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    明日叶多糖及寡糖的结构表征和抗氧化活性研究

    Structural Characterization and Antioxidant Activity of Polysaccharides and Oligosaccharides from Angelica Keiskei

      摘要
    • 摘要:
      目的  水提醇沉法制备明日叶水提多糖及寡糖，并对其理化性质、结构和抗氧化活性进行研究。
      方法 采用冷提、热提的序贯提取法提取明日叶总糖，分离纯化得到多糖和寡糖；化学分析法测定其总糖、蛋白、糖醛酸和氨基糖含量；利用高效凝胶渗透色谱法(high performance gel permeation chromatography，HPGPC)测定其分子量分布，PMP柱前衍生高效液相色谱法(PMP-HPLC)测定其单糖组成，结合红外光谱法初步表征其结构；通过1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl，DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)，ABTS自由基清除能力评价其体外抗氧化活性。
      结果 明日叶冷提及热提所得4种组分差异较大，其中多糖组分分子量主要集中于4~8 kDa，寡糖主要分布于1.5~2 kDa。冷提多糖和寡糖主要以半乳糖和阿拉伯糖为主，热提多糖和寡糖的单糖组成差异明显。热提多糖主要由半乳糖、半乳糖醛酸、木糖和阿拉伯糖构成，具有典型的果胶类多糖特点；热提寡糖以葡萄糖、半乳糖和鼠李糖为主。4种糖组分均表现出显著的抗氧化活性，其中冷提寡糖活性最佳，其IC50仅为0.11 mg·mL−1(DPPH)。
      结论 冷提及热提的序贯提取法所得明日叶多糖及寡糖组分的分子量分布及其单糖组成差异明显，但其均具有较好的体外抗氧化活性，具有开发为天然抗氧化剂的潜在价值。

       

      Abstract:
      OBJECTIVE  To extract the polysaccharides and oligosaccharides from Angelica keiskei by using the water extraction and alcohol precipitation methods, and to investigate their physicochemical properties, structure characterization and antioxidant activity.
      METHODS  The polysaccharides and oligosaccharides were prepared by the sequential extraction method of cold and hot extraction technique respectively, and fractioned following with membrane dialysis. Chemical analyses were used to determine the contents of total sugar, protein, uronic acid and amino sugar. The molecular weight distribution and monosaccharide composition was determined by high performance gel permeation chromatography(HPGPC) and PMP-HPLC, and the structure were preliminarily characterized by infrared spectroscopy. The antioxidant activity was determined by free radical scavenging activity1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS).
      RESULTS  The 4 fractions was found to be quite different by cold maceration and reflux extraction. The molecular weights of the fractions were mainly concentrated in the range of 4–8 kDa(polysaccharides) and 1.5–2.0 kDa(oligosaccharides). Cold-dipped fractionscold-extracted polysaccharide of Angelica keiskei(AKCP) and cold-extracted oligosaccharide of Angelica keiskei(AKCO) were mainly composed by Gal and Ara, while the composition of hot-extracted fractionshot-extracted polysaccharide of Angelica keiskei(AKHP) and hot-extracted oligosaccharide of Angelica keiskei(AKHO) were significantly different. AKHP predominantly constituted of Gal, GalA, Xyl and Ara, with the typical pectin polysaccharide structure. AKHO were mainly composed of Glc, Gal and Rha. All of the 4 fractions showed good antioxidant activity, among which AKCO showed the highest DPPH(IC50 0.11 mg·mL−1) radical scavenging activity.
      CONCLUSION  The molecular weight distribution and monosaccharide compositions of 4 components from Angelica keiskei extracted by the sequential extraction method of cold and hot extraction technique are significantly different. And the 4 components possess obvious antioxidant activity, with potential applications in natural antioxidants.

       

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