柱前衍生化HPLC分析蒲公英多糖的单糖组成

Analysis on Monosaccharide Composition of Taraxacum Polysaccharides by Pre-column Derivatization HPLC

目的合理优化衍生方法测定蒲公英多糖中单糖的组成及摩尔比。方法采用1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生,用反相高效液相色谱法进行测定。色谱条件为:ZORBAX-C18柱;流动相:0.1mol·L-1的磷酸盐缓冲液(pH6.7)-乙腈(83∶17);流速:1mL·min-1;检测波长:245nm;进样量:20μL;柱温:室温。结果蒲公英多糖由D-鼠李糖、葡萄糖、D-半乳糖、D-木糖和D-阿拉伯糖组成,以不同方法提取的各单糖含量的摩尔比为:水提取法,Rha∶Glc∶Gal∶Ara=0.088∶0.500∶0.190∶0.340;超声提取法,Rha∶Glc∶Gal∶Ara=0.050∶0.350∶0.080∶0.320;酶提取法,Rha∶Glc∶Gal∶Ara=0.270∶0.630∶0.330∶0.460;超声协提取同酶法,Rha∶Glc∶Gal∶Ara=0.078∶0.550∶0.130∶0.170。结论改进后的衍生化方法和等度洗脱的色谱方法具有操作简单、快速、重复性好等特点,可用于蒲公英多糖中单糖组成和摩尔比的测定。

OBJECTIVE To optimize the derivative method reasonably and detect monosaccharide components and molar ratio of taraxacum polysaccharides.METHODS Pre-column derivative reagent was 1-phenyl-3-methyl-5-pyrazolone（PMP）,monosaccharide composition and molar ratio of taraxacum polysaccharides were detected by reversed-phase high performance liquid chromatography（RP-HPLC）.Chromatographic conditions：ZORBAX-C18 column;mobile phase：0.1 mol·L^-1 phosphate buffer solution（pH 6.7）-acetonitrile（83 ：17）;velocity：1 mL·min^-1;detection wavelength：245 nm;injection volume：20 μL;column temperature：room temperature.RESULTS Taraxacum polysaccharides was composed of D-rhamnose,glucose,D-galactose D-xylose,D-Arabinose.The molar ratio of taraxacum polysaccharides which was extracted by different means was as forrows：water extraction method,Rha：Glc：Gal：Ara=0.088 ：0.500 ：0.190 ：0.340;ultrasonic extraction method,Rha：G lc：G al：A ra=0.050：0.350：0.080：0.320;cellulose extraction method,Rha：G lc：G al：A ra=0.270：0.630：0.330：0.460;synergistic ultrasonic extraction and cellulose extraction,Rha：Glc：Gal：Ara=0.078 ：0.550 ：0.130 ：0.170.CONCLUSION The improved derivative method and the chromatographic conditions of isocratic elution were simple,rapid and with a good reproducibility,and can be used to detect monosaccharide components and molar ratio of taraxacum polysaccharides.