Liquid Chromatograph-Triple Quadrupole Mass Spectrometer

描述（由申请人提供）：本资源的目标是继续使用最先进的方法和仪器为烟草生物标志物提供高质量的生物流体分析。该资源支持许多国家卫生研究院资助的生物医学研究项目，主要是烟草成瘾及其治疗的人类临床研究。它还将支持二手烟暴露的研究。本申请寻求资金购买过时仪器的替代品，并允许我们的设施为我们机构和整个美国的研究人员提供大量生物流体分析的承诺。由于分析灵敏度、特异性和能够分析大量样品（每年约5000-10,000）的原因，我们的大多数分析使用液相色谱-串联质谱（LC-MS/MS）进行。所有这些方法都是在我们的实验室开发的，有些方法在其他地方是不可用的。我们的两种方法需求量特别大：(1)同时测定尼古丁代谢物可替宁和反式-3′-羟基可替宁。这些代谢物在生物体液中的比例可用于尼古丁代谢酶CYP2A6的表型，并且与尼古丁从体内消除的速度高度相关(Dempsey等，尼古丁代谢物比例作为细胞色素P450 2A6代谢活性的指标。（临床药学杂志，2004;76:64-72）。该比值已被用于烟草依赖的个体化药物治疗（Lerman等人，尼古丁代谢物比值预测透皮尼古丁戒烟的效果）。[j] .临床药学杂志，2006;79:600-8。常规测定血浆、血清、尿液和唾液中的代谢物。(2)（4-（甲基亚硝胺）-1-（3-吡啶基）-1-丁醇），NNAL，一种烟草特异性致癌代谢物。它可以在吸烟者或暴露于二手烟（SHS）的非吸烟者的尿液中测量。这可能是SHS暴露的最佳生物标志物，因为它的生物半衰期很长，当暴露模式不稳定时，通过提供时间平均暴露测量（Eisner， MD, Jacob， P, 3rd, Benowitz， NL, Balmes， J, Blanc， PD）。长期暴露于二手烟和慢性阻塞性肺病的健康结果：尿4-（甲基亚硝胺）-1-（3-吡啶基）-1-丁醇（NNAL）尼古丁和烟草研究的影响。我们的资源支持的研究项目寻求通过进一步了解烟草成瘾和开发改进的治疗方式，包括使用药物遗传方法进行个体化治疗，来改善公众健康。一些受支持的项目涉及烟草毒理学，如接触二手烟和特定致癌物。我们的资源也可用于与烟草无关的项目，但我们资助的大多数研究和合作者的研究都在烟草领域。

DESCRIPTION (provided by applicant): The objective of this Resource is to continue to provide high quality biofluid analyses for tobacco biomarkers using state-of-the-art methods and instruments. This Resource supports numerous NIH- funded biomedical research projects, mostly human clinical studies of tobacco addiction and its treatment. It will also support studies of secondhand smoke exposure. This application seeks funds to purchase a replacement for an obsolete instrument, and allow our facility to support numerous commitments for biofluid analyses for investigators at our institution and throughout the US. For reasons of analytical sensitivity, specificity, and to be able to analyze large numbers of samples (about 5000-10,000 per year), most of our analyses are carried out using liquid chromatography - tandem mass spectrometry (LC-MS/MS). All of these methods have been developed in our laboratory and some are not available elsewhere. Two of our methods are in particularly high demand: (1) Simultaneous determination of the nicotine metabolites cotinine and trans-3'- hydroxycotinine. The ratio of these metabolites in biofluids can be used to phenotype for the nicotine- metabolizing enzyme CYP2A6, and is highly correlated with the rate of nicotine elimination from the body (Dempsey et al, Nicotine metabolite ratio as an index of cytochrome P450 2A6 metabolic activity. (Clin Pharmacol Ther. 2004;76:64-72.). This ratio has been used to individualize pharmacotherapies for tobacco dependence (Lerman et al., Nicotine metabolite ratio predicts efficacy of transdermal nicotine for smoking cessation. (Clin Pharmacol Ther. 2006;79:600-8.). The metabolites are routinely measured in plasma, serum, urine, and saliva. (2) (4-(Methylnitrosamino)-1- (3-pyridyl)-1-butanol), NNAL, a tobacco-specific carcinogen metabolite. It can be measured in urine of smokers or of non-smokers exposed to secondhand smoke (SHS). This is probably the best biomarker for SHS exposure, due to its long biological half-life, by providing a time-averaged measure of exposure when exposure patterns are erratic (Eisner, MD, Jacob, P, 3rd, Benowitz, NL, Balmes, J, Blanc, PD. Longer-term exposure to secondhand smoke and health outcomes in COPD: impact of urine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) Nicotine and Tobacco Research, in press). The research projects supported by our resource seek to improve public health by furthering our understanding of tobacco addiction and developing improved treatment modalities, including individualizing treatment using pharmacogenetic approaches. Some supported projects involve tobacco toxicology, such as exposure to secondhand smoke and specific carcinogens. Our resource is also available for non-tobacco related projects, but the majority our funded studies and collaborators studies are in the tobacco field. PUBLIC HEALTH RELEVANCE: About 500,000 Americans and about 5,000,000 people worldwide die each year from tobacco related diseases. The research projects supported by our resource are developing improved therapies for tobacco addiction, or are investigating exposure to secondhand tobacco smoke in order to reduce its adverse health consequences.