摘要
目的 建立同时测定全血中他克莫司(FK506)和西罗莫司(SRL)浓度的液相色谱-串联质谱(LC-MS/MS)法,检测使用四联免疫抑制方案的肾移植患者体内两种药物的浓度。方法 50 μL全血样品用甲醇-乙腈(90∶10)沉淀蛋白,上清液用纯水(1∶1)稀释后进样分析。色谱柱为Kinetex C18(3 mm×100 mm,2.6 μm),流动相为0.1%甲酸-水(含10 mmol·L^(-1)乙酸铵)和0.1%甲酸-甲醇,梯度洗脱,流速0.55 mL·min^(-1),进样量5 μL,分析时间5 min。采用电喷雾离子源,正离子多反应监测模式扫描,用于定量分析的离子对分别为:m/z 821.2→768.6(FK506)、m/z 936.4→409.3(SRL)。结果 FK506和SRL在1~40 ng·mL^(-1)与峰面积线性关系良好(r均>0.99),准确度在92.60%~114.41%,精密度RSD均≤15%,提取回收率为71.48%~94.88%,基质效应为72.36%~102.80%,稳定性均符合方法学要求。应用本方法测定37例接受四联免疫抑制方案的肾移植患者的血药浓度,FK506血药谷浓度(5.57±3.07)ng·mL^(-1),SRL血药谷浓度(4.41±2.36)ng·mL^(-1)。结论 本方法同时测定人全血中FK506和SRL浓度,操作简单快捷,准确度和灵敏度高,稳定性良好,可用于肾移植患者的临床血药浓度监测。
Objective To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of tacrolimus (FK506) and sirolimus (SRL) in the blood of renal transplant patients with quadruple immunosuppressive therapy.Methods The blood samples (50 μL) were precipitated with methanol-acetonitrile (90∶10,V/V).The supernatant was diluted with water (1∶1,V/V) and then injected for detection.The chromatographic separation was performed on a Kinetex C18 column (3 mm×100 mm,2.6 μm) with the mobile phase of 10 mmol·L^(-1) ammonium acetate solution and methanol,both containing 0.1% formic acid,at a flow rate of 0.55 mL·min^(-1).The injection volume was 5 μL and the total run time was 5 min.The detection of the analytes was performed by electrospray ionization in positive mode by multiple monitoring with the transition of m/z 821.2→768.6 (FK506),m/z 936.4→409.3 (SRL).Results The calibration curves for FK506 and SRL had good linearity at 1~40 ng·mL^(-1).The accuracy was 92.60%~114.41%.The intra and inter-day RSD did not exceed 15%.The recoveries were 71.48%~94.88% and the matrix effect was 72.36%~102.80%.The stability met the requirements.The blood concentrations of FK506 and SRL in 37 renal transplant patients were determined by this method.Trough concentration of FK506 was (5.57±3.07) ng·mL^(-1),and that of SRL was (4.41±2.36) ng·mL^(-1).Conclusion The method is simple,accurate,sensitive and stable,which is suitable for clinical therapeutic drug monitoring.
作者
杨旭萍
蒋振伟
蒋艳
凌静
董露露
邹素兰
胡楠
YANG Xu-ping;JIANG Zhen-wei;JIANG Yan;LING Jing;DONG Lu-lu;ZOU Su-lan;HU Nan(Department of Pharmacy,Changzhou First People’s Hospital,Changzhou Jiangsu 213003)
出处
《中南药学》
CAS
2022年第11期2514-2519,共6页
Central South Pharmacy
基金
江苏省卫生健康委员会2020年高层次卫生人才“六个一工程”拔尖人才科研项目(No.LGY2020033)
常州市科技计划(应用基础研究)指导性项目(No.CJ20190910,No.CJ20209011)
江苏省研究型医院学会精益化用药-石药专项科研基金项目(No.JY202043)。
