野西瓜低聚糖的提取分离、结构表征及功能活性研究

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

关键词:  野西瓜  低聚糖  结构表征  抗氧化活性  斑马鱼美白活性

Key words:  Capparis Spinosa L.  oligosaccharides  structural characterization  antioxidant activity  zebrafish-based whitening activity

摘要:

目的 初步研究野西瓜低聚糖的结构表征及功能活性。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-2两种野西瓜低聚糖。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、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+)、1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picrylhydrazyl，DPPH)、羟基自由基清除率和Fe2+还原能力实验评价其抗氧化能力，并基于斑马鱼评价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，二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671.37 g·mol-1；C-O-2为二糖，分子量约387.11 g·mol-1。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%，对黑色素的抑制率达35.2%，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 two fractions (C-O-1 and C-O-2). Their structures were characterized by PMP (1-Phenyl-3-methyl-5-pyrazolone) pre-column derivatization, methylation analysis, UV-Vis, FTIR, and HPLC. Antioxidant activities were evaluated via ABTS+(2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), DPPH(1,1-Diphenyl-2-picrylhydrazyl), hydroxyl radical scavenging assays, and Fe2+ reducing power. In vivo antioxidant and anti-melanogenic effects of C-O-1 were further assessed in zebrafish. 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(Terminal glucopyranose) connections. C-O-1 was hypothesized to be a tetrasaccharide with a molecular weight of approximately 671.37 g·mol-1;C-O-2 is a disaccharide with a molecular weight of approximately 387.11 g·mol-1 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 exhibit well-defined structures and potent bioactivities. C-O-1 demonstrates remarkable antioxidant and skin-whitening effects, suggesting its potential for functional applications.