LC-MS/MS法同时测定人血浆和尿液中_对乙酰氨基酚及其结合物的浓度

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(THE)为内标,采用高效液相色谱串联质谱法,Utilmate C₁₈(100 mm×2.1 mm,3.0 μm)色谱柱,流动相为乙腈-甲醇-水(含0.0875%甲酸)(4:4:92),流速0.3 mL·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:APAP m/z 151.6→109.4,PG m/z 349.9→173.8,THE m/z 180.9→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10~30000 ng·mL^-1(APAP)和20~15000 ng·mL^-1(PG)范围内线性良好。质控(QC)样品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均分别为10 ng·mL^-1和20 ng·mL^-1。尿样采取直接稀释进样,在100~6000 ng·mL^-1(APAP)和500~60000 ng·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,操作简便等特点,可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,用于药物代谢动力学的研究。

Objective: To develop an 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.Plasma sample was precipitated by acetonitrile of 10% propylen glycol,and urine sample was diluted with 8% methanol.LC was performed on an Utilmate C₁₈ (100 mm×2.1 mm,3.0 μm) column with the mobile phase composition of acetonitrile- methanol -0.0875% formic acid water solution (4:4:92,v/v/v).Flow rate was 0.3 mL·min^-1 and column temperature was 40 ℃;Positive ESI and MRM mode was adopted in MS 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 20-15000,500-60000 ng·mL^-1for PG.The average recovery rate in plasma was 93.1% for APAP and 93.7% for PG.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 fastness method with high sensitivity and good reproducibility which can be used for simultaneous determination of the paracetamol and its glucuronide conjugates in biological matrices and effectively facilitate the pharmacokinetic studies.