GC和HPLC分析石榴皮多糖的单糖组成方法比较
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摘要
用GC法和HPLC法对石榴皮多糖的单糖组成进行分析比较。石榴皮多糖经2.0 mol/L的三氟乙酸水解后,用还原乙酰化-GC法和1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生-HPLC法分别测定其单糖组成。结果表明,GC法操作步骤繁琐、试剂种类多、耗时长,不利于快速分析;而HPLC法操作简单、试剂种类少、耗时短,工作效率高;两种方法的分析结果在主要中性单糖组成方面基本一致,但对Gal A具有明显差异。总体认为两种方法对石榴皮多糖组成分析结果存在一定的差异。
The monosaccharide composition of pomegranate peel polysaccharides was analyzed and compared by GC and HPLC. Pomegranate peel polysaccharide was hydrolyzed by 2 mol · L-1 trifluoracetic acid. The monosaccharide composition was determined by reduction acetylation by GC method and 1-phenyl-3-methyl-5-pyrazolone( PMP)pre-column derivatization by HPLC method. The results showed that the GC method with tedious operation procedure,various for the needed reagents and long for the needed time,was not good for fast analysis,while the operation of HPLC with the high working efficiency was simple,few kind of reagents needed and short time needed. For the main neutral monosaccharide composition of the pomegranate peel polysaccharide,the result from the two methods was basically the same. But for Gal A,the result was obviously different. It was generally believed that there were some differences between the two methods in the analysis of polysaccharides in pomegranate peel.
The monosaccharide composition of pomegranate peel polysaccharides was analyzed and compared by GC and HPLC. Pomegranate peel polysaccharide was hydrolyzed by 2 mol · L-1 trifluoracetic acid. The monosaccharide composition was determined by reduction acetylation by GC method and 1-phenyl-3-methyl-5-pyrazolone( PMP)pre-column derivatization by HPLC method. The results showed that the GC method with tedious operation procedure,various for the needed reagents and long for the needed time,was not good for fast analysis,while the operation of HPLC with the high working efficiency was simple,few kind of reagents needed and short time needed. For the main neutral monosaccharide composition of the pomegranate peel polysaccharide,the result from the two methods was basically the same. But for Gal A,the result was obviously different. It was generally believed that there were some differences between the two methods in the analysis of polysaccharides in pomegranate peel.
引文
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[3]Sawardeker J S,Sloneker J,Jeanes Allene.Quantitative determination of monosaccharides as their alditol acetates by gas liquid chromatography[J].Anal.Chem.,1965,37(12):1602-1604.
[4]Wang X,Zhang L,Wu J,et al.Improvement of simultaneous determination of neutral monosaccharides and uronic acids by gas chromatography[J].Food Chemistry,2017,220(4):198-207.
[5]Wang W Z,Chen F,Wang Y Q,et al.Optimization of reactions between reducing sugars and 1-phenyl-3-methyl-5-pyrazolone(PMP)by response surface methodology[J].Food Chemistry,2018,254(7):158-164.
[6]王凤芹,张林,路则庆,等.高效液相色谱法分析新型细菌多糖中单糖和糖醛酸组成[J].分析科学学报,2016,34(1):32-36.
[7]张萍,贺茂萍,殷力,等.石榴皮多糖的Sevage法除蛋白工艺研究[J].食品科技,2013,38(12):219-231.
[8]张惟杰.糖复合物生化研究技术.2版[M].杭州:浙江大学出版社,2003:44-45.
[9]Wang C J,Fan W C,Zhang P,et al.One-pot non-reductive Oglycan release and labeling with 1-phenyl-3-methyl-5-pyrazolone(PMP)followed by ESI-MS analysis[J].Proteomics,2011,11(21):4229-4242.