摘要
建立了柱前PMP衍生高效液相色谱法分离检测8种中性糖、2种糖醛酸及2种糖胺的方法。筛选出适合PMP衍生物分离的色谱柱为Ec lipse XDB-C18,以0.1 mol/L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各种糖的PMP衍生物的保留及分离的影响,确定最佳pH值为6.7,最佳乙腈体积分数为17%;比较了不同衍生反应时间和萃取除杂前不同量盐酸中和反应产物时醛糖-PMP衍生物的峰面积大小变化,从而得到较佳的样品衍生化条件。应用优化的色谱条件和样品制备方法,测定了由杜氏盐藻提取纯化的多糖级分PD4 a和PD4b的单糖组成,其测定结果与高效阴离子交换色谱法的结果基本一致。
A simple and sensitive high performance liquid chromatographic method for simultaneous determination of eight kinds of neutral monosaccharides, two kinds of uronic acid and two kinds of glycosamine by pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone （PMP） has been developed. The effects of volume ratios of acetonitrile to phosphate buffer and pH of mobile phase on retention and separation of PMP derivatives of the ten monosaccharides were investigated with Eclipse XPB-C18 Column（4.6 mm×250 mm）. The results showed that the optimum volume ratio of acetonitrile to buffer was 17 ： 83 and that the pH was 6.7. The variations of the peak areas of monosaccharides derivatives due to different reaction times and amounts of HCl for neutralization of the reaction systems were compared to obtain better sample preparation conditions. The developed method was successfully applied to the analyses of monosaccharide compositions in the polysaccharide fractions （PD4a and PD4b） from Duanaliella Salina （D. Satina）. The analytical results obtained on the whole agreed with those obtained by high-performance anion-exchange chromatography with pulsed-amperometric detection.
出处
《分析测试学报》
CAS
CSCD
北大核心
2007年第2期206-210,共5页
Journal of Instrumental Analysis
基金
云南省省院省校科技合作项目(2001MAABA00A028)
关键词
杜氏盐藻
多糖
单糖
高效液相色谱
柱前衍生化
1-苯基-3-甲基-5-吡唑啉酮
Duanaliella salina
Polysaccharides
Monosaccharide
High performance liquid chromatography
Pre-column derivatization
1-Phenyl-3-methyl-5-pyrazolone
