基于高效液相色谱特征指纹图谱法探究不同品种苜蓿中黄酮抗氧化活性的谱-效关系

关淑文; 潘予琮; 寇伟; 年芳; 蒋林树

doi:10.3969/j.issn.1006-267x.2022.12.055

动物营养学报 >

2022 , Vol. 34 >Issue 12: 8086 - 8096

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.12.055

研究论文 RESEARCH PAPER

基于高效液相色谱特征指纹图谱法探究不同品种苜蓿中黄酮抗氧化活性的谱-效关系

关淑文 ,
潘予琮 ,
寇伟 ,
年芳 ,
蒋林树

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1. 北京农学院动物科学技术学院, 奶牛营养学北京市重点实验室, 北京 102206;
2. 甘肃农业大学动物科学技术学院, 兰州 730070

关淑文(1995—),女,黑龙江黑河人,硕士研究生,研究方向为反刍动物营养与免疫。E-mail:guan_shuwen@126.com

收稿日期: 2022-05-07

网络出版日期: 2022-12-15

基金资助

北京市教育委员会重点项目（KZ202010020029）；现代农业产业技术体系北京市家畜创新团队；2022分类发展定额项目-人才引进启动经费（5066516004/003）

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Exploring Spectrum-Effect Relationship of Flavonoid Antioxidant Activity in Different Kinds of Alfalfa Based on High Performance Liquid Chromatography Characteristic Fingerprint

GUAN Shuwen ,
PAN Yucong ,
KOU Wei ,
NIAN Fang ,
JIANG Linshu

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1. Beijing Key Laboratory of Dairy Cattle Nutrition, Institute of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China;
2. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China

Received date: 2022-05-07

Online published: 2022-12-15

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摘要

本试验旨在探究苜蓿中黄酮特征指纹图谱及其主效成分发挥抗氧化能力的谱-效关系。本试验在构建苜蓿中黄酮特征指纹图谱的基础上，采用体外抗氧化法测定14种苜蓿来源的黄酮对1，1-二苯基-2-三硝基苯肼（DPPH）、2，2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸（ABTS）自由基的清除力和还原力的影响，并通过半数抑制浓度系数（IC₅₀）和半数最大效应浓度系数（EC₅₀）评价14种苜蓿中黄酮的体外抗氧化活性；运用主成分分析、偏最小二乘回归模型和多变量间相关性分析构建谱-效关系。结果表明：1）成功构建14种苜蓿来源黄酮的指纹图谱，并确定9个共有峰，其相似度范围在0.471~0.976；指认出峰2为芹菜素，峰3为苜蓿素，峰4为山奈酚。2）14种苜蓿中的黄酮清除DPPH的IC₅₀为1.120~2.149 mg/mL，ABTS的IC₅₀为0.302~0.759 mg/mL，达到还原力的EC₅₀为0.035~0.143 mg/mL。3）经主成分分析、偏最小二乘回归模型和多变量之间相关性分析，峰1、峰6的面积与苜蓿中黄酮抗氧化活性呈显著正相关（P<0.05），峰2的面积与还原力的活性呈显著负相关（P<0.05）。综上所述，通过对苜蓿中黄酮的谱-效关系研究，初步确定了14种苜蓿来源黄酮中具有抗氧化活性的成分为峰1、峰3（苜蓿素）；峰6、峰8为DPPH自由基清除主效因子，峰1、峰2（芹菜素）、峰4（山奈酚）、峰6、峰7为ABTS自由基清除主效因子，峰1、峰4（山奈酚）、峰7、峰8为还原力的主效因子，本试验结果为苜蓿黄酮作为一种天然抗氧化饲料添加剂的研究应用提供理论依据。

关键词： 苜蓿黄酮; 抗氧化; 指纹图谱; 谱-效关系

本文引用格式

关淑文 , 潘予琮 , 寇伟 , 年芳 , 蒋林树 . 基于高效液相色谱特征指纹图谱法探究不同品种苜蓿中黄酮抗氧化活性的谱-效关系[J]. 动物营养学报, 2022 , 34(12) : 8086 -8096 . DOI: 10.3969/j.issn.1006-267x.2022.12.055

Abstract

The purpose of this paper was to explore the characteristic fingerprint of flavonoids in alfalfa and the spectrum-effect relationship of the main active components in their antioxidant capacity. In this experiment, based on the construction of alfalfa flavonoids characteristic fingerprint, the in vitro antioxidant method was used to determine the effects of flavonoids from 14 kinds of alfalfa on the scavenging power and reducing power of 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH) and 2,2'-azo-bis-3-ethyl benzothiazoline-6-sulfonic acid (ABTS) free radicals. The median inhibitory concentration coefficient (IC₅₀) and the half-maximal effect concentration coefficient (EC₅₀) was used to evaluate the antioxidant activities of flavonoids in 14 alfalfa species in vitro. Principal component analysis, partial least squares regression model and multivariate correlation analysis were used to construct a spectrum-effect relationship. The results showed as follows:1) the fingerprints of flavonoids from 14 alfalfa species were constructed, and 9 common peaks were identified, and their similarity ranged from 0.471 to 0.976; peak 2 was identified as apigenin, peak 3 was alfalfa and peak 4 was kaempferol. 2) Eventually, the IC₅₀ values of flavonoids from 14 alfalfa species for eliminating DPPH free radicals ranged from 1.120 to 2.149 mg/mL. The IC₅₀ values for scavenging ABTS free radicals were 0.302 to 0.759 mg/mL. Furthermore, the EC₅₀ values for achieving reducing force were 0.035 to 0.143 mg/mL. 3) Through principal component analysis, partial least squares regression and multivariate correlation analysis, the finding showed that the areas of peaks 1 and 6 were significantly positively correlated with the antioxidant activity of alfalfa flavonoids (P<0.05), and the area of peak 2 was prominently negatively correlated with the activity of reducing force (P<0.05). Through the study on the spectrum-effect relationship of flavonoids in alfalfa, the components with antioxidant activity in flavonoids from 14 alfalfa species are preliminarily determined, namely, peak 1, peak 3 (alfalfa), peak 6, and peak 8 are the main factors of DPPH free radical scavenging. Peak 1, peak 2 (apigenin), peak 4 (kaempferol), peak 6 and peak 7 are the main factors of ABTS free radical scavenging. Furthermore, peak 1, peak 4 (kaempferol), peak 7, and peak 8 are the mainsprings of reducing force. All the results testify a feasible basis for the application of alfalfa flavonoids is a natural antioxidant feed additive in animal production.

Key words： alfalfa flavonoids; antioxidant; fingerprint; spectrum-effect relationship

参考文献

[1] 李云鹤, 李月, 张巧娥, 等.苜蓿草饲用价值评定方法的研究进展[J].饲料研究, 2022, 45(6):154-157. LI Y H, LI Y, ZHANG Q E, et al.Research progress on evaluation methods of forage value of alfalfa[J].Feed Research, 2022, 45(6):154-157.(in Chinese)
[2] 杨蕾.黄酮类化合物在反刍动物营养上的研究进展[J].家畜生态学报, 2019, 40(2):8-12. YANG L.Advance in flavonoids on ruminant nutrition[J].Journal of Domestic Animal Ecology, 2019, 40(2):8-12.(in Chinese)
[3] 潘予琮.基于指纹图谱技术的苜蓿和燕麦草中黄酮类营养活性物质分析与评价[D].硕士学位论文.北京:北京农学院, 2021. PAN Y C.Analysis and evaluation of flavonoid nutricines in alfalfa and oat grass based on fingerprint technique[D].Master's Thesis.Beijing:Beijing University of Agriculture, 2021.(in Chinese)
[4] WU F, SHI S L, LI Y Z, et al.Physiological and biochemical response of different resistant alfalfa cultivars against thrips damage[J].Physiology and Molecular Biology of Plants, 2021, 27(3):649-663.

[5] 王梦竹, 刘艳丰, 王文奇, 等.苜蓿黄酮对绵羊生长性能和血清指标的影响[J].中国畜牧兽医, 2015, 42(9):2345-2351. WANG M Z, LIU Y F, WANG W Q, et al.Effect of alfalfa flavonoids on growth performance and serum indexes of sheep[J].China Animal Husbandry & Veterinary Medicine, 2015, 42(9):2345-2351.(in Chinese)
[6] ZHAN J S, LIU M M, SU X S, et al.Effects of alfalfa flavonoids on the production performance, immune system, and ruminal fermentation of dairy cows[J].Asian-Australasian Journal of Animal Sciences, 2017, 30(10):1416-1424.

[7] 占今舜, 陈小连, 詹康, 等.苜蓿黄酮对脂多糖诱导下奶牛乳腺上皮细胞凋亡的影响[J].草业学报, 2018, 27(1):187-194. ZHAN J S, CHEN X L, ZHAN K, et al.Effects of alfalfa flavonoids on apoptosis of bovine mammary epithelial cells induced by lipopolysaccharide[J].Acta Prataculturae Sinica, 2018, 27(1):187-194.(in Chinese)
[8] 孟祥君, 俞慧云, 付萍, 等.不同种植区陇东苜蓿营养价值的比较研究[J].中国饲料, 2021(17):95-99. MENG X J, YU H Y, FU P, et al.Comparative study on nutritional value of Medicago sativa.Longdong in different planting areas[J].China Feed, 2021(17):95-99.(in Chinese)
[9] 倪兴军, 康小红.河西走廊紫花苜蓿干草不同喂量对泌乳前期奶牛生产性能的效应研究[J].中国牛业科学, 2021, 47(6):24-26. NI X J, LANG X H.Effects of different feeding amount of alfalfa hay in Hexi corridor on production performance of dairy cows in early lactation[J].China Cattle Science, 2021, 47(6):24-26.(in Chinese)
[10] 李戎, 闫智勇, 李文军, 等.创建中药谱效关系学[J].中医教育, 2002, 21(2):62. LI R, YAN Z Y, LI W J, et al.Create a spectrum-activity relationship of traditional Chinese medicine[J].Education of Chinese Medicine, 2002, 21(2):62.(in Chinese)
[11] 吴思宇, 杨丹丹, 傅静, 等.谱效关系在中药质量控制中的应用[J].长春中医药大学学报, 2019, 35(2):399-402. WU S Y, YANG D D, FU J, et al.Application of spectrum-effect relationship in quality control of Chinese materia medica[J].Journal of Changchun University of Traditional Chinese Medicine, 2019, 35(2):399-402.(in Chinese)
[12] 黄纪念.迷迭香抗氧化活性及其作用机理研究和天然抗氧化保健品的研制[D].硕士学位论文.北京:中国农业大学, 2003. HUANG J N.Study on antioxidative activity and its mechanism of rosemary and development of natural functional food[D].Master's Thesis.Beijing:China Agricultural University, 2003.(in Chinese)
[13] 国家药典委员会.中华人民共和国药典[M].北京:中国医药科技出版社, 2020. Chinese Pharmacopoeia Commission.Pharmacopoeia of the People's Republic of China[M].Beijing:China Medical Science Press, 2020.(in Chinese)
[14] 郭新颖, 李晨阳, 王梦晗, 等.苜蓿总黄酮纯化及纯化前后对DPPH自由基清除能力比较[J].高师理科学刊, 2021, 41(3):48-51. GUO X Y, LI C Y, WANG M H, et al.Purification of total flavonoids from alfalfa and comparison of DPPH radical scavenging ability before and after purification[J].Journal of Science of Teachers' College and University, 2021, 41(3):48-51.(in Chinese)
[15] 康超, 刘凤听, 刘云芬, 等.不同品种芒果核多酚和黄酮含量及抗氧化活性评价[J].食品工业科技, 2021, 42(20):100-105. KANG C, LIU F T, LIU Y F, et al.Contents of polyphenols and flavonoids and its antioxidant activity evalution in different kinds of mango kernel seeds[J].Science and Technology of Food Industry, 2021, 42(20):100-105.(in Chinese)
[16] 钱松, 刘琴, 麻秀萍, 等.地稔醇提物抗氧化活性的谱效关系研究[J].中国药房, 2021, 32(16):1969-1974. QIAN S, LIU Q, MA X P, et al.Study on the spectrum-effect relationship of antioxidant activity of ethanol extract from melastoma dodecandrum[J].China Pharmacy, 2021, 32(16):1969-1974.(in Chinese)
[17] 李艳荣, 周维维, 王子怡, 等.山楂叶提取物抗氧化活性及谱效关系研究[J].中国药学杂志, 2020, 55(20):1673-1679. LI Y R, ZHOU W W, WANG Z Y, et al.Relationship between antioxidant activity and spectrum-effect of hawthorn leaf extracts[J].Chinese Pharmaceutical Journal, 2020, 55(20):1673-1679.(in Chinese)

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