Development of New Fluorescence Derivatization Reagents with 6-Aminoquinoxalines

体内有微量的生物活性物质，如羧酸、醇类和胺类。它们的测定对于阐明人体的生理功能、诊断疾病、阐明病因和医学治疗都有重要意义。因此，人们开发了许多荧光衍生试剂，用于高效液相色谱法（HPLS）测定这些物质。本课题研究了新型荧光6-氨基喹喔啉衍生物及其在羧酸荧光衍生试剂中的应用。以2，3-二氯-6-硝基喹喔啉为原料，与胺、醇、冠醚等亲核试剂反应，高产率地合成了1，2，3-二取代6-硝基喹喔啉衍生物。2.用10% Pd-C催化氢化得到荧光6-氨基喹喔啉。3.6-用二环己基碳二亚胺（DCC）-1-羟基苯并三唑（HOBt）法将氨基喹喔啉与长链羧酸偶联，得到相应的6-（酰基）氨基喹喔啉。4.6-采用配有荧光检测器（λ daex 330 nm，λ daem 395 nm）的反相HPLC，使用填充有Finepak SIL C_S. 5的色谱柱，对（酰基）氨基喹喔啉类化合物进行监测<12>。所有峰在40 min内完全分离。这些N-酰基衍生物的检测限为1.0 fmol/穆尔（进样体积）。

There are trace amounts of bioactive substances such as carboxylic acids, alcohols, and amines in the body. The determination of them is quite significant for elucidation of physiological function, diagnosis of sickness, explication of cause of disease, and medical treatment. Therefore, many fluorescence derivatization reagents have been developed for the determination of these substances by high-performance liquid chromatography (HPLS). In this project, I investigated new fluorescent 6-aminoquinoxalines and their application to fluorescence derivatization reagents for carboxylic acids.1.2,3-Disubstituted 6-nitroquinoxalines were obtained by treatment of 2,3-dichloro-6-nitroquinoxaline with some nucleophiles such as amine, alcochol, and crown ether in high yields. 2. The catalytic hydrogenation with 10% Pd-C afforded fluorescent 6-aminoquinoxalines. 3.6-Aminoquinoxalines were coupled with long-chain carboxylic acids by means of the dicyclohexylcarbodiimide (DCC)-1-hydroxybenzotriazol (HOBt) method t give the corresponding 6- (acyl) aminoquinoxalines. 4.6- (Acyl) aminoquinoxalines were monitored by reversed phase HPLC equipped with a fluorescence detector (lambdaex 330 nm, lambdaem 395 nm) by using a column packed with a Finepak SIL C_<12>S.5. All the peaks were completely separated within 40 min. 6. The detection limit of these N-acyl derivatives were found to be 1.0 fmol/mul (injection volume).In conclusion, 2,3-disubstituted 6-aminoquinoxalines have been demonstrated to be applicable to new fluorescence derivatization reagents for the carboxylic acids.