Quantitation of Endogeneous Retinoids

内源性类维生素A的定量

• 批准号: 6739982
• 负责人: MAUREEN A KANE
• 金额: $ 4.73万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6739982
• 关键词: alcoholism /alcohol abuse; diagnostic tests; ethanol; genetically modified animals; laboratory mouse; liquid chromatography mass spectrometry; nutrition related tag; postdoctoral investigator; retinoate; retinoids; technology /technique development; tissue /cell culture; toxicology; vitamin biosynthesis; vitamin metabolism; western blottings

DESCRIPTION (provided by applicant): This research proposes to develop an LC/MS assay to distinguish, identify, and quantitate endogeneous retinoids in biological tissues. Despite the importance of retinoic acid (RA) in development, differentiation, vision, reproduction, and immune response, no sensitive assay exists capable of measuring endogeneous levels. HPLC with various detection schemes lacks sensitivity, GC/MS requires derivitization and cannot resolve geometric isomers of RA, and LC/MS assays to date have not significantly improved detection limits of RA and/or have not been developed as an analytically vigorous assay. The measurement of endogeneous levels of RA and investigation of the hypothesis that RA will colocalize with enzymes that synthesize RA is of great interest towards the understanding of RA function. Specific aims are (1) to develop and validate a sensitive, specific assay for RA and its isomers in biological samples, (2) to demonstrate the utility of the assay by applying it to animal tissue studies consisting of (a) the measurement of retinoids in wt and CRBP -/- mice, (b) the assessment of the impact of ethanol intoxication on all-trans RA levels in wt mice, and (c) the investigation of whether IRA concentration colocalizes with the enzymatic apparatus that synthesizes it in mouse brain and established cell lines.

描述（由申请人提供）：本研究建议开发LC/MS测定法，以区分，识别和定量生物组织中的内差类维生素类似。尽管视黄酸（RA）在发育，分化，视力，繁殖和免疫反应中的重要性，但尚无敏感测定能力，能够测量内差异水平。具有各种检测方案的HPLC缺乏灵敏度，GC/MS需要衍生化，并且无法解析RA的几何异构体，而迄今为止，LC/MS分析尚未显着改善RA和/或尚未开发出作为分析性剧烈测定的检测限。对RA的内差异水平的测量以及RA将与酶共定位的假设的研究，即合成RA对RA功能的理解引起了极大的兴趣。 （1）的具体目的是开发和验证对RA及其在生物样品中的异构体的敏感，特异性测定，（2）通过将其应用于（a）测量类维生素类似的动物组织研究中，以证明测定的实用性。在WT和CRBP - / - 小鼠中，（b）评估乙醇中毒对WT小鼠全反射RA水平的影响，以及（c）研究IRA浓度是否与酶促仪合成小鼠中它合成小鼠中它是否合成小鼠中的酶浓度的研究大脑和建立的细胞系。