Bio-assay guided isolation of α-glucosidase inhibitory constituents from Hibiscus mutabilis leaves

Phytochem Anal. 2012 Sep-Oct;23(5):421-5. doi: 10.1002/pca.1375. Epub 2011 Dec 12.

Abstract

Introduction: The increasing demand for natural-product-based medicines and health-care products for the management of diabetes encouraged investigation of this commonly available Indian plant.

Objectives: To establish the anti-diabetic (α-glucosidase inhibitory) activity of H. mutabilis leaf extract, isolate and identify the constituents responsible for the activity, and validate a HPLC method for quantification of the active constituents for standardisation of the extract.

Material and methods: The methanolic extract of leaves was partitioned between water, n-butanol and ethyl acetate. Bio-assay guided fractionation, based on inhibition of α-glucosidase, allowed isolation and identification of the active components. The active components were quantified using RP-HPLC-DAD validated for linearity, limit of detection, limit of quantification, precision, accuracy and robustness for this plant extract and the partitioned fractions.

Results: Ferulic acid and caffeic acid were identified as the α-glucosidase inhibitors present in H. mutabilis. They were partitioned into an ethyl acetate fraction. The HPLC-DAD calibration curve showed good linearity (r² > 0.99). For the recovery studies the %RSD was less than 2%. The interday and intraday variations were found to be less than 4% RSD for retention time and response.

Conclusion: The identification of α-glucosidase inhibition activity in H. mutabilis supports further investigations into the possible use of the plant for the management of diabetes. The HPLC method validated for these extracts will be useful in future research with the plant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Caffeic Acids / chemistry
  • Caffeic Acids / isolation & purification
  • Chromatography, High Pressure Liquid / methods
  • Coumaric Acids / chemistry
  • Coumaric Acids / isolation & purification*
  • Enzyme Assays / methods*
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / isolation & purification*
  • Glycoside Hydrolase Inhibitors*
  • Hibiscus / chemistry*
  • Hypoglycemic Agents / chemistry
  • Hypoglycemic Agents / isolation & purification
  • Linear Models
  • Plant Extracts / chemistry
  • Plant Leaves / chemistry*
  • Plant Proteins / antagonists & inhibitors
  • Sensitivity and Specificity

Substances

  • Caffeic Acids
  • Coumaric Acids
  • Enzyme Inhibitors
  • Glycoside Hydrolase Inhibitors
  • Hypoglycemic Agents
  • Plant Extracts
  • Plant Proteins
  • ferulic acid
  • caffeic acid