SIMULTANEOUS DETERMINATION OF DERMATAN SULFATE AND OVERSULFATED DERMATAN SULFATE IN PLASMA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH POSTCOLUMN FLUORESCENCE DERIVATIZATION

A chemical method for the determination of dermatan sulfate (DS) and o versulfated dermatan sulfate has been developed and applied to the pha rmacokinetic study of these polysaccharides in experimental animals. T he analytical procedure includes a simple preparation step of administ ered DS and oversulfated DS from blood plasma, HPLC for the separation and detection of DS and oversulfated DS using an Asahipak NH2P-50 col umn, fluorometric reaction of the polysaccharides with guanidine in a strong alkaline medium. DS and oversulfated DS were extracted from pla sma by treating it with proteinase to remove plasma proteins and recov ered with endogenous plasma glycosaminoglycans by ethanol precipitatio n. Finally, DS and oversulfated DS were analyzed by fluorometric HPLC. The detection limits of DS and oversulfated DS were 10 and 20 ng, res pectively. Furthermore, we demonstrated that artificial oversulfation of DS increased its biological half-life after intravenous administrat ion to rats. (C) 1996 Academic Press, Inc.