柱前衍生HPLC分析银耳多糖的单糖组成

Analysis of monosaccharide compositions in tremella polysaccharides by pre-column derivatization high performance liquid chromatography

建立1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生高效液相色谱法分离检测5种单糖的方法。根据单糖PMP衍生物的峰面积大小,对衍生化反应中反应时间、PMP与NaOH的量、中和温度和盐酸用量进行了考察,从而得到较佳的衍生化样品制备条件。以20 mmol.L-1磷酸盐缓冲液-乙腈为流动相,考察了pH值和乙腈体积分数对糖衍生物分离的影响,确定最佳pH值为6.72,最佳乙腈体积分数为18%,并将该方法应用于银耳酸性多糖的单糖组成的测定。

A method of PMP pre-column derivatization high performance liquid chromatographic was established for analysis of five kind of monosaccharide.The optimum conditions of derivatization were determined by studying the quantity of PMP and sodium hydroxide,reaction time,reaction temperature and hydrochloric acid consumption during neutralization.Under the optimum mobile phase which was 18% acetonitrile-phosphate buffer（20 mmol·L-1,pH 6.72）,PMP derivatives can be separated successfully.This method was applied to monosaccharide compositions analysis of tremella polysaccharide（TPP2）.