论文部分内容阅读
建立一种同时测定人尿中茶碱及其代谢物1,3-二甲基尿酸(1,3-DMU)、3-甲基黄嘌呤(3-MX)和1-甲基尿酸(1-MU)的液相色谱-加热电喷雾电离源串联质谱法(LC-HESI/MS/MS),并用于人尿中茶碱及代谢物累积排泄量研究。本法采用d6-茶碱作为茶碱的内标,氟尿嘧啶作为1,3-DMU、3-MX和1-MU的内标。尿样用甲醇稀释并离心后,在Agilent XDB-Phenyl(150 mm×4.6 mm ID,5μm)色谱柱上进行色谱分离,以水-甲醇-甲酸(30∶70∶0.15)为流动相,流速为0.6 mL·min-1;采用加热电喷雾电离源(HESI),选择反应监测(SRM)负离子扫描检测。4种分析物的标准曲线的线性范围均为1.0~250μg·mL-1。该方法快速、简便,可用于人尿中茶碱及其3种代谢物浓度的测定。结果表明,中国受试者口服茶碱缓释片后,尿中茶碱、1,3-DMU、3-MX和1-MU的排泄量比值约为1.0∶3.4∶0.92∶1.6,与已报道的白种人受试者结果相近。
To establish a method for the simultaneous determination of theophylline and its metabolites 1,3-dimethylurea (1,3-DMU), 3-methylxanthine (3-MX) MU) by liquid chromatography-electrospray ionization-electrospray ionization tandem mass spectrometry (LC-HESI / MS / MS) and used in the study of cumulative excretion of theophylline and metabolites in human urine. This method uses d6-theophylline as the internal standard for theophylline and fluorouracil as the internal standard for 1,3-DMU, 3-MX and 1-MU. The urine sample was diluted with methanol and centrifuged, and then chromatographed on an Agilent XDB-Phenyl (150 mm × 4.6 mm ID, 5 μm) column using water-methanol-formic acid (30:70:0.15) 0.6 mL · min-1. The reaction was monitored by electrospray ionization (HESI) and negative ion scan (SRM). The calibration curve of the four analytes showed a linear range of 1.0-250 μg · mL -1. The method is rapid and simple and can be used for the determination of theophylline and its three metabolites in human urine. The results showed that the excretion ratio of theophylline, 1,3-DMU, 3-MX and 1-MU in urine of Chinese subjects after oral administration of theophylline sustained release tablets was about 1.0: 3.4: 0.92: 1.6, Of the Caucasian subjects had similar results.