Development and application of the analytical methods with spectrometry for bio-related compounds

Recent studies on the development and application of analytical methods in HPLC with spectrometric detection for bio-related compounds have been reviewed. First, new fluorescent derivatization reagents with benzofurazan skelton have been developed. In the course of the study, the relationship between fluorescent characteristics and substituent groups of benzofurazan compounds was investigated. The relationship enabled to design new fluorogenic reagents easily Secondly, the simultaneous determination method for the sequence and D/L configuration of peptides with fluorescent Edman reagent was developed. In this study, the racemization mechanism of amino acids in the cleavage/cyclization step was investigated and boron trifluoride was employed as cleavage/cyclization reagent in order to supress the racemization. Finally, sensitive and selective determination system for catecholamine with peroxyoxalate chemiluminescence detection after derivatization with ethylenediamine was developed. Utilizing this system, plasma catecholamine level of spontaneously hypertensive rats and Wistar-Kyoto rats after administration of calcium antagonists were measured, and the attenuation of the baroceptor reflex was investigated.