LC-MS/MS法测定大鼠血浆中的左旋紫堇达明及其药代动力学研究

Determination of L-corydalmine in rat plasma by LC-MS/MS and the pharmacokinetic studies

目的建立快速、灵敏的LC-MS/MS分析方法定量测定大鼠血浆中左旋紫堇达明（L-CDL）的浓度,经系统方法学验证后,应用于该药的大鼠药代动力学研究。方法以普萘洛尔为内标,以水（含0.1%甲酸与5 mmol/L的甲酸铵）和乙腈（含0.1%甲酸）为流动相,血浆样品中的L-CDL和内标经C18（2.1 mm×750 mm,3μm）柱分离后,质谱电喷雾离子化源（ESI）正离子多反应监测（MRM）方式分别对离子对m/z 342？192.1和m/z 260？116.2进行检测。方法验证包括专属性、标准曲线与定量下限、批内和批间精密度、准确度、基质效应、提取回收率、稳定性。结果 L-CDL在大鼠血浆样品的定量线性范围为15000 ng/ml,最低定量下限为1 ng/ml,批内和批间精密度（RSD）〈10%,回收率93.5%102.8%,基质效应108.3%112.5%。大鼠经胃给予LCDL 10 mg/kg后的主要药动学参数Cmax、T1/2、AUC0-24分别为：（557.8±330.1）ng/ml、（7.04±3.93）h、（2408±630）h·ng/ml。结论建立的LC-MS/MS方法简便、快速、灵敏,适用于L-CDL在大鼠体内的药代动力学研究。

Objective To develop a simple,rapid and accurate analysis method for determination of L-corydalmine（L-CDL）in rat plasma. Methods The plasma sample was determined with propranolol as the internal standard（IS）,the separation was accomplished in a Capcell PAK C18（2.1 mm×750 mm,3 μm）column,and the mobile phase consisted of water（including 0.1% formic acid and 5 mmol/L ammonium formate）and acetonitrile（including 0.1% formic acid）at a flow rate of 0.25 ml/min. The mass spectrometer consisted of an ESI interface operating at positive ionization mode and the detection was performed using multiple reaction monitoring（MRM）at the transitions of m/z 342？192.1 for L-CDL and m/z 260？116.2 for propranolol. Method validation included the evaluation of the linearity range,lower LOQ,within-run and between-run precision,accuracy and stability,matrix effect and extraction recovery. Results The calibration curve was linear across the concentration range of 1-5000 ng/ml for L-CDL with a lower LOQ of 1 ng/ml. The within-run and between-run precision（RSD%）was in the range 0.1%-10%. The extraction recovery was in 93.5%-102.8% and the matrix effect for three QC was 108.3%-112.5%. L-CDL reached the peak concentration at 0.5 h after dosing. The main pharmacokinetic parameters of rats after igl administration were as follows：T1/2：（7.04±3.93）h,Cmax：（557.8±330.1）ng/ml,AUC0~24：（2408±630）h·ng/ml. Conclusion A simple,rapid,accurate,high sensitivity and repeatability method has been successfully developed,which can analyze the concentration of L-CDL in rat plasma. The method can be used for the investigation of pharmacokinetics of L-CDL in rats.