试验采用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生-高效液相色谱法对木槿花多糖组分和含量进行分析.木槿花多糖经盐酸水解后,与标准单糖进行PMP进行衍生化.采用C18柱(4.6 mm × 250 mm,5 μm),以 0.1 mol/L的乙酸铵溶液(pH 6.9)和乙腈为流动相,在250 nm波长检测.分别采取3种不同比例的流动相,对木槿多糖溶液进行洗脱,最终选择流动相比例A-B=80+20(V/V)的洗脱机制.在该流动相比例下洗脱7种单糖,即甘露糖、无水葡萄糖、半乳糖、葡萄糖醛酸、木糖、鼠李糖、阿拉伯糖,建立77种单糖的线性方程,得出相关系数R2在0.95～0.99之间,并计算得出7种单糖含量,分别为0,0.3％,1.3％,1.4％,1.0％,1.8％和0.6％.该方法操作简单、精密度高,可为木槿花单糖组分特征评价与定量分析提供技术.
Analysis of Hydrolyzed Monosaccharide Components of Hibiscus Polysaccharide by HPLC-UV Detection
The l-phenyl-3-methyl-5-pyrazolone(PMP)derivatization-high performance liquid chromatography was used to analyze the composition and content of polysaccharide from Hibiscus.After being hydrolyzed by hydrochloric acid,hibiscus polysaccharide was derivatized with standard monosaccharide by PMP.C18 column(4.6 mm×250 mm,5 μm)was used,the mobile phase was 0.1 mol/L ammonium acetate solution(pH 6.9)and acetonitrile,and the detection was carried out at 250 nm.Three different mobile phases were used to elute the hibiscus polysaccharide solution,and finally the elution mechanism with the mobile phase ratio of A-B=80+20(V/V)was selected.Seven monosaccharides,namely mannose,anhydrous glucose,galactose,glucuronic acid,xylose,rhamnose and arabinose,were eluted at this mobile phase ratio,the linear equations of seven monosaccharides were established,the correlation coefficient R2 was between 0.95 and 0.99,and the contents of seven monosaccharides were calculated to be 0,0.3％,1.3％,1.4％,1.0％and 1.8,respectively.The method was simple and accurate,and can provide technology for the evaluation and quantitative analysis of monosaccharide composition characteristics of hibiscus.
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