Determination of ginsenoside Rb1 in rat plasma by LC-MS/MS and its application in pharmacokinetic study
LI Shu, SHI Chao, GUO Yushu
Department of Pharmacy, Medical Supplies Center of Chinese PLA General Hospital, Beijing 100048, China
摘要：采用液相色谱-串联质谱法快速、灵敏地测定大鼠血浆中人参皂苷Rb1(GRb1)的含量，并将该方法应用于大鼠口服GRb1后的代谢动力学研究。血浆样品采用96孔板进行液-液萃取后，应用Agilent SB-C18色谱柱(100 mm×2.1 mm，3.5 μm)进行分离，以甲醇-0.1%甲酸溶液(体积比为75∶25)为流动相进行洗脱，在正离子模式下对GRb1和内标人参皂苷Rg1(GRg1)进行检测，用于定量的离子反应分别为1131.5→365.1(GRb1)，823.3→643.4(GRg1)。人参皂苷Rb1血浆样品测定方法的定量线性范围为1～500 ng/mL，线性相关系数大于0.999，定量下限为1 ng/mL，批内和批间精密度(RSD)小于9.05%，回收率为79.7%～81.0%，基质效应为96.6%～99.3%。大鼠灌胃给予Rb1 5 mg/kg后，大鼠体内血药浓度到达高峰时间tmax为1.53 h，半衰期t1/2为13.54 h，药时曲线面积AUC0～72为16237.76 (ng·h)/mL。该方法快速、高效、灵敏，适用于人参皂苷Rb1的代谢动力学研究。
Abstract：A rapid and sensitive LC-MS/MS method for determination of ginsenoside Rb1 (GRb1) in rat plasma was developed to the pharmacokinetic study of ginsenoside Rb1 in rats. Sample preparation procedure utilized liquid-liquid extraction with n-butanol in 96-well format. The analysis was performed on an Agilent SB-C18 column (100 mm×3.5 mm, 3.5 μm) at 25℃with the mobile phase composed of methanol-0.1% formic acid soulution(Volume ratio was 75∶25). Multiple reaction monitoring of the precursor-to-product ion pairs for GRb1 (1131.5→365.1) and ginsenoside Rg1 (GRg1)(823.3→643.4) was used for quantitation. The limit of quantification (LLOQ) was 1 ng/mL, the linear range was 1-500 ng/mL, the linearly dependent coefficient was more than 0.999. The within and between-run precision was less than 9.05%. The recovery of GRb1 was 79.7%-81.0%, and average matrix effect was 96.6%-99.3%. The main pharmacokinetic parameters after oral administration of GRb1 at a single dose of 5 mg/kg were as follows: tmax was 1.53 h, t1/2 was 13.54 h, AUC0-72 was 16237.76 (ng·h)/mL. The method is rapid, effective and sensitive, which can be applied to the pharmacokinetic study of ginsenoside Rb1 in rats.
Keywords：Ginsenoside Rb1; LC-MS/MS; pharmacokinetics
LI S，SHI C，GUO Y S. Determination of ginsenoside Rb1 in rat plasma by LC-MS/MS and its application in pharmacokinetic study[J]. Chemical analysis and meterage，2020，29(3)：34-38.