BIOLOGICAL MARKERS OF CHRONIC EXCESS ETHANOL CONSUMPTION

这项提议的目的是开发一种气相色谱 气质联用(GC/MS)和一种新的筛选方法 血液中乙醇和乙醛(ALD)的测定 在血液中的结合物，并确定它们用作 酒精消费的标记物。 由于蛋白质，特别是红细胞蛋白质的寿命很长， 累积的血红蛋白-ALD-加合物的测量 已被提出作为生物标记。其他潜在的标记 是脂肪酸乙酯，存在于人体组织中。 最近，在尿液和血液中检测到乙醇结合物 戒酒。它被假设为并发 血液中乙醇和ALD结合物的测量可能 是最直接、最持久的生物标志物 乙醇消耗量。 目前乙醇偶联物的GC/MS方法可以是 进一步开发用于低气压的同时测量 浓度ALD共轭。在钠存在的情况下 一种还原剂--硼氢化钠，是酸化后产生的 水解应该会得到重氢乙醇，它可能是 经GC/MS测定。 此方法将用于测试同时使用这两种方法的可行性 乙醇和碱性磷酸酶结合物作为慢性阻塞性肺疾病的生物标志物 过量的乙醇消费，通过重新分析一个特殊的 采集酗酒者和对照组的血液样本。这些 样本是由生化主管伊斯雷尔博士收集的 临床成瘾研究所研究部 安大略省基金会为他开发了一种酶联 对Hb-ALD-加合物的免疫吸附试验(EL ISA)。 GC/MS程序也将用于开发一种新型血液 基于放射性白蛋白结合的筛查试验 乙醛和乙醇。

The aim of this proposal is to develop a gas-chromatographic mass-spectrometric (GC/MS) procedure and a novel screening blood test for measurement of ethanol and acetaldehyde (ALD) conjugates in blood and to establish the feasibility of their use as markers for ethanol consumption. Since proteins, especially red cell proteins, have a long life span, measurement of the accumulated hemoglobin-ALD-adducts have been proposed as biological markers. Other potential markers are ethyl esters of fatty acids, which are found in human tissues. Recently, ethanol conjugates were detected in urine and blood of abstaining alcoholics. It is hypothesized the concurrent measurements of both ethanol and ALD conjugates in blood may serve as the most direct and long-lasting biological markers for ethanol consumption. The present GC/MS procedure for ethanol conjugates can be further developed for simultaneous measurements of low concentration ALD conjugates. In the presence of sodium borodeuteride, a reducing agent, ALD produced after acid hydrolysis should give deuterated ethanol which could be determined by GC/MS. This method will be used to test the feasibility of using both ethanol and ALD conjugates as biological markers of chronic excessive ethanol consumption, by re-analysing a special collection of blood samples from alcoholics and controls. These samples have been collected by Dr. Israel, head of biochemical research, clinical Institute of The Addiction Research Foundation of Ontario for his development of an enzyme-linked immunosorbent assay (ELISA) against hemoglobin-ALD-adducts. The GC/MS procedure will also be used to develop a novel blood test for screening based on radio albumin binding of acetaldehyde and ethanol.