Highly Sensitive Detection in High Performance Liquid Chromatography

Abstract

Fluorogenic reagents for amines and thiols which are of value for their determination in high performance liquid chromatography (HPLC) are reviewed. Special attention was made on the development and application of 4-fluoro-7-nitrobenzo-2-oxa-1, 3-diazole (NBD-F) and ammonium 7-fluorobenzo-2-oxa-1, 3-diazole-4-sulfonate (SBD-F) which are respective reagents for amines and thiols. Finally, usage of a chemiluminescence reaction (TCPO-H₂O₂-fluorescent compounds) for the detection of fluorescent compounds separated by HPLC column was proposed. The proposed method allowed us to detect 2 fmol of dansylated amino acids.