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目的建立一种反相高效液相色谱法(RP-HPLC)分析大蒜多糖中的单糖组成及摩尔比。方法大蒜多糖样品经2mol·L-1硫酸降解为单糖后,经1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化,采用Shimadzu VP-ODS柱(150mm×4.6mm,5μm)分离,以乙腈-醋酸铵缓冲溶液(取醋酸铵7.708g加水溶解并稀释至1000mL,用醋酸调pH5.5(22:78)为流动相,流速为0.8mL·min-1,在245nm处检测单糖的衍生化产物。结果大蒜多糖中甘露糖(Man)、鼠李糖(Rha)、葡萄糖醛酸(GlcUA)半乳糖醛酸(GalUA)、葡萄糖(Glc)、半乳糖(Gal)、阿拉伯糖(Arab)7种单糖的摩尔比为4.31:4.23:4.19:9.13:27.4:4.99:3.81。结论本方法灵敏度高,结果准确可靠,可用于大蒜多糖的单糖组成测定及质量控制。
Objective To establish a reversed-phase high performance liquid chromatography (RP-HPLC) for the analysis of monosaccharide composition and molar ratio in garlic polysaccharides. Methods Garlic polysaccharide samples were degraded into monosaccharides by sulfuric acid (2mol·L-1) and derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP). Shimadzu VP-ODS column (150mm × 4.6) was used. Separated by mm, 5μm) in acetonitrile-ammonium acetate buffer solution (ammonium acetate 7.708g, dissolved in water and diluted to 1000mL, adjusted to pH 5.5 (22:78) as mobile phase with acetic acid, flow rate 0.8mL·min-1, The derivatization product of monosaccharide was detected at 245 nm.Results Garlic polysaccharide (Man), rhamnose (Rha), glucuronic acid (GlcUA), galacturonic acid (GalUA), glucose (Glc), galactose ( The molar ratios of the seven monosaccharides of Gal) and arabia were 4.31:4.23:4.19:9.13:27.4:4.99:3.81.Conclusion This method has high sensitivity and accurate and reliable results and can be used for the determination of monosaccharide composition of garlic polysaccharides. QC.