高效液相色谱法测定油茶饼粕多糖组成

Determination of polysaccharide composition of oil-tea camellia seed cake by high performance liquid chromatography

提出并对比了3种分析油茶饼粕多糖组成的高效液相色谱方法——氨基柱-折光示差检测器法、氨基柱-串级质谱法、1-苯基-3-甲基-5-吡唑啉酮柱前衍生-C18柱-紫外检测器法(柱前衍生法)。综合考虑分离效果、灵敏度、仪器成本等因素,最终选定柱前衍生法。柱前衍生法可同时测定油茶饼粕多糖中的鼠李糖、甘露糖、葡萄糖、半乳糖、木糖、阿拉伯糖、葡萄糖醛酸、半乳糖醛酸,线性关系良好(线性系数>0.99),检出限为0.02~0.29 mg/L,定量限为0.06~0.96 mg/L;单糖及糖醛酸的加标回收率分别为77.2%~111.9%、60.1%~87.0%;峰面积日内精密度为0.08%~3.77%,日间精密度为0.39%~17.15%。利用柱前衍生法检测油茶饼粕多糖样品,结果显示所检的4个样品中均含有5种或6种单糖及2种糖醛酸,且含量最高的组分均为葡萄糖。

Three high performance liquid chromatography methods for the determination of polysaccharide composition of oil-tea camellia seed cake were put up and compared,which were NH2 column-refractive index detector(RID)method,NH2 column-tandem mass spectrometry(MS/MS)method,1-phenyl-3-methyl-5-pyrazolone(PMP)pre-column derivation-C18 column-ultra-violet detector(UV)method(pre-column derivation method),respectively.Considering the separation effect,sensitivity,instrument cost and other factors,the pre-column derivation method was selected.The method could be used for determination of rhamnose,mannose,glucose,galactose,xylose,arabinose,glucuronic acid and galacturonic acid simultaneously.The correlation coefficients were higher than 0.99,with limits of detection 0.02-0.29 mg/L and limits of quantification 0.06-0.96 mg/L.The recoveries of standard addition of monosaccharides and uronic acids were 77.2%-111.9%and 60.1%-87.0%respectively.Intra-day relative standard deviations(RSD)of the peak area were 0.08%-3.77%while inter-day RSD were 0.39%-17.15%.The established method was then applied to detect four polysaccharide samples of oil-tea camellia seed cake.The results showed that all the polysaccharide samples consisted of five or six monosaccharides and two uronic acids,and glucose content was the highest.