Abstract:High-performance anion-exchange chromatography with pulsed amperometric detector (HPAEC-PAD) was widely used to analyze monosaccharide in botanical and fungal polysaccharides, but lightly used in marine animal polysaccharides. A method has been developed for determining monosaccharide composition and content in sea cucumber polysaccharide by HPAEC-PAD. Purifying sea cucumber polysaccharide were optimized by comparing three deproteinization methods. Optimum chromatographic conditions for the separation of monosaccharide were optimized by comparing eluent concentration on the separation of monosaccharide. The results indicated that the reagent of Sevag removed micro-molecule polypeptide more effectively than others from sea cucumber polysaccharides. The concentration of eluent had significant effect on separating monosaccharide. Eight monosaccharide, such as Fuc, Gal, Ara, GalN, GlcN, Man, Glc, and Lac, were separated using 20 mmol/L NaOH as eluent. GlcUA and GalUA were separated by 160 mmol/L NaOH and 200 mmol/L NaAC as eluent. The linear coefficients of monosaccharide standard were above 0.998. The detection limit of each monosaccharide was between 2.5 and 50 μg/L, and the recovery rates ranged from 83.6% to 113.1%. The method determining neutral sugar, amino sugar and uronic acid in sea cucumber polysaccharides could be used as analysis and quantitative method of monosaccharide in sea cucumber and its products. The composition and content of monosaccharide were significant indices for determining the biological activity of sea cucumber. The established method could provide basic monosaccharide information and promote the polysaccharides research progress in sea cucumber. It could provide references for the establishment of sea cucumber standards, and expand the application of active substances in medicine and other fields.
