Analysis Method for Relative Quantitation and Qualitation of Reductive Glycans via Aniline Stable Lsotopic Labeling and ESI-MS

Abstract

Abstract: An established method for qualitation and quantitation of reductive glycans based on aniline stable isotopic labeling was reported. Various conditions which affect the derivatization efficiency were optimized. Reducing oligosaccharide could be quantatively derivatized by aniline at 70 ℃ within 15 min, under the condition that the saccharide, aniline and sodium cyanoborohydride were in a molar ratio of 1∶1.2∶10. In order to test the linearity and dynamic range of this method, aliquots of standard oligosaccharide(maltodextrin) were labeled with d0/d5-aniline separately. The results indicate that the isotopic labeling method employed here could provide relative quantitative data with a dynamic range of 15-fold, adequate linearity(R=0.9986) and reproducibility with a coefficient of variation that is 10.30% on average. The established method was sensitive and reproducible for the candidate glycans and was successfully used for comparing the differences between free milk oligosaccharides for human(HMOs) and bovine(BMOs). The results show that the lactose amount in human milk is higher than that from bovine milk, and the varieties of oligosaccharides from human milk are more complex than that of bovine milk. The relative quantitative and qualitative method established here will provide an alternative option for studying the changes of the glycans and further understanding the mechanism of the glycosylation, which associate with a wide assortment of diseases intimately.

Key words: Aniline stable isotopic labeling, ESI-MS, Reductive amination, Quantitation of oligosaccharide

TrendMD:

XUE Xiang-Dong, ZHANG Ping, WANG Zhong-Fu, HUANG Lin-Juan* . Analysis Method for Relative Quantitation and Qualitation of Reductive Glycans via Aniline Stable Lsotopic Labeling and ESI-MS[J]. Chem. J. Chinese Universities, 2010, 31(11): 2173.

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