A rapid and sensitive LC-MS/MS method for the determination of vanillic acid in rat plasma with application to pharmacokinetic study
A rapid and sensitive LC-MS/MS method for the determination of vanillic acid in rat plasma with application to pharmacokinetic study
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DOI：
10.1002/bmc.5248
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发表时间：
2021-10-14
影响因子：
1.8
通讯作者：
Cai, Na
中科院分区：
文献类型：
--
作者：
Liang, Yuan;Ma, Tiancheng;Cai, Na
香草酸是从当归和绿茶中分离出来的酚类化合物，具有优异的抗氧化和抗炎活性。本研究建立并验证了快速、灵敏的超高效液相色谱串联质谱测定大鼠血浆中香草酸的方法。通过用乙腈沉淀蛋白质来制备血浆样品。在 Zorbax RRHD Eclipse Plus C-18 柱（2.1 x 100 mm，1.8 μm）上进行色谱分离，以 0.3 ml/min 的流速进行梯度洗脱，使用由 0.1% 甲酸（A）和乙腈（B）组成的流动相。通过负离子模式下的多重反应监测对香草酸和咖啡酸（内标，IS）进行定量。该方法根据美国食品和药物管理局指南得到了充分验证。校准曲线在 2-1,000 ng/ml 范围内呈线性，相关系数 >0.99。残留、基质效应、提取回收率、稀释效应、日内和日间精密度和准确度均在可接受的范围内。然后将该方法应用于香草酸在大鼠体内的药代动力学研究。口服2、5和10mg/kg剂量后，血药浓度分别在0.55-0.64小时时达到峰值，分别为0.42±0.09、0.73±0.21和0.92±0.28μg/ml。经计算，大鼠血浆中的口服生物利用度为25.3-36.2%。该结果为香草酸的进一步应用提供了临床前信息。
Vanillic acid, a phenolic compound isolated from Angelica sinensis and green tea, exhibits excellent antioxidant and anti-inflammatory activities. In this study, a rapid and sensitive ultra-high-performance liquid chromatography tandem mass spectrometry method was established and validated for the determination of vanillic acid in rat plasma. Plasma samples were prepared by protein precipitation with acetonitrile. Chromatographic separation was performed on a Zorbax RRHD Eclipse Plus C-18 column (2.1 x 100 mm, 1.8 mu m) with gradient elution at a flow rate of 0.3 ml/min, using mobile phase consisting of 0.1% formic acid (A) and acetonitrile (B). Vanillic acid and caffeic acid (internal standard, IS) were quantified by multiple reaction monitoring in negative ion mode. The method was fully validated according to the US Food and Drug Administration guidelines. The calibration curve was linear over the range of 2-1,000 ng/ml with a correlation coefficient of >0.99. The carryover, matrix effect, extraction recovery, dilution effect, intra- and interday precision and accuracy were within acceptable limits. The method was then applied to a pharmacokinetic study of vanillic acid in rats. After oral administration at doses of 2, 5 and 10 mg/kg, the plasma concentration reached peaks of 0.42 +/- 0.09, 0.73 +/- 0.21 and 0.92 +/- 0.28 mu g/ml at the time of 0.55-0.64 h, respectively. The oral bioavailability was calculated as 25.3-36.2% in rat plasma. The result provided pre-clinical information for further application of vanillic acid.