Evaluation of the antioxidant potentials of extract and fractions of Vitex doniana fruit and identification of the bioactive metabolites using HPLC-DAD-MS analysis

Keziah Uchechi  Ajah; Ozioma Anne  Asomugha; Chinazom Perpetua  Ifedigbo; Kosoluchukwu Chidubem  Umeh; Onyeka Chinwuba  Obidiegwu; Amara Anwuchaepe  Ajaghaku; Nkeoma Nkasi  Okoye; Moses Aziakpono  Omoriri; Festus Basden Chiedu  Okoye

doi:10.30574/gscbps.2021.17.1.0310

Authors

Keziah Uchechi Ajah Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Anambra State Nigeria.
Ozioma Anne Asomugha Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Anambra State Nigeria.
Chinazom Perpetua Ifedigbo Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Anambra State Nigeria.
Kosoluchukwu Chidubem Umeh Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Anambra State Nigeria.
Onyeka Chinwuba Obidiegwu Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Anambra State Nigeria.
Amara Anwuchaepe Ajaghaku Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Enugu State University of Science and Technology, Agbani, Enugu State Nigeria.
Nkeoma Nkasi Okoye Department of Pure and Industrial Chemistry, Faculty of Physical Sciences, Nnamdi Azikiwe University, Awka, Anambra State Nigeria.
Moses Aziakpono Omoriri Department of Pharmacology and Toxicology, Federal University Oye-Ekiti, Ekiti State, Nigeria.
Festus Basden Chiedu Okoye Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Anambra State Nigeria.

DOI:

https://doi.org/10.30574/gscbps.2021.17.1.0310

Keywords:

Vitex doniana, Antioxidant activity, HPLC-DAD, LC-MS, Isoflavones

Abstract

Isoflavones and flavonoids in general found in fruits and vegetable act as natural antioxidants. This study was aimed at screening the isoflavone-rich fractions of Vitex doniana fruit extract for antioxidant activity using in vitro and in vivo models. The ethyl acetate and butanol fractions were screened for in vitro antioxidant activity by DPPH free radical scavenging and FRAP (Fe3+ Reducing Antioxidant Power) assays using ascorbic acid as standard. In vivo antioxidant activity was also tested against carbon tetrachloride-induced toxicity in mice using carbon tetrachloride (4 mL/kg) and silymarin (100 mg/kg) as standard. The ethyl acetate fraction and butanol Vacuum Liquid Chromatography (VLC) sub-fractions were subjected to High Performance Liquid Chromatography–Diode Array Detector (HPLC-DAD) and Liquid Chromatography-Mass Spectrometry (LC-MS) analyses. The ethyl acetate and butanol fractions inhibited DPPH radicals with an IC₅₀ of 897 µg/mL and 2809.38 µg/mL respectively. Both fractions also showed mild FRAP result with EC₅₀ of 1401 µg/mL and 7051 µg/mL respectively. The fractions produced significant decrease (P < 0.05) in ALP levels only while ethyl acetate fraction caused a significant reduction in the value of only malondialdehyde. Both fractions also increased levels of superoxide dismutase (SOD). HPLC-DAD analysis led to the detection of cinnamic acid, protocatechuic acid and two isoflavones daidzein and genistein. Further LC-MS analysis confirmed the presence of the isoflavones tectorigenin, 5-O-methylgenistein, and 5-O-methyltectorigenin, among other flavonoids, peonidin, 5-methylpeonidin, methylmalvidin and kaempferol-3-O-(2''galloyl)-glucopyranoside. The detected isoflavones and other flavonoids may contribute to the observed mild in vitro and good in vivo antioxidant activity of V. doniana fruit extract and fractions.

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