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目的:建立测定网地藻多糖的单糖组成方法。方法:采用超声辅助提取、乙醇沉淀、Sevage法脱蛋白制备网地藻多糖,采用硫酸水解后,水解产物以1-苯基-3-甲基-5-吡唑酮(PMP)柱前衍生化后采用反相高效液相色谱法于248 nm处测定单糖组成。色谱柱为Diamonsil C18(250mm×4.6 mm,5μm);流动相为0.02 mol/L磷酸缓冲盐(A)-乙腈(B),洗脱梯度(0 min,0%B;10min,18%B;25 min,25%B;30 min,30%B)。结果:网地藻多糖含有甘露糖、核糖、鼠李糖、葡萄糖、半乳糖和木糖,其摩尔比为0.24:0.04:0.03:1:0.19:0.1。结论:该方法准确可靠、分离效果较好,可用于网地藻多糖的单糖组成测定。
OBJECTIVE: To establish a method for determining the monosaccharide composition of polysaccharides from Phytoplankton. Methods: Ultrasound-assisted extraction, ethanol precipitation and Sevage deproteinization were used to prepare polysaccharides from Mesorhizobium. Hydrolysis of the polysaccharides was carried out by sulfuric acid. The hydrolyzate was pre-column derivatized with 1-phenyl-3-methyl-5-pyrazolone After the determination of monosaccharide composition at 248 nm by reversed-phase high performance liquid chromatography. The column was Diamonsil C18 (250 mm × 4.6 mm, 5 μm). The mobile phase consisted of 0.02 mol / L phosphate buffered saline (A) -acetonitrile (B) with gradient elution (0 min, 0% 25 min, 25% B; 30 min, 30% B). Results: Mesophyll polysaccharide contained mannose, ribose, rhamnose, glucose, galactose and xylose in a molar ratio of 0.24: 0.04: 0.03: 1: 0.19: 0.1. Conclusion: The method is accurate and reliable with good separation effect and can be used for the determination of monosaccharide compositions of polysaccharides from.