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

在弱碱性介质中，邻乙酰基苯甲醛(OAB)与组胺快速反应生成高荧光加合物，建立了一种简单、高灵敏度的流动注射分析和高效液相色谱测定几种生物活性胺的方法。利用该荧光反应作为组胺的柱后荧光标记，建立了一种简便、灵敏的测定组织中组胺的高效液相色谱方法。另一方面，利用4-溴-1，2-苯甲醛(BBDA)与组胺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；在pH为9.3的条件下，谷胱甘肽与OAB迅速反应生成稳定的高荧光加合物。利用该反应作为柱前荧光标记，建立了测定人红细胞和…中谷胱甘肽的高效液相色谱方法，检出限(S/N=3)为50fmol/注射剂在碱性介质中与邻苯二甲醛反应产生稳定的荧光加合物，然后溶液酸化。该荧光反应对亚精胺具有很高的选择性，并用于柱前荧光标记高效液相色谱检测亚精胺。建立了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的方法。该方法适用于尿液、血清和血纸中苯丙氨酸的测定，对伯胺高度敏感的2，3-萘甲醛-KCN试剂体系进行荧光标记后，用高效液相色谱法测定生物素的代谢物之一--生物细胞素。较少

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