多糖是黄芪的重要成分，但多糖相对分子质量大、极性强，难以用色谱方法直接分析，导致目前缺乏能够反映黄芪多糖组成差异的质量评价方法。首先通过部分酸水解方法，将多糖水解成可供分析的寡糖，建立基于部分酸水解-亲水作用色谱的黄芪多糖指纹图谱。通过正交实验选取最佳水解条件：温度80℃、酸浓度1.5 mol／L，水解时间4 h。该方法重复性好，对20批黄芪药材的多糖指纹图谱分析显示相似度为0.258~0.949，反映出黄芪多糖组成的明显差异。同时建立了黄芪的反相液相色谱指纹图谱，用于控制除多糖以外的其他成分，对同样的20批黄芪药材进行分析，实现对黄芪全面的质量评价。实验表明，基于部分酸水解-亲水作用色谱的黄芪多糖指纹图谱可对黄芪多糖的质量进行有效评价，是对黄芪质量评价方法的重要补充。
Astragalus polysaccharides(APS)are important components in Astragalus membranaceus(A. membranaceus). However,for the huge relative molecular mass,strong polarity,and difficulty in chromato-graphic separation,there’s a lack of quality control finely reflecting the composition of APS up to now. To solve the problems,the key lies in degradation of polysaccharides and analysis of the hydrolyzates. A partial acid hydrolysis-hydrophilic interaction liquid chromatography( HILIC)fingerprint for APS and a comprehensive quality evaluation method for A. membranaceus combining with the traditional RPLC fingerprint were devel-oped. Orthogonal experiments were designed to investigate the influencing factors of partial hydrolysis and the conditions were optimized based on HILIC analysis. The hydrolyzates were successfully separated using HILIC-evaporative light scattering detection( ELSD)method and the optimal conditions of hydrolysis were selected as 80 ℃,1. 5 mol/L trifluoroacetic acid and 4 h,which proved to be stable and repeatable after the validation. This method was applied for the fingerprint analysis of 20 batches of A. membranaceus. Cosine method was used for similarity calculation using the average of samples Nos. 1-10 as reference. Moreover,RPLC fingerprint analysis was processed on other components and the data were compared for comprehensive evaluation. The similarities of ten samples with the reference ranged from 0. 258 to 0. 949,which were consistent with the chro-matograms reflecting the apparent disparity in APS,and could be as a supplement of the RPLC fingerprint. The new method,with the advantages of intuition,convenience,and quantifiable in similarity,could reflect the differences of the composition of polysaccharides after degradation and was successfully applied in profiling of APS,contributing to the quality control together with the RPLC fingerprint.
参考文献
| [1] | Fu, Q.;Liang, T.;Li, Z.;Xu, X.;Ke, Y.;Jin, Y.;Liang, X..Separation of carbohydrates using hydrophilic interaction liquid chromatography[J].Carbohydrate research,2013:13-17. |
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