SIMULTANEOUS DETERMINATION OF DERMATAN SULFATE AND OVERSULFATED DERMATAN SULFATE IN PLASMA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH POSTCOLUMN FLUORESCENCE DERIVATIZATION
Y. Huang et al., SIMULTANEOUS DETERMINATION OF DERMATAN SULFATE AND OVERSULFATED DERMATAN SULFATE IN PLASMA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH POSTCOLUMN FLUORESCENCE DERIVATIZATION, Analytical biochemistry, 240(2), 1996, pp. 227-234
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.