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Master's Dissertation
DOI
https://doi.org/10.11606/D.46.2000.tde-23012015-143743
Document
Author
Full name
Erick Leite Bastos
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2000
Supervisor
Committee
Baader, Josef Wilhelm (President)
Campa, Ana
Moreno, Paulo Roberto Hrihorowitsch
Title in Portuguese
Desenvolvimento de um ensaio para determinação da capacidade antioxidante de produtos naturais através da quimiluminescência do luminol
Keywords in Portuguese
Biofísica
Produtos naturais
Abstract in Portuguese
Na última década, o estudo de espécies ativas de oxigênio e nitrogênio e o seus papeis em um grande número de patologias revelou que substâncias antioxidantes são capazes de prevenir os efeitos do estresse oxidativo. A reação quimiluminescente de luminol e peróxido de hidrogênio, na presença de hemina como catalisador, tem sido utilizada como método de avaliação de atividade antioxidante, uma vez que a emissão pode ser suprimida por este tipo de substância. Foi realizado um estudo cinético para estabelecer as condições experimentais do ensaio. As concentrações dos reagentes foram variadas em diferentes condições experimentais na ausência e na presença de antioxidantes, e os resultados obtidos levaram ao melhor entendimento acerca do sistema. Um novo método para o tratamento de dados foi também utilizado, permitindo a correlação entre o efeito antioxidante e o número de fótons suprimidos. Vários antioxidantes conhecidos (trolox, ácido ascórbico e ácido úrico) foram utilizados para estabelecer a metodologia. A atividade antioxidante foi calculada a partir da correlação entre a área de supressão obtida e a concentração de antioxidante, usando trolox como composto de referência. Empregando esta metodologia foi possível determinar a atividade antioxidante dos extratos e do produto majoritário isolado de Photomorphe umbellata, o 4-nerolidicatecol.
Title in English
Development of an assay to determine the antioxidant capacity of natural products by chemiluminescence of luminol
Keywords in English
Biophysics
Natural products
Abstract in English
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.
 
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Publishing Date
2015-01-23
 
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