along the length of the column.

  3. Development and validation of a repharsed phase- HPLC method for simultaneous determination of rosiglitazone and glimepiride in combined dosage forms and human plasma

    Directory of Open Access Journals (Sweden)

    El-Enany Nahed M

    2012-01-01

    Full Text Available Abstract Background Rosiglitazone (ROZ and glimepiride (GLM are antidiabetic agents used in the treatment of type 2 diabetes mellitus. A survey of the literature reveals that only one spectrophotometric method has been reported for the simultaneous determination of ROS and GLM in pharmaceutical preparations. However the reported method suffers from the low sensitivity, for this reason, our target was to develop a simple sensitive HPLC method for the simultaneous determination of ROZ and GLM in their combined dosage forms and plasma. Results A simple reversed phase high performance liquid chromatographic (RP-HPLC method was developed and validated for the simultaneous determination of Rosiglitazone (ROS and Glimepiride (GLM in combined dosage forms and human plasma. The separation was achieved using a 150 mm × 4.6 mm i.d., 5 μm particle size Symmetry® C18 column. Mobile phase containing a mixture of acetonitrile and 0.02 M phosphate buffer of pH 5 (60: 40, V/V was pumped at a flow rate of 1 mL/min. UV detection was performed at 235 nm using nicardipine as an internal standard. The method was validated for accuracy, precision, specificity, linearity, and sensitivity. The developed and validated method was successfully used for quantitative analysis of Avandaryl™ tablets. The chromatographic analysis time was approximately 7 min per sample with complete resolution of ROS (tR = 3.7 min., GLM (tR = 4.66 min., and nicardipine (tR, 6.37 min. Validation studieswas performed according to ICH Guidelines revealed that the proposed method is specific, rapid, reliable and reproducible. The calibration plots were linear over the concentration ranges 0.10-25 μg/mL and 0.125-12.5 μg/mL with LOD of 0.04 μg/mL for both compounds and limits of quantification 0.13 and 0.11 μg/mL for ROS and GLM respectively. Conclusion The suggested method was successfully applied for the simultaneous analysis of the studied drugs in their co-formulated tablets and human

  4. European Analytical Column

    DEFF Research Database (Denmark)

    Karlberg, B.; Grasserbauer, M.; Andersen, Jens Enevold Thaulov

    2009-01-01

    The European Analytical Column has once more invited a guest columnist to give his views on various matters related to analytical chemistry in Europe. This year, we have invited Professor Manfred Grasserbauer of the Vienna University of Technology to present some of the current challenges...... for European analytical chemistry. During the period 2002–07, Professor Grasserbauer was Director of the Institute for Environment and Sustainability, Joint Research Centre of the European Commission (EC), Ispra, Italy. There is no doubt that many challenges exist at the present time for all of us representing...... a major branch of chemistry, namely analytical chemistry. The global financial crisis is affecting all branches of chemistry, but analytical chemistry, in particular, since our discipline by tradition has many close links to industry. We have already noticed decreased industrial commitment with respect...

  5. Column: File Cabinet Forensics

    Directory of Open Access Journals (Sweden)

    Simson Garfinkel

    2011-12-01

    Full Text Available Researchers can spend their time reverse engineering, performing reverse analysis, or making substantive contributions to digital forensics science. Although work in all of these areas is important, it is the scientific breakthroughs that are the most critical for addressing the challenges that we face.Reverse Engineering is the traditional bread-and-butter of digital forensics research. Companies like Microsoft and Apple deliver computational artifacts (operating systems, applications and phones to the commercial market. These artifacts are bought and used by billions. Some have evil intent, and (if society is lucky, the computers end up in the hands of law enforcement. Unfortunately the original vendors rarely provide digital forensics tools that make their systems amenable to analysis by law enforcement. Hence the need for reverse engineering.(see PDF for full column

  6. Analysis of flame retardant additives in polymer fractions of waste of electric and electronic equipment (WEEE) by means of HPLC-UV/MS and GPC-HPLC-UV.

    Science.gov (United States)

    Schlummer, Martin; Brandl, Fritz; Mäurer, Andreas; van Eldik, Rudi

    2005-01-28

    An HPLC-UV/MS method has been developed to identify and quantify flame retardants in post-consumer plastics from waste of electric and electronic equipment (WEEE). Atmospheric pressure chemical ionisation spectra of 15 brominated and phosphate-based flame retardants were recorded and interpreted. The method was applied to detect flame retardant additives in polymer extracts obtained from pressurised liquid extraction of solid polymers. In addition, a screening method was developed for soluble styrene polymers to isolate a flame retardant fraction through the application of gel permeation chromatography (GPC). This fraction was transferred to an online-coupled HPLC column and detected by UV spectroscopy, which allowed a reliable qualitative and quantitative analysis of brominated flame retardants in the polymer solutions.

  7. A new HPLC method for pidotimod plasma levels determination.

    Science.gov (United States)

    Dal Bo, L; Broccali, G P; Silingardi, S; Coppi, G

    1993-04-01

    This paper describes a HPLC method for the determination of Pidotimod (3-L-pyroglutamyl-L-thiazolidine-4-carboxylic acid; PGT/1A), a new biological response modifier, in plasma. The column was an Aminex Ion Exclusion HPX 874 with a PRP precolumn, the mobile phase was 0.05% sulfuric acid-acetonitrile (88:12, v/v), the flow rate was 0.6 ml/min, the detection wavelength was 210 nm. Plasma (1 ml) was added with internal standard (Oxiracetam, concentration 400 micrograms/ml) (50 microliters) and 35% perchloric acid (100 microliters). The supernatant (0.5 ml) was added with mobile phase (0.5 ml) and, after centrifugation, injected into the column. The retention times of Pidotimod and Oxiracetam were 16.5 and 13.8 min. respectively. The method was validated for recovery, accuracy and reproducibility. The results after oral administration of 800 mg of Pidotimod in male volunteers were also given. This method is better than that previously described because it utilizes an internal standard and reaches a lower detection limit.

  8. HPLC-ELSD analysis of spectinomycin dihydrochloride and its impurities.

    Science.gov (United States)

    Zhou, Junyi; Zhang, Lin; Wang, Yan; Yan, Chao

    2011-08-01

    A simple, rapid and reliable reversed-phase ion-pair chromatography method by HPLC coupled to an evaporative light scattering detector (ELSD) has been developed to simultaneously determine chloride, spectinomycin and its related substances in a sample. The column was a TSKgel ODS-100V. The mobile phase was ACN/aqueous solution of 15 mM ammonium acetate adjusted with TFA to pH 3.0 (2:98 v/v), in an isocratic mode. The drift tube temperature was set at 50°C and the nebulizing gas flow rate of air was 3.5 L/min for ELSD detection. Almost all of the reported degradation compounds of spectinomycin such as actinamine, actinospectinoic acid and biosynthesis intermediates such as dihydrospectinomycin diastereoisomers were baseline separated. MS was utilized for the identification of spectinomycin and its seven related substances. The method for the assay of spectinomycin was successfully validated with respect to accuracy, precision (RSD less than 2%), linearity (throughout the linear range 0.025-3 mg/mL, r=0.9993), sensitivity (LOD: 100 ng on column) and robustness. The experimental results demonstrated that the simultaneous determination of chloride, spectinomycin and related substances is feasible in a single run, which suggests applicability in routine assays. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Effect of Mobile Phase Additives on the Resolution of Four Bioactive Compounds by RP-HPLC

    Directory of Open Access Journals (Sweden)

    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.

  10. Stability-indicating HPLC Method for Simultaneous Determination of Aspirin and Prasugrel

    OpenAIRE

    Patel, Shital M.; Patel, C. N.; Patel, V. B.

    2013-01-01

    A simple, sensitive, specific, accurate, and stability-indicating reversed phase high performance liquid chromatographic method was developed for the simultaneous determination of aspirin and prasugrel, using a Kromasil 100 C 18 (150×4.6 mm, 5 μ) column and a mobile phase composed of acetonitrile:methanol:water (30:10:60, v/v), pH 3.0 adjusted with o-phosphoric acid. The retention times of aspirin and prasugrel were found to be 3.28 min and 6.61 min, respectively. Linearity was established fo...

  11. Determination of the solvent density profiles across mesopores of silica-C18 bonded phases in contact with acetonitrile/water mixtures: A semi-empirical approach.

    Science.gov (United States)

    Gritti, Fabrice

    2015-09-04

    The local volume fractions of water, acetonitrile, and C18-bonded chains across the 96Åmesopores of 5μm Symmetry particles were determined semi-empirically. The semi-empirical approach was based on previous molecular dynamics studies, which provided relevant mathematical expressions for the density profiles of C18 chains and water molecules, and on minor disturbance experiments, which measured the excess amount of acetonitrile adsorbed in the pores of Symmetry-C18 particles. The pore walls of the Symmetry-C18 material were in thermodynamic equilibrium with a series of binary mixtures of water and acetonitrile. The results show that C18 chains are mostly solvated by acetonitrile molecules, water is excluded from the C18-bonded layer, and acetonitrile concentrates across a 15-25Åthick interface region between the C18 layer and the bulk phase. These actual density profiles are expected to have a direct impact on the retention behaviour of charged, polar, and neutral analytes in RPLC. They also provide clues to predict the local mobility of analytes inside the pores and a sound physico-chemical description of the phenomenon of surface diffusion observed in RPLC. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. trans-C18:1 isomers in cheeses enriched in unsaturated fatty acids and manufactured with different milk fat globule sizes.

    Science.gov (United States)

    Briard-Bion, Valerie; Juaneda, Pierre; Richoux, Romain; Guichard, Elisabeth; Lopez, Christelle

    2008-10-22

    Increasing the knowledge on dietary fat composition, mainly the minor components, will improve the nutritional value of foods and their labeling. In this study, we examined the trans-octadecenoic acid (C18:1) composition of Emmental cheeses enriched in unsaturated fatty acids (FA) and manufactured with milks produced by cows selected to produce small and large fat globules. The FA composition of the milks was not significantly ( P > 0.05) different from the FA composition of the corresponding Emmental cheeses. Increasing the unsaturated FA content of the cheeses using dietary manipulations lead to an increase in the trans-C18:1 and changed their isomeric profiles. In milk fat produced with the linseed-enriched diet, the trans-10 C18:1 concentration was greater than trans-11 C18:1 (vaccenic acid), which is classically the major trans-C18:1 in milk fat. The content in trans-C18:1 and more particularly in trans-10 C18:1 was negatively correlated with the size of fat globules ( r (2) = 0. 82 and 0.87, respectively) and related to milk fat depression. The trans-C18:1 content was negatively correlated with the saturated FA (slope = -0.35; r (2) = 0.81) and positively correlated with the unsaturated (slope = 0.29; r (2) = 0.85) and monounsaturated (slope = 0.32; r (2) = 0.81) FA. Focusing on the health-related considerations of fat in food products, further nutritional studies are needed to elucidate the role of trans-C18:1 isomers.

  13. L'Introduction et la diffusion de la variété d'igname C18 en région ...

    African Journals Online (AJOL)

    Ces facteurs sont constitués par la réduction de la pénibilité du travail et des coûts de production liée à l'adoption cette variété; la productivité élevée de C18 en comparaison des variétés traditionnelles d'igname et l'aptitude de C18 à la confection d'un foutou de bonne qualité. La résistance de C18 à une virose qui attaque ...

  14. Identification of a potential biomarker panel for the intake of the common dietary trans fat elaidic acid (trans∆9-C18:1)

    DEFF Research Database (Denmark)

    Kroager, Toke Peter; Nielsen, Lone Vendel; Bak, Steffen

    2012-01-01

    Trans fatty acid intake has been correlated to an unfavorable plasma lipoprotein profile and an increased cardiovascular disease risk. The present study aimed to identify a plasma protein biomarker panel related to human intake of elaidic acid. The human liver cell line HepG2-SF was used as a model...... system, and the cells were maintained for seven days in serum-free medium containing 100 μM elaidic acid (trans∆9-C18:1), oleic acid (cis∆9-C18:1) or stearic acid (C18:0). The secretomes were analyzed by stable isotope labeling of amino acids in cell culture (SILAC), difference in gel electrophoresis...

  15. Seamless Combination of High-Pressure Chip-HPLC and Droplet Microfluidics on an Integrated Microfluidic Glass Chip.

    Science.gov (United States)

    Gerhardt, Renata F; Peretzki, Andrea J; Piendl, Sebastian K; Belder, Detlev

    2017-12-05

    We introduce an approach for the integration of high performance liquid chromatography and droplet microfluidics on a single high-pressure resistant microfluidic glass chip. By coupling these two functionalities, separated analyte bands eluting from the HPLC column are fractionated into numerous droplets in a continuous flowing oil phase. The compartmentalization of the HPLC-eluate in a segmented flow was performed with droplet sizes of approximately 1 nL and with droplet frequencies reaching up to 45 Hz. This approach prevents peak dispersion and facilitates post column processing of chromatographic fractions on chip. A reliable generation of droplets is also possible in reversed phase gradient elution mode as demonstrated by applying a solvent gradient from 20% to 100% acetonitrile. A chip design with an incorporated dosing unit enabled the directed postcolumn addition of reagents to individual droplet fractions. The capability of this dosing function was successfully evidenced by post column addition of a reagent which quenches the fluorescence signal of the analytes. The chip-integration of gradient HPLC, fractionation, detection and post column addition of reagents opens up new avenues to perform multistep chemical processes on a single lab-on-a-chip device.

  16. High-Throughput Parallel Proteomic Sample Preparation Using 96-Well Polyvinylidene Fluoride (PVDF) Membranes and C18 Purification Plates.

    Science.gov (United States)

    Bennike, Tue Bjerg; Steen, Hanno

    2017-01-01

    Meaningful proteomic-based biomarker discovery projects using primary human-derived specimens require the analysis of hundreds of samples in order to address the issue of interpersonal variability. Thus, robust high-throughput methods for the digestion of plasma samples are a prerequisite for such large clinical proteomic studies with hundreds of samples. Commonly used sample preparation methods are often difficult to parallelize and/or automate. Herein we describe a method for parallel 96-well plate-based sample preparation. Protein digestion is performed in 96-well polyvinylidene fluoride (PVDF) membrane plates and the subsequent purification in 96-well reversed phase C18 purification plates, enabling the usage of multichannel pipettes in all steps. The protocol can be applied using neat or depleted plasma/serum samples, but has also proven effective with other sample types.

  17. Anticancer Activities of C18-, C19-, C20-, and Bis-Diterpenoid Alkaloids Derived from Genus Aconitum.

    Science.gov (United States)

    Ren, Meng-Yue; Yu, Qing-Tian; Shi, Chun-Yu; Luo, Jia-Bo

    2017-02-13

    Cancer is one of the most common lethal diseases, and natural products have been extensively studied as anticancer agents considering their availability, low toxicity, and economic affordability. Plants belonging to the genus Aconitum have been widely used medically in many Asian countries since ancient times. These plants have been proven effective for treating several types of cancer, such as lung, stomach, and liver cancers. The main effective components of Aconitum plants are diterpenoid alkaloids-which are divided into C18-, C19-, C20-, and bis-diterpenoid alkaloids-are reportedly some of the most promising, naturally abundant compounds for treating cancer. This review focuses on the progress of diterpenoid alkaloids with different structures derived from Aconitum plants and some of their derivatives with potential anticancer activities. We hope that this work can serve as a reference for further developing Aconitum diterpenoid alkaloids as anticancer agents.

  18. Isolation of lactoperoxidase using different cation exchange resins by batch and column procedures.

    Science.gov (United States)

    Fweja, Leonard Wt; Lewis, Michael J; Grandison, Alistair S

    2010-08-01

    Lactoperoxidase (LP) was isolated from whey protein by cation-exchange using Carboxymethyl resin (CM-25C) and Sulphopropyl Toyopearl resin (SP-650C). Both batch and column procedures were employed and the adsorption capacities and extraction efficiencies were compared. The resin bed volume to whey volume ratios were 0.96:1.0 for CM-25C and 0.64:1.0 for SP-650 indicating higher adsorption capacity of SP-650 compared with CM-25C. The effluent LP activity depended on both the enzyme activity in the whey and the amount of whey loaded on the column within the saturation limits of the resin. The percentage recovery was high below the saturation point and fell off rapidly with over-saturation. While effective recovery was achieved with column extraction procedures, the recovery was poor in batch procedures. The whey-resin contact time had little impact on the enzyme adsorption. SDS PAGE and HPLC analyses were also carried out, the purity was examined and the proteins characterised in terms of molecular weights. Reversed phase HPLC provided clear distinction of the LP and lactoferrin (LF) peaks. The enzyme purity was higher in column effluents compared with batch effluents, judged on the basis of the clarity of the gel bands and the resolved peaks in HPLC chromatograms.

  19. Method Development and Validation for Determination of Febuxostat from Spiked Human Plasma Using RP-HPLC with UV Detection

    OpenAIRE

    Monita Gide; Pankaj Sharma; Ravindra Saudagar; Birendra Shrivastava

    2014-01-01

    A rapid, simple, selective, and specific reverse phase high performance liquid chromatography (RP-HPLC) method with UV detection (315 nm) was developed and validated for estimation of febuxostat from spiked human plasma. The analyte and internal standard (diclofenac) were extracted using LLE with diethyl ether. The chromatographic separation was performed on Shodex C-18-4E (5 μm; 250×4.6 mm) with a mobile phase comprised of methanol : acetate buffer pH 4, 20 mM (90 : 10 v/v), at a flow rate o...

  20. Determination of the Antibiotic Oxytetracycline in Commercial Milk by Solid-Phase Extraction: A High-Performance Liquid Chromatography (HPLC) Experiment for Quantitative Instrumental Analysis

    Science.gov (United States)

    Mei-Ratliff, Yuan

    2012-01-01

    Trace levels of oxytetracylcine spiked into commercial milk samples are extracted, cleaned up, and preconcentrated using a C[subscript 18] solid-phase extraction column. The extract is then analyzed by a high-performance liquid chromatography (HPLC) instrument equipped with a UV detector and a C[subscript 18] column (150 mm x 4.6 mm x 3.5 [mu]m).…