Abstract
The flavonoids were extracted from alfalfa using ethanol assisted with ultrasonic extraction and purified by D101 macroporous resin column chromatography. The chemical composition and content of ethanol elution fractions (EEFs) were assessed by ultrahigh-performance liquid chromatography and hybrid quadrupole time of flight mass spectrometry (UHPLC-Q-TOF-MS) and aluminum nitrate-sodium nitrite-sodium hydroxide colorimetric method. The in vitro antioxidant activity of two EEFs was conducted by scavenging DPPH free radical, and the main antioxidants of 75% EEFs were screened using DPPH-UHPLC. Moreover, the in vivo antioxidant activity of 75% EEFs and the growth performance of broilers were studied. The results showed that the content of 30% and 75% EEFs was 26.20% and 62.57%. Fifteen compounds were identified from 75% EEFs, and five of them were reported in alfalfa for the first time. The scavenging activity of 75% and 30% EEFs (200 μg/mL) against DPPH was 95.51% and 78.85%. The peak area of 5,3′,4′-trihydroxyflavone and hyperoside was decreased by 82.69% and 76.04%, which exhibited strong scavenging capacities. The total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) level of three treated groups against the normal control group (NC) fed with basal diet significantly increased by 3.89-24.49%, 0.53-7.39%, and 0.79-11.79%, respectively. While the malondialdehyde (MDA) decreased by 0.47-18.27%. Compared with the NC, the feed to gain ratio (F : G) of three treated groups was lowered by 2.98-16.53% and survival rate of broilers significantly increased. Consequently, 75% EEFs extracted from alfalfa exhibited powerful antioxidant activities and might be a potential feed additive to poultry and livestock.
Highlights
Oxidation is an essential biological process central to the functioning of living organism for energy production
The aim of our work was to select a representative DPPH free radical to determine the antioxidant capacity of 30% and 75% ethanol elution fractions (EEFs), and vitamin c (Vc) was used as the positive control
It can be seen that the DPPH⋅ scavenging rate increased with the increase of the content of alfalfa flavonoids
Summary
Oxidation is an essential biological process central to the functioning of living organism for energy production. Antioxidant plays an important role in delaying, preventing, and removing peroxide products and has been used to lessen the impact on oxidative damage [2]. With the characteristics of low cost, high performance, and broad bioactivity, have long been in common usage in food, pharmaceutical and cosmetics, health products, and animal feed industry. There has been more and more interests in developing and using natural, effective, safe, and multiple biological antioxidants to replace synthetic antioxidants in many industries, such as natural plant polyphenol compounds [4, 5]. The flavonoids are a group of polyphenol compounds with a basic structure of C6-C3-C6 carbon skeleton and known for powerful antioxidant capacities based on their polyhydroxyl structures [6, 7]
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