柱前衍生化-高效液相色谱法分析缅甸刺梧桐胶多糖的单糖组成

Analysis of Monosaccharide Composition of Myanmar’s Karaya gum Polysaccharides by HPLC with Pre-column Derivatization

利用1-苯基-3-甲基-5-吡唑啉酮柱前衍生化-高效液相色谱法,优化10种常见单糖的色谱分离条件,并用于不同等级的自制刺梧桐胶多糖的单糖组成分析。对自制刺梧桐胶多糖的单糖组成分析结果表明,自制刺梧桐胶多糖由葡萄糖醛酸(GlcUA)、鼠李糖(Rha)、半乳糖醛酸(GalUA)、半乳糖(Gal)、阿拉伯糖(Ara)和岩藻糖(Fuc)6种单糖组成,平均物质的量比为1∶3.30∶1.62∶2.45∶0.08∶0.03。对不同等级的刺梧桐胶多糖的分析鉴定结果表明,不同等级的刺梧桐胶多糖的单糖组成均含Rha、GalUA、GlcUA、Gal 4种单糖组分,其中Ara在缅甸A、B级中没有检测出来,Fuc在缅甸D级中没有检测出来。方法学结果显示:10种单糖在各自线性范围内线性关系良好,相关系数r≥0.9992,6种单糖加标回收率在96.80%~101.26%之间,相对标准偏差小于3%。该方法简单、准确灵敏,可用于刺梧桐胶多糖的单糖组成分析。

A method for separation of ten monosaccharides was established by using high performance liquid chromatography(HPLC)with 1-phenyl-3-methyl-5-pyrazolone(PMP)pre-column derivatization,and used to analyze the monosaccharide composition of polysaccharides of seven different grades from Karaya gum.The results showed there were good linear relationships(r≥0.9992)between the peak area values and concentrations of ten monosaccharides tested within their linear ranges.The relative standard deviations of precision were less than 3%,and the relative standard deviations of repeatability and stability were less than 5%.The recoveries of six monosaccharides were 96.80%-101.26%.This method was simple,sensitive,suitable for the analysis of Karaya gum Polysaccharides’monosaccharide composition.The results of monosaccharide composition analysis of self-made polysaccharides from Karaya gum showed that Karaya gum polysaccharide was composed of glucuronic acid,rhamnose,galacturonic acid,galactose,arabinose and fucose with the average molar ratio of 1∶3.30∶1.62∶2.45∶0.08∶0.03.The results of monosaccharide composition analysis of polysaccharides from Karaya gum showed that the monosaccharide composition of different grades of polysaccharides contained rhamnose,galacturonic acid,glucuronic acid and galactose,among which arabinose was not detected in Myanmar A and B,fucose was not detected in Myanmar D.