人尿中对乙酰氨基苯甲酸和N,N-二甲氨基-2-丙醇的液质联用法测定及尿药排泄特征研究

Determination of p-Acetaminobenzoic Acid and N,N-Dimethylamino-2-propanol in Human Urine by LC-MS and Study on Their Urinary Excretion Profiles

目的:建立分别测定人尿中对乙酰氨基苯甲酸和N,N-二甲氨基-2-丙醇的液质联用法,考察二者在中国健康受试者尿液中的排泄特征。方法:以LC-MS法测定尿样中对乙酰氨基苯甲酸,色谱采用Amethyst C18柱(150 mm×2.1 mm,5μm),流动相为甲醇-0.1%甲酸水溶液(25∶75),流速为0.4 mL.min-1;质谱采用气动辅助电喷雾离子化和正离子选择性离子检测。以LC-MS/MS法测定尿样中N,N-二甲氨基-2-丙醇,色谱采用Hedera CN柱(150 mm×2.1 mm,5μm),流动相为乙腈-5 mmol.L-1醋酸铵水溶液(含0.03%甲酸)(55∶45),流速为0.35 mL.min-1;质谱采用气动辅助电喷雾离子化和正离子多反应检测。10名受试者单次口服异丙肌苷片(1.0 g),测定对乙酰氨基苯甲酸和N,N-二甲氨基-2-丙醇的尿药排泄参数。结果:对乙酰氨基苯甲酸的尿药浓度在0.202 0～202.0 mg.L-1范围内线性关系良好,平均回收率大于97.7%;N,N-二甲氨基-2-丙醇尿药浓度在0.797 8～398.9 mg.L-1范围内线性关系良好,平均回收率大于99.1%。受试者服药后,对乙酰氨基苯甲酸和N,N-二甲氨基-2-丙醇分别在6和12 h后基本随尿排泄完全,36 h内其平均尿药累积排泄百分率分别为(30.7±5.7)%和(49.0±8.6)%。结论:本法适用于人尿中对乙酰氨基苯甲酸和N,N-二甲氨基-2-丙醇的测定及其尿药排泄特征研究。

Objective： To establish LC-MS methods for the separate determination of p-acetaminobenzoic acid（PAcBA） and N,N-dimethylamino-2-propanol（DIP） in human urine,and to investigate their urinary excretion profiles in Chinese healthy volunteers.Methods：PAcBA in urine was determined by LC-MS using Amethyst C18-P column,with methanol-0.1% formic acid（25 ∶75） as mobile phase at a flow rate of 0.4 mL ·min-1 and positive ion SIM detection;DIP in urine was determined by LC-MS/MS using Hedera CN column,with acetonitrile-5 mmol ·L-1 ammonium acetate（55 ∶45） containing 0.03% formic acid as mobile phase at a flow rate of 0.35 mL ·min-1 and positive ion MRM detection.The urinary excretion parameters of PAcBA and DIP were measured after a single oral administration of isoprinosine tablets（1.0 g） in 10 volunteers.Results： The calibration curve was linear in the range of 0.202 0-202.0 mg ·L-1 for PAcBA in urine or in the range of 0.797 8-398.9 mg ·L-1 for DIP in urine.The average recoveries of PAcBA and DIP in urine were more than 97.7% and 99.1%,respectively.After taking the tablets,PAcBA and DIP were almost completely excreted in urine and the average cumulative excretion percentages of PAcBA and DIP in urine within 36 h were（30.7±5.7）% and（49.0±8.6）%,respectively.Conclusion：The methods are suitable for the determination of PAcBA and DIP in human urine and for the investigation on their urinary excretion profiles.