Journal of Chromatography B 2011-04-15

Microdetermination of hyaluronan in human plasma by high-performance liquid chromatography with a graphitized carbon column and postcolumn fluorometric detection.

Hidenao Toyoda, Fumie Muraki, Toshio Imanari, Akiko Kinoshita-Toyoda

Index: J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci. 879(13-14) , 950-4, (2011)

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Abstract

A chemical method for the determination of hyaluronan (hyaluronic acid, HA) has been developed and applied to the human blood plasma. Human blood plasma HA was converted to the ΔDi-HA by digestion with hyaluronidase SD and determined by a sensitive and selective high-performance liquid chromatography (HPLC). The HPLC includes the separation and detection of ΔDi-HA using a graphitized carbon column and fluorometric reaction with 2-cyanoacetamide in an alkaline eluent. The calibration graph for ΔDi-HA was linear over the range 0.2 ng-1 μg. It was revealed that the concentration of HA in normal human blood plasma is very low levels (about 24 ng/ml) in comparison to low-sulfated chondroitin 4-sulfate (about 13 μg/ml).Copyright © 2011 Elsevier B.V. All rights reserved.