Stereospecific determination of cis- and trans-resveratrol in rat plasma by HPLC:: application to pharmacokinetic studies
Stereospecific determination of cis- and trans-resveratrol in rat plasma by HPLC:: application to pharmacokinetic studies
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DOI：
10.1002/bmc.747
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发表时间：
2007-03-01
影响因子：
1.8
通讯作者：
Yu, Qiaoling
中科院分区：
文献类型：
--
作者：
Chen, Xijing;He, Hui;Yu, Qiaoling
建立了一种简单、准确、精密、特异和重现性好的高效液相色谱（HPLC）方法，用于同时测定大鼠血浆中的白藜芦醇异构体。顺式白藜芦醇的制备方法是将反式白藜芦醇溶液暴露在阳光下 5 天，然后通过 HPLC 分离并通过质谱 (MS) 鉴定。测定程序涉及从少量血浆中将白藜芦醇异构体和内标（IS，咖啡因）直接液-液萃取到乙腈中。向上清液中加入等体积的水，注入Hypersil ODS2 C-18柱(5μm，4.6×250mm)上。使用由甲醇和蒸馏水组成的流动相，流速为 1.0 mL/min，有效分离顺式、反式白藜芦醇和咖啡因 (IS)。通过使用设置在 303 nm 的 UV 检测器监测洗脱液来实现分析物峰的检测。分析物的峰面积与 IS 的比率用于血浆样品的定量。顺式、反式白藜芦醇和内标的标称保留时间分别为 3.2 分钟、4.3 分钟和 6.1 分钟。校准曲线在0.066至6.64和0.134至13.4μg/mL范围内呈线性，反式和顺式异构体的相关系数分别为0.9998和0.9997。两种异构体的绝对回收率均超过85%。质量控制 (QC) 样品（0.066、0.664 和 6.64 μg/mL 反式白藜芦醇）测量的日间和日内精密度分别在 2.37-6.95% 相对标准偏差 (RSD) 和 0.77-6.97% RSD 范围内。质量控制 (QC) 样品（0.134、1.34 和 13.4 μg/mL 顺式白藜芦醇）测量的日间和日内精密度分别在 1.93-3.72% 相对标准偏差 (RSD) 和 1.13-6.57% RSD 范围内。分析物和内标在一系列稳定性研究和冻融循环中都很稳定。发现白藜芦醇异构体在 -20 摄氏度下储存 30 天是稳定的。描述了应用该测定法来确定大鼠单次口服剂量后的药代动力学分布。版权所有 (c) 2007 John Wiley & Sons, Ltd.
A simple, accurate, precise, specific and reproducible high-performance liquid chromatography (HPLC) method was developed for simultaneous determination of resveratrol isomers in rat plasma. Cis-resveratrol was made by exposure of a trans-resveratrol solution to sunlight for 5 days followed by separation by HPLC and identification by mass spectrometry (MS). The assay procedure involved simple liquid-liquid extraction of resveratrol isomers and internal standard (IS, caffeine) from a small plasma volume directly into acetonitrile. The supernatant liquid was added an equal volume of water and injected onto a Hypersil ODS2 C-18 column (5 mu m, 4.6 x 250 mm). Mobile phase consisting of methanol and distilled water was used at a flow rate of 1.0 mL/min for the effective separation of cis-, trans-resveratrol and caffeine (IS). The detection of the analyte peak was achieved by monitoring the eluate using a UV detector set at 303 nm. The ratio of peak area of analyte to IS was used for quantification of plasma samples. Nominal retention times of cis-, trans-resveratrol and IS were 3.2, 4.3 and 6.1 min, respectively. The calibration curve was linear ranging from 0.066 to 6.64 and 0.134 to 13.4 mu g/mL with correlation coefficients of 0.9998 and 0.9997 for trans and cis isomers, respectively. The absolute recovery of both isomers was more than 85%. The inter- and intra-day precisions in the measurement of quality control (QC) samples, 0.066, 0.664 and 6.64 mu g/mL of trans-resveratrol, were in the range 2.37-6.95% relative standard deviation (RSD) and 0.77-6.97% RSD, respectively. The inter- and intra-day precisions in the measurement of quality control (QC) samples, 0.134, 1.34 and 13.4 mu g/mL of cis-resveratrol, were in the range 1.93-3.72% relative standard deviation (RSD) and 1.13-6.57% RSD, respectively. Both analytes and IS were stable in the battery of stability studies and freeze-thaw cycles. Resveratrol isomers were found to be stable for a period of 30 days on storage at -20 degrees C. The application of the assay to determine the pharmacokinetic disposition after a single oral dose to rats is described. Copyright (c) 2007 John Wiley & Sons, Ltd.