Trace determination and characterization of some proteins in blood serum by high-performance liquid chromatography with a time-resolved laser fluorimetric detector

Protein in human serum is measured by high-performance liquid chromatography with a time- resolved laser fluorimetric detector. The detection limit is 2.3 pmol for bovine serum albumin; the value is governed by fluctuation of the background fluorescence from free 1-anilino-8-naphthalene sulfonate (ANS) used as the fluorescence probe. Micropolarity of the binding site in protein is estimated by measuring the fluorescence lifetime of ANS bound to protein; the micropolarity for albumin is lower than those for α- and γ-globulins.