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目的建立一种同时测定库拉索芦荟中多糖分子量和含量的高效凝胶渗透色谱(HPGPC)分析方法。方法芦荟多糖采用水提醇沉透析法制备。以葡聚糖为标样,采用Agilent PL aquagel-OH 60(8μm,300 mm×7.5mm)和Agilent PL aquagel-OH40(8μm,300 mm×7.5mm)色谱柱串联,纯化水为流动相,流速为0.6 m L·min~(-1),柱温30℃;示差折光检测器(RID)检测,检测温度为40℃,进样量50μL。结果在一定范围内,葡聚糖分子量与保留时间、含量与峰面积之间均呈现良好的线性关系。对5批库拉索芦荟汁样品进样分析,均可检测到4个多糖峰,其中以t_R≈20 min和t_R≈25 min两个峰为主。综合分析,样品中多糖相对分子质量大于1110 k D占24%~66%、500 k D~1110 k D占8%~14%、300 k D~500 k D占15%~23%、5 k D~300 k D占7%~46%;其多糖含量为0.10%~0.16%。结论该方法简便、准确、重复性好,适用于库拉索芦荟多糖的分子量和含量测定。
OBJECTIVE To establish a high performance gel permeation chromatography (HPGPC) method for simultaneous determination of polysaccharide molecular weight and content in Aloe vera leaves. Methods Aloe polysaccharides were prepared by water precipitation alcohol dialysis. Using dextran as a standard sample, the purified water was used as the mobile phase with Agilent PL aquagel-OH 60 (8μm, 300 mm × 7.5mm) and Agilent PL aquagel-OH40 (8μm, 300 mm × 7.5mm) 0.6 m L · min -1, the column temperature was 30 ℃. The refractive index detector (RID) was used to detect the temperature at 40 ℃ and the injection volume was 50μL. Results Within a certain range, dextran molecular weight and retention time, content and peak area showed a good linear relationship. Four batches of polysaccharides were detected in five batches of Aloe vera juice samples, of which two peaks at t_R≈20 min and t_R≈25 min were dominant. The results showed that the relative molecular weight of polysaccharides in samples was 24% ~ 66% when the molecular weight was 1110 kD, 8% ~ 14% at 500 kD ~ 1110 kD, 15% ~ 23% at 300 kD ~ 500 kD, D ~ 300 k D accounted for 7% ~ 46%; the polysaccharide content of 0.10% ~ 0.16%. Conclusion The method is simple, accurate and reproducible. It is suitable for the determination of molecular weight and content of polysaccharides in Aloe vera.