Liang Yuan, Ma Tiancheng, Li Yuwei, Cai Na
Department of Genetics, Northwest Women's and Children's Hospital, China.
Department of Orthopedics, The First Affiliated Hospital of Air Force Medical University, China.
Biomed Chromatogr. 2022 Jan;36(1):e5248. doi: 10.1002/bmc.5248. Epub 2021 Oct 14.
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 column (2.1 × 100 mm, 1.8 μm) 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 μg/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.
香草酸是一种从当归和绿茶中分离出的酚类化合物，具有出色的抗氧化和抗炎活性。在本研究中，建立并验证了一种快速灵敏的超高效液相色谱串联质谱法，用于测定大鼠血浆中的香草酸。血浆样品通过乙腈蛋白沉淀法制备。采用Zorbax RRHD Eclipse Plus C18柱（2.1×100 mm，1.8μm）进行色谱分离，梯度洗脱，流速为0.3 ml/min，流动相由0.1%甲酸（A）和乙腈（B）组成。香草酸和咖啡酸（内标，IS）在负离子模式下通过多反应监测进行定量。该方法根据美国食品药品监督管理局的指南进行了全面验证。校准曲线在2-1000 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、日内和日间精密度及准确度均在可接受范围内。然后将该方法应用于大鼠香草酸的药代动力学研究。分别以2、5和10 mg/kg的剂量口服给药后，血浆浓度在0.55-0.64 h时分别达到峰值0.42±0.09、0.73±0.21和0.92±0.28μg/ml。大鼠血浆中的口服生物利用度计算为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。
