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Development of an assay to determine the antioxidant capacity of natural products by chemiluminescence of luminol

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Author(s):
Erick Leite Bastos
Total Authors: 1
Document type: Master's Dissertation
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ)
Defense date:
Examining board members:
Josef Wilhelm Baader; Ana Campa; Paulo Roberto Hrihorowitsch Moreno
Advisor: Josef Wilhelm Baader
Abstract

In the last decade the study of active oxygen and nitrogen species and their role in a large number of chronicle diseases, including cancer, heart disease and even aging itself, revealed that natural and synthetic antioxidants are able to prevent the effects caused by oxidative stress. Several methods can be used to evaluate the total antioxidant activity in body fluids, complex mixtures and isolated substances. Simple trapping assays can quantify the total antioxidant content in a sample, which is expressed as TRAP (total radical-trapping potential) or TEAC (trolox equivalent antioxidant capacity) and these indexes are well accepted due to its high sensitivity and operational facilities. The determination of the antioxidant potential of plant extracts and isolated natural products may constitute a simple tool to evaluate the potential biological activity of plant constituents. The chemiluminescence reaction of luminol and hydrogen peroxide in the presence of hemin as catalyst has been used as the method to evaluate the antioxidant activity, since the chemiluminescence emission can be suppressed by antioxidants, and a linear relationship between antioxidant concentration and the observed induction time is obtained. A kinetic study was performed to establish the ideal experimental conditions for the assay. The reactant concentrations were varied in the absence and the presence of antioxidants, and the results lead to a better understanding of the system. A new method for data treatment is also used, which allows the correlation of the antioxidant effect to the number of photons suppressed. Several well-known antioxidants (trolox, ascorbic and uric acid) were used to establish the methodology. TRAP values were calculated from the correlations between the number of photons suppressed and the antioxidant concentration, using trolox as reference compound. Using this methodology we were able to determine the antioxidant activity of Photomorphe umbellata extracts, and of its isolated major compound, 4-nerolidylcatechol. (AU)