Identification of 2″-galloylated flavonol 3-o-glycosides accumulating in developing leaves of persimmon

Phytochem Anal. 2011 Sep-Oct;22(5):403-10. doi: 10.1002/pca.1295. Epub 2011 Mar 17.

Abstract

Introduction: Dried leaves of persimmon, traditionally consumed as a herbal tea in Japan, contain bioactive galloylated flavonol glycosides. Investigation of the seasonal compositional changes in these compounds is important for determining the optimum harvest time.

Objective: The three objectives were: (1) to elucidate the compositional changes of the flavonol glycosides in persimmon leaves collected in different seasons; (2) to develop an efficient method for separation of the constituents; and (3) to compare their antioxidant activities.

Methodology: Fresh persimmon leaves were collected at different growth stages in 2007 and 2008 in Niigata, Japan. Total flavonols were extracted with 70% aqueous acetone followed by ethyl acetate partitioning, and were analysed by high-performance liquid chromatography. Enzymatic transformation and chromatographic fractionation was performed to isolate the individual flavonols. Two antioxidant assays were performed.

Results: Four nongalloylated flavonol glycosides were detected at the leaf-shooting stage, and four additional galloylated flavonol glycosides accumulated during leaf development, resulting in a total of eight constituents. Isolation of nongalloylated and galloylated constituents was successfully achieved through enzymatic transformation of the flavonol mixture using combinations of tannase, β-glucosidase and β-galactosidase, followed by chromatographic fractionation. The gallates were identified as regiospecific 2″-galloylated galactosides and glucosides of kaempferol and quercetin. A mixture of 2″-galloylated flavonol glycosides had a two-fold stronger antioxidant activity than the nongalloylated mixture.

Conclusion: This study showed that four 2″-galloylated flavonol glycosides had accumulated in developing leaves of persimmon by the end of May through a rapid 2″-galloylation of the corresponding nongalloylated flavonol glycosides.

MeSH terms

  • Biphenyl Compounds / chemistry
  • Chromatography, High Pressure Liquid / methods*
  • Diospyros / chemistry*
  • Flavonols / chemistry
  • Flavonols / isolation & purification*
  • Free Radical Scavengers / chemistry
  • Free Radical Scavengers / isolation & purification*
  • Glycosides / chemistry
  • Glycosides / isolation & purification*
  • Nuclear Magnetic Resonance, Biomolecular
  • Picrates / chemistry
  • Plant Leaves / chemistry
  • Seasons
  • Tandem Mass Spectrometry

Substances

  • Biphenyl Compounds
  • Flavonols
  • Free Radical Scavengers
  • Glycosides
  • Picrates
  • 1,1-diphenyl-2-picrylhydrazyl