Simultaneous quantification of eight bioactive secondary metabolites from Codonopsis ovata by validated high performance thin layer chromatography and their antioxidant profile

J Pharm Biomed Anal. 2014 Nov:100:300-308. doi: 10.1016/j.jpba.2014.07.034. Epub 2014 Aug 7.

Abstract

Chemical investigation of Codonopsis ovata resulted in the isolation and identification of β-sitosterol-3-O-glycoside, luteolin, apigenin, gentiacaulein, swertiaperenine, β-sitosterol, taraxeryl-3-acetate, and 3β-acetoxyoleanane-12-one. A rapid, precise, sensitive and validated HPTLC method for simultaneous quantification of these natural products (NPs) was developed on silica-gel 60F254 plate using ternary solvent system, n-hexane:ethyl acetate:formic acid (10.5:3.5:0.43, v/v/v). Markers were quantified after post chromatographic derivatization with cerric ammonium sulfate reagent. The method was validated for accuracy, precision, LOD, LOQ and all calibration curves showed a good linear relationship (r>0.9924) within test range. Precision was evaluated by intra- and inter-day tests with RSDs <2.59%, accuracy validation recovery 92.43-99.50% with RSDs <1.00%. Apigenin was found major component (natural abundance: 1.103%) and β-sitosterol the least (0.0263%). The NPs displayed antioxidant activity with luteolin exhibiting maximum effect at 1μg/mL concentration (75.9% for DPPH and 43.7% for ABTS) and others at 10 and 25μg/mL, suggesting thereby their apparent potential use for the prevention of free radical induced diseases or as an additive element to food and pharmaceutical industry.

Keywords: Antioxidant; Codonopsis ovata; HPTLC; Quantification; Validation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Acetates / chemistry
  • Ammonium Sulfate / chemistry
  • Benzothiazoles / chemistry
  • Biphenyl Compounds / chemistry
  • Calibration
  • Chromatography, Thin Layer / methods*
  • Codonopsis / metabolism*
  • Formates / chemistry
  • Free Radical Scavengers / analysis*
  • Free Radical Scavengers / pharmacology
  • Hexanes / chemistry
  • Limit of Detection
  • Linear Models
  • Phytotherapy
  • Picrates / chemistry
  • Plant Extracts / analysis*
  • Plant Extracts / pharmacology
  • Plants, Medicinal
  • Reference Standards
  • Reproducibility of Results
  • Silica Gel / chemistry
  • Solvents / chemistry
  • Sulfonic Acids / chemistry

Substances

  • Acetates
  • Benzothiazoles
  • Biphenyl Compounds
  • Formates
  • Free Radical Scavengers
  • Hexanes
  • Picrates
  • Plant Extracts
  • Solvents
  • Sulfonic Acids
  • formic acid
  • 2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid
  • n-hexane
  • Silica Gel
  • ethyl acetate
  • 1,1-diphenyl-2-picrylhydrazyl
  • Ammonium Sulfate