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

关键词:  反相高效液相色谱法  环孢素A  血药浓度监测

Key words:  RP-HPLC  cyclosporin A  therapeutic drug monitoring

摘要:

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，3.5 μm) 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。

Abstract:

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 .