Use of a Binary Mixture of Quaternary Ammonium Salts in Fluorometric Determination of Glycosaminoglycans

Abstract

The concentration of glycosaminoglycans in an aqueous solution was determined by observing the fluorescent intensity of the ion of 2-hexadecyl-9H-pyrido[4,3b]indole at 450 nm by irradiating 253-nm light after dissolution of the insoluble salt in ethanol, which was formed by the reaction between a solution of a sample of glycosaminoglycan and an aqueous solution of the binary mixture of fluorescent 2-hexadecyl-9H-pyrido[4,3b]indolium bromide and hexadecyl pyridinium chloride. The fluorescent quaternary ammonium salt, which was slightly soluble in water, was solubilized in water by forming mixed micelles with a nonfluorescent soluble quaternary ammonium salt. The present method showed good linearity for the calibration curves between 5 and 1000 μg/ml in all samples of glycosaminoglycan except keratan sulfate. The relative standard deviation in determination was less than 3% for the whole calibration range.