期刊
ANALYTICA CHIMICA ACTA
卷 972, 期 -, 页码 102-110出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2017.03.029
关键词
Phosphorylated sugars; HILIC-MS; Methylphosphonic acid; Ion-pairing; BEH amide column
资金
- National Science Foundation [NSF CHE-1213845, IOS-1121626]
- USDA [NIFA-AFRS 2011-04015]
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-phosphate, ribose-5-phosphate, sucrose-6-phosphate, and threhalose-6-phosphate. Fructose-1-phosphate and fructose-6-phosphate were not resolved but quantification of total fructose-phosphate is possible. (C) 2017 Elsevier B.V. All rights reserved.
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