[Analytical chemical studies on high-performance recognition and detection of bio-molecules in life]

In order to understand the mechanism for maintaining life of animals based on the search of dynamics of biomolecules, I have developed several sensitive and selective methods for their quantification. Using the methods of derivatization with the developed benzofurazan fluorogenic reagents (4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), ammonium 7-fluoro-2,1,3-benzoxadiazole 4-sulfonate (SBD-F) and etc.) followed by high-performance liquid chromatography (HPLC)--fluorescence detection, a certain kind of biological and clinical importance was demonstrated of chiral bio-molecules (D-amino acids, D-lactic acid and so on), peptides and proteins. The proposed method (derivatization with SBD-F, isolation of the fluorescent proteins by two-dimensional HPLC, enzymatic digestion and identification of the altered proteins by HPLC-mass spectrometry (MS)/MS with database-searching algorithm) for proteomics studies revealed the changed proteins in the islets of Langerhans of the dexamethazone-induced diabetic rats. An importance of catecholamine metabolism on the blood pressure regulation was also suggested by the method of HPLC-chemiluminescence detection of catecholamines and their 3-O-methylmetabolites. A new field of Analytical Chemistry, i.e., Bio-Analytical Chemistry, was also proposed.