摘要
目的:建立人尿液中替芬泰(Y101)及其代谢物M8和M9浓度的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法并用于Y101在人尿液中排泄的研究。方法:采用甲醇沉淀法进行尿液前处理并稀释后分析,色谱柱为ACQUITY UPLC BEH C18(50 mm×2.1 mm,1.7μm),流动相为5 mmol·L^-1醋酸铵水溶液(含0.1%甲酸)-甲醇=57∶43(v∶v),流速为0.3 mL·min^-1,柱温为40℃,进样量为2μL。质谱采用电喷雾离子源(electrospray ionization,ESI),正离子模式,多反应监测(multiple reaction monitoring,MRM)方式。Y101、代谢物M8、M9和帕拉米韦(内标)的监测离子对为m/z490.1→339.0,m/z357.2→105.2,m/z373.1→105.1和m/z329.1→270.2。结果:该方法选择性良好,人尿液中原型Y101、代谢物M8和M9分别在00200~20.0,0.200~100和0.0300~10.0μg·mL^-1范围内线性良好,平均回收率分别为95.9%~102.2%,973%~99.3%和98.1%~98.5%,内标归一化基质因子分别为095~104,098~102和099~100,批内、批间精密度均<15%。同时,对Y101、代谢物M8和M9浓度分别为1500,4000和3000μg·mL^-1的样本采用空白基质进行10倍稀释的稀释可靠性考察,结果表明分析物准确度为920%~1070%,精密度<5%。结论:所建立的UPLC-MS/MS分析方法专属性强、灵敏度高、重现性好,适用于人尿液样本Y101及其代谢物M8和M9的定量分析。
Objective:To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determination of Y101 and its metabolites M8 and M9 in human urine for the study ofits excretion.Methods:Urine samples were pretreated by protein precipitation and analyzed after dilution.Thechromatography was performed on an ACQUITY UPLC BEH C18(50 mm×2.1 mm,1.7μm,Waters)column usinga mixture of 0.1%formic acid water,5 mmol·L^-1 um acetate and methanol(57∶43,v∶v)as the mobilephase at a flow rate of 0.3 mL·min^-1 ammoni.The injection volume was 2μL.The column temperature was 40℃.Positiveelectrosmpray ionization(ESI)detection was performed in multiple reaction monitoring(MRM)mode using targetions mat/z490.1→339.0 for Y101,m/z357.2→105.2 for metabolite M8,m/z373.1→105.1 for metabolite M9 and/z329.1→270.2 for peramivir.Results:The specificity of the method was good.The calibration curve exhibited excellent linearity over the range of 0.0200~20.0,0.200~100 and 0.0300~10.0μg·mL-1 forY101,metabolite M8,and M9,respectively.The average recovery rate was 95.9%~102.2%,97.3%~99.3%and 98.1%~98.5%and the internal standard normalized matrix factors were 0.95~1.04,0.98~1.02 and 0.99~1.00 for Y101,metabolite M8,and M9,respectively.The intra-assay and inter-assay precision RSDs were lessthan 15%.Meanwhile,the dilution i n1 tegrity of 10 times dilution of samples with Y101,M8 and M9 concentrationof 150.0,400.0 and 30.00μg·mL^-1 was investigated.The results showed that the accuracy of the analytes was 92.0%~107.0%,and the precision was less than 5%.Conclusion:The established UPLC-MS/MS method hashigh specificity,sensitivity and reproducibility.It is suitable for the quantitative analysis of Y101 and its metabolitesM8 and M9 in human urine.
作者
刘筱雪
黄晨蓉
薛领
许青青
张诗超
张华
钱丽芳
蒋坤
吴贵辉
李星
夏文
缪丽燕
LIU Xiao-xue;HUANG Chen-rong;XUE Ling;XU Qing-qing;ZHANG Shi-chao;ZHANG Hua;QIAN Li-fang;JIANG Kun;WU Gui-hui;LI Xing;XIA Wen;MIAO Li-yan(Department of Clinical Pharmacology,The First Affliated Hospital of Soochou University,Suzhou 215006,China;Bailing Enterprise Group Pharmaceutical Co.,Ld.,Anshun 561000,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2020年第14期1628-1635,共8页
Chinese Journal of New Drugs
基金
国家科技重大专项课题资助项目(2015ZX09101012)
国家自然科学基金青年科学基金资助项目(81703619)
贵州省科技计划课题资助项目[黔科合重大专项字(2015)6002号]
江苏省药学会奥赛康医院药学基金资助项目(A201723)。
