HPLC测定大鼠血浆中1,8-二川芎嗪基大黄酸及其药代动力学研究

Determination of 1,8-TMP rhein and its pharmacokinetics in rat plasma by HPLC

目的建立测定大鼠血浆中1,8-二川芎嗪基大黄酸浓度的HPLC方法,并对大鼠尾静脉注射给药1,8-二川芎嗪基大黄酸后,研究其在大鼠体内的药代动力学。方法采用大黄素为内标,血浆样品用甲醇沉淀蛋白后进行HPLC分析,流动相为甲醇-0.1%甲酸水(78∶22,V/V),流速为1.0m L·min-1,检测波长为275 nm。大鼠单剂量静脉注射2、4、8 mg·kg-11,8-二川芎嗪基大黄酸后,测定给药后不同时间点大鼠血浆中1,8-二川芎嗪基大黄酸的浓度,并对其血药浓度-时间曲线采用DAS 2.1软件拟合,计算药动学参数。结果 1,8-二川芎嗪基大黄酸在0.05-10.00 mg·L-1血浆浓度范围内线性关系良好(r=0.996 2),定量下限为0.05mg·L-1,提取回收率均大于88%,日内和日间差异RSD均小于6%,方法学考察均符合要求。1,8-二川芎嗪基大黄酸按2、4和8 mg·kg-1静脉注射给药后,在大鼠体内的T1/2分别为(68.35±1.36)、(69.32±2.21)、(69.32±2.03)min,AUC0-t分别为(101.03±24.9),(144.79±3.29),(231.92±19.30)min·mg·L-1。结论本实验所建立的HPLC方法操作简便、快速、专属性强,可用于1,8-二川芎嗪基大黄酸血药浓度分析及其药代动力学研究。

Aim To develop a HPLC method for the determination of the concentration of 1,8-TMP rhein in rat plasma and study the pharmacokinetics of 1,8-TMP rhein in rat plasma after single dose i. v. administration of 1,8-TMP rhein( 2,4,8 mg·kg- 1). Methods Emodin was used as an internal standard. Plasma samples were extracted with methanol and analyzed by HPLC. The mobile phase was methanol- 0. 1% formic acid water( 78 ∶ 22,V / V),with a flow rate of1. 0 mL ·min- 1and UV 275 nm as the detection wavelength. The plasma concentration of 1,8-TMP rhein in rats was determined by HPLC after single-dose intravenous injection in rats with 2,4 and 8 mg·kg- 1of 1,8-TMP rhein,and the pharmacokinetic parameters were caclulated by DAS 2. 1. Results The result of calibration curve was linear over the range of 0. 05 ~ 10. 00mg·L- 1( r = 0. 996 2). The lower limit of quantification was 0. 05 mg · L- 1. The intra-day and inter-day precision( RSD%) were both lower than 6%,and the extraction recoveries were higher than 88%,respectively. The validated method was successfully applied to a pharmacokinetic study after i. v. administration of1,8-TMP rhein in rats with a dose of 2,4 and 8 mg·kg- 1. The T1 /2was( 68. 35 ± 1. 36),( 69. 32 ± 2. 1)and( 69. 32 ± 2. 03) min,respectively. The AUC0- t was( 101. 03 ± 24. 90),( 144. 79 ± 3. 29) and( 231. 92 ± 19. 30) min · mg · L- 1,respectively.Conclusion A simple and specific HPLC method for the analysis of 1,8-TMP rhein is successfully developed and applied to a pharmacokinetic study in rat plasma.