改良RP-HPLC测定全血中环孢素A的浓度

Determination of Cyclosporin A in Human Whole Blood by Improved RP-HPLC

目的建立一种简便的RP-HPLC法测定全血中环孢素A（CsA）的浓度,为临床合理应用CsA提供依据。方法全血样品1 mL经乙醚提取,正己烷洗涤去干扰后,采用Zorbax SB-C18柱（4.6 mm×150 mm,3.5μm）分离,流动相为乙腈-甲醇-水-异丙醇（60∶10∶29∶1）,流速1.5 mL.min^-1,柱温70℃,检测波长210 nm;环孢素D（CsD）为内标。结果CsA全血浓度在25-1 600 ng.mL^-1内线性关系良好（r=0.999 8）,平均方法回收率99.51%,日内、日间RSD均小于5%,最低检测限5 ng.mL^-1。结论本法简便快速,灵敏度高,重复性好,线性范围广。用于患者全血中CsA浓度监测,效果良好。

OBJECTIVE To set up a practical RP-HPLC method to determine the concentration of cyclosporin A（CsA）in the whole blood.METHODS CsA was extracted from whole blood with diethyl and removed the endogenous interference in blood with n-hexane ether and separated using Zorbax SB-C18 column（4.6 mm×150 mm,3.5 μm）.Mobile phase was acetonitrile-methanol-deionized water-isopropyl（60∶10∶29∶1）.The flow rate was 1.5 mL.min^-1.The temperature of column was set at 70 ℃.The detecting wavelength was 210 nm.The internal standard was cyclosporine D.RESULTS A good linear relationship was obtained between the concentration of CsA from 25 ng.mL^-1 to 1 600 ng.mL^-1（r=0.999 8）.The average method recovery rate of CsA was 99.51%.The intra-and inter-day RSD were less than 5%.The detection limit of CsA was 5 ng.mL^-1.CONCLUSION The established method was simple,accurate,sensitive,reproducible and large-scale of detection range and suitable for monitoring blood-drug concentration of CsA.