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Development of FIA and HPLC Methods for the Determination of Bioactive Amines by Using Highly Selective Fluorogenic Reagents.

开发使用高选择性荧光试剂测定生物活性胺的 FIA 和 HPLC 方法。

基本信息

批准号：
06672134
负责人：
MIURA Toshiaki
金额：
$ 1.34万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

SImple and highly sensitive FIA and HPLC methods were developed for the determination of several bioactive amines by using highly selective fluorogenic reagents for each the amine.Orthoacetylbenzaldehyde (OAB) reacts rapidly with histamine in weekly alkaline medium to form a highly fluorescent adduct. Simple and sensitive HPLC method for the assay of histamine in tissues was developed by employing this fluorescence reaction as postcolumn fluoresence labeling of histamine. On the other hand, FIA method for the rapid assay of histamine was established by using 4-bromo-1,2-benzenedicarbaldehyde (BBDA) which reacts specifically with histamine to form a stable fluorescent adduct.Glutathione also reacts rapidly with OAB at pH 9.3 to form a stable and highly fluorescent adduct. By employing this reaction as precolumn fluorescence labeling, Simple and ultrasensitive HPLC method with detection limit (S/N=3) of 50 fmol/injection was developed for the assay of glutathione in human erythrocytes an … More d sera.Spermidine gives a stable fluorescent adduct in the reaction with orthophthalaldehyde in alkaline medium followed by acidification of the solution. This fluorescence reaction was highly selective for spermidine and was used to precolumn fluorescence labeling for HPLC measuremment of spermidine. Serum spermidine was determined by this HPLC method in combination with solid phase extraction using ion exchage minicolumn.Micromethod for the determination of phenylalamine was established, which consisted of conversion of phenylalanine into trans-cinnamic acid with phenylalanine ammonia-lyase, and measurement of trans-cinnamic acid by reverse phase HPLC with UV detection. This method was applicable to the assay of phenylalanine in urine, serum and blood paper for screening of phenylketourea.Biocytin, one of metabolites of biotin, was determined by HPLC method after fluorescence labeling with the reagent system of 2,3-naphthalenedicarbaldehyde and KCN,which is highly sensitive for primary amines. Less

在弱碱性介质中，邻乙酰苯甲醛（OAB）与组胺反应生成高荧光加合物。利用该荧光反应作为组胺的柱后荧光标记，建立了组织中组胺的高效液相色谱测定方法。另一方面，利用4-溴-1，2-苯二甲醛（BBDA）能与组胺形成稳定的荧光加合物，谷氨酸在pH9.3时能与邻氨基苯甲酸（OAB）形成稳定的荧光加合物，建立了快速测定组胺的流动注射分析方法。利用该反应作为柱前荧光标记，建立了一种简便、灵敏的测定人红细胞中谷胱甘肽的高效液相色谱法，检测限（S/N=3）为50 fmol/injection， ...更多信息亚精胺在碱性介质中与邻苯二甲醛反应，然后酸化溶液，得到稳定的荧光加合物。该荧光反应对亚精胺具有高选择性，并用于柱前荧光标记，用于亚精胺的HPLC测定。采用离子交换微柱固相萃取-高效液相色谱法测定血清中的亚精胺，建立了微量测定苯丙氨酸的方法，即用苯丙氨酸解氨酶将苯丙氨酸转化为反式肉桂酸，反相高效液相色谱-紫外检测法测定反式肉桂酸。该方法适用于尿、血清和血纸片中苯丙氨酸的检测，可用于苯酮脲类药物的筛选。以2，3-萘二甲醛-KCN为试剂体系，荧光标记后，用高效液相色谱法测定生物素的代谢产物生物胞素。少