RRLC-MS/MS法测定大鼠血浆中咖啡酸四聚体的含量及药代动力学研究  

Determination of caffeic acid tetramers in rat plasma by RRLC-MS/MS and pharmacokinetic study

作　　者：毛艳 张瑞萍[2] 贺金华 王新堂 王中华[2] 王惠青[2] 周帜 

机构地区：[1]新疆维吾尔自治区药物研究所,乌鲁木齐830004 [2]天然药物活性物质与功能国家重点实验室、中国医学科学院北京协和医学院药物研究所,北京100050

出　　处：《药物分析杂志》2014年第10期1771-1776,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：新疆维吾尔自治区自然科学基金(201233146-8)

摘　　要：目的：建立大鼠血浆中咖啡酸四聚体（CAT）的测定方法,并研究其药代动力学。方法：大鼠尾静脉注射CAT后不同时间经眼静脉采血,血浆样品经液-液萃取后,采用RRLC-MS/MS法,色谱柱为Agilent SB-C18（4.6 mm×100 mm,1.8μm）,流动相为0.1%甲酸溶液（A）-乙腈（B）,梯度洗脱（0~3 min,68%A;3~6.5 min,68%A→2%A;6.5~10 min,2%A）,流速0.4 mL·min-1,柱温30℃,进样量10μL;质谱采用电喷雾离子源（ESI源）,正离子模式检测,扫描方式为多反应监测（MRM）。结果：咖啡酸四聚体检测限和最低定量限分别为5.7和11.4 ng·mL-1;在11.4~1420 ng·mL-1浓度范围内线性关系良好（r=0.9991）;提取回收率为85.2%~97.1%;药动学参数：AUC0-t为（1.25±0.25）mg·L-1·h,t1/2为（4.66±1,07）h-1,K为（0.15±0.04）h-1,CLz为（3.19±0.52）L·h-1·kg-1,Vz为（1.87±0.59）L·kg-1。结论：本方法专属性强、灵敏度高、准确性好,可用于咖啡酸四聚体的血药浓度测定。Objective ：To establish a method to determine the plasma concentration of caffeic acid tetramer（CAT） in rats by using RRLC - MS/MS, and study the pharmacokinetics of CAT. Methods： CAT in rat tail vein and plasma samples were collected at different time points through vena ophthalmica. After a liquid - liquid extraction process with methl tert - mutyl ether, the HPLC separation for plasma samples was performed on a Agilent SB - Cj8 column （4. 6 mm × 100 mm,1.8 μm）in gradient elute mode（0 -3 rain,68% A;3 -6. 5 min,68% A→2% A;6. 5 - 10 rain, 2% A）with water containing 0. 1% formic acid （A）and acetonitrile （B）as the mobile phase at a flow rate of 0. 4 mL · min-1. MS detection was performed with multiple reaction monitoring（MRM） mode using positive electrospray ioniza- tion. Results ：The linearity was achieved in the range of 11.4 - 1420 ng · mL -1 （r =0. 9991 ）for CAT. The extraction recovery of CAT was from 85.2% - 97. 1%. The lowest limit of detection （LOD） and quantification （ LOQ ） of caffeic acid tetramer were 5.7 ng · mL - 1 and 11.4 ng · mL - 1. The pharmacokinetic parameters of AUC0-t, t1/2, K, CLz and Vz were as follows：（1.25 ±0.25）mg · L-1 · h,（4. 66 ± 1,07）h-1,（0. 15 ±0.04）h-1,（3. 19 ±0.52）L · h-1 · kg-1, and （ 1.87± 0. 59） L · kg - 1. Conclusion： A sensitive, accuracy and suitable RRLC - MS/MS method for determining CAT was developed.

关 键 词：紫草 咖啡酸四聚体 含量测定 液相色谱-串联质谱 药动学 

分 类 号：R917[医药卫生—药物分析学]