Antioxidant activity evaluation of rosemary ethanol extract and their cellular antioxidant activity toward HeLa cells

J Food Biochem. 2019 Jul;43(7):e12851. doi: 10.1111/jfbc.12851. Epub 2019 Apr 14.


Rosemary ethanol extract (REE) from Rosmarinus officinalis was identified by LC-ESI-MS/MS and 12 compounds were found. Among them, rosmarinic acid (389.78 μg/mg in REE), luteolin-3'-O-glucuronide (325.58 μg/mg), luteolin-5-O-glucuronide (120.92 μg/mg), and geniposide (120.83 μg/mg) are the major components. The antioxidant activity evaluation of REE by off-line HPLC methods indicated that among the 12 compounds, rosmarinic acid had the strongest scavenging activities in both DPPH· and ·OH. The cytotoxicity experiment showed that REE with the concentration ranges from 1 to 100 µg/ml did not significantly affect the cell viability of HeLa, while inhibitory rate reduced to 62.3% when the concentration was increased to 1,000 µg/ml. The results of intracellular antioxidation assay showed that the ability of REE in reducing the reactive oxygen species (ROS) in HeLa cells was higher than rosmanol, and lower than rosmarinic acid without cell toxicity. PRACTICAL APPLICATIONS: Plant polyphenols are essential components of functional foods, due to their antioxidant and enzyme inhibition activities. This paper is the first study about the quantification of antioxidant compounds, antioxidant activity evaluation, and their cellular antioxidant activity of polyphenols extract from R. officinalis toward HeLa cells. We aimed to elucidate the chemical composition and recognition of antioxidant components with DPPH and OH free radicals scavenging activity. In addition, the polyphenols dose-response correlations with cellular antioxidant activity were also determined. These results indicated that off-line HPLC method with DPPH and OH free radicals as markers is available for screening antioxidant activity of polyphenols from the mixture.

Keywords: Rosmarinus officinalis; antioxidant activity evaluation; free radicals; intracellular antioxidation; rosemary ethanol extract (REE).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abietanes / metabolism
  • Abietanes / toxicity
  • Antioxidants* / analysis
  • Antioxidants* / pharmacology
  • Antioxidants* / toxicity
  • Chromatography, High Pressure Liquid
  • Cinnamates / metabolism
  • Cinnamates / toxicity
  • Depsides / metabolism
  • Depsides / toxicity
  • Ethanol
  • Free Radical Scavengers
  • HeLa Cells
  • Humans
  • Plant Extracts* / analysis
  • Plant Extracts* / pharmacology
  • Plant Extracts* / toxicity
  • Plant Leaves / chemistry
  • Polyphenols* / analysis
  • Polyphenols* / pharmacology
  • Polyphenols* / toxicity
  • Reactive Oxygen Species / metabolism
  • Rosmarinus / chemistry*


  • Abietanes
  • Antioxidants
  • Cinnamates
  • Depsides
  • Free Radical Scavengers
  • Plant Extracts
  • Polyphenols
  • Reactive Oxygen Species
  • Ethanol
  • rosmanol
  • rosmarinic acid