食品科学 ›› 2022, Vol. 43 ›› Issue (20): 18-24.doi: 10.7506/spkx1002-6630-20210809-114

• 食品化学 • 上一篇    下一篇

苦丁茶提取物的分离纯化、鉴定及不同萃取部位活性分析

邹成梅,厉莉,史硕硕,胡婷   

  1. (武汉工程大学环境生态与生物工程学院,湖北 武汉 430205)
  • 出版日期:2022-10-25 发布日期:2022-10-26
  • 基金资助:
    国家自然科学基金青年科学基金项目(21702156);武汉工程大学第十二届研究生教育创新基金项目(CX2020348); 武汉工程大学科学研究基金项目(K201759)

Fractionation, Identification and Biological Activities of Ethanol Extract of Kuding Tea (Ilex latifolia Thunb.)

ZOU Chengmei, LI Li, SHI Shuoshuo, HU Ting   

  1. (School of Environmental Ecology and Biological Engineering, Wuhan Institute of Technology, Wuhan 430205, China)
  • Online:2022-10-25 Published:2022-10-26

摘要: 以苦丁茶为原料,对其乙醇提取物和不同溶剂萃取部位的抗氧化活性、抗补体活性、抑制癌细胞活性进行探究,并对活性最佳的萃取部位进一步分离纯化,分析鉴定出单体化合物。抗氧化活性以总还原力、清除1,1-二苯基-2-三硝基苯肼自由基和超氧阴离子自由基的能力为指标,通过溶血实验评价抗补体活性,并采用MTT法测定对人乳腺癌细胞MCF-7的存活率,结合多种分离技术获得不同单体化合物,通过对质谱、核磁共振波谱的数据分析鉴定其化学结构。结果表明,苦丁茶乙醇提取物及各部位均具有良好的抗氧化效果,其中乙酸乙酯部位不仅在实验质量浓度范围内有较强的抗氧化活性(0.8 mg/mL均达到90%以上),还具有良好的抗补体活性(79.67±0.99)%,能很好抑制人乳腺癌MCF-7细胞增殖活性(54.8±0.26)%,对乙酸乙酯部位进行分离纯化,总共分离得到11 个单体化合物。由此可见,苦丁茶乙醇提取物和各部位均具有抗氧化、抗补体活性和人乳腺癌MCF-7细胞抑制活性,乙酸乙酯作为萃取剂时活性最佳。分离的单体化合物鉴定后多为酚类、黄酮类物质。

关键词: 苦丁茶;化合物;分离纯化;活性评价;结构鉴定

Abstract: In this study, an ethanol extract of Ilex latifolia Thunb. was fractionated by successive solvent extraction, and the antioxidant, anti-complement and anti-cancer activity of the ethanol extract and its fractions were explored. Furthermore, the fractions with the highest bioactivity activity were further fractionated into monomers, which were subsequently identified. The antioxidant activity was measured by total reducing power, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity and superoxide anion scavenging capacity. The anti-complement activity was evaluated by hemolysis assay, and the inhibitory effect on the proliferation of MCF-7 cells was determined by MTT assay. The chemical structures of the purified monomers were identified by mass spectrometry (MS) and nuclear magnetic resonance (NMR). The results showed that the ethanol extract and its fractions all had good antioxidant effect. Among them, the ethyl acetate fraction not only had a strong antioxidant activity in the experimental concentration range, but also had a good anti-complement activity with an inhibition percentage of (79.67 ± 0.99)%. It could inhibit the proliferation of human breast cancer MCF-7 cells with an inhibition percentage of (54.8 ± 0.26)%. A total of 11 monomer compounds were isolated from the ethyl acetate fraction, and most of them were identified as phenols and flavonoids. In conclusion, the ethanol extract of I. latifolia Thunb. and its fractions have antioxidant and anti-complement activities and an inhibitory effect on the proliferation of MCF-7 cells, and the ethyl acetate fraction has the strongest biological activity.

Key words: Ilex latifolia Thunb.; compound; isolation and purification; activity evaluation; structural identification

中图分类号: