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Analytical method for determination of disaccharides derived from keratan sulfates in human serum and plasma by high‐performance liquid chromatography/turbo‐ionspray ionization tandem mass spectrometry
Author(s) -
Oguma Toshihiro,
Tomatsu Shunji,
Okazaki Osamu
Publication year - 2007
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.760
Subject(s) - chemistry , chromatography , tandem mass spectrometry , mass spectrometry , elution , keratan sulfate , selected reaction monitoring , glycosaminoglycan , biochemistry , chondroitin sulfate
We established a highly sensitive LC/MS/MS method for the analysis of the disaccharides produced from keratan sulfates (KS). It was revealed that the disaccharides produced by keratanase II enzymatic digestion of KS could be determined with high sensitivity by negative ion mode of multiple reaction monitoring. Furthermore, monosulfated and disulfated disaccharides can be separated using a Hypercarb (2.0 mm i.d. × 150 mm, 5 µm) with a gradient elution of acetonitrile–0.01 m ammonium bicarbonate (pH 10). This method was applied to the determination of KS in serum and plasma of control subjects. The intra‐day precision expressed as %CV was within 6.8% for five replicate analyses with three different control serum. The inter‐day (overall, n = 15) precision was within 7.3% for three days. This method is sensitive, reproducible and would be useful for clinical analysis. Copyright © 2007 John Wiley & Sons, Ltd.