Development of a Quantitative and Sensitive Liquid Chromatographic/Mass Spectrome

定量、灵敏的液相色谱/质谱仪的开发

• 批准号: 7891310
• 负责人: DERRICK J SWINTON
• 金额: $ 40.22万
• 依托单位: LINCOLN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-07 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7891310
• 关键词: Acids; African American; Area; Cancer Etiology; Cigarette; Clinic; Clinical Research; Cotinine; Detection; Development; Development Plans; Doctor of Philosophy; Economically Deprived Population; Environmental Tobacco Smoke; Exposure to; Faculty; Funding; Glucuronides; Goals; Graduate Education; Historically Black Colleges and Universities; Institution; Kinetics; Laboratories; Learning; Liquid substance; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Medicine; Metabolic; Methods; Minority; Morbidity - disease rate; Nicotine; Oxides; Peer Review; Pennsylvania; Population; Preparation; Procedures; Relative (related person); Research; Research Activity; Research Infrastructure; Research Personnel; Research Project Grants; Sampling; Smoke; Smoker; Students; Technology; Time; Tobacco; Training; United States National Institutes of Health; Universities; Urine; anticancer research; base; career; career development; caucasian American; college; experience; gang; health disparity; improved; insight; liquid chromatography mass spectrometry; mass analyzer; mass spectrometer; member; non-smoker; programs; skills; smoking cessation; stem

Studies conclude that the cessation rates amongst African Americans are lower than White Americans despite the fact that this group smokes fewer cigarettes per day than White Americans (1-4). Furthermore, African Americans experience disproportionately higher rates of tobacco-related morbidity (5-7). I hypothesize that African Americans may have a more difficult time with quitting smoking because of their inability to metabolize nicotine and its metabolites efficiently. As a result, this research will attempt to determine if there is a correlation between smoking cessation rates amongst African Americans and their nicotinic metabolic profile. This research project will use LC/MS to measure the nicotinic metabolic profile of African Americans participating in smoking cessation clinics. An improved LC/MS method will be developed enhancing the sensitivity and accuracy of detection of nicotine and its major metabolites, cotinine, trans-3'-OH-cotinine, nicotine N'-oxide, 4-OH-4-(3-pyridyl)-butanoid acid, nicotine-A/-glucuronide, cotinine-A/-glucuroninde and trans- 3'-OH-cotinine-O-glucuronide, in urine samples. The improved LC/MS method will incorporate a q-TOF mass analyzer extending the sensitivity and accuracy of the developed method relative to the current LC/MS (ESIquadrupole) methods in practice to date. The project will focus on sequestering urine samples from African Americans participating in smoking cessation clinics. The samples will be analyzed and the levels of nicotine and its major metabolite, cotinine, quantified. Again, the research project will establish a metabolic profile of African Americans participating in a smoking cessation clinic and investigate whether or not there is a correlation between cessation rates and metabolic profile. Studies also conclude that exposure to environmental tobacco smoke (ETS) contributes to a majority of the cancer cases amongst non-smokers (8,9). Because of the low-levels of cancer causing tobacco metabolites in the urine sample of non-smokers, a reliable and simple extraction method is needed to measure nicotine metabolites. Current extraction methods are too cumbersome and time-consuming. Furthermore, because of the kinetics of degradation of nicotine and the time-consuming preparation times, the extraction methods can induce errors in quantitation of nicotine in urine samples. This research project will expand the studies conducted in the laboratory of Dr. Gang Chen at PSCOM, as well as focus on developing a robust, reliable method to extract nicotine and its metabolites from the urine of non-smokers. The main purpose of this component of the research project is so that the researcher can become better acquainted with the procedures and practices in place related to conducting clinical research and cancer related research. The specific aims of this research project are as follows: Specific Aims: 1. To develop a quantitative LC/MS method using a quadrupole time-of-flight mass spectrometer (qTOF) to detect low levels of nicotine and its major metabolites, in urine samples. 2. To apply the developed qTOF method to investigate the metabolic profile of African-Americans attempting to quit smoking. 3. To develop better extraction methods of nicotine and its metabolites from urine samples of SHS. 4. To continue existing research and technology developed by Dr. Gang Chen to measure low levels of nicotine and its metabolites in non-smokers (SHS) exposed to environmental tobacco smoke (ETS)

研究得出结论，非裔美国人的戒烟率低于白色美国人， 事实上，这一群体每天吸烟少于白色美国人（1-4）。此外，非洲 美国人的烟草相关发病率不成比例地高（5-7）。我假设 非裔美国人可能有一个更困难的时间与戒烟，因为他们无法代谢 尼古丁及其代谢产物。因此，本研究将试图确定是否存在 非裔美国人的戒烟率与他们的尼古丁代谢特征之间的相关性。 这项研究项目将使用LC/MS来测量非裔美国人的尼古丁代谢概况 参加戒烟诊所。将开发一种改进的LC/MS方法， 检测尼古丁及其主要代谢产物可替宁、反式-3 '-OH-可替宁 尼古丁N '-氧化物、4-OH-4-（3-吡啶基）-丁酸、尼古丁-A/-葡糖苷酸、可替宁-A/-葡糖苷酸和反式- 尿液样本中的3 '-OH-可替宁-O-葡糖苷酸。改进的LC/MS方法将结合q-TOF质量 分析仪相对于当前LC/MS（ESI四极杆）扩展了所开发方法的灵敏度和准确度 实践中的方法。该项目的重点是隔离非洲人的尿样， 参加戒烟诊所的美国人。将对样本进行分析， 及其主要代谢物可替宁定量。同样，该研究项目将建立以下代谢特征： 非裔美国人参加戒烟诊所，并调查是否有一个 戒烟率与代谢特征之间的相关性。 研究还得出结论，暴露于环境烟草烟雾（ETS）是大多数人吸烟的原因。 非吸烟者中的癌症病例（8，9）。由于烟草中致癌代谢物的含量很低， 非吸烟者的尿液样本中，需要一种可靠和简单的提取方法来测量尼古丁 代谢物。目前的提取方法过于繁琐和耗时。而且因为 尼古丁降解的动力学和耗时的制备时间，提取方法可以 导致尿液样品中尼古丁定量误差。这项研究项目将扩大研究 在PSCOM的Gang Chen博士的实验室中进行，并专注于开发一种强大，可靠的 从非吸烟者的尿液中提取尼古丁及其代谢物的方法。的主要目的 研究项目的组成部分是为了使研究人员能够更好地了解程序 与进行临床研究和癌症相关研究相关的实践。的具体目标 该研究项目如下： 具体目标： 1.开发使用四极杆飞行时间质谱仪（qTOF）的定量LC/MS方法 检测尿液样本中低水平的尼古丁及其主要代谢物。 2.应用开发的qTOF方法研究非裔美国人的代谢特征 尝试戒烟。 3.目的：研究SHS患者尿液中尼古丁及其代谢产物的提取方法。 4.继续现有的研究和技术开发的陈刚博士，以衡量低水平的 暴露于环境烟草烟雾（ETS）的非吸烟者（SHS）中的尼古丁及其代谢物