管花肉苁蓉多糖超声波辅助提取工艺及其多糖分离、结构特征与抗疲劳作用

段继华; 韩海霞; 赵文燕; 贾明玺; 孙婧婧; 周吉祥; 宋素琴; 朱东亮; 包晓玮

doi:10.13386/j.issn1002-0306.2025070372

管花肉苁蓉多糖超声波辅助提取工艺及其多糖分离、结构特征与抗疲劳作用

Ultrasonic-assisted Extraction Process of Polysaccharides from Cistanche tubiflora and Its Polysaccharide Isolation, Structural Characterization and Anti-fatigue Effect

摘要

摘要: 本研究旨在优化管花肉苁蓉多糖（Polysaccharides of Cistanche tubulosa，CP）提取工艺，并对其结构和抗疲劳作用进行研究。以CP得率为指标，通过单因素实验结合响应面法优化提取工艺；CP经DEAE-52纤维素柱分离纯化得到均一多糖，基于体外抗氧化活性筛选多糖组分，采用红外光谱、高效凝胶渗透色谱和离子色谱进行结构表征，并评估其抗疲劳作用。结果显示，响应面法优化的最佳提取工艺是：超声时间20 min，料液比为1:20，超声功率220 W。在此条件下，CP得率为8.55%。CP分离纯化得到1个中性多糖（CPN）和3个酸性多糖（CPA-1、CPA-2、CPA-3），经测定，CPN、CPA-1、CPA-2、CPA-3得率依次为40.3%、22.6%、13.7%、9.2%，多糖组分回收率分别为78.3%、89.6%、65.4%、52.7%。四个组分抗氧化活性强弱顺序依次为：CPA-2>CPN>CPA-1>CPA-3。鉴于CPN得率高、多糖组分回收率较高，且抗氧化活性强（与CPA-2相近），故选定CPN进行后续研究。CPN具备多糖特征吸收峰，主要由葡萄糖和甘露糖构成，可能存在两种分子量差异显著的组分；动物实验表明其可显著延长小鼠负重游泳时间，降低血尿素氮与血乳酸水平，并升高肝糖原含量，证实其抗疲劳作用。本研究为肉苁蓉多糖的深度开发及抗疲劳应用奠定基础。

Abstract: This study aimed to optimize the extraction process of Cistanche tubulosa polysaccharides (CP) and investigate their structural characteristics and anti-fatigue effects. Single-factor experiments combined with response surface methodology were employed to optimize extraction conditionswith the polysaccharide yield as the evaluation index. Subsequently, CP was separated and purified through DEAE-52 cellulose column chromatography to obtain homogeneous polysaccharide fractions. On the basis of in vitro antioxidant activity screening, the structural properties were characterized via infrared spectroscopy, high-performance gel permeation chromatography, and ion chromatography, and animal experiments were performed to evaluate the anti-fatigue capacity of the optimal fraction. The results revealed that the optimal extraction parameters were 20 min of ultrasonication, a solid-to-liquid ratio of 1:20, and an ultrasonic power of 220 W, under which the maximum CP yield reached 8.55%. After purification, one neutral polysaccharide (CPN) and three acidic polysaccharides (CPA-1, CPA-2, CPA-3) were isolated. The yields of CPN, CPA-1, CPA-2, and CPA-3 were 40.3%, 22.6%, 13.7%, and 9.2% respectively, with their corresponding polysaccharide component recoveries being 78.3%, 89.6%, 65.4%, and 52.7%. The antioxidant activity of the four fractions followed the order: CPA-2>CPN>CPA-1>CPA-3. Given its high yield, favorable recovery efficiency and antioxidant activity comparable to CPA-2, CPN was selected for further investigation. Structural analysis confirmed that CPN possessed characteristic absorption peaks of polysaccharides, was mainly composed of glucose and mannose, and might contain two components with significantly different molecular weights. In vivo experiments demonstrated that CPN could significantly prolong the weight-bearing swimming time of mice, reduce the levels of blood urea nitrogen and blood lactic acid, and increase hepatic glycogen content, thereby verifying its anti-fatigue activity. These findings lay a solid foundation for the in-depth development and anti-fatigue application of Cistanche tubulosa polysaccharides.