摘要
目的：建立LC-MS/MS法同时测定人全血中环孢素A、依维莫司、他克莫司和西罗莫司的浓度。方法：人全血样品用乙腈沉淀蛋白后,上清液用叔丁基甲醚提取后选用ZORBAX SB C18（2.1 mm×100 mm,3.5μm）色谱柱,流动相采用甲醇-10 mmol/L乙酸铵溶液（均含0.1%甲酸）梯度洗脱,采用电喷雾离子化源,正离子方式,多反应监测（MRM）扫描方式进行监测,用于定量分析的离子反应分别为m/z 1 202.6→m/z 425.1（环孢素A）、m/z 975.8→m/z 908.6（依维莫司）、m/z 821.6→m/z 768.5（他克莫司）、m/z 9 31.8→m/z 864.2（西罗莫司）和m/z 366.2→m/z 132.0（内标吲达帕胺）。结果：全血中环孢素A的线性范围为10~2 000 ng/m L（r=0.996 2）,定量下限为10 ng/m L,依维莫司、他克莫司和西罗莫司的线性范围为0.50~100 ng/m L（r〉0.99）,定量下限为0.50 ng/m L;四种药物的平均回收率95.1%~110.6%;日内、日间RSD均小于15%;提取回收率在78.6%~85.9%之间。结论：该方法快速、灵敏、准确,专属性强,重复性好,适用于人全血中同时测定环孢素A、依维莫司、他克莫司和西罗莫司浓度。
AIM： To develop an LC-MS / MS method for simultaneous determination of cyclosporin A,everolimus,tacrolimus,and sirolimus concentration in human blood. METHODS： The blood samples were precipitated by acetonitrile,then the supernatant were processed with tert-butyl methyl ether extraction. Chromatographic separation was performed on ZORBAX SB C18column（ 2. 1 mm ×100 mm,3. 5 μm） and eluted with linear gradient using methanol and 10 mmol / L ammonium acetate,both of which were acidified with 0. 1% formic acid.Detection of the analytes were achieved using positive ion electrospray ionization（ ESI） in the multiple reaction monitoring（ MRM） mode. The monitored MS / MS ion transitions were m / z 1 202. 6 → m / z425. 1,m / z 975. 8→m / z 908. 6,m / z 821. 6 →m / z768. 5,m / z 931. 8 → m / z 864. 2 and m / z 366. 2 →m / z 132. 0 for cyclosporin A,everolimus,tacrolimus,sirolimus and internal standard,respectively.RESULTS： Cyclosporin A showed good linearity over the concentration range of 10-2000 ng / m L（ r =0. 9962） and its lower limit of quantification was 10 ng / m L,while everolimus,tacrolimus and sirolimus were obtained concentration range of 0. 50-200 ng /m L（ r 〉0. 99）,with the lower limit of quantitation（ LLOQ） of 0. 50 ng /m L. The average recovery was between 95. 1%-110. 6%,Intra-day and inter-day relative standard deviations were both below 15%.The extraction recoveries of low,middle and high concentrations were 78. 6%-85. 9%. CONCLUSION： The established method is rapid,sensitive,accurate,specific and reliable,and suitable for simultaneous determination of cyclosporin A,everolimus,tacrolimus,and sirolimus in human blood.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2015年第10期1126-1130,共5页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
国家科技重大专项项目-老年病新药临床技术平台(2013ZX09303005)
