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林业科学 ›› 2015, Vol. 51 ›› Issue (9): 96-105.doi: 10.11707/j.1001-7488.20150913

• 论文与研究报告 • 上一篇    下一篇

纯化作用对五味子木脂素抗氧化性的影响

谷艳菲1, 闫伯前2, 丁轲1, 王宗义1, 张忠杰3, 韩涛1   

  1. 1. 北京农学院食品科学与工程学院 农产品有害微生物及农残安全检测与控制北京市重点实验室 北京 102206;
    2. 北京农学院植物科学技术学院 北京 102206;
    3. 国家粮食局科学研究院 北京 100037
  • 收稿日期:2014-09-15 修回日期:2014-10-14 出版日期:2015-09-25 发布日期:2015-10-16
  • 通讯作者: 韩涛
  • 基金资助:

    功能(保健)食品产品开发及其生物活性研究(PXM2013_014207_000048); 农产品加工及贮藏工程北京市重点建设学科项目(PXM2013-014207-000057)。

Effect of Purification on Antioxidative Activity of Lignan Fractions from Schisandra chinensis

Gu Yanfei1, Yan Boqian2, Ding Ke1, Wang Zongyi1, Zhang Zhongjie3, Han Tao1   

  1. 1. Beijing Key Laboratory of Agricultural Product Detection and Control of Spoilage Organisms and Pesticide Residue College of Food Science and Engineering, Beijing University of Agriculture Beijing 102206;
    2. College of Plant Science and Technology, Beijing University of Agriculture Beijing 102206;
    3. Academy of State Administration of Grain Beijing 100037
  • Received:2014-09-15 Revised:2014-10-14 Online:2015-09-25 Published:2015-10-16

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摘要：

[目的] 研究五味子木脂素在不同纯化程度下的抗氧化性能,为开发富集木脂素的五味子功能食品提供理论依据。[方法] 采用微波-超声波联用法提取五味子木脂素,得木脂素粗提液; 首先用AB-8大孔吸附树脂对粗提液初步纯化,用pH10、体积分数100%的乙醇进行洗脱; 采用高效液相色谱进行定性分析,检测方法为: Venusil XBP C18 柱(150 mm×4.6 mm,5 μm),流动相为甲醇(B)-水(A),梯度洗脱条件: 0~7 min (67%B)、7~13 min(67%~75%B)、13~22 min (75%~80%B)、22~30 min(80%~67%B),检测波长220 nm,流速0.8 mL·min-1,进样量10 μL,柱温为室温; 面积归一法计算木脂素纯度,粗提液中木脂素纯度为29.33%,大孔树脂初步纯化后的木脂素纯度为58.95%; 再采用制备液相色谱进一步纯化粗提液,检测方法为: Adsorbosphere XL C18柱 (250 mm×22 mm,5 μm),流动相为甲醇(A)-水(B),梯度洗脱条件: 0~10 min (75%A)、10~15 min(75%~78%A)、15~38 min (75%~80%A),检测波长220 nm,流速10 mL·min-1,进样量3 mL,柱温为室温; 经制备液相纯化后,得到4个木脂素单体,五味子醇甲纯度为95.40%,五味子酯甲纯度为98.85%,五味子甲素纯度为94.76%,五味子乙素纯度为96.44%; 比较不同纯度组分木脂素的还原能力,对·O2-、·OH、H2O2、DPPH自由基、ABTS自由基的清除能力,评价纯化作用对五味子木脂素抗氧化能力的影响。[结果] 在0.1~0.5 mg·mL-1范围内,不同纯度组分的五味子木脂素均表现出一定的抗氧化能力,但均低于抗坏血酸; 纯化后的木脂素抗氧化能力均低于未纯化的木脂素粗提液,制备液相得到的单体中,五味子乙素的抗氧化能力较强。[结论] 五味子木脂素粗提液中含有多糖等成分,五味子多糖具有抗氧化能力; 木脂素、多糖等成分协同作用,抗氧化能力强; 经大孔树脂初步纯化后,除去了多糖等大分子物质,单独的木脂素抗氧化能力相对变弱。 此结果说明,随着纯化程度增大,物质组分单一程度高,物质的生物活性不一定随之增强; 制备液相得到的单体中,五味子乙素的抗氧化能力较强,原因可能与其联苯环、辛烯环上的取代基有关。

关键词: 五味子, 木脂素, 抗氧化活性, 纯化, 制备液相

Abstract:

[Objective] Schisandra chinensis is the dried ripe fruit of Schisandra chinensis. Lignans are the major active ingredients of S. chinensis, which have many pharmacological actions. So far in China, except for the raw of S.chinensis, existing products are mostly Chinese patent drugs. Studies on antioxidant activities under different purities will provide theoretical references for developing lignan-rich Schisandra functional foods. [Method] Microwave-ultrasound extraction methods were used in extracting lignans from S.chinensis, and purify the crude extracts of lignan from S.chinensis. It is preliminarily purified with AB-8 macroporous resin and then eluted with pH10.0, 100% ethanol. This step can remove macromolecular substances, playing a role in preliminarily purifying the crude extracts of lignan from S.chinensis.Qualitive analyses were made by high performance liquid chromatography (HPLC), the separated conditions of Venusil XBP C18 (150 mm×4.6 mm,5 μm), methanol(B)-water(A) was used as gradient mobile phase (0-7 min 67%B, 7-13 min 67%-75%B, 13-22 min 75%-80%B, 22-30 min 80%-67%B),the detection wavelength was set at 220 nm,the flow rate was 0.8 mL·min-1, sample solution was 10 μL, the column temperature was as room temperature. And the purity of lignans was calculated by area normalization: in the crude extracts, the purity of lignans was 29.33% and the purity of lignans preliminarily purified by macroporous resin was 58.95%. Then the crude extracts were further purified by preparative liquid chromatography (PLC), the separated conditions of Adsorbosphere XL C18 (250 mm ×22 mm,5 μm), methanol(A)-water(B) was used as gradient mobile phase (0-10 min 75%A, 10-15 min 75%-78%A, 15-38 min 75%-80%A),the detection wavelength was set at 220 nm,the flow rate was 10 mL·min-1, sample solution was 3 mL, the column temperature was as room temperature.Four lignan monomers were obtained by PLC; purities of schisandrin, schisantherin A, deoxyschizandrin and schisandrin B were 95.40%, 98.85%, 94.76% and 96.44%, respectively. The reducing capacities and free radical scavenging capacities (·O2-,·OH, H2O2, DPPH and ABTS) of lignans from S.chinensis of different purities were compared, and an evaluation was made for effect of purification on antioxidant activities of lignans.[Result] In the range of 0.1 to 0.5 mg·mL-1, purified lignan extracts showed weaker antioxidant activity than that of unpurified crude lignan extracts; in the monomers obtained by PLC, schisandrin B showed a stronger antioxidant activity.[Conclusion] The crude extracts of lignan from S.chinensis contains polysaccharides and other ingredients, polysaccharide has antioxidant capacity. The synergistic reactions among lignans, polysaccharides and other ingredients make a stronger antioxidant capacity.Yet after purifying the crude extracts with AB-8 macroporous resin, the polysaccharides was removed, the antioxidant capacity of individual lignan is relatively weaker. These results indicate that as the degree of purification is increased, the single degree of material's component is higher, but biological activity of the substance is not necessarily increased as well. And in the monomers obtained by PLC, schisandrin B showed a stronger antioxidant activity, the reason may be related to the substituent on its biphenyl ring and octylene ring.

Key words: Schisandra chinensis, lignan, antioxidation, purification, PLC

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