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Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma
A simple, accurate, precise, sensitive, and reproducible high-performance liquid chromatography method was developed for the determination of Resveratrol (trans-3, 4’,5-trihydroxystilbene) in human plasma using liquid-liquid extraction. Caffeine was employed as an internal standard (IS). However, li...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Medknow Publications & Media Pvt Ltd
2012
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3401675/ https://www.ncbi.nlm.nih.gov/pubmed/22837962 http://dx.doi.org/10.4103/2231-4040.97296 |
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author | Singh, Gurinder Pai, Roopa S. Pandit, Vinay |
author_facet | Singh, Gurinder Pai, Roopa S. Pandit, Vinay |
author_sort | Singh, Gurinder |
collection | PubMed |
description | A simple, accurate, precise, sensitive, and reproducible high-performance liquid chromatography method was developed for the determination of Resveratrol (trans-3, 4’,5-trihydroxystilbene) in human plasma using liquid-liquid extraction. Caffeine was employed as an internal standard (IS). However, little information is known about its distribution in the organism generally because of the lack of accurate and precise detection methods. The chromatographic separation was achieved on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm) at room temperature in isocratic mode, and the column effluent was monitored by UV detector at 306 nm. The mobile phase used was methanol: phosphate buffer (pH 6.8 adjusted with 0.5% (v/v) orthophosphoric acid solution in Milli-Q water) (63:37%, v/v) at a flow rate of 1.0 ml/min. Nominal retention times of trans-resveratrol and IS were 3.94 and 7.86 minutes, respectively. Limits of detection and Limits of quantification of trans-resveratrol were 0.006 μg/ml and 0.008 μg/ml, respectively. This method was linear over the range of 0.010 to 6.4 μg/ml with regression coefficient greater than 0.9998. The inter- and intra-day precisions in the samples, 0.010, 3.2 and 6.4 μg/ml of trans-resveratrol was in the range 0.63 to 2.12% relative standard deviation (RSD) and 0.46 to 1.02% RSD, respectively. Resveratrol was found to be stable for a period of 15 days on storage at -20°C. The method was found to be precise, accurate, and specific during the study. |
format | Online Article Text |
id | pubmed-3401675 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Medknow Publications & Media Pvt Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-34016752012-07-26 Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma Singh, Gurinder Pai, Roopa S. Pandit, Vinay J Adv Pharm Technol Res Original Article A simple, accurate, precise, sensitive, and reproducible high-performance liquid chromatography method was developed for the determination of Resveratrol (trans-3, 4’,5-trihydroxystilbene) in human plasma using liquid-liquid extraction. Caffeine was employed as an internal standard (IS). However, little information is known about its distribution in the organism generally because of the lack of accurate and precise detection methods. The chromatographic separation was achieved on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm) at room temperature in isocratic mode, and the column effluent was monitored by UV detector at 306 nm. The mobile phase used was methanol: phosphate buffer (pH 6.8 adjusted with 0.5% (v/v) orthophosphoric acid solution in Milli-Q water) (63:37%, v/v) at a flow rate of 1.0 ml/min. Nominal retention times of trans-resveratrol and IS were 3.94 and 7.86 minutes, respectively. Limits of detection and Limits of quantification of trans-resveratrol were 0.006 μg/ml and 0.008 μg/ml, respectively. This method was linear over the range of 0.010 to 6.4 μg/ml with regression coefficient greater than 0.9998. The inter- and intra-day precisions in the samples, 0.010, 3.2 and 6.4 μg/ml of trans-resveratrol was in the range 0.63 to 2.12% relative standard deviation (RSD) and 0.46 to 1.02% RSD, respectively. Resveratrol was found to be stable for a period of 15 days on storage at -20°C. The method was found to be precise, accurate, and specific during the study. Medknow Publications & Media Pvt Ltd 2012 /pmc/articles/PMC3401675/ /pubmed/22837962 http://dx.doi.org/10.4103/2231-4040.97296 Text en Copyright: © Journal of Advanced Pharmaceutical Technology & Research http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Singh, Gurinder Pai, Roopa S. Pandit, Vinay Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma |
title | Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma |
title_full | Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma |
title_fullStr | Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma |
title_full_unstemmed | Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma |
title_short | Development and validation of a HPLC method for the determination of trans-resveratrol in spiked human plasma |
title_sort | development and validation of a hplc method for the determination of trans-resveratrol in spiked human plasma |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3401675/ https://www.ncbi.nlm.nih.gov/pubmed/22837962 http://dx.doi.org/10.4103/2231-4040.97296 |
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