LC-MS-MS and GC-MS analyses of biologically active extracts and fractions from Tunisian Juniperus phoenice leaves

Pharm Biol. 2017 Dec;55(1):88-95. doi: 10.1080/13880209.2016.1230139. Epub 2016 Sep 22.

Abstract

Context: Despite some studies related to Juniperus phoenicea L. (Cupressaceae), phytochemical and biological investigations of this plant remain unexplored.

Objective: This work is the first report dealing with the identification and characterization of volatile components and flavonoids in hexane and methanol extracts from J. phoenicea leaves Materials and methods: Antioxidant activity of hexane, and methanol extracts from J. phoenicea leaves were determined by DPPH-radical scavenging assay. α-Amylase inhibitory activity was evaluated by enzyme inhibition using in vitro assay (each extract was dissolved in DMSO to give concentrations of 50, 100 and 200 mg/mL). The chemical composition of fractions (Fr1-Fr3) from methanol extract was determined by high-performance liquid chromatography coupled with mass spectroscopy (HPLC-MS) analysis.

Results and discussion: The hexane extract was analyzed by GC-MS technique which allowed the identification of 32 compounds. The main constituents were α-humulene (16.9%), pentadecane (10.2%) and α-cubebene (9.7%). Fraction Fr 2 exhibited a strong DPPH radical-scavenging activity (IC50 = 20.1 μg/mL) compared to that of BHT as well as the highest α-amylase inhibitory activity (IC50 = 28.4 μg/mL). Three flavonoids were identified in these fractions using HPLC-MS analysis: Quercetin 3-O-glucoside, isoscutellarein 7-O-pentoside and quercetin 3-O-pentoside. In addition, the more active fraction (Fr 2) was purified with semi-preparative HPLC affording one pure compound (amentoflavone) using 1H NMR analysis. This compound exhibited powerful DPPH radical-scavenging (IC50 = 14.1 μg/mL) and α-amylase inhibition (IC50 = 20.4 μg/mL) effects.

Conclusion: This study provides scientific support to some medicinal uses of J. phoenicea found in North Africa.

Keywords: HPLC-DAD–ESI/MS; biological activities; semi-prep HPLC.

MeSH terms

  • Antioxidants / isolation & purification
  • Antioxidants / pharmacology
  • Biphenyl Compounds / chemistry
  • Chromatography, High Pressure Liquid*
  • Enzyme Inhibitors / isolation & purification
  • Enzyme Inhibitors / pharmacology
  • Flavonoids / isolation & purification*
  • Flavonoids / pharmacology
  • Gas Chromatography-Mass Spectrometry*
  • Hexanes / chemistry
  • Juniperus / chemistry*
  • Methanol / chemistry
  • Phytotherapy
  • Picrates / chemistry
  • Plant Extracts / isolation & purification*
  • Plant Extracts / pharmacology
  • Plant Leaves / chemistry*
  • Plants, Medicinal
  • Solvents / chemistry
  • Tandem Mass Spectrometry*
  • Tunisia
  • Volatile Organic Compounds / isolation & purification*
  • Volatile Organic Compounds / pharmacology
  • alpha-Amylases / antagonists & inhibitors
  • alpha-Amylases / metabolism

Substances

  • Antioxidants
  • Biphenyl Compounds
  • Enzyme Inhibitors
  • Flavonoids
  • Hexanes
  • Picrates
  • Plant Extracts
  • Solvents
  • Volatile Organic Compounds
  • 1,1-diphenyl-2-picrylhydrazyl
  • alpha-Amylases
  • Methanol