Characterization of phenolic compounds using UPLC–HRMS and HPLC–DAD and anti-cholinesterase and anti-oxidant activities of Trifolium repens L. leaves

Sultan Ahmad, Alam Zeb*, Muhammad Ayaz, Michael Murkovic

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


This study was aimed to investigate the phenolic profile of methanolic extract of Trifolium repens L. leaves using LC–HRMS and LC–DAD and its anti--cholinesterase and anti-oxidant activities. The n-Hexane (Tr.Hex), chloroform (Tr.Chf), ethanol (Tr.Et), methanol (Tr.Cme), and aqueous (Tr.Aq) extracts were tested for anti-cholinesterases and anti-oxidant potentials. Twenty-nine phenolic compounds were quantified and identified. The highest amount present were of tyrosol (11.1 mg/g), quercetin-3-glucuronide (5.36 mg/g), formononetin-7-glucoside (5.03 mg/g), quercetin-3-O-glucoside (4.71 mg/g), 3,4-di-O-caffeoylquinic acid (4.69 mg/g), formononetin-7-glucoside-acetate (4.29 mg/g), quercetin-3-O-glucoside (4.26 mg/g), and formononetin (3.64 mg/g). Tr.Chf and Tr.Et were most active against AChE, BChE, and free radicals. Tr.Chf exhibited IC50 of 15 and 21 µg/mL against AChE and BChE, respectively. In the anti-oxidant study, Tr.Chf showed IC50 of 42 and 25 µg/mL against ABTS and DPPH radicals, respectively. The current results of anti-cholinesterase and anti-radical studies on T. repens further signify its potential application in neurodegenerative disorders.
Original languageEnglish
Pages (from-to)485-496
Number of pages12
JournalEuropean Food Research & Technology
Issue number3
Publication statusPublished - 1 Mar 2020


  • Anti-cholinesterase activity
  • Anti-oxidant activity
  • Phenolic compounds
  • Trifolium repens

ASJC Scopus subject areas

  • Food Science
  • Chemistry(all)
  • Biochemistry
  • Biotechnology
  • Industrial and Manufacturing Engineering

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