液－固萃取高效液相色谱测定全血环孢菌素A

Determination of cyclosporine A in whole blood using liquid-solid extraction by high performance chromatograph

为了达到省时经济、简便敏感、准确快速的检测全血汗孢菌素Ａ，我们根据液－固萃取的原理，用自制Ｃ＿（１８）小柱进行样本的预处理，乙睛、水、三氟乙酸体系为洗脱剂、Ｃ＿（１８）分析柱（柱温７０℃），检测波长２１０ｎｍ，以环孢菌素Ｄ为内标的全血环孢菌素反相高效液相色谱测定法。本方法的线性范围为５０～１５００ｎｇ／ｍｌ，回收率为９９％～１０４％，批内变异系数＜４．７％，批间变异系数＜６．５％，临床上与ＣｙｓＡ常同时使用的３５种药物的病人全血按上述前处理，再使用ＨＰＬＣ分析表明在ＣｙｓＡ和ＣｙｓＤ的保留时间内均未见干扰峰；本方法使用了自制Ｃ＿（１８）小柱替代了进口小柱，效果满意，因而适合在国内临床上推广使用。

n liquid-solid extracted and HPLC analysis of cyclosporine A in whole blood was reported,1ml whole blood specimens mixed with 3ml extraction/internal standard solution,after centrifugation the su-pernate containing the cyclosporine is pass through a C_(18) minicolumn. The cyclosporines are eluted with 400μl 95% ethanol and collecting the elution,add 200μl water and 500μl hexane to it,vortex well and allow vials stand several minutes for phase separation;5 0μl ethanol layer was injected into reversed-phase column (33mm× 4. 6mm, 3μm; ODS)at 70℃,which was eluted with a mobile phase of acetoni-trite/water(70%/30%,and trifluoroacetic acid(25μl/L mobile phase ) at flow rate 1. 0ml/min and monitored at 210nm. The whole time of chromatograph is 12 min.The analystical recovery were 93%～104%and the inter and intra CV was below 4.7%and 6. 5%respectively. The detection limit was 20ng/ml and linearity was within 50～1500ng/ml. No interfer-ance was observed.