基于LC-MS/MS同时测定小鼠血浆中对乙酰氨基酚及其毒性代谢产物对乙酰氨基酚半胱氨酸的浓度  

Simultaneous determination of acetaminophen and its toxicity related metabolite acetaminophen-cysteine in mouse plasma by LC-MS/MS

作　　者：何慧 柴媛媛 徐云霞 俞沁玮 黄鑫[1] 张陆勇[1,2] 江振洲[1,3] HE Hui;CHAI Yuanyuan;XU Yunxia;YU Qinwei;HUNAG Xin;ZHANG Luyong;JINAG Zhenzhou(New Drug Screening Center,China Pharmaceutical University,Nanjing Jiangsu 210009,China;Center for Drug Research and Development,Guang­dong Pharmaceutical University,Guangzhou Guangdong 510006,China;Jiangsu Center for Pharmacodynamics Research and Evaluation,Nanjing Jiangsu 210009,China)

机构地区：[1]中国药科大学新药筛选中心,江苏南京210009 [2]广东药科大学新药研发中心,广东广州510006 [3]江苏省药效研究与评价服务中心,江苏南京210009

出　　处：《中国药物警戒》2023年第3期266-272,共7页Chinese Journal of Pharmacovigilance

基　　金：国家自然科学基金资助项目(82074114);“双一流”团队:药物安全预警关键技术研究创新团队资助项目(CPU2018G-Y33)。

摘　　要：目的建立一种同时测定小鼠血浆中对乙酰氨基酚(APAP)及其毒性相关代谢产物对乙酰氨基酚半胱氨酸(APC)的液相色谱-串联质谱(LC-MS/MS)方法。方法以茶碱为内标,采用蛋白沉淀后取上清液挥干复溶的方法进行血浆样品前处理。采用Agilnet Zorbax SB-C_(18)(3.0 mm×100 mm,3.5μm)色谱柱进行梯度洗脱,水相为含0.1%甲酸的水溶液,有机相为含0.1%甲酸的甲醇溶液。选择三重四极杆质谱仪(TSQ Quantum Ultra),采用电喷雾离子化方式(ESI),在选择反应离子监测(SRM)模式下进行正离子检测,同时对APAP(152.1行正离子检测)和APC(271.0行正离子检测)进行定量分析。结果APAP在60~3000 ng·mL^(-1)的范围内线性关系良好,APC在20~1000 ng·mL^(-1)的范围内线性关系良好,该方法选择性、批内和批间准确度、批内和批间精密度、基质效应、提取回收率均符合生物样品分析要求。结论本研究所建立的LC-MS/MS方法可应用于APAP及APC在小鼠体内药代动力学研究。Objective To establish a liquid chromatography-mass spectrometry(LC-MS/MS)method for simultaneous determination of acetaminophen(APAP)and its toxicity related metabolite acetaminophen-cysteine(APC)in mouse plasma.Methods Theophylline was taken as the internal standard(IS)while plasma samples were pretreated by volatilizing and re-dissolving the supernatant after protein precipitation.The gradient was eluted by Agilnet Zorbax SBC_(18)(3.0 mm×100 mm,3.5μm).The aqueous phase was an aqueous solution containing 0.1%formic acid while the organic phase was a methanol solution containing 0.1%formic acid.The contents of APAP and APC were detected under positive ions detection with electrospray ionization(ESI)in the selective ion monitoring(SRM)mode of m/z 152.1→110.0 and 271.0→140.0 in the triple quagdrupole tandem mass spectrometer(TSQ Quantum Ultra).Results There were good linear relationships when concentrations ranged from 60 ng·mL^(-1)to 3000 ng·mL^(-1)for APAP and from 20 ng·mL^(-1)to 1000 ng·mL^(-1)for APC.The selectivity,accuracy and precision,matrix effect and extraction recovery of the method met the requirements of biological sample analysis.Conclusion The LC-MS/MS method established in this study could be applied to the pharmacokinetics of APAP and APC in mice.

关 键 词：对乙酰氨基酚 对乙酰氨基酚半胱氨酸 N-乙酰半胱氨酸 液相色谱-串联质谱 药代动力学 小鼠 毒性 

分 类 号：R927.2[医药卫生—药学] R994.11