基于手性整体柱全自动二维高效液相色谱技术的体内D-型酸性氨基酸的分布及生理机能研究

项目名称： 基于手性整体柱全自动二维高效液相色谱技术的体内D-型酸性氨基酸的分布及生理机能研究

项目编号： No.81202499

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 药物学、药理学

项目作者： 韩海

作者单位： 暨南大学

项目金额： 21万元

中文摘要： 自D-型酸性氨基酸在哺乳动物体内的微量存在被发现以来，它们的重要生理机能及疾病标志物作用已逐渐成为作用于神经及内分泌系统的新型药物开发及新诊断方法开发的研究热点。然而，因体内大量存在的L-型氨基酸和内源性干扰物的影响，体内样品中酸性氨基酸D，L体的高选择性同时快速定量分析一直难以实现，其生理作用也难以进一步阐明。本研究旨在申请人过去的2D-HPLC研究基础上，开发新型高效整体手性色谱柱及ODS柱，并配合多样品环柱切换技术建立高选择性的迅速手性拆分的2D-HPLC全自动化系统，以实现哺乳动物组织和体液中微量的D-型酸性氨基酸及其L-异构体的高专属性同时测定。并应用该系统研究D-天门冬氨酸氧化酶基因敲除型鼠和多种疾病模型鼠体内组织及体液中D及L型酸性氨基酸的分布，为揭示其在神经，内分泌及免疫系统中的生理药理作用及疾病诊断标志物作用；开发如D-型氨基酸类保健食品和DDO抑制剂等药物提供科学依据。

中文关键词： D-型酸性氨基酸；二维高效液相色谱；体内分析；生理机能；手性整体柱

英文摘要： Since the presence of naturally occurring D-acidic amino acids in mammals has been observed, their physiological importance and roles of disease bio-marks and has gradually become the focus of the discovery of new drugs which act on neuron and endocrine system, as well as the discovery of new diagnostic methods of the neuron and endocrine disease. However, the sensitivity quantitative analysis of D-acidic amino acids and its L- isomers in mammalian tissues and physiological fluids is quite difficult because of interference of large amount L-amino acids and unknown intrinsic substances in the complex biological matrices. How to separate the trace D-acidic amino acids from their L-forms and determine the both of them simultaneously with high speed and selectivity is becoming a problem. In this study, based on the 2D-HPLC methods for the analysis of amino acids established in previous work, we plan to discover some new type of high efficiency monolithic enantiomer separation column and ODS column to establish a new fully-automatic highly selectivity monolithic 2D-HPLC system using multi-loop technique so that the simultaneous determination of trace amounts of D-acidic amino acids and their L- isomers in the biological samples can be carried out with high selectivity. Moreover, by applying this monolithic 2D-HPLC sy

英文关键词： D-acidic amino acids；2D-HPLC；bio-analysis；physiological function；chiral monolithic column