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

摘要

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1．0 mL·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 =0．9962),定量下限为0．05 mg·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 sam-ples were extracted with methanol and analyzed by HPLC. The mobile phase was methanol - 0. 1% for-mic acid water (78 ∶ 22, V/ V), with a flow rate of 1. 0 mL·min - 1 and UV 275 nm as the detection wave-length. The plasma concentration of 1,8-TMP rhein in rats was determined by HPLC after single-dose intrave-nous injection in rats with 2,4 and 8 mg·kg - 1 of 1,8-TMP rhein, and the pharmacokinetic parameters were caclulated by DAS 2. 1. Results The result of cali-bration curve was linear over the range of 0. 05 ~ 10. 00 mg·L - 1 (r = 0. 996 2). The lower limit of quantifica-tion 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% , respec-tively. The validated method was successfully applied to a pharmacokinetic study after i. v. administration of 1,8-TMP rhein in rats with a dose of 2,4 and 8 mg· kg - 1 . The T1 / 2 was (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.