摘要
建立了柱前1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生-高效液相色谱法,分离检测新型细菌多糖中5种单糖及糖醛酸组成的分析方法。新型细菌多糖中加入单糖内标木糖后,经3mol/L三氟乙酸水解,再用PMP进行衍生化,采用XBridgeTMC18柱(250×4.6mm,5μm),以50mmol/L KH2PO4溶液(pH=5.5)和乙腈为流动相,在245nm波长检测。定量分析结果表明,该多糖由甘露糖、葡萄糖醛酸、葡萄糖、半乳糖和岩藻糖组成,其质量比为1.77∶0.44∶8.52∶1.32∶1.67。该方法重现性良好,能够快速对多糖样品进行组成分析。
High performance liquid chromatography(HPLC)was employed for the analysis of monosaccharides and uronic acid in novel bacterial polysaccharides with pre-column derivatization.After being degradated by 3 mol/L trifluoroacetic acid and derivatized with 1-phenyl-3-methyl-5-pyrazolone(PMP),monosaccharides and uronic acid were separated by reversed-phase HPLC with isocratic elution,and detected by ultraviolet detector at 245 nm.The result showed that the bacterial polysaccharides consisted of 4 kinds of monosaccharides and 1 kind of galacturonic acid.The mass ratio of mannose,glucuronic acid,glucose,galactose and fucose was 1.77:0.44:8.52:1.32:1.67.The developed method was suitable for the rapid analysis of polysaccharide with good reproducibility.
作者
王凤芹
张林
路则庆
汪以真
WANG Feng—qin;ZHANG Lin;LU Ze—qing;WANG Yi—zhen(Feed Science Institute of Zhejiang University,Key Laboratory of Animal Nutrition and Feed Science,Ministry of Agriculture(East China),Zhjiang Provincial Laboratory of Feed and Animal Nutrition,Hangzhou 310058)
出处
《分析科学学报》
CAS
CSCD
北大核心
2018年第1期32-36,共5页
Journal of Analytical Science
关键词
细菌多糖
PMP
衍生化
高效液相色谱法
Bacterial polysaccharides
1-Phenyl-3-methyl-5-pyrazolone
Pre-column derivatization
High performante liquid chromatography
