Development of an extensive linkage library for characterization of carbohydrates

Ace Gita Galermo, Eshani Nandita, Juan Jose Castillo, Matthew Joseph Amicucci, Carlito B. Lebrilla

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The extensive characterization of glycosidic linkages in carbohydrates remains a challenge due to the lack of known standards and limitations in current analytical techniques. This study encompasses the construction of an extensive glycosidic linkage library built from synthesized standards. It includes an improved liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantitation of glycosidic linkages derived from disaccharides, oligosaccharides, and polysaccharides present in complicated matrices. We present a method capable of the simultaneous identification of over 90 unique glycosidic linkages using ultrahigh-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC/QqQ MS) operated in dynamic multiple reaction monitoring (dMRM) mode. To build the library, known monosaccharides commonly found in plants were subjected to partial methylation to yield partially derivatized species representing trisecting, bisecting, linear, and terminal structures. The library includes glycosidic linkage information for three hexoses (glucose, galactose, and mannose), three pentoses (xylose, arabinose, and ribose), two deoxyhexoses (fucose and rhamnose), and two hexuronic acids (glucuronic acid and galacturonic acid). The resulting partially methylated monosaccharides were then labeled with 1-phenyl-3-methyl-5-pyrazolone (PMP) followed by separation and analysis by UHPLC/dMRM MS. Validation of the synthesized standards was performed using disaccharide, oligosaccharide, and polysaccharide standards. Accuracy, reproducibility, and robustness of the method was demonstrated by analysis of xyloglucan (tamarind) and whole carrot root. The synthesized standards represent the most comprehensive group of carbohydrate linkages to date.

Original languageEnglish (US)
JournalAnalytical chemistry
DOIs
StateAccepted/In press - Jan 1 2019

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Carbohydrates
Monosaccharides
Disaccharides
Liquid chromatography
Oligosaccharides
Mass spectrometry
Polysaccharides
Hexuronic Acids
Pentoses
Rhamnose
Glucuronic Acid
Arabinose
Methylation
Hexoses
Ribose
Fucose
Monitoring
Xylose
Mannose
Galactose

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Galermo, A. G., Nandita, E., Castillo, J. J., Amicucci, M. J., & Lebrilla, C. B. (Accepted/In press). Development of an extensive linkage library for characterization of carbohydrates. Analytical chemistry. https://doi.org/10.1021/acs.analchem.9b03101

Development of an extensive linkage library for characterization of carbohydrates. / Galermo, Ace Gita; Nandita, Eshani; Castillo, Juan Jose; Amicucci, Matthew Joseph; Lebrilla, Carlito B.

In: Analytical chemistry, 01.01.2019.

Research output: Contribution to journalArticle

Galermo, Ace Gita ; Nandita, Eshani ; Castillo, Juan Jose ; Amicucci, Matthew Joseph ; Lebrilla, Carlito B. / Development of an extensive linkage library for characterization of carbohydrates. In: Analytical chemistry. 2019.
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