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Analytical method of chondroitin/dermatan sulfates using high performance liquid chromatography/turbo ionspray ionization mass spectrometry: application to analyses of the tumor tissue sections on glass slides
Author(s) -
Oguma Toshihiro,
Toyoda Hidenao,
Toida Toshihiko,
Imanari Toshio
Publication year - 2001
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.74
Subject(s) - chemistry , chromatography , dermatan sulfate , disaccharide , chondroitin , reagent , chondroitin sulfate , mass spectrometry , glycosaminoglycan , biochemistry , organic chemistry
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 ‐galactoseΔDi‐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 ‐glucoseTBAOH tetrabutylammonium hydroxideTPAOH tetrapropionylammonium hydroxide