RP-HPLC法测定人全血中环孢素的浓度

Determination of Cyclosporine A Concentration in Whole Human Blood by RP-HPLC

目的:建立测定人全血中环孢素浓度的方法。方法:采用反相高效液相色谱法,血样经乙醚萃取、正己烷洗涤后进样测定。色谱柱为Nova-pakC18,流动相为乙腈-甲醇-水(30.8∶51.2∶18),流速为1mL·min-1,柱温为55℃,检测波长为210nm。结果:CsA血药浓度在50～1550ng·mL-1范围内线性关系良好(r=0.9999);平均方法回收率为99.90%～102.13%,平均提取回收率为64.27%～66.58%;日内RSD为2.86%～6.08%,日间RSD为2.95%～6.27%。结论:本法快速、灵敏、准确、实用,适合临床环孢素治疗药物监测的需要。

OBJECTIVE： To establish a method to determine the concentration of cyclosporine A （CsA） in whole human blood. METHODS： RP-HPLC was adopted. CsA was extracted from whole blood with diethyl ether and the endogenous interference was removed from blood sample with n-hexane. The separation was performed on Nova-pak C18 column with mobile phase consisted of acetonitrile-methanol-water （30.8：51.2：18）at the flow rate of 1 mL.min-1. The temperature of column was set at 55 ℃, and the detection wavelength was 210 nm. RESULTS： The linear range of CsA was 50-1 550 ng.mL-t （r=0.999 9）. The average method recovery was 99.90%~102.13% ; the average extraction recovery was 64.27%-66.58%. The intra-day RSD was 2.86%~ 6.08%, and the inter-day RSD was 2.95% ~6.27%. CONCLUSION： The established method is rapid, sensitive, accurate, and practical, and it is suitable for the drug monitoring of CsA in the clinic.