Free radical scavenging activity and comparative metabolic profiling of in vitro cultured and field grown Withania somnifera roots

PLoS One. 2015 Apr 14;10(4):e0123360. doi: 10.1371/journal.pone.0123360. eCollection 2015.


Free radical scavenging activity (FRSA), total phenolic content (TPC), and total flavonoid content (TFC) of in vitro cultured and field grown Withania somnifera (Ashwagandha) roots were investigated. Withanolides analysis and comprehensive metabolic profiling between 100% methanol extracts of in vitro and field grown root tissues was performed using high performance thin layer chromatography (HPTLC) and gas chromatography-mass spectrometry (GC-MS), respectively. Significantly higher levels of FRSA, TPC, and TFC were observed in in-vitro cultured roots compared with field grown samples. In addition, 30 day-cultured in vitro root samples (1 MIR) exhibited a significantly higher FRSA (IC50 81.01 μg/mL), TPC (118.91 mg GAE/g), and TFC (32.68 mg CE/g) compared with those in 45 day-cultured samples (1.5 MIR). Total of 29 metabolites were identified in in vitro cultured and field grown roots by GC-MS analysis. The metabolites included alcohols, organic acids, purine, pyrimidine, sugars, and putrescine. Vanillic acid was only observed in the in vitro cultured root samples, and higher level of the vanillic acid was observed in 1 MIR when compared to 1.5 MIR. Therefore, it is suggested that 1 MIR might serve as an alternative to field grown roots for the development of medicinal and functional food products.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Chromatography, Thin Layer
  • Flavonoids / analysis
  • Flavonoids / metabolism
  • Free Radical Scavengers / chemistry*
  • Gas Chromatography-Mass Spectrometry
  • Metabolomics*
  • Phenols / analysis
  • Phenols / metabolism
  • Plant Roots / chemistry
  • Plant Roots / metabolism
  • Principal Component Analysis
  • Withania / chemistry
  • Withania / metabolism*
  • Withanolides / analysis
  • Withanolides / metabolism


  • Flavonoids
  • Free Radical Scavengers
  • Phenols
  • Withanolides

Grant support

This work was supported by Mid-career Researcher Program (NRF-2012R1A2A2A02011748) through NRF grant funded by the MSIP of Republic of Korea and Major Research Project by University Grants Commission Government of India (F.No.34-278/2008 (SR)).