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Abstract

We established a highly sensitive quantitative analytical method for chondroitin/dermatan sulfates by LC/MS method. By this method, the unsaturated disaccharides produced after the enzymatic digestion of chondroitin/dermatan sulfates can be determined in the amounts as low as 0.5 pmol levels. The use of tetrabutylammonium hydroxide as an ion-pair reagent for LC/MS allowed us to separate unsaturated 4-sulfated disaccharide and unsaturated 6-sulfated disaccharide. Furthermore, the peak areas of unsaturated disaccharides were increased almost 10 times by the postcolumn addition of acetonitrile. We applied this LC/MS method to the analyses of unsaturated disaccharides from chondroitin/dermatan sulfates in the tissues sections on glass slides, which were prepared from MethA tumor-bearing mice. This method brought about considerable reduction in the time distance from sample collection to preparation of analytical results. Copyright © 2001 John Wiley & Sons, Ltd.

Abbreviations used:
ΔDi-4S

2-acetamido-2-deoxy-3-O-(β-D-gluco-4-enopyranosyluronic acid)-4-O-sulfo-D-galactose

ΔDi-6S

2-acetamido-2-deoxy-3-O-(β-D-gluco-4-enopyranosyluronic acid)-6-O-sulfo-D-galactose, ΔDiHS-NS, 2-deoxy-2-sulfamino-4-O-(4-deoxy-α-L-threo-hex-4-enopyranosyluronic acid)-D-glucose

TBAOH

tetrabutylammonium hydroxide

TPAOH

tetrapropionylammonium hydroxide