Research On New Highly Sensitive Fluorescent Reagents And Their Applications In HPLC

HPLC is one of the fastest developing and the most widely used fields in analytical chemistry. Due to its separation characteristic, the enthusiastic adoption of RPLC method has had a great impact on environmental chemistry, biochemistry, toxicology, clinical chemistry and many other fields. However, not all compounds are accessible to direct analysis by chromatography due to the limitations of properties of the compounds themselves and the detect means. Therefore, using chemistry derivatization reaction to alter the chemical physics properties of target compounds to make them easy to be analyzed by 1-IPLC is the purpose of this thesis. Although many kinds of fluorescent reagents have been developed, there are still disadvantages in their applications. A systematic research on the optimum derivatization condition of several new kinds of highly sensitive reagents such as 9-(2-hydroxylethyl)-acridone and 9-acridone-(ct-methyl)-acetic acid etc. for the determination of amines, alcohols and carboxylic acids as well as HPLC separation of their derivatives were investigated for the first time in this thesis. In this study, the composition of the optimum mobile phase and the influences of many other factors (reaction time, reaction temperature, volume of condensing agent and catalyst) were investigated, the optimization of derivatization conditions was also studied. In addition, the applications of these fluorescent reagents in the separation of more complicated samples such as amino acids and bile acids and determination of composition from a real sample were carried out for the first time with satisfactory results. It can be seen from our experiments that, compared with some existing fluorescent reagents, the derivatization reactions of these new reagents with amines, oxy-compounds and carboxylic acids are mild and easy to be carried out; The fluorescence responses of their derivatives are high enough to be detected, the products are stable and easy to be separated; At the same time, the by-products didn抰 interfere with the separation and determination of the derivatives. It shows that these fluorescent reagents can be widely used in the determination of many kinds of compounds by HIPLC with fluorescence detection.