黄芩多糖的提取及结构表征

doi:10.16431/j.cnki.1671-7236.2023.06.036

摘要/Abstract

摘要： 【目的】优化水提醇沉法提取黄芩多糖最佳工艺条件，提高黄芩多糖提取率，并探究黄芩多糖的结构，为工业化提取黄芩多糖提供依据。【方法】采用单因素试验考察提取时间、提取温度、料液比和提取次数对多糖提取率的影响。运用Box-Behnken中心组合试验设计法，选取提取时间、提取温度、料液比和提取次数为优化的4个因素，以多糖提取率为响应值，设计4因素3水平试验方案，通过响应面分析确定水提醇沉法提取黄芩多糖的最佳条件；采用柱层析法对多糖进行分离纯化，采用红外光谱、紫外光谱、高效液相色谱（HPLC）、凝胶色谱对黄芩多糖的基团、纯度、分子质量和单糖组成进行分析。【结果】经响应面分析确定当提取温度为98 ℃，料液比为1：12.40（g/mL），提取时间为138 min，提取次数为4次时，黄芩粗多糖提取率为12.23%。用DEAE-52纤维素柱层析分离和葡聚糖G-100凝胶柱层析进行纯化，得到Hq-3-1组分。凝胶渗透色谱分析表明，其Hq-3-1为均一多糖，分子质量为58 794 u。HPLC法柱前衍生化分析Hq-3-1组分的单糖组成结果显示，其包含果糖、甘露糖、鼠李糖、葡萄糖醛酸、半乳糖、阿拉伯糖和岩藻糖。紫外吸收图谱显示，Hq-3-1在260和280 nm处均无吸收峰，说明其不含核酸和蛋白质。红外光谱分析显示，Hq-3-1具有多糖特征的吸收峰，是α-型D-甘露糖吡喃型杂多糖。【结论】采用水提醇沉法在最佳提取条件下黄芩多糖提取率为12.23%，该方法操作简便且多糖提取率高，同时本研究证明黄芩多糖是一种分子质量为58 794 u的杂多糖。

关键词: 黄芩多糖; 提取工艺; 分离纯化; 结构分析

Abstract: 【Objective】 The purpose of the test was to explore the optimal extraction conditions of Scutellaria baicalensis polysaccharides by water extraction and alcohol precipitation, improve the extraction rate of Scutellaria baicalensis polysaccharide, and explore the structure of Scutellaria baicalensis polysaccharide, so as to provide the basis for industrial extraction of Scutellaria baicalensis polysaccharide.【Method】 The effects of extraction time, extraction temperature, material-to-liquid ratio and extraction times on the extraction yield of polysaccharide were investigated by single factor test.Box-Behnken central combination experimental design method was used to select the optimal extraction time, extraction temperature, material-to-liquid ratio and extraction times, the extraction rate of polysaccharide was the response value (the design of four factors and three levels test), through the response surface analysis to determine the optimal extraction conditions of Scutellaria baicalensis polysaccharide extraction by water extraction and alcohol precipitation method.The polysaccharide was separated and purified by column chromatography.The group, purity, molecular weight and monosaccharide composition of Scutellaria baicalensis polysaccharide were analyzed by infrared spectrum, ultraviolet spectrum, high performance liquid chromatography (HPLC) and gel chromatography.【Result】 Response surface analysis determined that the extraction temperature was 98 ℃, the material-to-liquid ratio was 1:12.40 (g/mL), the extraction time was 138 min, and the extraction times were 4, the extraction yield of crude polysaccharide was 12.23%.Hq-3-1 was purified by DEAE-52 cellulose column chromatography and dextran G-100 gel column chromatography.Gel permeation chromatography analysis showed that Hq-3-1 was a homogeneous polysaccharide with a molecular weight of 58 794 u.The monosaccharides of Hq-3-1 components were analyzed by pre-column derivatization by HPLC.The results showed that the monosaccharides contained fructose, mannose, rhamnose-glucuronic acid, galactose, arabinose and fucose.The UV absorption map showed that Hq-3-1 had no absorption peak at 260 and 280 nm, indicating that it did not contain nucleic acid and protein.Infrared spectrum analysis showed that Hq-3-1 had a polysaccharide characteristic absorption peak, which was α-D-mannose-pyranoid heteropoly.【Conclusion】 Under the optimal extraction conditions, the extraction rate of Scutellaria baicalensis polysaccharides by water extraction and alcohol precipitation was 12.23%.This method was simple, convenient, and had a high extraction rate.This study proved that Scutellaria baicalensis polysaccharide was a heteropolysaccharide with a molecular weight of 58 794 u.

Key words: Scutellaria baicalensis polysaccharide; extraction process; separation and purification; structural analysis

中图分类号:

S859.3

刘鹏, 李静静, 乔彦良, 朱连勤, 朱风华. 黄芩多糖的提取及结构表征[J]. 中国畜牧兽医, 2023, 50(6): 2518-2530.

LIU Peng, LI Jingjing, QIAO Yanliang, ZHU Lianqin, ZHU Fenghua. Extraction and Structure Characterization of Scutellaria baicalensis Polysaccharide[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2518-2530.

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