Separation of ten phosphorylated mono-and disaccharides using HILIC and ion-pairing interactions

Analytica Chimica Acta
Caroline MathonCynthia K Larive

Abstract

Phosphorylated carbohydrates are indispensable cogs in several key metabolic wheels for all forms of life. Here, a straightforward liquid chromatography method coupled to mass spectrometry detection was developed for phosphorylated sugars. For separation of the targeted compounds, hydrophilic interaction chromatography (HILIC) was used with a bridged-ethylene hybrid amide column under alkaline conditions using triethylamine as a mobile phase modifier. Methylphosphonic acid was added to the aqueous mobile phase to reduce the tailing of compounds containing phosphate groups, which are known to interact with stainless steel components of the separation system. Under alkaline conditions and addition of methylphosphonic acid, the retention behavior can be attributed to both conventional HILIC mechanisms as well as ion-pairing interactions in the mobile phase. This hypothesis is supported by comparing the retention behavior of phosphorylated sugars and unmodified sugars. The HILIC method resolved eight biologically important phosphorylated sugars and thereby enables simultaneous detection and quantification of these compounds: fructose-1,6-bisphosphate, glucose-1-phosphate, glucose-6-phosphate, lactose-1-phosphate, mannose-6-phosphat...Continue Reading

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Citations

Apr 3, 2019·Annual Review of Analytical Chemistry·Yu LiangYukui Zhang
Oct 18, 2019·Critical Reviews in Food Science and Nutrition·Supradip SahaKaushik Banerjee
Jun 26, 2018·Journal of Proteome Research·Corey M GriffithCynthia K Larive
Oct 16, 2021·Analytical and Bioanalytical Chemistry·Salomé PoyerRégis Daniel

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