A new liquid chromatography/mass spectrometry method for 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine.
A new liquid chromatography/mass spectrometry method for 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine.
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尿液中 4-(甲基亚硝基氨基)-1-(3-吡啶基)-1-丁醇 (NNAL) 的新液相色谱/质谱分析方法。
DOI：
10.1002/rcm.4824
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Blair,IanA
中科院分区：
文献类型：
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作者：
Bhat,ShowketH;Gelhaus,StacyL;Mesaros,Clementina;Vachani,Anil;Blair,IanA
4-（Methylnitrosamino）-1-（3-pyridyl）-1-butanone（NNK）是一种致癌的亚硝胺，在烘烤烟草时产生。它存在于烟草烟雾中，并在肺中代谢为4-（甲基亚硝胺）-1-（3-吡啶基）-1-丁醇（NNAL）。NNAL进一步经历尿苷二磷酸葡萄糖醛酸转移酶（UGT）介导的代谢，产生N-和O-葡萄糖醛酸苷代谢产物，然后与游离（非结合）NNAL一起经尿液排泄。对人尿液中游离和结合NNAL进行经验证分析的能力对于评估暴露于香烟烟雾的肺癌风险的个体间差异非常重要。稳定同位素稀释（SID）方法与液相色谱/多反应监测/质谱（LC/MRM-MS）联用可为此类分析提供最高的生物分析特异性。我们描述了一种新的衍生化方法，该方法导致形成预电离的N-丙基-NNAL衍生物。由此衍生物产生的LC/MS灵敏度增加，使得仅在0.25 mL尿液中分析游离NNAL成为可能。与已开发的其他方法相比，这种尿量的大幅减少将有助于保存从正在进行的纵向生物标志物研究中获得的有限数量的储存尿液样本。采用新的高灵敏度SID LC/MRM-MS测定法测定60名无疾病吸烟者的尿样中游离和结合NNAL浓度。然后评估了尿肌酐清除率个体间差异对NNAL浓度的影响，并根据尿NNAL葡萄糖醛酸苷/游离NNAL的比值在60例受试者中确定了3种代谢物表型。NNAL葡萄糖醛酸苷/游离NNAL比值<2（平均值= 1.3）的弱代谢者（PM，14例受试者）、比值在2 - 5之间（平均值= 3.4）的中间代谢者（IM，36例受试者）和比值>5（平均值= 11.1）的快代谢者（EM，10例受试者）。版权所有© 2010约翰威利父子有限公司.
4‐(Methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanone (NNK) is a carcinogenic nitrosamine produced upon curing tobacco. It is present in tobacco smoke and undergoes metabolism to 4‐(methylnitrosamino)‐1‐(3‐pyridyl)‐1‐butanol (NNAL) in the lungs. NNAL undergoes further uridine diphosphate glucuronosyltransferase (UGT)‐mediated metabolism to give N‐ and O‐glucuronide metabolites, which together with free (non‐conjugated) NNAL are then excreted in the urine. The ability to conduct validated analyses of free and conjugated NNAL in human urine is important in order to assess inter‐individual differences in lung cancer risk from exposure to cigarette smoke. The use of stable isotope dilution (SID) methodology in combination with liquid chromatography/multiple reaction monitoring/mass spectrometry (LC/MRM‐MS) provides the highest bioanalytical specificity possible for such analyses. We describe a novel derivatization procedure, which results in the formation of a pre‐ionizedN‐propyl‐NNAL derivative. The increased LC/MS sensitivity arising from this derivative then makes it possible to analyze free NNAL in only 0.25 mL urine. This substantial reduction in urine volume when compared with other methods that have been developed will help preserve the limited amounts of stored urine samples that are available from on‐going longitudinal biomarker studies. The new high sensitivity SID LC/MRM‐MS assay was employed to determine free and conjugated NNAL concentrations in urine samples from 60 individual disease‐free smokers. Effects of inter‐individual differences in urinary creatinine clearance on NNAL concentrations were then assessed and three metabolizer phenotypes were identified in the 60 subjects from the ratio of urinary NNAL glucuronides/free NNAL. Poor metabolizers (PMs, 14 subjects) with a ratio of NNAL glucuronides/free NNAL <2 (mean = 1.3), intermediate metabolizers (IMs, 36 subjects) with a ratio between 2 and 5 (mean = 3.4), and extensive metabolizers (EMs, 10 subjects) with a ratio >5 (mean = 11.1). Copyright © 2010 John Wiley & Sons, Ltd.