高效液相色谱法分析新型细菌多糖中单糖和糖醛酸组成

Detection of Monosaccharides and Uronic Acid in a Novel Bacterial Polysaccharides Using High Performance Liquid Chromatography

建立了柱前1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生-高效液相色谱法,分离检测新型细菌多糖中5种单糖及糖醛酸组成的分析方法。新型细菌多糖中加入单糖内标木糖后,经3mol/L三氟乙酸水解,再用PMP进行衍生化,采用XBridgeTMC18柱(250×4.6mm,5μm),以50mmol/L KH2PO4溶液(pH=5.5)和乙腈为流动相,在245nm波长检测。定量分析结果表明,该多糖由甘露糖、葡萄糖醛酸、葡萄糖、半乳糖和岩藻糖组成,其质量比为1.77∶0.44∶8.52∶1.32∶1.67。该方法重现性良好,能够快速对多糖样品进行组成分析。

High performance liquid chromatography （HPLC） was employed for the analysis of monosaccharides and uronic acid in novel bacterial polysaccharides with pre-column derivatization. After being degradated by 3 mol/L trifluoroacetic acid and derivatized with 1-phenyl-3-methyl-5-pyrazolone （PMP）, monosaccharides and uronic acid were separated by reversed-phase HPLC with isocratic elution, and detected by ultraviolet detector at 245 nm. The result showed that the bacterial polysaccharides consisted of 4 kinds of monosaccharides and 1 kind of galacturonic acid. The mass ratio of mannose, glucuronic acid,glucose,galactose and fucose was 1.77：0.44：8.52：1.32：1.67. The developed method was suitable for the rapid analysis of polysaccharide with good reproducibility.