微波消解-1-苯基-3-甲基-5-吡唑啉酮柱前衍生-高效液相色谱法测定枸杞多糖含量及组成

Determination of content and composition of Lycium barbarum polysaccharides by microwave digestion-1-phenyl-3-methyl-5-pyrazolone pre-column derivatization-high performance liquid chromatography

目的建立微波消解-1-苯基-3-甲基-5-吡唑啉酮(1-phenyl-3-methyl-5-pyrazolone,PMP)柱前衍生-高效液相色谱(high performance liquid chromatography,HPLC)法测定枸杞多糖(Lycium barbarum polysaccharides,LBPs)含量及组成的方法。方法采用微波消解技术水解LBPs进行样品前处理,利用PMP进行柱前衍生,建立结合HPLC检测LBPs的方法,并利用该方法测定4个不同枸杞产地LBPs的含量及单糖组成。结果经方法学评价,8种单糖均具有良好的线性关系,相关系数均大于0.98;平均加标回收率为94.95%~99.22%,相对标准偏差小于等于1.90%;检出限和定量限良好。结论该方法可用于LBPs含量及组成的测定,分析表明LBPs主要由葡萄糖、阿拉伯糖、半乳糖、甘露糖、鼠李糖、木糖、葡萄糖醛酸、半乳糖醛酸组成,不同产地LBPs含量存在差异,其中甘肃玉门含量较高。

Objective To establish a method for the determination of the content and composition of Lycium barbarum polysaccharides(LBPs)by microwave digestion-1-phenyl-3-methyl-5-pyrazolone(PMP)pre-column derivatization and high performance liquid chromatography(HPLC).Methods Microwave digestion technology was used to hydrolyze LBPs for sample pretreatment,PMP was used for pre-column derivatization,and a method combining HPLC was established to detect LBPs.The content and monosaccharide composition of LBPs from four different Lycium barbarum producing areas were determined by this method.Results After methodological evaluation,8 kinds of monosaccharides had good linear relationship,the correlation coefficients were all greater than 0.98,the average recoveries were 94.95%‒99.22%,the relative standard deviations were less than or equal to 1.90%,and the limits of detection and limits of quantification were good.Conclusion This method can be used to determine the content and composition of LBPs.The analysis shows that LBPs mainly consist of glucose,arabinose,galactose,mannose,rhamnose,xylose,glucuronic acid and galacturonic acid.The content of LBPs in different regions is different,and the content in Yumen City of Gansu Province region is higher.