LC-MS/MS法测定大鼠血浆中人参皂苷Rb1的含量及其药代动力学研究

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,3.5μm)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Rb15 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。

Determination of ginsenoside Rb1 in rat plasma by LC-MS/MS and its application in pharmacokinetic study

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,3.5μm)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.