液质联用法同时测定干血斑中环孢菌素A、西罗莫司、他克莫司和依维莫司药物浓度

建立了液相色谱-串联质谱法同时定量检测干血斑中4种临床常用免疫抑制剂的方法。打孔裁取直径8mm的干血斑样本,经甲醇-乙腈混合溶液提取和沉淀蛋白后取上清液氮气吹干,甲醇溶液复溶后进样分析。选用C18色谱柱分离,甲醇与水(均含2mM乙酸铵和0.1%甲酸)为流动相梯度洗脱,在电喷雾正离子电离(ESI+)和多反应监测(MRM)模式下检测。结果表明:环孢菌素A在20~1000ng/mL范围内,西罗莫司、他克莫司和依维莫司在2.0~100ng/mL范围内线性良好。该方法对病患友善(样本采血量仅30μL),提取回收率高(73.6%~110.8%),其准确度、精密度(CV)、残留效应和基质效应均符合生物样品的分析要求。该方法快速、高效、灵敏度高、精确度好,适用于干血斑中同时测定环孢菌素A、西罗莫司、他克莫司和依维莫司浓度。

LC-MS/MS method for simultaneous determination of cyclosporin A,sirolimus,tacrolimus and everolimus in dried blood spots

A dried blood spot sample was extracted by methanol-acetonitrile,and protein precipitated,the sample was evaporated by nitrogen and measured by methanol reconstitution.C 18 column(100mm×2.1mm,3μm)was used for separation methanol and water(containing 0.1%formic acid and 2mM of ammonium acetate)were used as mobile phase gradient elution.The measurement was carried out under electrospray positive ionization(ESI+)and multiple reaction monitoring(MRM)mode.The linear range of cyclosporin A was 20—1000 ng/mL,and that of sirolimus,tacrolimus and everolimus was 2.0—100 ng/mL.The blood sample volume of this method was only 30μL,and the extraction recovery rate was high(73.6%—110.8%).The accuracy,precision,residual effect of the methed and matrix effect meet the requirements of biological samples analysis.