柱前衍生化HPLC法分离不同聚合度的壳寡糖

Separation of Chitosan Oligosaccharides with Different Degrees of Polymerization Using Pre-column Derivatization HPLC

目的:建立一种可以同时分离聚合度为2~6的壳寡糖柱前衍生化高效液相测定方法。方法:以1-苯基-3甲基-5-吡唑啉酮(PMP)为衍生剂对壳寡糖进行衍生化,采用C18柱(250mm×4.5mm,5μm),乙腈-乙酸铵水溶液为流动相,梯度洗脱,在254nm处进行测定,建立一种同时分析聚合度为2~6的壳寡糖柱前衍生化HPLC法。结果:聚合度为2~6的壳寡糖能得到较好的分离。结论:该方法简便可行,灵敏度高,重复性好,可为壳寡糖组分分析提供新的方法。

Objective: To establish a pre-column derivatization HPLC method that allows simultaneous separation of chitosan oligosaccharides with the degrees of polymerization of 2 ~ 6. Methods: Chitosan oligosaccharides were derivatized using 3-methyl-1-phenyl-2-pyrazolin-5-one (PMP) as the derivatization reagent;gradient elution was performed using a C18 column (250 mm×4.5 mm, 5 μm) and acetonitrile-ammonium acetate in water as the mobile phase;and the detection was performed at 254 nm, in an effort to establish a pre-column derivatization HPLC method that allows simultaneous analysis of chitosan oligosaccharides with the degrees of polymerization of 2 ~ 6. Results: Chitosan oligosaccharides with the degrees of polymerization of 2 ~ 6 were well separated. Conclusions: With simple operations, good applicability, high sensitivity and good repeatability, this method is of certain application value for component analysis of chitosan oligosaccharides.