Rapid Analysis of Serum Albumin by High-Performance Liquid Chromatography Using N-Methylpyridinium Polymer

Abstract

The isocratic elution of serum albumin on a column containing N-methylpyridinium polymer was investigated and compared with the gradient elution reported previously. Separation of mercaptalbumin and nonmercaptalbumin was markedly affected by the pH, buffer system and salt concentration in the eluent. Good peak resolution of mercaptalbumin and nonmercaptalbumin was obtained with tris (hydroxymethyl)aminomethane-acetic acid buffer (pH 7.0) containing sodium acetate. This method was applied to the analysis of human serum: f_HMA (human mercaptalbumin fraction in albumin) was determined with a high precision.