High throughput liquid chromatography-tandem mass spectrometry assay for mercapturic acids of acrolein and crotonaldehyde in cigarette smokers' urine
High throughput liquid chromatography-tandem mass spectrometry assay for mercapturic acids of acrolein and crotonaldehyde in cigarette smokers' urine
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DOI：
10.1016/j.jchromb.2013.07.004
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发表时间：
2013-09-15
影响因子：
3
通讯作者：
Hecht, Stephen S.
中科院分区：
文献类型：
--
作者：
Carmella, Steven G.;Chen, Menglan;Hecht, Stephen S.
3-羟丙基巯基尿酸（3-HPMA）和3-羟基-1-甲基丙基巯基尿酸（HMPMA）分别是毒物丙烯醛和巴豆醛的尿液代谢产物。几乎所有的人类尿液样本都含有这些代谢物，这是由谷胱甘肽-S-转移酶对丙烯醛和巴豆醛的作用产生的，丙烯醛和巴豆醛是脂质过氧化产物、环境和饮食污染物以及香烟烟雾的成分。我们已经开发了一种高通量的液相色谱-串联质谱法定量分析3-HPMA和HMPMA在大量的小尿样，将需要在分子流行病学和临床研究这些代谢物的水平与癌症的风险。混合模式反相-阴离子交换96孔板上的固相萃取为LC-MS/MS分析提供了足够的纯化，使用96孔格式通过自动进样进行LC-MS/MS分析，得到了干净、易于解释的色谱图，3-HPMA的检测限为4.5 pmol/mL尿液，HMPMA的检测限为3.5 pmol/mL尿液。3-HPMA和HMPMA的准确度分别为92%和97%，日间精密度分别为9.1%和11.0%（变异系数）。该方法应用于2600多个吸烟者的尿液样本; 3-HPMA和HMPMA的平均水平为4800+/-5358（S.D.）pmol/mL和3302 +/- 3341 pmol/mL。(C)2013爱思唯尔有限公司版权所有。
3-Hydroxypropylmercapturic acid (3-HPMA) and 3-hydroxy-1-methylpropylmercapturic acid (HMPMA) are urinary metabolites of the toxicants acrolein and crotonaldehyde, respectively. Virtually all human urine samples contain these metabolites, resulting from the action of glutathione-S-transferases on acrolein and crotonaldehyde, which are lipid peroxidation products, environmental and dietary contaminants, and constituents of cigarette smoke. We have developed a high throughput liquid chromatography-tandem mass spectrometry method for quantitative analysis of 3-HPMA and HMPMA in large numbers of small urine samples, as would be required in molecular epidemiology and clinical studies relating levels of these metabolites to cancer risk. Solid-phase extraction on mixed mode reverse phase-anion exchange 96-well plates provided sufficient purification for LC-MS/MS analysis, which was performed by auto-injection using a 96-well format, and resulted in clean, readily interpretable chromatograms, with detection limits of 4.5 pmol/mL urine for 3-HPMA and 3.5 pmol/mL urine for HMPMA. Accuracy was 92% for 3-HPMA and 97% for HMPMA while inter-day precision was 9.1% (coefficient of variation) for 3-HPMA and 11.0% for HMPMA. The method was applied to more than 2600 urine samples from smokers; mean levels of 3-HPMA and HMPMA were 4800+/-5358 (S.D.) pmol/mL and 3302 +/- 3341 pmol/mL, respectively. (C) 2013 Elsevier B.V. All rights reserved.