HPLC测定人全血中的环孢素A

Determination of Ciclosporin A in human blood by HPLC

目的采用RP-HPLC分析人全血中环孢素A（Cs A）的浓度。方法全血样本经乙醚两步萃取后,采用Diamonsil RP-C18柱（150 mm×4.6 mm,5μm）,流动相为乙腈-甲醇-水（70∶5∶25）,流速1.0 m L·min^-1,柱温60℃,检测波长215nm,内标为熊果酸。结果全血中Cs A的线性范围为0.0101~5.030μg·m L-1（r=0.9996）,检测限为4.02 ng·m L^-1,方法回收率为96.0%~106%,平均提取回收率为74.4%~77.5%,批内、批间的RSD不大于6.0%;室温（26℃）、8 h时稳定性的RSD不大于4.0%。结论所用方法灵敏、准确、精密,适宜于临床上Cs A血药浓度的监测。

OBJECTIVE To establish an HPLC method for the analysis of the concentration of Ciclosporin A （ CsA ） in human blood. METHODS The blood sample was extracted by ether with two steps and then analyzed by HPLC. Diamonsil RP - C18 column （ 150 mm× 4.6 mm ,5 μm） was used with the mobile phase consisted of acetonitrile - methanol - water（ 70 ： 5 ： 25 ） at the flow rate of 1.0 mL· min^-1. The column temperature was maintained at 60 ℃. The detected wavelength was 215 nm. Ursolic acid was used as the internal standard. RESULTS A good linear relationship was obtained between the concentration of CsA from 0. 0101 μg· mL^-1 to 5. 030 μg· mL^-1（ r2 = 0. 9996 ）. The limit of detection for the analysis of CsA was 4.02 ng· mL^-1. The recovery of method was 96.0% - 106%. The average extraction recovery rate was 74.4% - 77.5%. The intra - day and inter - day RSD were all less than 6. 0%. The human blood sample of CsA maintained at room temperature（ 26 ℃） for 8 h was stability with the RSD was less than 4.0%. CONCLUSION The method is sensitive, accurate, precision and suitable for therapeutic drug monitoring of CsA.