Analytical Chemical Studies on High‐Performance Recognition and Detection of Bio‐Molecules in Life

Abstract

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 quantiˆcation. Using the methods of derivatization with the developed benzofurazan ‰uorogenic reagents (4-‰uoro-7-nitro-2,1,3-benzoxadiazole (NBD F), ammonium 7-‰uoro-2,1,3-benzoxadiazole 4-sulfonate (SBD F) and etc.) followed by high-performance liquid chromatography (HPLC)-‰uorescence detection, a certain kind of biological and clinical importances 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 ‰uorescent proteins by two-dimensional HPLC, enzymatic digestion and identiˆca-tion of the altered proteins by HPLC mass spectrometry (MS)/MS with database-searching algorithm) for proteomics studies revieled 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 newˆeld of Analytical Chemistry, i.e., Bio-Analytical Chemistry, was also proposed.

Determination of fluvoxamine in rat plasma by HPLC with pre-column derivatization and fluorescence detection using 4-fluoro-7-nitro-2,1,3-benzoxadiazole

“…4-Fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) was synthesized as a fluorescent reagent by Imai et al, and selectively reacts toward the primary or secondary amino group Imai, 1981, 1983;Imai, 2003). Simultaneous determination of amino acids as well as amino glycoside was possible using NBD-F Imai, 1981, 1983;Honda et al, 2000;Aoyama et al, 2004;Fukushima et al, 2004).…”