Validation of a colorimetric assay for the in vitro screening of inhibitors of angiotensin-converting enzyme (ACE) from plant extracts.
Anew 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-tri...
| Autores: | , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2005 |
| País: | Brasil |
| Institución: | Universidade Federal de Ouro Preto (UFOP) |
| Repositorio: | Repositório Institucional da UFOP |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.ufop.br:123456789/1215 |
| Acceso en línea: | http://www.repositorio.ufop.br/handle/123456789/1215 |
| Access Level: | acceso abierto |
| Palabra clave: | In vitro colorimetric assay Plant extracts Screening Angiotensin-converting enzyme inhibitors |
| Sumario: | Anew 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 415nm 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 ¼ 7.63%) and inter-day precision (CV ¼ 13.61%), recovery (97–102.1%), sensitivity (IC50 ¼ 14.1 nmol/l) and linearity in the range 7.5–120 mmol/l of glycyl-glycine (r2 ¼ 0:9921). Besides, the method showed good correlation with a HPLC assay already established for the screening of ACE inhibitors (r ¼ 0:9935 and 0:9034; respectively, for captopril solutions and for plant extracts). The method involves only inexpensive reagents and apparatus. |
|---|