Liquid Chromatograph-Triple Quadrupole Mass Spectrometer

描述（由申请人提供）：本资源的目的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NIH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。它还将支持对二手烟暴露的研究。该申请寻求资金来购买过时仪器的替代品，并允许我们的设施支持我们机构和整个美国的研究人员的生物流体分析的许多承诺。出于分析灵敏度、特异性以及能够分析大量样品（每年约5000- 10，000份）的原因，我们的大多数分析都使用液相色谱-串联质谱法（LC-MS/MS）进行。所有这些方法都是在我们的实验室开发的，有些是其他地方没有的。我们的两个方法是特别高的需求：（1）同时测定尼古丁代谢物可替宁和反式-3 '-羟基可替宁。生物流体中这些代谢物的比率可用于尼古丁代谢酶CYP 2A 6的表型，并且与尼古丁从身体中消除的速率高度相关（Dempsey等人，Nicotine metabolite ratio as an index of cytochrome P450 2A 6 metabolic activity. (Clin药理学治疗2004;76：64-72.）。该比率已用于使烟草依赖的药物治疗个体化（Lerman et al.，尼古丁代谢物比率预测尼古丁透皮戒烟的疗效。(Clin药理学治疗2006;79：600-8.）。代谢物在血浆、血清、尿液和唾液中进行常规测定。(2)（4-（甲基亚硝胺基）-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，因为其生物半衰期长，当暴露模式不稳定时，通过提供暴露的时间平均测量（Reinner，MD，Jacob，P，3rd，Benowitz，NL，Balmes，J，Blanc，PD.长期暴露于二手烟和COPD的健康结果：尿4-（甲基亚硝胺）-1-（3-吡啶基）-1-丁醇（NNAL）的影响尼古丁和烟草研究，出版中）。 我们的资源支持的研究项目旨在通过进一步了解烟草成瘾和开发改进的治疗方式，包括使用药物遗传学方法进行个性化治疗，来改善公共卫生。一些受资助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非烟草相关的项目，但我们资助的研究和合作者的研究大多是在烟草领域。 公共卫生相关性：每年约有50万美国人和全世界约500万人死于烟草相关疾病。我们的资源支持的研究项目正在开发改进的烟草成瘾疗法，或正在调查二手烟草烟雾的暴露情况，以减少其对健康的不良影响。

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.