Bioassay-guided fractionation and identification of α-amylase inhibitors from Syzygium cumini leaves

Pharm Biol. 2017 Dec;55(1):206-211. doi: 10.1080/13880209.2016.1257031.

Abstract

Context: Pancreatic α-amylase and α-glucosidase inhibitors serve as important strategies in the management of blood glucose. Even though Syzygium cumini (L.) Skeels (Myrtaceae) (SC) is used extensively to treat diabetes; scientific evidence on antidiabetic effects of SC leaves is scarce.

Objective: SC leaf extract was investigated for α-amylase inhibitory effect and continued with isolation and identification of α-amylase inhibitors.

Materials and methods: Bioassay-guided fractionation was conducted using in vitro α-amylase inhibitory assay (with 20-1000 μg/mL test material) to isolate the inhibitory compounds from ethyl acetate extract of SC leaves. Structures of the isolated inhibitory compounds were elucidated using 1H NMR and 13C NMR spectroscopic analysis and direct TLC and HPLC comparison with authentic samples. Study period was from October 2013 to October 2015.

Results: An active fraction obtained with chromatographic separation of the extract inhibited porcine pancreatic α-amylase with an IC50 of 39.9 μg/mL. Furthermore, it showed a strong inhibition on α-glucosidase with an IC50 of 28.2 μg/mL. The active fraction was determined to be a 3:1 mixture of ursolic acid and oleanolic acid. Pure ursolic acid and oleanolic acid showed IC50 values of 6.7 and 57.4 μg/mL, respectively, against α-amylase and 3.1 and 44.1 μg/mL respectively, against α-glucosidase.

Discussion and conclusions: The present study revealed strong α-amylase and α-glucosidase inhibitory effects of ursolic acid and oleanolic acid isolated from SC leaves for the first time validating the use of SC leaves in antidiabetic therapy.

Keywords: oleanolic acid; ursolic acid; α-Glucosidase.

MeSH terms

  • Acetates / chemistry
  • Biological Assay*
  • Carbon-13 Magnetic Resonance Spectroscopy
  • Chromatography, High Pressure Liquid
  • Chromatography, Thin Layer
  • Glycoside Hydrolase Inhibitors / isolation & purification
  • Glycoside Hydrolase Inhibitors / pharmacology*
  • Hypoglycemic Agents / isolation & purification
  • Hypoglycemic Agents / pharmacology*
  • Molecular Structure
  • Oleanolic Acid / isolation & purification
  • Oleanolic Acid / pharmacology
  • Pancreas / drug effects*
  • Pancreas / enzymology
  • Plant Extracts / isolation & purification
  • Plant Extracts / pharmacology*
  • Plant Leaves / chemistry*
  • Proton Magnetic Resonance Spectroscopy
  • Solvents / chemistry
  • Syzygium / chemistry*
  • Triterpenes / isolation & purification
  • Triterpenes / pharmacology
  • Ursolic Acid
  • alpha-Amylases / antagonists & inhibitors*
  • alpha-Amylases / metabolism

Substances

  • Acetates
  • Glycoside Hydrolase Inhibitors
  • Hypoglycemic Agents
  • Plant Extracts
  • Solvents
  • Triterpenes
  • Oleanolic Acid
  • ethyl acetate
  • alpha-Amylases