Validation of a colorimetric assay for the in vitro screening of inhibitors of angiotensin-converting enzyme (ACE) from plant extracts

Phytomedicine. 2005 Jun;12(6-7):424-32. doi: 10.1016/j.phymed.2004.07.002.

Abstract

A new method for the in vitro screening of plant extracts with potential angiotensin-converting enzyme (ACE) inhibitory activity is proposed. The method is based on the cleavage of the substrate hippuryl-glycyl-glycine by ACE and subsequent reaction with trinitrobenzenesulfonic acid to form 2,4,6-trinitrophenyl-glycyl-glycine, whose absorbance is determined at 415 nm in a microtitre plate reader. Rabbit lung dehydrated by acetone was employed as an enzyme source. Validation of the method showed satisfactory intra-day (CV = 7.63%) and inter-day precision (CV = 13.61%), recovery (97-102.1%), sensitivity (IC50 = 14.1 nmol/l) and linearity in the range 7.5-120 mmol/l of glycyl-glycine (r2 = 0.9921). Besides, the method showed good correlation with a HPLC assay already established for the screening of ACE inhibitors (r = 0.9935 and 0.9034, respectively, for captopril solutions and for plant extracts). The method involves only inexpensive reagents and apparatus.

Publication types

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

MeSH terms

  • Angiotensin-Converting Enzyme Inhibitors / chemistry*
  • Animals
  • Brazil
  • Colorimetry
  • Dose-Response Relationship, Drug
  • Humans
  • Medicine, Traditional
  • Oligopeptides / chemistry
  • Phytotherapy*
  • Plant Extracts / chemistry*
  • Plants, Medicinal*
  • Rabbits
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Trinitrobenzenesulfonic Acid / chemistry

Substances

  • Angiotensin-Converting Enzyme Inhibitors
  • Oligopeptides
  • Plant Extracts
  • hippuryl-glycyl-glycine
  • Trinitrobenzenesulfonic Acid