UPLC(超高效液相色谱)/MS/MS联用技术测定全血中的西罗莫司

Determination of sirolimus in whole blood by UPLC/MS/MS

目的建立全血中西罗莫司的UPLC(超高效液相色谱)/MS/MS测定方法。方法全血样品中加入他克莫司(FK506)作内标,用叔丁基甲醚进行提取。以乙腈与含千分之五甲酸的水溶液为流动相(40∶50),色谱柱为Acquity UPLCTMBEHC8(50mm×2.1mm,1.7μm),流速0.5ml/min。三重四极杆质谱采用正离子模式,离子采集方式为多反应监测模式(MRM),离子源温度105℃,离子源电离电压为3300V,雾化气流速500L/h。采集离子(母离子/子离子)西罗莫司为931.2/864.1,FK506为822.0/577.0。结果西罗莫司在1～240μg/L浓度范围内呈良好的线性(r=0.9995)。日内、日间精密度均在15%以内,提取回收率大于75.0%,方法回收率为95.0%～98.5%,最低检测限为0.2μg/L。结论本方法灵敏、准确,适合临床西罗莫司的全血分析。

Objective To establish a UPLC/MS/MS method for the detemination of sirolimus in whole blood. Methods The blood sample was extracted with methyl tert-butyl ether and separated over an ACQUITY UPLCTM BEH C8 （50mm ×2. 1mm, 1.7pro）. The mobile phase consisted of a mixture of water （0.5% formic acid） and acetonitrile （50 ： 40）. The flow rate was 0.5ml/min. Tacrolimus （FK506） was used as an internal standard. The triple quadrupole mass spectrometer with the turbo ion spray interface was operated in the positive ion mode and the temperature of the turbo was set at 105℃ and the ionization voltage was 3300V. The multi-reaction monitoring （MRM） mode was employed to scan the ions. The selected ions （precursor-ion/product ion） were 931.2/864.1 for sirolimus and 822.0/577.0 for FK506. Results The calibration linear range of blood-sirolimus concentrations was 1 to 240 pg/L （r=0. 9995）. The between-day and within-day RSD% were less than 15%. The extraction recoveries were more than 75.0% and the method recoveries were between 95.0% and 98. 5%. The low limit of detection was 0.2μg/L. Conclusion The UPLC/MS/MS method is rapid, sensitive and reliable and can be applied to monitor sirolimus of whole blood.