LC-MS/MS法研究中药莲必治注射液在大鼠体内的药动学

Pharmacokinetics of Chinese Medicine Lianbizhi Injection in Rats by LC-MS/MS

目的建立LC-MS/MS法测定大鼠血浆中莲必治注射液主要有效成分亚硫酸氢钠穿心莲内酯（ASB）浓度,研究大鼠静脉注射莲必治注射液的体内药动学。方法利用甲醇提取血浆中药物,以脱水穿心莲内酯（DAG）为内标,在Hypersil Gold C18色谱柱（50mm×2.1mm,1.9μm）上以流动相甲醇-水梯度洗脱,流速0.2mL·min^-1,柱温35℃,分析时间6min。在电喷雾离子化电离源上以选择反应监测方式进行负离子检测,用于定量分析的离子反应分别为m/z 413.2→287.2（ASB）和m/z 331.2→303.3（DAG）。大鼠静脉注射80mg·kg^-1莲必治注射液,收集不同时间点血样,测定药物浓度。结果ASB血浆标准样品的线性范围为10-1000ng·mL-1,方法定量下限为10ng·mL-1。方法的日内和日间精密度（RSD）在6.9%和10.8%以内,日内和日间准确度分别在96.4%-103.2%和93.7%-98.0%,提取回收率为71.2%-98.9%,基质效应为88.9%-99.9%。莲必治的末端消除半衰期t1/2λ为1.72±1.24h,表观分布容积Vd为2.33±1.98L·kg^-1,表观清除率CL为0.87±0.39L·h-1·kg^-1。结论本方法专属性好、准确度高,灵敏度高,适用于莲必治体内药动学研究。莲必治大鼠药动学研究可为莲必治注射剂安全性再评价提供参考。

Objective To establish a liquid chromatography-tandem mass spectrometry（LC-MS/MS）method for the quantification of the major effective component of Lianbizhi injection（andrographolide sodium bisulphite,ASB）in rat plasma,and to investigate pharmacokinetics of ASB in rats by following single intravenous administration. Methods ASB and an internal standard dehydroandrographolide（DAG） were extracted from plasma by using methanol and were separated on a Hypersil Gold C18column（50mm×2.1mm,1.9μm）by using gradient mobile phase of methanol and water at a flow rate of 0.2mL·min^-1 with the temperature of 35℃.The analysis lasted for 6min.At negative electrospray ionization mode,selected reaction monitoring of the precursor-product transitions m/z 413.2→287.2for ASB and m/z 331.2→303.3for DAG was used for the quantification.Plasma was collected after rats were intravenously administered with ASB at a single dose of 80 mg· kg^-1 at differenttime,and the quantification of the medicine is investigated.Results The linear calibration curve was obtained in a concentration range of 10-1 000ng·mL-1 with a lower limit of quantification of 10ng·mL-1.Intra-and inter-day precision was less than6.9% and 10.8% respectively.Intra-and inter-day accuracy fell in the ranges of 96.4%-103.2% and93.7%-98.0% respectively.The recovery ranged from 71.2% to 98.9%,and the matrix effects from88.9%to 99.9%.Pharmacokinetic parameters were obtained,t1/2λof 1.72±1.24 h,V of 2.33±1.98L·kg^-1,CL of 0.87±0.39L·h-1·kg^-1.Conclusion The study on specific,accurate,sensitive and suitable for in vivo pharmacokinetic experiment of Lianbizhi.pharmacokinetics of Lianbizhi injection in rats provides valuable information for safety reevaluation of Chinese medicine injection.