苗药黑骨藤中绿原酸在大鼠体内药动学研究

Pharmacokinetic study of chlorogenic acid in rats administered Miao medicine Periploca forrestii Schltr.

目的采用UPLC-MS/MS法建立大鼠血浆中绿原酸含量测定的方法,并将其应用于黑骨藤中绿原酸在大鼠体内的药动学研究。方法采用沉淀蛋白法处理血浆样品,以葛根素为内标,色谱柱:ACQUITY UPLC CSH C18(150 mm×3.0 mm,1.7μm),流动相:乙腈-水(0.2%甲酸)(80:20)等度洗脱,流速:0.3 mL/min,柱温:25℃,进样量:2μL,采用电喷雾离子源(ESI),以正离子扫描,选择离子监测模式进行检测。结果血浆中绿原酸在0.018~6.750μg/mL范围内线性良好,定量限为:0.018μg/mL,日内和日间精密度RSD小于6.51%,准确度在96.73%~115.55%范围内,提取回收率在88.76%~113.28%范围内,基质效应在88.01%~99.83%范围内。绿原酸在高、中、低剂量组的药动学参数为:Cmax分别为0.177、0.370、0.757 mg/L;AUC(0-t)分别为0.584、1.717、2.857 mg·h/L;消除半衰期(t1/2z)分别为3.553,5.507,5.148 h。结论所建立的大鼠体内绿原酸分析方法灵敏度高、快速、选择性好;在6.4~25 mg/kg剂量范围内,黑骨藤中绿原酸在大鼠体内表现为线性动力学特征,各剂量下绿原酸药动学数据存在一定差异。

Objective To establish a method for the determination of chlorogenic acid in rat plasma by UPLC-MS/MS method,and apply it to the pharmacokinetic study of chlorogenic acid in rats admisnistered Periploca forrestii Schltr.Methods Plasma samples were processed by protein precipitation method,and puerarin was employed as an internal standard--column:ACQUITY UPLC CSH C18(150 mm×3.0 mm,1.7μm),mobile phase:acetonitrile-water(0.2%formic acid)(80:20)Isocratic elution,flow rate:0.3 mL/min,column temperature:25℃,injection volume:2μL.The electrospray ion source(ESI)was used to scan with positive ions and select the ion monitoring mode for detection.Results The linearity of chlorogenic acid in plasma was good,within the range of 0.018-6.750μg/mL;the lower limit of quantification was 0.018μg/mL;the intraday and interday precision RSD was less than 6.51%;the accuracy was within the range of 96.73%-115.55%;the recovery rate was in the range of 88.76%-113.28%;and the matrix effect was in the range of 88.01%-99.83%.The pharmacokinetic parameters of chlorogenic acid in the high,medium and low dose groups were:Cmax--0.177,0.370,0.757 mg/L;AUC(0-t)--0.584,1.717,2.857 mg/(L·h);Elimination half-life(t1/2z)--3.553,5.507,5.148 h,respectively.Conclusion The established method is sensitive,rapid and selective for the determination of chlorogenic acid in rats.The chlorogenic acid in Periploca forrestii Schltr.shows linear kinetic characteristics in rats within the dose range of 6.4-25 mg/kg,yet there are some differences in the pharmacokinetic data of chlorogenic acid at different doses.