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
Penta-O-galloyl-β-D-glucose hydrate, ≥96% (HPLC)
Sigma-Aldrich
L-Ascorbic acid, tested according to Ph. Eur.
Sigma-Aldrich
Chlorotrimethylsilane, ≥98.0% (GC)
Sigma-Aldrich
L-Ascorbic acid, BioUltra, ≥99.5% (RT)
Sigma-Aldrich
Chlorotrimethylsilane, puriss., ≥99.0% (GC)
Supelco
Sodium carbonate, reference material for titrimetry, certified by BAM, >99.5%
Supelco
Sulfuric acid, for the determination of nitrogen, ≥97.0%
Sigma-Aldrich
Aluminum chloride, 99.99% trace metals basis
Sigma-Aldrich
Aluminum chloride, powder, 99.99% trace metals basis
Sigma-Aldrich
Methoxyamine hydrochloride, 98%
Sigma-Aldrich
Sodium carbonate, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Aluminum chloride, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Sodium carbonate, BioUltra, anhydrous, ≥99.5% (calc. on dry substance, T)
Sigma-Aldrich
Chlorotrimethylsilane, Wacker Chemie AG, ≥99.0% (GC)
Sigma-Aldrich
L-Ascorbic acid, puriss. p.a., ACS reagent, reag. ISO, Ph. Eur., 99.7-100.5% (oxidimetric)
Sigma-Aldrich
Chlorotrimethylsilane, purified by redistillation, ≥99%
Sigma-Aldrich
Methanol, anhydrous, 99.8%
Sigma-Aldrich
L-Ascorbic acid, 99%
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, derivatization grade (GC derivatization), LiChropur, contains 10% TMCS, 98% (excluding TMCS)
Sigma-Aldrich
Sulfuric acid, 99.999%
Sigma-Aldrich
L-Ascorbic acid, reagent grade, crystalline
Sigma-Aldrich
L-Ascorbic acid, powder, suitable for cell culture, γ-irradiated
Sigma-Aldrich
L-Ascorbic acid, reagent grade
Sigma-Aldrich
L-Ascorbic acid, suitable for cell culture, suitable for plant cell culture, ≥98%
Sigma-Aldrich
L-Ascorbic acid, meets USP testing specifications
Sigma-Aldrich
Sodium carbonate, BioXtra, ≥99.0%
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, with 1% trimethylchlorosilane, derivatization grade (GC derivatization), LiChropur
Sigma-Aldrich
L-Ascorbic acid, BioXtra, ≥99.0%, crystalline
Sigma-Aldrich
L-Ascorbic acid, ACS reagent, ≥99%
Sigma-Aldrich
Methoxylamine hydrochloride solution, 25-30 wt. % in H2O