液相色谱电喷雾质谱联用测定器官移植患者全血中环孢素A

Determination of cyclosporine A in the whole blood of organ transplantation patients by HPLC/electrospray ionization-MS(HPLC/ESI^+-MS)

目的 :建立液相色谱 -电喷雾质谱联用 (HPLC/ESI+ MS)测定器官移植患者全血中环孢素A(CsA)方法。方法 :0 .5mL患者全血样品用乙醚单步提取 ,色谱柱为岛津C18柱 ,流动相为乙腈 :醋酸铵水溶液 (90∶10 ) ,流速为 0 .95mL·min-1,分流比为 3∶1;质谱条件采用电喷雾正离子源 (ESI+ ) ,毛细管电压为 3.96kV ,一级锥孔电压为 130V ,选择离子监测 (SIR)通道CSA采用 12 0 3+12 2 5m/z,内标环孢素D(CsD)采用 12 17+12 39m/z。结果 :在 9～ 12 0 0 μg·L-1浓度范围内 ,CsA线性关系良好 ,最低检测浓度为 1μg·L-1,日内RSD(n =7)与日间RSD(n =5 )均低于 10 %。方法平均回收率为 98%。结论 :本方法灵敏 ,色谱图几乎无杂质干扰 ,操作步骤简单 ,适于器官移植术后CsA的血药浓度监测 ,也可用于CSA药动学的研究。

ABSTRACT:OBJECTIVE To develop an HPLC/ESI + MS method for quantifying cyclosporine A (CsA)in the whole blood of renal transplantation patients.METHODS Blood sample ( 0.5 mL ) was extracted with ether.The extracted sample was analyzed on liquid chromatograph using a C 18 column with acetonitrile NH 4AC ( 20 mmol ·L -1 ) (90∶10) as mobile phase.The flow rate was 0.95 mL ·min -1 .The split ratio was 3∶1. Mass spectrometry conditions:ESI +.The voltages of capillary and cone were 3.96 kV and 130 V respectively.The ions for quantification were m/z 1203+1225 for CsA,m/z 1217+1239 for cyclosporine D(CsD)(internal standard).RESULTS The linear calibration curve was obtained in the concentration range of 9～ 1200 μg ·L -1 .The limit of detectable concentration was 1 μg ·L -1 .The intra ( n =7) and inter day RSD ( n =5) were less than 10%.The average recovery was 98%.CONCLUSIONS The method is sensitive,specific and easy to operate.It is suitable for determination of CsA in the whole blood of organ transplantation patients and pharmacokinetic study of CsA.