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

• Published in: Analytical biochemistry Vol. 252; no. 1; pp. 56 - 61
• Main Authors: Kakehi, Kazuaki, Maeya, Yu-ko, Miki, Yasuyoshi, Oda, Yasuo, Hayase, Shozo
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.1997
• Subjects: Chondroitin Sulfates - chemistry; Fluorescent Dyes; Fluorometry - methods; Glycosaminoglycans - analysis; Glycosaminoglycans - chemistry; Molecular Weight; Quaternary Ammonium Compounds - chemistry; Reproducibility of Results; Solutions; Sulfates - chemistry
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1006/abio.1997.2306

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.