摘要
目的:建立一种检测小鼠血浆中对乙酰氨基酚(APAP)及其5个代谢产物的液相色谱-串联质谱(LC-MS/MS)方法,将之应用于对乙酰氨基酚的代谢研究。方法:血浆样品经甲醇沉淀蛋白后,以对氨基苯甲酸(PABA)为内标,采用LC-MS/MS法,C18色谱柱,流动相为甲醇-5 mmol·L^(-1)乙酸铵水溶液(含0.1%甲酸)(55∶45),流速为0.5 ml·min^(-1),柱温为25℃,采用电喷雾离子化(ESI),正离子多反应监测(MRM)检测方式进行定量分析:APAP,[M-H]^-,m/z 152.0→110.0;APAP-cys,[MH]^-,m/z 271.2→140.1;APAP-glut,[M-H]^-,m/z 457.0→328.0;APAP-NAC,[M-H]-,m/z 313.4→208.0;APAP-sulf,[MH]^-,m/z 232.4→152.1;APAP-gluc,[M-H]^-,m/z 328.2→152.1;IS,[M-H]^-,m/z 138.2→120.0。结果:在0.2~10μg·ml^(-1)(APAP)、1.0~20μg·ml^(-1)(APAP-gluc)、1.0~20μg·ml^(-1)(APAP-sulf)、1.0~20μg·ml^(-1)(APAP-glut)、0.4~15μg·ml^(-1)(APAP-NAC)和0.2~10μg·ml^(-1)(APAP-cys)范围内线性良好(r≥0.990 0)。各测定物的日内精确度均在85%~115%间,精密度均小于15%,加样回收率均在85%~115%之间,RSD均低于15%。结论:该方法快速、灵敏、准确,适用于小鼠血浆中对乙酰氨基酚及其5个代谢产物的定量测定。
Objective : To establish an LC-MS/MS method for the simuhaneous determination of acetaminophen and its five metabolites in mice plasma, and inveatigate the metabolism of acetaminophen by using the method. Methods: Para aminobenzoic acid was used as the internal standard. The plasma samples were precipitated by methanol, and then separated on a C18 column with the mobile phase of methanol and 5 mmol·ml^-1 ammonium acetate buffer solution containing 0.1% formic acid ( 55 : 45 ). The flow rate was 0.5 ml· min^-1, and the column temperature was 25℃. An electrospray ionization source was applied and operated in a positive ion mode using MRM : APAP, [ M-H ]^-, m/z 152.0→110.0; APAP-cys, [ M-H ]^ -, m/z 271.2 → 140.1 ; APAP-glut, [ M-H ]^ -, m/z 457.0→ 328.0 ; APAP-NAC, [ M-H ]^- , m/z 313.4 →208.0 ; APAP-sulf, [ M-H ]^ -, m/z 232.4→152.1 ; APAP-gluc, [ M-H ] ^-, m/z 328.2→152.1 ;IS, [ M-H]^ - m/z 138.2→120.0. Results: The method exhibited good linearity over the concentration range of 0.2-10μg·ml^-1 for APAP, 1.0-20μg·ml^-1 for APAP-glue, 1.0-20 μg·ml^-1 for APAP-sulf, 1.0-20μg·ml^-1 for APAP-glut, 0.4-15μg·ml^-1 for APAP-NAC and 0.2-10μg·ml^-1 for APAP-eys (r ≥0. 990 0). The inter-day accuracy and precision of acetaminophen and its five metabolites were all below 15%. The average recovery was between 85% and 115% , and RSDs were all below 15%. Conclusion: The LC-MS/MS method is proved to be quick, sensitive and accurate, and suitable for the determination of acetaminophen and its five metabolites in mice plasma.
出处
《中国药师》
CAS
2017年第4期597-602,616,共7页
China Pharmacist
基金
国家自然科学基金项目(编号:81274171)