RESEARCH PAPER

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

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.

Cite this article

GUAN Shuwen , PAN Yucong , KOU Wei , NIAN Fang , JIANG Linshu . Exploring Spectrum-Effect Relationship of Flavonoid Antioxidant Activity in Different Kinds of Alfalfa Based on High Performance Liquid Chromatography Characteristic Fingerprint[J]. Chinese Journal of Animal Nutrition, 2022 , 34(12) : 8086 -8096 . DOI: 10.3969/j.issn.1006-267x.2022.12.055

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