中国烟草科学2025，Vol.46Issue(2)：52-59,8.DOI:10.13496/j.issn.1007-5119.2025.02.007
柱前衍生HPLC法分析烟叶水溶性多糖的单糖组成
Analysis of Monosaccharide Composition of Tobacco Water-soluble Polysaccharides by Pre-column Derivatization HPLC
摘要
Abstract
A determination method was established for the monosaccharide composition and content of tobacco water-soluble polysaccharides by 1-phenyl-3-methyl-5-imidazolinone(PMP)pre-column derivatization high performance liquid chromatography.Complete acid hydrolysis method was employed to hydrolyze tobacco water-soluble polysaccharides,and the hydrolysis products were subjected to PMP derivatization reaction.The signal of monosaccharide derivatives of tobacco water-soluble polysaccharides after acid hydrolysis was determined by reverse phase high-performance liquid chromatography.The method was used to explore the monosaccharide composition and content of tobacco water-soluble polysaccharides of different regions and types.The optimized detection conditions were as follows:0.05 mol/L phosphate(KH2PO4-NaOH,pH 7.0)buffer(solvent A)and acetonitrile(solvent B)were chosen as the mobile phase of elution system at the isocratic elution condition(84∶16),with Agilent Eclipse XDB-C18 column(250 mm×4.6 mm,5 μm);the UV detection wavelength was 250 nm.Results showed that the linear relationship is good for the nine monosaccharides in the concentration range of 20.0-800 μg/mL,with the correlation coefficient(R2)>0.9927.The limits of detection and quantification were 0.07-0.17 and 0.22-0.56 μg/mL,respectively.The average recoveries ranged 82.3%-104.0%in spiked samples.The tobacco water-soluble polysaccharides were composed of mannose(Man),rhamnose(Rha),galactose(Gal),arabinose(Ara),glucose(Glc),glucuronic acid(GlcUA),and galacturonic acid(GalUA),with Glc and GalUA accounting for a higher proportion.There were significant differences in the monosaccharide composition of tobacco water-soluble polysaccharides from various regions and types.The method is fast,simple,highly sensitive,and has good repeatability,thus can be used for monosaccharide composition analysis of tobacco polysaccharides.At the same time,it provides a basis for further research on the structure of tobacco polysaccharides.关键词
烟叶水溶性多糖/柱前衍生化/液相色谱/单糖组成Key words
tobacco water-soluble polysaccharides/pre-column derivatization/liquid chromatography/monosaccharide composition分类
轻工业引用本文复制引用
赵妍,李耀群,黄延俊,刘秀彩,黄华发,刘江生,叶仲力,张廷贵,刘泽春,谢卫..柱前衍生HPLC法分析烟叶水溶性多糖的单糖组成[J].中国烟草科学,2025,46(2):52-59,8.基金项目
福建中烟工业有限责任公司博士后项目(FJZYKJJH2023BH002) （FJZYKJJH2023BH002）
