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    野西瓜低聚糖的提取分离、结构表征及功能活性研究

    Analysis of Extraction, Isolation, Structural Characterization, and Functional Activities of Oligosaccharides from Capparis Spinosa L.

      摘要
    • 摘要:
      目的 初步研究野西瓜低聚糖的结构表征与功能活性。
      方法 采用水提醇沉法提取低聚糖，经大孔树脂AB-8、DEAE-650M纤维素柱及HW-40F尺寸排阻柱纯化，制备C-O-1和C-O-2 2种野西瓜低聚糖。运用1-苯基-3-甲基-5-吡唑啉酮(1-phenyl-3-methyl-5-pyrazolone，PMP) 柱前衍生化和甲基化法结合紫外光谱、傅里叶变换红外光谱、高效液相色谱对C-O-1和C-O-2的进行结构表征。通过2,2’-联氮-双-3-乙基苯并噻唑啉-6-磺酸2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)，ABTS阳离子自由基(ABTS+·)、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl，DPPH)自由基(DPPH·)、羟基自由基清除率和Fe2+还原能力试验评价其抗氧化能力，并基于斑马鱼模型评价C-O-1的抗氧化活性以及美白活性。
      结果 C-O-1和C-O-2均含O-H、C-H键，C-O-1主要由葡萄糖和甘露糖组成(89.4∶10.6)，C-O-2主要由葡萄糖组成，两者均以末端葡萄糖吡喃环糖苷键(T-Glcp)连接，推测C-O-1为四糖，分子量约为671.37；C-O-2为二糖，分子量约为387.11。C-O-1和C-O-2在体外对ABTS+·、DPPH·、羟基自由基清除能力显著，且高于阳性对照水苏糖。在斑马鱼模型中，0.25~2 mg·mL–1的安全浓度范围内，C-O-1对活性氧生成的抑制率达43.8%，黑色素含量达22.6%，显示出良好的抗氧化效果和美白潜力。
      结论 C-O-1和C-O-2具有明确的结构特征，C-O-1具有显著的抗氧化能力和良好的美白效果，具有潜在的应用价值。

       

      Abstract:
      OBJECTIVE To investigate the structural characterization and functional activities of oligosaccharides from Capparis spinosa L.
      METHODS The oligosaccharides were extracted by water-ethanol extraction and purified sequentially with AB-8 macroporous resin, DEAE-650M cellulose column, and HW-40F gel filtration to obtain 2 fractions(C-O-1 and C-O-2). Their structures were characterized by 1-phenyl-3-methyl-5-pyrazolone(PMP) pre-column derivatization, methylation analysis, ultraviolet-visible spectroscopy(UV-Vis), Fourier transform infrared spectroscopy(FTIR), and high performance liquid chromatography(HPLC). Antioxidant activities were evaluated via 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) cationic radical(ABTS+·), 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical(DPPH·), hydroxyl radical scavenging assays, and Fe2+ reducing power. In vivo antioxidant and anti-melanogenic effects of C-O-1 were further assessed in zebrafish model.
      RESULTS Both C-O-1 and C-O-2 contained O-H and C-H bonds. C-O-1 was composed of glucose and mannose(89.4 : 10.6), while C-O-2 consisted solely of glucose. Glycosidic linkage analysis indicated terminal glucopyranose(T-Glcp) linkages in both samples. C-O-1 was hypothesized to be a tetrasaccharide with a molecular weight of approximately 671.37; whereas C-O-2 was a disaccharide with a molecular weight of approximately 387.11. In vitro, C-O-1 and C-O-2 exhibited significantly stronger scavenging activities against ABTS+, DPPH, and hydroxyl radicals compared to the positive control(stachyose). At safe concentrations(0.25–2 mg·mL–1), C-O-1 reduced ROS generation by 43.8% and inhibited melanin synthesis by 35.2% in zebrafish.
      CONCLUSION  C-O-1 and C-O-2 both exhibit well-defined structures. C-O-1 demonstrates remarkable antioxidant and skin-whitening effects, suggesting its potential for functional applications.

       

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