固相萃取-液相色谱／质谱法测定全血中环孢素A的血药浓度

Rapid Determination of Cyclosporine A in Whole Blood by LC/MS with SPE Sample Preparation

目的:建立高效液相色谱/质谱联用测定全血中环孢素A的血药浓度的分析方法。方法:样品用固相萃取小柱提取处理。液相色谱:流动相为乙腈-甲醇-0.01 mol·L-1乙酸铵水溶液(47:33:20),流速0.5 mL·min-1;色谱柱为ZORBAX SB-C18(4.6 mm×12.5 mm,5μm),柱温60℃。质谱:电喷雾电离源(ESI),选择性监测质荷比(m/z)为1203(环孢素A),1217(内标,环孢素D)带正电荷的分子离子峰。结果:线性范围为25-500μg·L-1,回收率94.7%-105%,日内、日间的RSD均小于5%。结论:本方法取样量少(0.2 mL全血),灵敏度高,操作简便,适用于临床上环孢素A的血药浓度监测及药动学研究。

Objective:To establish an LC/MS method for the rapid determination of cyclosporine A(CsA)in whole blood. Method: Whole blood samples were prepared by C18 solid phase extraction(SPE). LC mobile phase:acetoni-trile -methanol -0. 01 mol·L-1 ammonium acetate(47: 33: 20) ,flow rate:0. 5 mL· min-1;chromatography column: ZORBAX SB - C18 ( 4. 6 mm× 12. 5 mm, 5μm ) , column temperature: 60℃. MS: electrospray ( ESI) with multi -ion monitoring(selected ion monitoring, SIM ) :positive ions,m/z 1203 for cyclosporine A and m/z 1217 for cyclosporine D(CsD,internal standard). Results:Linear range for the calibration curve was between 25 -500μg·L-1 (r =0.9997). Spike recoveries were between 94.7% - 105% with within - day and day - to - day RSD lessthan 5%. Conclusion: The method is sensitive, rapid and reliable for monitoring cyclosporin A concentration inblood,and is suitable for clinics and pharmacokinetics studies.