Development of Ultra-sensitive HPLC Determination Method for Histamine Based on Intramolecular Excimer-forming Fluorescence Derivatization and Its Applications to Clinical and Neuroscience Investigations

基于分子内准分子形成荧光衍生化的超灵敏 HPLC 测定组胺方法的开发及其在临床和神经科学研究中的应用

• 批准号: 14572043
• 负责人: YAMAGUCHI Masatoshi
• 金额: $ 2.24万
• 依托单位: Fukuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14572043/
• 关键词: Histamine; Excimer fluorescence; HPLC; Fluorescence derivatization; Sensitive determination; Histidine; 臨床研究; 神経薬理; 神経科学

Sensitive and selective HPLC determination methods for histamine related compounds based on intramolecular excimer-forming fluorescence derivatization have been developed. At first, the suitable reagent was screened including the design of novel reagents, and derivatization and HPLC conditions were optimized for the preliminary examinations using histamine standard. By the validation tests, the proposed method is more selective and sensitive than the previous methods with tedious pretreatment procedure. Therefore, the following application studies were carried out.(1) Human urine ; the amounts of histamine in only 1 μL of human urine were determined by the established method with high accuracy without any pretreatment. (2) Rat plasma and tissues; the histamine levels in rat plasma and tissues (abdominal skin, lung, stomach, heart and liver) were measured using excimer fluorescence derivatization method after a simple pretreatment. The obtained values in plasma and tissues were in good agreements with those reported by other workers. (3) Microdialysis samples from rat brain ; real-time monitoring of histamine in rat brain could be carried out by combined with microdialysis techniques. The relationships between the amounts of histamine in some regions of brain (hypothalamus, prefrontal cortex, hippocampus and striatum) and the influence of drugs were investigated.At last, simultaneous determination method of histamine and histidine (precursor of histamine) based on intramolecular excimer-forming fluorescence derivatization was developed. This method was highly sensitive and selective, and useful for clinical and food sanitary fields.In conclusion, the present derivatization technique allows simple, selective and sensitive determination of histamine related compounds.

基于分子内准分子形成荧光衍生化的组胺相关化合物的灵敏和选择性 HPLC 测定方法已经开发出来。首先筛选合适的试剂，包括新型试剂的设计，并优化衍生化和HPLC条件，以使用组胺标准品进行初步检查。通过验证测试，该方法比以前预处理过程繁琐的方法更具选择性和敏感性。因此，开展了以下应用研究。(1)人尿液；仅用所建立的方法即可测定1μL人尿液中的组胺含量，且无需任何预处理，准确度高。 (2) 大鼠血浆和组织；简单预处理后，采用准分子荧光衍生法测定大鼠血浆和组织（腹部皮肤、肺、胃、心脏和肝脏）中的组胺水平。血浆和组织中获得的值与其他工作人员报告的值非常一致。 (3) 大鼠脑微透析样品；结合微透析技术可以对大鼠脑组织胺进行实时监测。研究了大脑某些区域（下丘脑、前额皮质、海马和纹状体）组胺含量与药物影响的关系。最后建立了基于分子内准分子形成荧光衍生化的组胺和组氨酸（组胺前体）同时测定方法。该方法具有高灵敏度和选择性，可用于临床和食品卫生领域。 总之，本衍生技术可以简单、选择性和灵敏地测定组胺相关化合物。

项目成果

T.Yoshitake, F.Ichinose, H.Yoshida, K.Todoroki, J.Kehr, O.Inoue, H.Nohta, M.Yamaguchi: "A Sensitive and Selective Determination Method of Histamine by HPLC with Intramolecular Excimer-forming Derivatization and Fluorescence Detection"Biomedical Chromatogr

T.Yoshitake、F.Ichinose、H.Yoshida、K.Todoroki、J.Kehr、O.Inoue、H.Nohta、M.Yamaguchi：“采用分子内准分子形成衍生化和 HPLC 技术灵敏、选择性测定组胺的方法

T.Yoshitake, M.Yamaguchi, H.Nohta, F.Ichinose, H.Yoshida, S.Yoshitake, K.Fuxe, J.Kehr: "Determination of Histamine in Microdialysis Samples from Rat Brain by Microbore Column Liquid Chromatography Following Intramolecular Excimer-forming Derivatization wi

T.Yoshitake、M.Yamaguchi、H.Nohta、F.Ichinose、H.Yoshida、S.Yoshitake、K.Fuxe、J.Kehr：“分子内准分子后采用微孔柱液相色谱法测定大鼠脑微透析样品中的组胺

H.Yoshida, F.Ichinose, T.Yoshitake, Y.Nakano, K.Todoroki, H.Nohta, M.Yamaguchi: "Simultaneous Determination of Histamine and Histidine by Liquid Chromatography Following Intramolecular Excimer-forming Fluorescence Derivatization with Pyrene-labeling Reage

H.Yoshida、F.Ichinose、T.Yoshitake、Y.Nakano、K.Todoroki、H.Nohta、M.Yamaguchi：“用芘标记试剂进行分子内准分子形成荧光衍生化后，通过液相色谱同时测定组胺和组氨酸

H.Yoshida, F.Ichinose, T.Yoshitake, Y.Nakano, K.Todoroki, H.Nohta, M.Yamaguchi: "Simultaneous Determination of Histamine and Histamine by Liquid Chromatography Following Intramolecular Excimer-forming Fluorescence Derivatization with Pyrene-labeling Reage

H.Yoshida、F.Ichinose、T.Yoshitake、Y.Nakano、K.Todoroki、H.Nohta、M.Yamaguchi：“用芘标记试剂进行分子内准分子形成荧光衍生化后，通过液相色谱同时测定组胺和组胺