Isolation and analysis of sugar nucleotides using solid phase extraction and fluorophore assisted carbohydrate electrophoresis

摘要

class="kwd-title">Abbreviations: HPLC, high performance liquid chromatography; SPE, solid phase extraction; FACE, fluorophore assisted carbohydrate electrophoresis; UDP, uridine diphosphate; GDP, guanosine diphosphate; CMP, cytosine monophosphate; TEAA, triethylamine acetate; APS, ammonium persulfate; Gal, galactose; GalNAc, N-acetylgalactosamine; GlcNAc, N-acetylglucosamine; Man, Mannose; NeuAc, sialic acid; GlcUA, glucuronic acid; AMAC, 2-aminoacridone; TEMED, N′,N′,N′N′-tetramethylenediamine class="kwd-title">Method name: Sugar nucleotide analysis by SPE and FACE class="kwd-title">Keywords: Sugar nucleotides, Face, HPLC, Carbohydrate, Electrophoresis class="head no_bottom_margin" id="abs0010title">AbstractThe building blocks of simple and complex oligosaccharides, termed sugar nucleotides, are often overlooked for their role in metabolic diseases and may hold the key to the underlying disease pathogenesis. Multiple reasons may account for the lack of analysis and quantitation of these sugar nucleotides, including the difficulty in isolation and purification as well as the required expensive instrumentation such as a high performance liquid chromatography (HPLC), mass spectrometer, or capillary electrophoresis. We have established a simple yet effective way to purify and quantitate sugar nucleotides using solid phase extraction (SPE) chromatography combined with fluorophore assisted carbohydrate electrophoresis (FACE). The simplicity of use, combined with the ability to run multiple samples at one time, give this technique a distinct advantage over the established methods for isolation and analysis of sugar nucleotides from cell culture models. class="first-line-outdent">

• Sugar nucleotides can be easily purified with solid phase extraction chromatography.

• FACE can be used to analyze multiple nucleotide sugar extracts with a single run.

• The proposed method is simple, affordable, and uses common everyday research labware.