Skip to Content
Merck
  • Free radical scavenging activity and comparative metabolic profiling of in vitro cultured and field grown Withania somnifera roots.

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

PloS one (2015-04-16)
Kalaiselvi Senthil, Pankajavalli Thirugnanasambantham, Taek Joo Oh, So Hyun Kim, Hyung Kyoon Choi
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ethyl acetate
USP
Dimethyl sulfoxide, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Sodium nitrite, anhydrous, Redi-Dri, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Ethyl acetate, ACS reagent, ≥99.5%
Sigma-Aldrich
Toluene, ACS reagent, ≥99.5%
Sigma-Aldrich
Formic acid, ACS reagent, ≥96%
Sigma-Aldrich
Sodium nitrite, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Toluene, ACS reagent, ≥99.5%
Sigma-Aldrich
Dimethyl sulfoxide, ReagentPlus®, ≥99.5%
Supelco
Ethyl Acetate, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Pyridine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Dimethyl sulfoxide, ACS reagent, ≥99.9%
Sigma-Aldrich
Pyridine, biotech. grade, ≥99.9%
Sigma-Aldrich
Ethyl acetate, biotech. grade, ≥99.8%
Sigma-Aldrich
Sodium nitrite, ACS reagent, ≥97.0%
Sigma-Aldrich
Toluene, ACS reagent, ≥99.5%
Sigma-Aldrich
Pyridine, ACS reagent, ≥99.0%
Sigma-Aldrich
Formic acid, reagent grade, ≥95%
Sigma-Aldrich
Ethyl acetate, ACS reagent, ≥99.5%
Sigma-Aldrich
Formic acid, ACS reagent, ≥88%
Sigma-Aldrich
Toluene, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%
Sigma-Aldrich
Dimethyl sulfoxide, suitable for HPLC, ≥99.7%
Sigma-Aldrich
Ethyl acetate, suitable for HPLC, ≥99.8%
Sigma-Aldrich
Ethyl acetate, suitable for HPLC, ≥99.7%
Sigma-Aldrich
Ethyl acetate, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%
Sigma-Aldrich
Pyridine, suitable for HPLC, ≥99.9%
Supelco
Sodium carbonate concentrate, Na2CO3 72 mM in water, IC eluent concentrate (20x) for Metrosep A Supp 7
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
Supelco
Sodium carbonate concentrate, 0.1 M Na2CO3 in water, eluent concentrate for IC
Sigma-Aldrich
Ethyl acetate, ≥99%, FCC, FG