梁图
,
傅青
,
辛华夏
,
李芳冰
,
金郁
,
梁鑫淼
色谱
doi:10.3724/SP.J.1123.2014.08023
来自中药的水溶性多糖具有广谱治疗和低毒性特点，是天然药物及保健品研发中的重要组成部分。针对中药多糖结构复杂、难以表征的问题，本文以中药黄芪中的多糖为研究对象，采用“自下而上”法完成对黄芪多糖的表征。首先使用部分酸水解方法水解黄芪多糖，分别考察了水解时间、酸浓度和温度的影响。在适宜条件（4 h、1.5 mol／L三氟乙酸、80℃）下，黄芪多糖被水解为特征性的寡糖片段。接下来，采用亲水作用色谱与质谱联用对黄芪多糖部分酸水解产物进行分离和结构表征。结果表明，提取得到的黄芪多糖主要为1→4连接线性葡聚糖，水解得到聚合度4~11的葡寡糖。本研究对其他中药多糖的表征具有一定的示范作用。
关键词:
部分酸水解
,
亲水作用色谱
,
质谱
,
多糖
,
黄芪
,
表征
王惠
,
辛华夏
,
蔡剑锋
,
李芳冰
,
金郁
,
傅青
色谱
doi:10.3724/SP.J.1123.2016.03006
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。
关键词:
部分酸水解
,
亲水作用色谱
,
黄芪多糖
,
指纹图谱
,
质量评价
Hui WANG
,
Huaxia XIN
,
Jianfeng CAI
,
Fangbing LI
,
Yu JIN
,
Qing FU
色谱
doi:10.3724/SP.J.1123.2016.03006
Astragalus polysaccharides (APS) are important components in Astragalus membranaceus (A. membranaceus). However, for the huge relative molecular mass, strong polarity, and difficulty in chromatographic 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 developed. 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 chromatograms 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.
关键词:
partial acid hydrolysis
,
hydrophilic interaction liquid chromatography (HILIC)
,
Astragalus polysaccharides (APS)
,
fingerprint
,
quality evaluation
