反相高效液相色谱法测定全血环孢霉素A的含量

目的　探讨反相高效液相色谱法 (RP -HPLC)测定全血中环孢霉素A(CyA)的含量。方法　以液 -液提取法萃取、C8(4.6mm× 15 0mm)色谱柱 乙腈 甲醇 水 =6 4 10 2 6作流动相 ,在 6 5℃、2 14nm波长下、CyD为内标分离 ,检测全血中CyA的含量。 结果　在 4 5～ 10 0 3ng·ml-1范围内线性关系良好 ,回归方程及相关系数分别为 :cCyA=0 .5 75ACyA/ACyD+0 .0 12 6 ,r=0 .9998。结果的相对标准偏差 (RSD)均在 10 %以内 ,平均回收率为 99.9% ,最低检测浓度为 2 0ng·ml-1。结论　本方法简便、快速、准确 ,线性范围宽 ,可用于临床和科研工作中CyA浓度的快速检测。

Determination of cyclosporine A in human whole blood by RP-HPLC

Objective To exproe the method of measurement of the content of cyclosporine A(CyA) in human whole blood by reverse phase high-performance liquid chromatography(RP-HPLC) method.Method CyA and internal standard-cyclosporine D (CyD) were extracted from whole blood by liquid-liquid extraction, then separated on a C8 column at 65℃, 214nm. The mobile phase was acetonitrile-methanol-water(64∶10∶26).Results The calibration curve was linear in the range of 45～1003?ng·ml -1 with regression equation and correlation coefficient:c CYA =0.575 A CYA /A CYD +0..0126,r=0.9998.The relative standard deviation values of within-day and between-day were all lower then 10%. The average recoveries of CyA was 99.9%. The lowest limit of detection of CyA was 20?ng·ml -1 .Conclusion The method is rapid, simple, accurate and could be used for the determination of cyclosporine A in the clinical and scientific research.