Skip to Content
MilliporeSigma
  • 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

Supelco
Sulfuric acid, for the determination of nitrogen, ≥97.0%
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, with 1% trimethylchlorosilane, for GC derivatization, LiChropur
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Sodium carbonate, powder, ≥99.5%, ACS reagent
Sigma-Aldrich
Sodium carbonate, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium carbonate, BioUltra, anhydrous, ≥99.5% (calc. on dry substance, T)
Sigma-Aldrich
Chlorotrimethylsilane, purified by redistillation, ≥99%
Sigma-Aldrich
Chlorotrimethylsilane, ≥98.0% (GC)
Sigma-Aldrich
Chlorotrimethylsilane, puriss., ≥99.0% (GC)
Sigma-Aldrich
Chlorotrimethylsilane, produced by Wacker Chemie AG, Burghausen, Germany, ≥99.0% (GC)
Supelco
Chlorotrimethylsilane, for GC derivatization, LiChropur, ≥99.0% (GC)
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, for GC derivatization, LiChropur, contains 1% TMCS, 99% (excluding TMCS)
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, for GC derivatization, LiChropur, contains 10% TMCS, 98% (excluding TMCS)
Sigma-Aldrich
Methanol, puriss., meets analytical specification of Ph Eur, ≥99.7% (GC)
Sigma-Aldrich
Methoxyamine hydrochloride, 98%
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Sigma-Aldrich
Methanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)
Sigma-Aldrich
Methanol, BioReagent, ≥99.93%
Sigma-Aldrich
Methanol, ACS spectrophotometric grade, ≥99.9%
Supelco
Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Methanol, suitable for HPLC, ≥99.9%
Supelco
Methanol, analytical standard
Sigma-Aldrich
Aluminum chloride, reagent grade, 98%
Sigma-Aldrich
Aluminum chloride, ReagentPlus®, 99%
Sigma-Aldrich
Aluminum chloride, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Aluminum chloride solution, 1.0 M in nitrobenzene
Sigma-Aldrich
Aluminum chloride, anhydrous, powder, 99.99% trace metals basis
Sigma-Aldrich
Aluminum chloride, 99.99% trace metals basis
Sigma-Aldrich
Sulfuric acid solution, puriss. p.a., ≥25% (T)