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目的:建立一种简便可行的RP-HPLC法测定全血中环孢素(CsA)的浓度,为临床合理应用CsA提供参考。方法:全血样品1ml经乙醚单步提取,正已烷洗涤去干扰后采用C_(18)柱(200mm×4.6mm,5μm)分离;流动相为乙腈-甲醇-水(72:6:22);流速为1.5ml·min~(-1);柱温为60℃;检测波长为210nm;环孢素D(CsD)为内标。结果:CsA全血浓度在50～1000ng·ml~(-1)范围内呈良好的线性关系(r=0.9996),平均方法回收率为96.6%～99.2%,平均萃取回收率为59.5%～65.3%;日内、日间RSD均小于6%,最低检测限为10ng·ml~(-1)。结论:本法简便、快速、准确可靠,用于移植术后患者及再生障碍性贫血患者的CsA血药浓度监测,效果良好。
OBJECTIVE: To establish a simple and feasible RP-HPLC method for the determination of CsA in whole blood and to provide a reference for the clinical application of CsA. Methods: 1 ml of whole blood samples were extracted by diethyl ether, separated by C18 column (200 mm × 4.6 mm, 5 μm) after the n-hexane was washed to remove interference. The mobile phase consisted of acetonitrile-methanol- water (72: 6: 22) ; The flow rate was 1.5ml · min -1; the column temperature was 60 ℃; the detection wavelength was 210nm; CsD was the internal standard. Results: The linear range of CsA whole blood was 50 ~ 1000ng · ml ~ (-1) (r = 0.9996). The average recoveries were 96.6% ~ 99.2% and the average recoveries were 59.5% ~ 65.3 %. The intra-day and inter-day RSD were less than 6%, the lowest detection limit was 10ng · ml -1. Conclusion: The method is simple, rapid, accurate and reliable, and is suitable for the monitoring of CsA blood concentration in patients after transplantation and in patients with aplastic anemia.