UPLC/MS/MS法同时测定人血浆中人参皂苷Re和Rg1含量及对参附冻干粉针剂的药代动力学评价

Quantitative Determination of Ginsenoside Re and Rg1 in Human Plasma by Ultra Performance Liquid Chromatography-tandem Mass Spectrometry

目的在人血浆中建立一种快速灵敏的同时测定人参皂苷Re和Rg1含量的超高效液相色谱串联质谱(UPLC/MS/MS)检测方法。方法采用Acquity UPLCTM BEHC18分析柱(1.7μm,2.1×100mm),三重四级杆质谱检测器,在多反应检测模式(MRM)下分别选择质荷比(m/z)969.6→789.3、m/z823.5→643.2和m/z803.5→387.1对Re、Rg1和内标地高辛进行检测。18名健康志愿者提供的324份血浆样品用该方法进行检测,并进行参附冻干粉针剂的药代动力学研究。结果每个样品仅需要50μL的血浆,洗脱时间仅需4min。Re和Rg1的最低检测限均为0.025ng/mL。在0.1～200ng/mL内线性关系良好(r2>0.999)。人参皂苷Re及Rg1在人血浆中的药动学参数均符合三室开放模型。结论本法具有灵敏、快速的特点,适用于人参皂苷Re、Rg1血药浓度的同时测定以及静脉滴注参附冻干粉针剂药代动力学的研究。

Objective To develop a rapid and sensitive method for the pharmacokinetie study of ginsenoside Re and Rgl in SHEN-FU injective powder in human plasma by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry （UPLC/ESI/MS/MS）. Methods A Waters C18 column （1.7μm, 2.1× 100 mm） was used in this study. The detection of Re and Rgl was performed on a triple-quadruple mass spectrometer with multiplereaction monitoring （MRM） mode using the following transitions., m/z 969.6→89. 3 for Re; m/z 823.5→643.2 for Rg1 and m/z 803.5→387. 1 for digoxin. A total of 324 plasma samples from 18 healthy volunteers were tested. Results A total run could be accomplished in 4 minutes. Only 50 μL plasma sample was needed to detect Re and Rgl. The lowest detectable concentration for both Re and Rg1 was 0. 025 ng/mL. Good linearity appeared from 0. 1 to 200 ng/mL （r^2〉0. 999）. The decline of Re and Rgl in plasma could be described by a triple-compartment model. Conclusion The proposed method provides a rapid and sensitive method for the quantification of Re and Rgl in human plasma, which has been successfully applied to the pharmaeokinetic study on intravenous infusion of SHENFU powder.