Isolation, Purification, Monosaccharide Composition and Antioxidant Activity of Polysaccharides from Cortex Periplocae
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摘要: 目的：分离纯化香加皮多糖（Cortex Periplocae Polysaccharides，CPP），并对其进行单糖组成和抗氧化活性研究，以期为香加皮多糖在食品领域的开发和应用提供参考。方法：通过水提醇沉和Sevag法除蛋白得到香加皮粗多糖，经DEAE-52纤维素柱分离纯化得到4种多糖组分CPP0、CPP1、CPP2和CPP3，并对其进行化学成分检测、分子量测定、单糖组成分析、红外光谱和抗氧化活性分析。结果：4种多糖的糖含量分别为82.20%、77.13%、75.23%和72.85%，且都含有糖醛酸；相对分子量分别为685、477、411和572 kDa。4种多糖均为甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖和阿拉伯糖按照不同摩尔比组成的杂多糖。红外光谱表明4种多糖含有β-糖苷键且具有呋喃环。抗氧化实验结果显示4种多糖均具有一定的抗氧化性，总体抗氧化能力顺序为：CPP0>CPP3>CPP1>CPP2。结论：从香加皮中提取得到的4种酸性多糖，糖含量高、相对分子量大且均具有抗氧化活性，其中CPP0组分抗氧化活性最好。Abstract: Objective: In this experiment, polysaccharides from cortex periplocae (CPP) were isolated and purified, and their monosaccharide composition and antioxidant activity were investigated, so as to provide a reference for the development and application of CPP in the food industry. Methods: The crude polysaccharides were obtained by aqueous extraction, alcoholic precipitation and protein removal by Sevag method, and four polysaccharide fractions CPP0, CPP1, CPP2 and CPP3 were separated and purified by DEAE-52 cellulose column. The chemical composition, molecular weight, monosaccharide composition, infrared spectrum and antioxidant activity were analyzed. Results: The sugar contents of these four polysaccharides were 82.20%, 77.13%, 75.23% and 72.85%, respectively, and all of them contained glyoxylate. The relative molecular weights were 685, 477, 411 and 572 kDa, respectively. The four polysaccharides were all heteropolysaccharides composed of mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose in different molar ratios. The infrared spectra showed that four polysaccharides contained β-glycosidic bonds and with furan rings. The antioxidant experiments showed that all four polysaccharides had certain antioxidant properties, and the overall antioxidant capacity was in the following order: CPP0>CPP3>CPP1>CPP2. Conclusion: The four acidic polysaccharides extracted from cortex periplocae have high sugar content, high relative molecular weight, and antioxidant activity, among which CPP0 fraction has the best antioxidant activity.
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图 1 香加皮多糖DEAE-52纤维素柱洗脱曲线
Figure 1. Elution curve of polysaccharides on DEAE-52 cellulose column of CCPP
图 4 单糖标准品HPLC分析图谱
Figure 4. HPLC analysis spectrum of monosaccharide standards
注：1.PMP；2.甘露糖；3.鼠李糖；4.半乳糖醛酸；5.葡萄糖；6.半乳糖；7.阿拉伯糖；图5同。
图 5 CPP0（A）、CPP1（B）、CPP2（C）、CPP3（D）的HPLC色谱图
Figure 5. HPLC chromatograms of CPP0 (A), CPP1 (B), CPP2 (C), CPP3 (D)
表 1 香加皮多糖的化学成分分析
Table 1. Chemical composition analysis of CPPS
编号 糖含量（%） 蛋白含量（%） 糖醛酸含量（%） CPP0 82.20±0.48a − 31.06±0.70a CPP1 77.13±0.71b − 28.29±0.37b CPP2 75.23±0.63c − 23.82±0.24c CPP3 72.85±1.06d 2.5±0.8 29.11±0.37b 注：平均值±标准差；同列不同小写字母表示不同处理间差异显著（P<0.05）；“−”表示未检测出。 
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表 2 香加皮多糖各组分单糖的摩尔比
Table 2. Molar ratios of monosaccharides of CPPS
编号 甘露糖 鼠李糖 半乳糖醛酸 葡萄糖 半乳糖 阿拉伯糖 CPP0 1.65 1.61 1.25 4.71 1.74 0.99 CPP1 0.76 0.53 0.43 1.90 1.42 0.46 CPP2 0.48 0.57 0.15 1.11 0.31 0.17 CPP3 1.41 1.32 0.58 2.30 1.02 0.48
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