Study on Isolation ，Purification and in vitro Antioxidant Activity of the Polysaccharides from Cucumis satiuus

ZHANG Zhihong; XING Na; PENG Donghui; WANG Qiuhong; KUANG Haixue

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Study on Isolation ，Purification and in vitro Antioxidant Activity of the Polysaccharides from Cucumis satiuus

更新时间：2022-06-21

- Study on Isolation ，Purification and in vitro Antioxidant Activity of the Polysaccharides from Cucumis satiuus
- China Pharmacy Vol. 32, Issue 4, (2021)
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- Published：2021，
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ZHANG Zhihong, XING Na, PENG Donghui, et al. Study on Isolation ，Purification and in vitro Antioxidant Activity of the Polysaccharides from Cucumis satiuus. [J]. China Pharmacy 32(4).(2021)
DOI：

ZHANG Zhihong, XING Na, PENG Donghui, et al. Study on Isolation ，Purification and in vitro Antioxidant Activity of the Polysaccharides from Cucumis satiuus. [J]. China Pharmacy 32(4).(2021) DOI：

摘要

目的：对黄瓜子多糖进行分离、纯化，并考察其体外抗氧化活性。方法：采用水提醇沉法提取黄瓜子粗多糖（CCL），然后以AmberliteFPA90Cl阴离子和AmberliteFPC3500H阳离子串联交换树脂柱和离子交换纤维素DEAECellulose52柱对CCL进行分离、纯化得到CCL-M2、CCL-M3、CCL-M6等3种多糖。结合高效液相色谱-蒸发光散射法、超高效液相色谱-四极杆飞行时间质谱法和紫外、红外光谱等多种技术对3种均一多糖的纯度、分子量、结构和单糖组成等进行分析。以维生素C（VC）为阳性对照，考察CCL-M2、CCL-M3、CCL-M6（质量浓度均为0.4、0.8、1.2、1.6、2.0mg/mL）对1，1-二苯基-2-三硝基苯肼自由基（DPPH·）、羟基自由基（·OH）、超氧阴离子自由基（O2－·）的清除能力，并计算其半数清除浓度（IC50）。结果：CCL-M2、CCL-M3、CCL-M6均为吡喃糖形式的均一多糖，分子量分别为9.614×106、2.024×107、3.343×107Da。CCL-M2的单糖组成为鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖，其摩尔比为5.1∶2.6∶1.7∶1.4∶31.6；CCL-M3和CCL-M6的单糖组成均为甘露糖、鼠李糖、葡萄糖、半乳糖、阿拉伯糖，其摩尔比分别为1.7∶3.8∶6.4∶3.3∶1.3和1.3∶3.0∶3.2∶4.4∶8.9。体外抗氧化活性结果显示，3种均一多糖对DPPH·、·OH、O2－·均具有一定的清除活性，其对DPPH·的清除活性强弱顺序为VC＞CCL-M2＞CCL-M3＞CCL-M6（IC50依次为0.309、1.240、1.489、1.713mg/mL），对·OH的清除活性强弱顺序为VC＞CCL-M2＞CCL-M6＞CCL-M3（IC50依次为0.968、1.032、1.233、1.356mg/mL），对O2－·的清除活性强弱顺序为VC＞CCL-M2＞CCL-M3＞CCL-M6（IC50依次为0.335、0.379、0.812、1.662mg/mL）。结论：从黄瓜子中分离得到3种吡喃糖形式的均一多糖均具有一定的体外抗氧化活性，其中以CCL-M2的活性最强。

Abstract

OBJECTIVE：To isolate and purify the polysaccharides from Cucumis satiuus ，and to investigate its in vitro antioxidant activity. METHODS ：The crude polysaccharides （CCL）were extracted from C. satiuus by water extraction and alcohol precipitation. CCL was separated and purified on Amberlite FPA 90Cl anion and Amberlite FPC 3500H cation exchange resin column and DEAE cellulose 52 column，so as to obtain 3 kinds of polysaccharides as CCL-M 2，CCL-M3，CCL-M6. The purity ， molecular weight ，structure and monosaccharide composition of 3 kinds of polysaccharides were analyzed by HPLC-ELSD ， UPLC-QTOF/MS，UV and IR. Using vitamin C （VC）as positive control ，the scavenging activity of CCL-M 2，CCL-M3 and CCL-M6（0.4，0.8，1.2，1.6，2.0 mg/mL）to 1，1-dibenzene-2-trinitrobenzene free radicals （DPPH·），hydroxyl free radicals （·OH） and superoxycyclic anion free radicals （O2－·）were investigated ，and IC50 was calculated. RESULTS ：CCL-M2，CCL-M3 and CCL-M6 were homogeneous polysaccharides in the form of pyranose ，with molecular weights of 9.614×106，2.024×107 and 3.343× 107 Da，respectively. CCL-M 2 monosaccharides were composed of rhamnose ，semi-lactose aldehyde acid ，glucose， semi-lactose and arabic sugar ，with a molar ratio of 5.1∶2.6∶1.7∶1.4∶31.6. CCL-M 3 and CCL-M 6 monosaccharides were composed of mannose， rhamnose，glucose，semi-lactose，arabic sugar ，with a molar ratio of 1.7 ∶ 3.8 ∶ 6.4 ∶ 3.3 ∶ 1.3 and 1.3 ∶ 3.0 ∶ 3.2 ∶ 4.4 ∶ 8.9. The results of antioxidant activity in vitro showed that 3 kinds of homogeneous polysaccharides had certain scavenging activity on DPPH ·，·OH and O 2－·；the order of scavenging activity to DPPH ·was VC ＞CCL-M2＞CCL-M3＞CCL-M6（IC50 was 0.309， 1.240， 1.489 and 1.713 mg/mL）； the order of scavenging activity to ·OH was VC ＞CCL-M2＞CCL-M6＞CCL-M3（IC50 was 0.968，1.032，1.233，1.356 mg/mL）；the order of scavenging activity to O 2 －· was VC ＞CCL-M2＞CCL-M3＞CCL-M6 （IC50 was 0.335， 0.379， 0.812， 1.662 mg/mL）. CONCLUSIONS：3 kinds of monosaccharides in the form of pyranose are isolated from C. satiuus ，and all of them have antioxidant activity in vitro ，and CCL-M 2 is the best.

关键词

黄瓜子均一多糖分离纯化抗氧化活性

Keywords

Cucumis satiuusHomogeneous polysaccharidesSeparationPurificationAntioxidant activity

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