【作者】 谭亲友； 陈根姩； 李焕德； 朱荣华； 方平飞； 颜苗； 王峰；

【Author】 TAN Qin-you 1,2,3 ,CHEN Gen-nian3,LI Huan-de ,ZHU Rong-hua 1,ZHANG Qi-zhi 1,2,FANG Ping-fei1 ,YAN Miao1,PENG Wen-xing1 (1. The Clinical Pharmacy & Pharmacology Research Institute,The Second Xiangya Hospital,Central South University,Changsha 410011,China; 2.The Pharmaceutical Science of Centre South University,Changsha 410013,China; 3. Affiliated Hospital of Guilin Medical University,Guilin 541001,China)

【机构】 中南大学湘雅二医院临床药学教研室； 中南大学药学院；

【摘要】 目的:建立同时测定人血浆及尿液中对乙酰氨基酚(APAP)和其葡糖醛酸结合物(PG)的LC-MS/MS法。方法 :血浆经10%丙二醇乙腈溶液沉淀蛋白处理,尿样则采取直接稀释进样,以茶碱为内标,采用高效液相串联质谱法,以UtilmateC18柱(100mm2.1mm,3.0μm)为色谱柱,流动相乙腈-甲醇-水(含0.0875%甲酸)(4:4:92),流速0.3mL·min-1,柱温40℃;采用电喷雾电离(ESI+),多反应离子检测(MRM)方式进行定量分析:APAPm/z151.6→109.4PGm/z349.9→173.8,THEm/z180.9→123.4。结果 :经10%丙二醇乙腈溶液处理后,血浆内源性物质无干扰,APAP和PG血浆平均萃取回收率分别为93.1%和93.7%,在10-30000ng·mL-1(APAP),20-15000ng·mL-1(PG)范围内线性良好。质控样品(QC)APAP的日内、日间RSD分别为3.2%-10.8%和8.3%-10.6%,PG的日内、日间RSD分别为2.1%-9.3%和6.2%-11.2%。在血浆中APAP和PG的LLOQ均分别10ng·mL-1和20ng·mL-1。尿样采取直接稀释进样,在100-6000ng·mL-1(APAP),500-60000ng·mL-1(PG)范围内线性良好,APAP和PG的尿样平均萃取回收率分别为89.1%和92.3%,准确度在85%-115%之间。结论 :本方法具有灵敏度高,重现性好,操作简便等特点,可用于同时测定对乙酰氨基酚及其葡萄糖醛酸结合物在生物样品中的药物浓度,方便于药物代谢动力学的研究。更多还原

【Abstract】 Objective:To develop a LC-MS/MS method for the simultaneous determination of paracetomal and its glucuronide in human plasma and urine.Methods:Theophylline was used as the internal standard.The plasma sample was injected directly for determination after being precipitated by acetonitrile of 10% propylen glycol and urine sample was diluted with 8% methanol for direct injection.LC conditions: Utilmate C18 ( 100 mm×2.1 mm,3.0 um)column was used and the mobile phase composition was a mixture of acetonitrile- methanol -water containing (0.0875% formic acid) in a ratio 4:4:92 (v/v/v).Flow rate was 0.3 mL.min-1 and column temperature was 40 ℃ ;MS conditions:using positive ESI and MRM mode for quantitative analysis.Results:The calibration of curves in plasma and urine was linear within the range of 10-30000,100-6000 ng·mL-1 for APAP,and 10-15000,200-60000 ng·mL-1for PG.The average recovery rate in plasma was 93.1% for APAP and 93.7% for PG.Meantime,the recovery rate in human urine was 89.1% for APAP and 92.3% for PG.The RSD of intra-day and inter-day in QC samples was 3.2%～10.8% and 8.3%-10.6% for APAP,2.1%-9.3% and 6.2%-11.2% for PG .Conclusions:This is a high sensitivity,good reproducibility and fastness method.It can be simultaneously determinated the paracetamol and its glucuronide conjugates in the biological matrices and effectively utilized for pharmacokinetic studies.更多还原