异丁酰衍生化结合液质联用法定量分析人尿液中的5-氨基水杨酸  

Quantitative Analysis of 5-Aminosalicylic Acid in Human Urine by Isobutyryl Derivatization Combined with UPLC-MS/MS

作　　者：刘筱雪[1] 马晟[1] 张诗超[1] 许青青[1] 缪丽燕[1] LIU Xiao-xue;MA Sheng;ZHANG Shi-chao;XU Qing-qing;MIAO Li-yan(Department of Clinical Pharmacology, The First Affiliated Hospital of Soochow University, Suzhou 215006,China)

机构地区：[1]苏州大学附属第一医院临床药理室,苏州215006

出　　处：《中国药学杂志》2018年第13期1104-1109,共6页Chinese Pharmaceutical Journal

摘　　要：目的建立人尿液样品中5-氨基水杨酸(5-ASA)的液质联用(UPLC-MS/MS)定量分析方法用于临床药物排泄研究。方法尿液样本加入异丁酸酐进行异丁酰衍生化后,加入乙腈(1∶3,V/V)并涡旋、离心取上清液进行UPLC-MS/MS分析。色谱柱为ACQUITY UPLC~?BEH-C18(2.1 mm×100 mm,1.7μm),流动相为5 mmol·L^(-1)醋酸铵水溶液(含0.075%甲酸)-乙腈=80∶20(V/V),流速为0.4 mL·min^(-1),柱温为40℃,进样量为3μL。质谱分析采用电喷雾离子源(ESI),负离子模式,多反应监测(MRM)方式测定,异丁酰化5-ASA和异丁酰化3-氨基苯甲酸(3-ABA,内标)的监测离子分别为m/z 222.0→178.1和m/z 206.1→162.0。结果方法选择性良好,人尿液中5-ASA的线性范围为0.100~30.0μg·mL^(-1),定量下限为0.100μg·mL^(-1),平均回收率为101.7%~115.4%,内标归一化基质因子为1.000~1.025,批内、批间精密度均小于15%。结论所建立的异丁酰衍生化结合UPLC-MS/MS的分析方法专属性强、灵敏度高、重现性好,适用于人尿液样本中5-ASA的定量分析。OBJECTIVE To establish an isobutyryl derivatization combined with UPLC-MS/MS method for the quantitative analysis of 5-aminosalicylic acid （5-ASA） in human urine.METHODS Urine sample was analyzed by UPLC-MS/MS after pretreatment with isobutyryl derivatization and precipitation with acetonitrile. The separation was performed on the ACQUITY UPLC BEH-C18 column （2.1 mm×100 mm,1.7 μm） at 40 ℃ with injection volume of 3 μL. The mobile phase was acetonitrile-5 mmol·L^-1 ammonium formate containing 0.075% formic acid （20∶80） at a flow rate of 0.4 mL·min^-1. Electrospray ionization （ESI） source was applied and operated in multiple reaction monitoring （MRM） mode with negative ionization using the transitions of m/z 222.0→178.1 and m/z 206.1→162.0 for isobutyl 5-ASA and isobutyl 3-amino benzoic acid （3-ABA, IS）, respectively.RESULTS The method was specific and sensitive. It exhibited good linearity over the concentration of 0.100-30.0 μg·mL^-1 for 5-ASA with the LLOQ of 0.100 μg·mL^-1, extraction recovery of 101.7%-115.4% and normalized matrix factor of 1.000-1.025. The intra- and inter-day precisions were both less than 15%.CONCLUSION The method is specific, simple, accurate and stable for quantitative analysis of 5-ASA in human urine.

关 键 词：异丁酰化 液质联用法 5-氨基水杨酸 定量分析 

分 类 号：R969[医药卫生—药理学]