This study evaluated the antioxidant activity and phenolic content of red propolis samples from the state of Sergipe, Brazil. All propolis samples showed antioxidant activity in DPPH radical scavenging tests, which was confirmed by more sensitive lipid peroxidation and chemiluminescence assays. Lipid peroxidation inhibition ranged from 48.08% to 93.37%, higher than previous studies. Extracts with the highest antioxidant activity also had the highest total phenolic content, though not the highest flavonoid levels. The presence of flavonoids catechin, epicatechin and formononetin was confirmed in all samples by UFLC.
การวิเคราะห์ฤทธิ์ต้านอนุมูลอิสระที่ระยะต่างกันในกล้วยเล็บมือนาง
Analysis of Antioxidant Activity at Different Stage in Musa (AA group) ‘Kluai Leb Mu Nang’
อดิศร จำรูญ
Evaluation of antioxidant and antiradical properties of pomegranate (punica g...Pritam Kolge
Evaluation of antioxidant and antiradical properties
of Pomegranate (Punica granatum L.) seed and defatted
seed extracts
This is Journal Club activity Presentation with the reference of various research papers.
This Presentation Contain following...
#Info about Paper
#Abstract
#Materials
#Methods
#Results
#Discussion
#Conclusion
#References
*Important Methods used
#Moisture content
#Fat content
#Acid value
#Peroxide value
#Oxidative stability index
#Total phenols content
#Preparation of Pomegranate seed extracts and calculate extract yield
#Evaluation of antioxidant properties of Pomegranate seed extracts using
-DPPH radicals scavenging activity
-FRAP assay
#Antioxidant efficiency of seed extract (Oxidative stability extract)
#Statistical analysis
Journal Club Presentation at Bharati Vidyapeeth College of Pharmacy, Kolhapur.
Thanks for Help and Guidance of Dr. P. B. Choudhari (Assistant Professor, Pharmaceutical Chemistry) and Dr. A. J. Shinde (Assistant Professor, Department of Pharmaceutics)
การวิเคราะห์ฤทธิ์ต้านอนุมูลอิสระที่ระยะต่างกันในกล้วยเล็บมือนาง
Analysis of Antioxidant Activity at Different Stage in Musa (AA group) ‘Kluai Leb Mu Nang’
อดิศร จำรูญ
Evaluation of antioxidant and antiradical properties of pomegranate (punica g...Pritam Kolge
Evaluation of antioxidant and antiradical properties
of Pomegranate (Punica granatum L.) seed and defatted
seed extracts
This is Journal Club activity Presentation with the reference of various research papers.
This Presentation Contain following...
#Info about Paper
#Abstract
#Materials
#Methods
#Results
#Discussion
#Conclusion
#References
*Important Methods used
#Moisture content
#Fat content
#Acid value
#Peroxide value
#Oxidative stability index
#Total phenols content
#Preparation of Pomegranate seed extracts and calculate extract yield
#Evaluation of antioxidant properties of Pomegranate seed extracts using
-DPPH radicals scavenging activity
-FRAP assay
#Antioxidant efficiency of seed extract (Oxidative stability extract)
#Statistical analysis
Journal Club Presentation at Bharati Vidyapeeth College of Pharmacy, Kolhapur.
Thanks for Help and Guidance of Dr. P. B. Choudhari (Assistant Professor, Pharmaceutical Chemistry) and Dr. A. J. Shinde (Assistant Professor, Department of Pharmaceutics)
Evaluation of antioxidant properties of pomegranate peel extract in compariso...Pritam Kolge
Evaluation of antioxidant properties of pomegranate peel extract in comparison with pomegranate pulp extract.....
This is Journal Club activity Presentation with the reference of various research papers.
This Presentation Contain following...
#Info about Paper
#Highlights
#Introduction
#Materials
#Methods
#Results
#Discussion
#Conclusion
#References
Important Methods used
1)Antioxidants extraction
2)Determination of total phenolics content
3)FRAP Assay
4)Determination of content of phenolic compounds in the
extract
5)Superoxide radical scavenging activity (DPPH)
6)Hydroxyl radical (OH) prevention activity
Journal Club Presentation at Bharati Vidyapeeth College of Pharmacy, Kolhapur.
Thanks for Help and Guidance of Dr. P. B. Choudhari (Assistant Professor, Pharmaceutical Chemistry) and Mr. D. P. Mali Assistant Professor, Pharmaceutical Chemistry
Various human diseases have oxidative stress as one of their component. Many herbs have been reported to exhibit properties that combat oxidative stress through their active constituents such as flavonoids, tannins, phenolic compounds etc. Different Plants of Dillenicea family has been shown in in vitro experiments to be endowed with antioxidant activity. Therefore this study was carried out to evaluate Dillenicea family for its antioxidant activity.
Proximate analysis, antimicrobial and antioxidant activities of aloe vera.Kojo Ashiadey
It's a project describing the nutritional composition of aloe vera barbadensis, its antimicrobial properties on selected microbes and the ability of aloe vera to scavenge free radicals.
DETERMINATION OF MIANSERIN USING TROPAEOLIN-OOO BY ION PAIR FORMATIONRatnakaram Venkata Nadh
Objective: The present study was aimed at the development of a simple visible spectrophotometric method for the assay of mianserin, a drug used for the treatment of depression.
Methods: The method was developed using tropaeolin-ooo (TPooo) as an ion associative complex forming a chromophore. Developed the chromophore by sequential mixing of aqueous solutions of mianserin, hydrochloric acid, and TPooo. Chromophore was extracted into an organic solvent (chloroform) and absorbance values of organic layers were measured. As per the existing guidelines of an international conference on harmonization (ICH), various parameters of the method were tested for validation.
Results: At the optimized reaction conditions, the formed chromophore (λ max
Conclusion: Due to lack of pre-treatment process for this method, it was simple. All the tested parameters of the method were validated as per ICH guidelines.
Isolation and Characterization of Colchicine from Physostigma VenenosiumIOSR Journals
A unique anti-inflammatory drug Colchicine has been isolated from a Nigeria medicinal plant called
Physostigma venenosium The structure was established using NMR spectroscopy of 1H and 13C, Cosy, and
DEPT In combination with IR. The ethanol extract of the plant seed was partitioned to get the chloroform,
water, methanol and pet-either fractions. The chloroform fraction, from which colchicines was isolated was
discovered to contain the highest number of compounds with wide range of Rf values on TLC result.. The result
supports the use of the plant seeds for the treatment of phantom tumor and conjunctival inflammation.
Phyto pharmaceutical - TOCOPHEROLS AND TOCOTRIENOLS (Vitamin E )SudhindraKini
Vitamin E (tocopherol) is a naturally occurring antioxidant. Biochemical functions of vitamin E. applications of vitamin E. symptoms of vitamin E deficiency. Global scenario of production and consumption of natural vitamin E and mixed tocopherols
Determination of Chemical Groups and Investigation of Anthelmintic, Cytotoxic...Syed Masudur Rahman Dewan
The present study was conducted for the characterization of possible chemical groups,
evaluation of anthelmintic, cytotoxic and antibacterial activities of crude methanolic extract
of leaves of Cinnamomum tamala. The study revealed the presence of alkaloids, reducing
sugar, tannin, amino acids, glycosides and steroid in the crude extract. The extract showed
very potent anthelmintic activity while compared with the standard albendazole. To
investigate the cytotoxic activity, brine shrimp lethality bioassay was conducted, and the
extract showed significant activity while compared with the standard vincristine sulphate
(LC50 value 1.007 and 0.839μg/ml respectively). To evaluate the antibacterial activity, disc
diffusion method was followed, and the extract showed activity against Bacillus subtilis,
Staphylococcus aureus, Bacillus cereus, and Vibrio cholera, and resistant to Escherichia coli
and Salmonella typhi.
LABORATORY MANUAL ON
QUALITY CONTROL OF ANIMAL FEEDS by Dr. G. DEVEGOWDA, PROFESSOR & HEAD, DEPARTMENT OF POULTRY SCIENCE
UNIVERSITY OF AGRI. SCIENCES
HEBBAL, BANGALORE
Evaluation of antioxidant properties of pomegranate peel extract in compariso...Pritam Kolge
Evaluation of antioxidant properties of pomegranate peel extract in comparison with pomegranate pulp extract.....
This is Journal Club activity Presentation with the reference of various research papers.
This Presentation Contain following...
#Info about Paper
#Highlights
#Introduction
#Materials
#Methods
#Results
#Discussion
#Conclusion
#References
Important Methods used
1)Antioxidants extraction
2)Determination of total phenolics content
3)FRAP Assay
4)Determination of content of phenolic compounds in the
extract
5)Superoxide radical scavenging activity (DPPH)
6)Hydroxyl radical (OH) prevention activity
Journal Club Presentation at Bharati Vidyapeeth College of Pharmacy, Kolhapur.
Thanks for Help and Guidance of Dr. P. B. Choudhari (Assistant Professor, Pharmaceutical Chemistry) and Mr. D. P. Mali Assistant Professor, Pharmaceutical Chemistry
Various human diseases have oxidative stress as one of their component. Many herbs have been reported to exhibit properties that combat oxidative stress through their active constituents such as flavonoids, tannins, phenolic compounds etc. Different Plants of Dillenicea family has been shown in in vitro experiments to be endowed with antioxidant activity. Therefore this study was carried out to evaluate Dillenicea family for its antioxidant activity.
Proximate analysis, antimicrobial and antioxidant activities of aloe vera.Kojo Ashiadey
It's a project describing the nutritional composition of aloe vera barbadensis, its antimicrobial properties on selected microbes and the ability of aloe vera to scavenge free radicals.
DETERMINATION OF MIANSERIN USING TROPAEOLIN-OOO BY ION PAIR FORMATIONRatnakaram Venkata Nadh
Objective: The present study was aimed at the development of a simple visible spectrophotometric method for the assay of mianserin, a drug used for the treatment of depression.
Methods: The method was developed using tropaeolin-ooo (TPooo) as an ion associative complex forming a chromophore. Developed the chromophore by sequential mixing of aqueous solutions of mianserin, hydrochloric acid, and TPooo. Chromophore was extracted into an organic solvent (chloroform) and absorbance values of organic layers were measured. As per the existing guidelines of an international conference on harmonization (ICH), various parameters of the method were tested for validation.
Results: At the optimized reaction conditions, the formed chromophore (λ max
Conclusion: Due to lack of pre-treatment process for this method, it was simple. All the tested parameters of the method were validated as per ICH guidelines.
Isolation and Characterization of Colchicine from Physostigma VenenosiumIOSR Journals
A unique anti-inflammatory drug Colchicine has been isolated from a Nigeria medicinal plant called
Physostigma venenosium The structure was established using NMR spectroscopy of 1H and 13C, Cosy, and
DEPT In combination with IR. The ethanol extract of the plant seed was partitioned to get the chloroform,
water, methanol and pet-either fractions. The chloroform fraction, from which colchicines was isolated was
discovered to contain the highest number of compounds with wide range of Rf values on TLC result.. The result
supports the use of the plant seeds for the treatment of phantom tumor and conjunctival inflammation.
Phyto pharmaceutical - TOCOPHEROLS AND TOCOTRIENOLS (Vitamin E )SudhindraKini
Vitamin E (tocopherol) is a naturally occurring antioxidant. Biochemical functions of vitamin E. applications of vitamin E. symptoms of vitamin E deficiency. Global scenario of production and consumption of natural vitamin E and mixed tocopherols
Determination of Chemical Groups and Investigation of Anthelmintic, Cytotoxic...Syed Masudur Rahman Dewan
The present study was conducted for the characterization of possible chemical groups,
evaluation of anthelmintic, cytotoxic and antibacterial activities of crude methanolic extract
of leaves of Cinnamomum tamala. The study revealed the presence of alkaloids, reducing
sugar, tannin, amino acids, glycosides and steroid in the crude extract. The extract showed
very potent anthelmintic activity while compared with the standard albendazole. To
investigate the cytotoxic activity, brine shrimp lethality bioassay was conducted, and the
extract showed significant activity while compared with the standard vincristine sulphate
(LC50 value 1.007 and 0.839μg/ml respectively). To evaluate the antibacterial activity, disc
diffusion method was followed, and the extract showed activity against Bacillus subtilis,
Staphylococcus aureus, Bacillus cereus, and Vibrio cholera, and resistant to Escherichia coli
and Salmonella typhi.
LABORATORY MANUAL ON
QUALITY CONTROL OF ANIMAL FEEDS by Dr. G. DEVEGOWDA, PROFESSOR & HEAD, DEPARTMENT OF POULTRY SCIENCE
UNIVERSITY OF AGRI. SCIENCES
HEBBAL, BANGALORE
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay
Evaluation of Anti-oxidant Activity of Elytraria acaulis Aerial ExtractsIJERA Editor
Elytraria acaulis, a stem less perennial herb of Acantheceae family has many medicinal and therapeutic properties. Anti oxidative activity of the aerial parts of this Elytraria acaulis were assessed in the present study. The aerial parts of the plant (Stem & Leaves) were extracted in different organic solvents such as n-Hexane, Ethanol, Methanol, Ethyl Acetate and Chloroform. Initially, Total Phenolic & Total Flavonoids content in different solvent plant extracts were estimated. The free radical scavenging and antioxidant activity of the Elytraria acaulis aerial extracts in different organic solvents were also assayed by DPPH assay, FRAP assay. The aerial extracts of Elytraria acaulis have shown significant anti oxidant activity. Hence, further studies on this plant will enable elucidation of its therapeutic properties and medicinal applications
Adsorption of Phenol from Aqueous Solution using Algal BiocharSagar Sonkar
Although the food and beverage industries are not as polluting as some other sectors like metal or leather industries, but they have been responsible for air, water and soil pollution by emitting dust and unpleasant odor in the air.
If the effluents from the food and beverage industry are contaminated with toxic metals, these can affect adversely on human health.
Phenolic compounds which are present in various concentrations in several of these waste streams cause toxic effects and are reported as Cancer causing and may also cause long-term ecological damage.
Microalgae have been reported to accumulate pollutants such as heavy metals, hexachlorobenzene, herbicides, insecticides and even Phenol.
Spirulina Platensis, that was commonly used as nutritional supplements, could be easily cultured, and the species were shown to thrive in municipal and agricultural wastewater effluents for removal of contaminants by production of biochar.
The most common method for the removal of this dissolved organic material is the adsorption with activated carbon, a product that is produced from a variety of carbonaceous materials and biochar is one of it.
Phytochemical screening and antioxidant activity of clove mistletoe leaf extr...iosrphr_editor
Clove mistletoe (Dendrophthoe pentandra (L.) Miq) is one of the semi-parasitic plants belonging to the Loranthaceae family. Clove mistletoe leaf extracts have many biological activities such as antibacterial, antioxidant and antidiabetes. The purpose of this study was to determine the content of secondary metabolites in clove mistletoe leaf extracts through phytochemical screening and determine its antioxidant activity through DPPH free radical scavenging. Samples were tested include water and ethanol 70 % extracts, as well as n-hexane, ethyl acetate and ethanol fractions. Phytochemical screening showed that all samples containing tannins and flavonoids but no alkaloids. The highest total phenol contents was ethyl acetate fraction namely 358.4 mg GAE/ g. The best antioxidant activity was water extract, ethanol 70 % extract and ethyl acetate fraction. Therefore, clove mistletoe leaf extracts are potential source for antioxidant.
Phytochemical screening and antioxidant activity of clove mistletoe leaf extr...iosrphr_editor
Clove mistletoe (Dendrophthoe pentandra (L.) Miq) is one of the semi-parasitic plants belonging to the Loranthaceae family. Clove mistletoe leaf extracts have many biological activities such as antibacterial, antioxidant and antidiabetes. The purpose of this study was to determine the content of secondary metabolites in clove mistletoe leaf extracts through phytochemical screening and determine its antioxidant activity through DPPH free radical scavenging. Samples were tested include water and ethanol 70 % extracts, as well as n-hexane, ethyl acetate and ethanol fractions. Phytochemical screening showed that all samples containing tannins and flavonoids but no alkaloids. The highest total phenol contents was ethyl acetate fraction namely 358.4 mg GAE/ g. The best antioxidant activity was water extract, ethanol 70 % extract and ethyl acetate fraction. Therefore, clove mistletoe leaf extracts are potential source for antioxidant.
Phytochemical composition and antiradical activity of Sakersia africana Hook....Open Access Research Paper
The valorization of the medicinal plants of our country and determination of their impact on health due to their abundance of substances with various pharmacological effects are our principal objective. This study was evaluated the phytochemical screening and radical 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging activity of different extracts of Sakersia africana Hook. f.. The results revealed that Sakersia africana Hook. f. is rich in phenols compounds, sterols, triterpenes, alkaloids and reducing compound. The values in total phenols and proanthocyanidines are ranging respectively from 391.58 ± 0.04 to 777 ± 0.03 mg/100 g of drugs and 113.5 ± 3.17 to 653.5 ± 36.83 mg/100 g of drugs. Results also show that different extracts tested present antiradical activity with values of IC50 ranging from 164.21± 0.014 to 195.54± 0.012 % and abundance in bioactive compounds. This study could justify the use of Sakersia africana of some chronic diseases.
The Role of Cell Wall-Degrading Enzymes in the Development of Anthracnose Dis...Agriculture Journal IJOEAR
— The ability of Colletotrichumtruncatum CP2 in producing pectinolytic and cellulolytic enzymes was evaluated by shake flask fermentations. The results of enzymatic activity experiment indicated that PG was the first cell wall-degrading enzymes detected and the activities obtained were higher (0.24±0.10 U/mL) than other enzymes, which appeared later and in lower amount. After the cell wall was degraded by the action of PG, further degradation of the cell wall was affected by pectin methylesterases, pectin lyase, pectate lyase and cellulases. The disparity in enzymatic activity at different intervals may suggest their specific role for pathogenesis at proper timings.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
1. REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228
Volume 16 - Número 2 - 2º Semestre 2016
ATIVIDADE ANTIOXIDANTE E COMPOSTOS FENÓLICOS DE PRÓPOLIS
VERMELHA DO ESTADO DE SERGIPE, BRASIL.
Yzila Liziane Farias Maia de Araújo1
; Lucyana Santos de Mendonça1
; Nayjara Carvalho Gualberto2
;
Sara Cuadros Orellana3
; Juliana Cordeiro Cardoso4
; Sona Jain5
; Edilson Divino de Araújo1
; Narendra Narain2
RESUMO
A própolis é uma resina natural extraída das plantas e modificada pelas abelhas. Este estudo teve
como objetivo avaliar a atividade antioxidante da própolis vermelha brasileira por DPPH,
peroxidação lipídica e métodos de quimiluminescência, juntamente com a análise de seu conteúdo
fenólico por calorimetria e cromatografia líquida ultrarrápida. Todas as amostras de própolis
analisadas neste estudo mostraram atividade antioxidante por DPPH ∙ teste que foi confirmado por
peroxidação lipídica e quimiluminescência métodos mais sensíveis. A peroxidação lipídica mostrou
inibição na gama de 48,08 % a 93,37 %, valores melhores do que os citados em trabalhos
anteriores. Os extratos com maior atividade antioxidante também mostram o melhor conteúdo de
fenólicos totais, no entanto, não apresentaram melhores níveis de flavonoides. Presença de
flavonoides (catequina, epicatequina e formononetina) foi confirmado em todas as amostras
analisadas.
Palavras-chave: Resina, Peroxidação lipídica, Quimiluminescência, Flavonoides.
ANTIOXIDANT ACTIVITY AND PHENOLIC CONTENT OF THE RED PROPOLIS
FROM THE STATE OF SERGIPE, BRAZIL.
ABSTRACT
Propolis is a natural resin extracted from the plants and modified by the honeybees. This study
aimed to evaluate the antioxidant activity of Brazilian red propolis by DPPH, lipid peroxidation and
chemiluminescence methods, together with the analysis of its phenolic content by calorimetry and
ultra fast liquid chromatography. All the propolis samples analyzed in this study showed
antioxidant activity by DPPH∙ test which was confirmed by more sensitive lipid peroxidation and
chemiluminescence methods. Lipid peroxidation showed inhibition in the range of 48.08% to
93.37%, values better than those cited in previous report. The extracts with the highest antioxidant
activity also showed the best total phenolic content, however, did not show better levels of
flavonoids. Presence of flavonoids (catechin, epicatechin and formononetin) was confirmed in all
the analyzed samples.
Keywords: Bee glue, Lipid peroxidation, Chemiluminescence, Flavonoids.
73
2. 1. Introduction
Propolis is a complex mixture of resinous
substances collected by the bees from various
plant parts (LOTTI et al., 2010). The resin
collected by the bees contains a variety of
chemical compounds such as flavonoid
aglycones, phenolic acids and their esters,
phenolic aldehydes, alcohols and ketones,
sesquiterpene, quinones, coumarins, steroids,
amino acids and inorganic compounds
(PIERINI et al., 2013) which have diverse
biological properties, including the anti-
inflammatory, antimicrobial, antioxidant,
antitumor, antiulcer and anti-HIV activities
(HUANG et al., 2014).
Brazilian propolis was divided into 12
types by Park et al., (2000). In 2007, a new type
of propolis called the red propolis was found
along the sea and rivers of the Northeastern
Brazil and was classified as propolis belonging
to group 13 (DAUGSCH et al., 2007). Red
propolis of Sergipe has been reported to have
antitumoral and immunomodulatory properties
(FROZZA et al., 2013) and wound healing
activities (ALBUQUERQUE JUNIOR et al.,
2009).
Studies with the Brazilian red propolis
have shown a great potential for this product
including its use in the prevention and treatment
of cardiovascular diseases such as
atherosclerosis (ALBUQUERQUE JUNIOR et
al., 2009; IIO et al., 2012).
Several methods can be utilized for the
extraction of natural chemical compounds from
propolis. One such method includes the use of
ultrasound. The use of ultrasound in the
extraction processes has drawn increasing
attention due to its higher efficiency and
reduced extraction time, compared to
conventional extraction methods, such as in
Soxhlet and maceration (LUQUE-GARCÍA;
CASTRO, 2003). Using this method, the
antioxidant activity and chemical composition
of the red propolis samples collected from the
Northeast Brazil was determined, which is
detailed in this manuscript.
2. Materials and Methods
2.1. Propolis
The red propolis samples were collected
from two different apiaries located in the lower
Sao Francisco region, Sergipe, Brazil. Five
samples were collected from the apiary CJ (S
10°26'04'', W 36°30'34'') and four samples from
the apiary CP (S 10°28'25'', W 36°26'12”). The
samples were stored at -20°C.
2.2. Chemicals
All the commercially available chemicals
such as 1,1-diphenyl-2-picrylhydrazylradical
(DPPH), thiobarbituric acid (TBA), luminol and
xanthine, xanthine oxidase (XOD) were
purchased from Sigma-Aldrich
2.3. Preparation of hydroalcoholicic
extracts (HAE) and Yied determination
1g of propolis sample was extracted with
12.5 ml of 70% (v/v) ethanol using ultrasound
for 60 min and then the mixture was centrifuged
and filtered to obtain its hydroalcoholic extract
(HAE). The dry weight was obtained from
respective hydroalcoholic extracts and used for
yield determination. All the tests were carried
out using 5% HAE.
2.4. Total polyphenol and flavonoid
contents
The total polyphenols were determined
using the Folin-Ciocalteau method (LEE et al.,
2003; FUNARI; FERRO, 2006), and expressed
as gallic acid equivalents in mg phenol/g of
propolis.
The flavonoid content was determined
using the colorimetric method of aluminum,
using 1.0 mL of the 1:10 diluted sample, 3.0 mL
95% ethanol, 0.5 ml of 10% aluminum nitrate,
0.2 mL potassium acetate and 5.6 mL of
distilled water. The blank was prepared with 1.0
mL diluted sample, 3.0 mL 95% ethanol, 0.2
mL of 1 M potassium acetate and 5.8 mL of
distilled water. After 20 minutes of incubation
at RT, the absorbance was measured at 415 nm.
Results were expressed as mg of flavonoids / g
of propolis using quercetin as a reference.
2.5. Ultra Fast Liquid Chromatography –
UFLC
Chromatographic analysis was carried out
using ultra fast liquid chromatography – UFLC
(Shimadzu Co.), equipped with a quat pump, a
degasser, a diode array detector (DAD) and a
reversed phase column (XP-ODS 50 x 3 mm;
particle size, 2.2 micrometers).
3. The binary mobile phase was composed of
water (solvent A) and methanol (solvent B)
according to the method described by Alencar et
al., (2007) and Cabral et al., (2009) with
modifications. The HAE was dissolved in
methanol (50 mg /mL) and filtered with a 0.45
μm filter (Millipore). Aliquots of 2 μL of 5% of
HAE (w/v) were injected into the UFLC system.
UFLC was performed using a linear gradient
elution as follows: 40% B increased to 60% B
after 22.5 min, a hold at 90% B for 37.3- 42.3
min, followed by a decrease to 30% B after 42.3
min with a solvent flow rate of 0.4 mL/min. The
following authentic standards of flavonoids and
phenolic acids were used: formononetin,
quercetin, 3-hydroxy-7-methoxyflavone,
catechin, epicatechin and propyl gallate. All the
tests were performed in triplicates using three
samples from each apiary
2.6. DPPH radical scavenging activity
The effect of DPPH radical was evaluated
by the method described by Brand-Williams et
al., 1995 and Tominaga et al., 2005 with
adaptations. The assay mixture contained 1.0
mL of 100mM sodium acetate buffer, 50 μL of
HAE, 1.0 ml of 95% ethanol, and 500 μL of 200
mM DPPH solution. Absorbance was recorded
at 517 nm after 10 min of incubation at room
temperature, in the dark. Blank containing the
same amount of ethanol and DPPH solution was
used as the negative control. The percentage
inhibition was plotted against phenol content
and IC50 (concentration of total phenol able to
scavenge 50% of DPPH free radical) was
determined.
2.7. Lipid peroxidation assay
Reaction mixture was prepared by adding
10 µL of each sample to 1 mL of a reaction
medium containing 130 mM KCl, and 10 mM
TrisHCl pH7.4, and mitochondria was added to
yield a final concentration of 1 mg of protein.
Later 50 µL ferrous ammonium sulfate and
2mM sodium citrate were added and the
samples were incubated at 37°C for 30 minutes.
Mitochondria was isolated from male Wistar
rats livers by differential centrifugation as
described by Pedersen et al., 1978, and the
mitochondrial protein content was determined
by the biuret reaction (CAIN; SKILLETER,
1987). For the determination of TBA-reactive
compound, 1 mL of 1% thiobarbituric acid
(TBA), 0.1 mL of 10M NaOH and 0.5 mL of
H3PO4 were added, followed by incubation in a
water bath for 20 minutes at 80°C. The TBA-
reactive compounds were extracted with 2 mL
of n-butanol and the samples were then
centrifuged at 9800 g for 10 min. The readings
were taken at 535 nm, using a Hitachi U2001
spectrophotometer. The experiments were
performed in triplicates together with blank
(without mitochondria), positive control
(without sample) and negative control (without
iron). The IC50 values were determined using
the GraphPad Prism ®.
2.8. Xanthine/luminol/XOD system
Samples of red propolis extract were
diluted in hydro alcoholic solution to a
concentration of 5 mg/mL and subsequent
dilutions were prepared in 0.1 M glycine buffer
pH 9.4. To verify free radical scavenging
activity of the propolis extract, a solution
containing 400 μL of 1 mM EDTA and 0.1 M
glycine pH 9.4 were added to a test tube
together with 150 μL of 6 mM xanthine, 10 μL
of test sample and 10 μL of 0.6 mM luminol
solution. The reaction was initiated with the
addition of 100 μL of freshly prepared cold 20
mU/mL xanthine oxidase solution. The
scavenging activity was determined after an
incubation of 5 minutes at 25 °C in an
Autolumat LB 953 luminometer. The results
were expressed according to the concentration
of propolis extract in the reaction medium
(MARQUELE-OLIVEIRA et al., 2005). The
IC50 values were determined using GraphPad
Prism® software.
2.9. Statistics
All the measurements were performed in
triplicates. The average was determined with
Assistat 7.7 Beta (registre INPI 0004051-2), and
used for the analysis of variance (ANOVA) and
Tukey test for means comparison, when
necessary.
3. Results and Discussion
The hydroalcoholic extracts of all the
propolis samples presented yield ranging from
9.45% to 74.62% (w/w) with the exception of
two samples that showed lower yield than the
value determined by Ministry of Agriculture
4. (less than 11%), as shown in Table 1 (BRASIL, 2001).
Table 1. Yield, flavonoid content (mg quercetina/g própolis), total phenols (mg fenol/g propolis), DPPH∙
and lipid
peroxidation of the hydroalcoholic extracts of red propolis. Same letters in the same column represent statistically
identical values for the Tukey test (p<0,05)
Funari et al., (2007) obtained a yield of
38.34% (w/w) using cold ethanol extracts of
propolis from the state of São Paulo, while
work done by Longhine et al., (2007) using
ethanol extracts of propolis from Paraná using
turboextractor showed an yield of 3.68% to
14.90% (w / w). Alencar et al., (2007) acquired
a yield of 58.20% (w / w) with ethanol extracts
of propolis using water bath at 70 ° C from the
state of Alagoas.
The propolis has different characteristics
according to their geographical location, and its
chemical composition is related to the climate,
the genetic characteristics of bees, the
methodology for conducting the tests and the
time of year that propolis was produced
(LASKAR et al.,2010; MIGUEL et al., 2010).
However, the results of this study suggest that
the forging activity of the honey bees can differ
between the hives of the same apiary This
suggests that the genetic characteristics of the
queen and foraging habits of the workers may
be related to this issue, facts also commented
by Silva e Azevedo, (2006). Propolis samples
from different boxes of the same apairy differed
in their yield, confirming the fact that the choice
of the plant to collect the resin varies between
the bees of the same apiary. This variable is
therefore important in determining the final
yield of propolis samples. Yield can also be
related to the type and volume of extracting
solvent, the method and the extraction time
(GRIGORIS et al., 2005).
3.1. Total polyphenol, flavonoid, and free
radical scavenging activity of DPPH ▪
The amount of total phenols in the
propolis samples analyzed in this study ranged
from 1.35% to 2.27% exceeding the minimum
limit of 0.50%. 70% of propolis samples
analyzed met the minimum value of 0.25%
(w/w) of flavonoid (Table 1) required by the
Ministry of Agriculture (BRASIL, 2001).
The content of phenolic compounds
ranged from 13.52 to 22.78 mg phenol/g
propolis, values comparable to those cited in the
literature (CASTRO et al., 2007; DAUGSCH et
al., 2008). The values expressed in mg phenol
equivalent to gallic acid/g propolis acid can be
seen in Table 1.
Despite the importance of classifying the
content of polyphenols and flavonoids in
propolis samples, it is important to note that in
natural extracts, which are a mixture of different
polyphenols and flavonoids, antioxidant activity
can not be directly related to these chemical
compounds, as two different polyphenols can
interact with the Folin-Ciocalteau forming
products with similar absorbance, but with very
different antioxidant activity (MARQUELE-
OLIVEIRA et al., 2008).
The flavonoid content (mg propolis/g
quercetin) of propolis samples analyzed in this
study presented values well below the values
reported in the literature with other types of
Brazilian propolis. The colorimetric method
utilized in this study can also be an interfering
agent for the evaluation of flavonoids in these
samples. Other analytical methods may exhibit
Extracts
Yield
(%m/m)
Flavonoids
(mg/g)
Total phenol
(mg/g)
DPPH▪
IC50 (μg/mL)
Lipid
peroxidation (%)
CP 03 74.6 ± 0.00 g
0.00 ± 0.00 d
13.52 ± 0.28d
324.3 ± 17.8 e
60.77 ± 0.43 e
CP 14 11.7 ± 0.00 g
0.03 ± 0.01 c
22.70 ± 0.17a
127.2 ± 7.90 c,f
91.74 ± 0.37 a
CP 18 11.8 ± 0.00 d
0.04 ± 0.01 b,c
21.91 ± 0.28a
133.3 ± 0.70 f
69.53 ± 2.55 d
CP 19 20.4 ± 0.00 b
0.03 ± 0.01 c
21.54 ± 0.46b
194.3 ± 9.00 d
92.63 ± 0.51 a
CP 29 15.5 ± 0.00 cd
0.00 ± 0.00 d
22.78 ± 0.28a
138.4 ± 0.00 f
93.37 ± 0.75 a
CJ 04 9.4 ± 0.00 a
0.04 ± 0.02 b,c
17.70 ± 0.40c
405.3 ± 20.0 a
48.08 ± 0.68 g
CJ 06 9.3 ± 0.00 f
0.07 ± 0.02 a
21.06 ± 0.44b
225.3 ± 4.60 b
83.33 ± 0.77 b
CJ 09 18.6 ± 0.00 f
0.00 ± 0.00 d
20.92 ± 0.54b
101.6 ± 1.30 c
75.37 ± 0.34 c
CJ 10 24.3 ± 0.00 c
0.05 ± 0.01 b
20.39 ± 0.57b
218.4 ± 5.20 b,d
91.15 ± 0.43 a
5. different results than those observed in this
study.
Samples that showed the best IC50 values
in DPPH test were CJ 09 (101.6 mg/mL), CP 14
(127.2 mg/mL), CP 18 (133.6 mg/mL) and CP
29 (138.4 mg/mL), and these samples also
showed higher levels of total phenols 4.12, 3.93,
2.50 and 3.60% respectively. However, these
samples did not show better levels of
flavonoids, and the sample CJ 09 that showed
the best antioxidant activity showed no
flavonoid content by colorimetric method that
was used here (Table 1). It is well established
that phenolic compounds, such as the ones
present in propolis, work as antioxidants by
breaking the chain reaction of lipids (TOREL et
al., 1986), inhibiting chemiluminescence
reactions (GEORGETTI et al., 2003),
scavenging reactive oxygen species (BORS et
al., 1990), and so forth. Nevertheless, no
correlation between phenolic and flavonoid
contents and antioxidant activity has been
confirmed. Reports published till date suggests
that the antioxidant properties arise from
complex mechanisms or synergistic interactions
between compounds (RAMANAUSKIENE et
al., 2007; MARQUELE-OLIVEIRA et al.,
2008).
Red propolis extract analyzed in this study
showed similar DPPH results to those obtained
by Pinheiro (2009) and Frozza et al., (2013)
with red propolis from the same geographic
region, with IC50 of 294μg/mL and IC50 of
270.13μg/mL, respectively.
3.2. Lipid peroxidation assay and
Xanthine/luminol/XOD system
According to Halliwell e Gutteridge
(1990), the detection and measurement of lipid
peroxidation is evidence most cited in the
literature that demonstrates the involvement of
free radical reactions in toxicology and human
diseases. Use of mitochondria as the source of
lipids may mimic human physiological
conditions. From the results of DPPH test,
concentration of 0.25 mg/mL which showed the
best antioxidant activity was chosen to for lipid
peroxidation and chemiluminescence test.
The red propolis extracts showed
inhibitory properties for lipid peroxidation by
preventing the formation of reactive species
against 2-thiobarbitric acid (TBA). The extracts
showed an inhibition of 48.08% to 93.37% at a
concentration of IC50 0.95 µg/mL, which were
better than those cited in the literature.
Argentina propolis extracts showed inhibition
from 20 to 80% (ISLA et al., 2001). According
to Jasprica et al., (2007), a decrease in the
concentration of malondialdehyde (an indicator
of lipid peroxidation) by 23.2% was observed in
volunteer men patients in Croatia after treatment
with propolis, demonstrating its antioxidant
action. Marquele-Oliveira et al., (2005) found in
their study IC50 values equivalente to 16µg/mL
and 12µg/mL for fluid and glycolic extract of
brown propolis.
As for chemiluminescence technique,
Dodeigene et al., (2000) argued that within the
techniques of antioxidant activity, this is an
advantageous method because of its high
sensitivity and speed. When compounds like
propolis are added to a chemiluminescent
solution, the light emission is reduced,
indicating antioxidant activity by removing any
radical generated in this system. The
chemiluminescence activity of the propolis
samples was analysed using one extract from
each apiary (CP29 and CJ10). Samples that
showed the best lipid peroxidation inhibition
were chosen for this assay. Luminol was used as
a detector in the chemiluminiscence test, where
it is oxidized by the superoxide anion.
The inhibition of the luminiscence by the
reduction of the superoxide was measured and
the extracts of red propolis from Sergipe were
shown to possess inhibiting activity for the
enzyme XOD, revealing IC50 of 0.096 µL/ml
(approximately 96 µg/ml) for sample CP29 and
IC50 of 0.089 µL/ml (approximately 89 µg/ml)
for sample CJ10 and inhibiting concentration
ranging between 15.66 to 89.80% as shown in
Figure1. As can bee seem in Figure 1, no
significant difference was found between
samples CP29 and CJ10. Pascual et al., (1994)
found similar values of IC50 ranging from 50 to
95 µg/mL in propolis samples from Cuba by the
method of xanthine-xanthine oxidase.
6. Figure 1. Inhibition of light emission from
xanthine/luminol/XOD luminescent reactions with
luminol found for different concentrationsof: (■) CP 29
and (♦) CJ 10. Results are mean±SD of three experiments
run in parallel.
3.3. Ultra Fast Liquid Chromatography –
UFLC
The hydroalcoholic extracts of propolis
showed a complex composition, with several
peaks at different retention times as analyzed by
UFLC-DAD. The chromatographic profiles of
the hydroalcoholic extracts of red propolis
indicated the presence of approximately 20
compounds with totally distinct profiles.
Presence of flavonoids such as catechin,
epicatechin and formononetin was confirmed as
shown in Figure 2.
Figure 2. Chromatograms of red propolis samples from two different regions of the river São Francisco
basin with three samples from apiary Cajuipe (CJ) and three from capivara (CP). 1- Catechin, 2-
Epicatechin and 3- Formononetin.
7. And the amount of catechin was
quantified to 0.26 to 0.88 mg/ml extract of
propolis. Catechins belong to a group of
polyphenols that have high antioxidant activity
and presents possible mechanisms of action
against cancer (HO et al., 1992; RODGERS et
al., 1998; PINHEIRO et al., 2014). The
flavonoid that appeared with the highest
intensity in the red propolis samples is
formononetin which is a chemical marker
present in the samples of red propolis (JAIN et
al., 2014; MENDONÇA et al., 2015). The
presence of formononetin in propolis shows
estrogenic and antifungal activity. According to
Moraes (2009), it has been shown that when
mammals consume this isoflavin it is
metabolized into daidzein, which is one of
isoflavins aglycones present in soy, and is used
in the prevention and treatment of menopausal
symptoms, and treatment of prostate cancer and
breast cancer.
4. Conclusion
Propolis extracts analyzed in this study
showed yield and total phenols comparable to
other similar reports with red propolis. IC50
values for lipid peroxidation were infact better
than previous reports suggesting higher
potential of the propolis samples from the state
of Sergipe. Presence of flavonoids such
catechin, epicatechin and formononetin known
for their medicinal properties was confirmed by
UFLC and it was possible to quantify the
catechin content.
REFERENCES
Albuquerque Junior, R. L. C., Barreto, A. L. S.,
Pires, J.A., Reis, F. P., Lima, S. O., Ribeiro,
M.A.G., Cardoso, J. C. Effect of bovine type-I
collagen-based films containing red propolis on
dermal wound healing in rodent model.
International Journal of Morphology, v. 27,
n.4, p. 1105-1110, 2009.
Alencar, S.M., Oldoni, T.L.C., Castro, M.L.,
Cabral, I.S.R., Costa-Neto, C.M., Cury, J.A.,
Rosalen, P.L., Ikegakid, M. Chemical
composition and biological activity of a new
type of Brazilian propolis: red propolis. Journal
of Ethnopharmacology, v. 113, n. 2, p. 278-
283, 2007.
Bors, W., Heller, W., Michel, C., Saran, M.
Flavonoids as antioxidants: determination of
radical-scavenging efficiencies.
Methods in Enzymology, v.186, p.343-355,
1990.
Brand-Williams, W., Cuvelier, M.E., Berset, C.
Use of a free radical method to evaluate
antioxidant activity. Food Science and
Technology, v. 28, p. 25–30, 1995.
Brasil. Ministério da Agricultura, Pecuária e do
Abastecimento. (2001). Instrução Normativa nº
3, de 19 de janeiro de 2001. Aprova os
regulamentos Técnicos de Identidade e
Qualidade de Apitoxina, Cera de Abelha,
Geléia Real, Geléia Real Liofilizada, Pólen
Apícola, Própolis e Extrato de Própolis,
conforme consta dos Anexos desta Instrução
Normativa. Diário Oficial da União,
Seção 1, Página 18.
Cabral, I.S.R., Oldoni, T.L.C., Prado, A.,
Bezerra, R. M. N., Alencar, S. M. Composição
fenólica, atividade antibacteriana e antioxidante
da própolis vermelha brasileira. Química Nova,
v. XY, p.1-5, 2009.
Cain, K, Skilleter, D.N. Preparation and use of
mitochondria in toxicological research. In Snell,
K.; Mullock , B. (Eds.), Biochemical
Toxicology, IRL Press, Oxford. p. 217–254,
1987.
Castro, M.L., Cury, J.A., Rosalen, P.L.,
Alencar, S.M., Ikegaki, M., Duarte, S., Koo, H.
Própolis do Sudeste e Nordeste do Brasil:
influencia da sazonalidade na atividade
antibacteriana e composição fenólica. Química
Nova, v. 30, n.7, p. 1512-1516, 2007.
Daugsch, A., Moraes, C.S., Fort, P., Park, Y.K.
Brazilian Red Propolis - Chemical Composition
and Botanical Origin. Evidence-Based
Complementary and Alternative Medicine,
v.5, n.4, p.435-441, 2007.
Daugsch, A., Moraes, C.S., Fort, P., Park, Y.K.
Brazilian red propolis - chemical composition
and botanical origin. Evidence-Based
Complementary and Alternative Medicine, v.
5, p. 435–441, 2008.
Dodeigene, L., Thumus, R., Lejeune.
Chemiluminescence as a diagnostic tool. A
review. Talanta, v.51, p.415–439, 2000.
8. The binary mobile phase was composed of
water (solvent A) and methanol (solvent B)
according to the method described by Alencar et
al., (2007) and Cabral et al., (2009) with
modifications. The HAE was dissolved in
methanol (50 mg /mL) and filtered with a 0.45
μm filter (Millipore). Aliquots of 2 μL of 5% of
HAE (w/v) were injected into the UFLC system.
UFLC was performed using a linear gradient
elution as follows: 40% B increased to 60% B
after 22.5 min, a hold at 90% B for 37.3- 42.3
min, followed by a decrease to 30% B after 42.3
min with a solvent flow rate of 0.4 mL/min. The
following authentic standards of flavonoids and
phenolic acids were used: formononetin,
quercetin, 3-hydroxy-7-methoxyflavone,
catechin, epicatechin and propyl gallate. All the
tests were performed in triplicates using three
samples from each apiary
2.6. DPPH radical scavenging activity
The effect of DPPH radical was evaluated
by the method described by Brand-Williams et
al., 1995 and Tominaga et al., 2005 with
adaptations. The assay mixture contained 1.0
mL of 100mM sodium acetate buffer, 50 μL of
HAE, 1.0 ml of 95% ethanol, and 500 μL of 200
mM DPPH solution. Absorbance was recorded
at 517 nm after 10 min of incubation at room
temperature, in the dark. Blank containing the
same amount of ethanol and DPPH solution was
used as the negative control. The percentage
inhibition was plotted against phenol content
and IC50 (concentration of total phenol able to
scavenge 50% of DPPH free radical) was
determined.
2.7. Lipid peroxidation assay
Reaction mixture was prepared by adding
10 µL of each sample to 1 mL of a reaction
medium containing 130 mM KCl, and 10 mM
TrisHCl pH7.4, and mitochondria was added to
yield a final concentration of 1 mg of protein.
Later 50 µL ferrous ammonium sulfate and
2mM sodium citrate were added and the
samples were incubated at 37°C for 30 minutes.
Mitochondria was isolated from male Wistar
rats livers by differential centrifugation as
described by Pedersen et al., 1978, and the
mitochondrial protein content was determined
by the biuret reaction (CAIN; SKILLETER,
1987). For the determination of TBA-reactive
compound, 1 mL of 1% thiobarbituric acid
(TBA), 0.1 mL of 10M NaOH and 0.5 mL of
H3PO4 were added, followed by incubation in a
water bath for 20 minutes at 80°C. The TBA-
reactive compounds were extracted with 2 mL
of n-butanol and the samples were then
centrifuged at 9800 g for 10 min. The readings
were taken at 535 nm, using a Hitachi U2001
spectrophotometer. The experiments were
performed in triplicates together with blank
(without mitochondria), positive control
(without sample) and negative control (without
iron). The IC50 values were determined using
the GraphPad Prism ®.
2.8. Xanthine/luminol/XOD system
Samples of red propolis extract were
diluted in hydro alcoholic solution to a
concentration of 5 mg/mL and subsequent
dilutions were prepared in 0.1 M glycine buffer
pH 9.4. To verify free radical scavenging
activity of the propolis extract, a solution
containing 400 μL of 1 mM EDTA and 0.1 M
glycine pH 9.4 were added to a test tube
together with 150 μL of 6 mM xanthine, 10 μL
of test sample and 10 μL of 0.6 mM luminol
solution. The reaction was initiated with the
addition of 100 μL of freshly prepared cold 20
mU/mL xanthine oxidase solution. The
scavenging activity was determined after an
incubation of 5 minutes at 25 °C in an
Autolumat LB 953 luminometer. The results
were expressed according to the concentration
of propolis extract in the reaction medium
(MARQUELE-OLIVEIRA et al., 2005). The
IC50 values were determined using GraphPad
Prism® software.
2.9. Statistics
All the measurements were performed in
triplicates. The average was determined with
Assistat 7.7 Beta (registre INPI 0004051-2), and
used for the analysis of variance (ANOVA) and
Tukey test for means comparison, when
necessary.
3. Results and Discussion
The hydroalcoholic extracts of all the
propolis samples presented yield ranging from
9.45% to 74.62% (w/w) with the exception of
two samples that showed lower yield than the
value determined by Ministry of Agriculture
9. Fonseca, M. J. V. Assessment of the
antioxidant activities of Brazilian extracts of
propolis alone and in topical pharmaceutical
formulations. Journal of Pharmaceutical and
Biomedical Analysis, v.39, p.455-462, 2005.
Marquele-Oliveira, F., Fonseca, Y.M.,
Georgetti, S. R., Vicentini, F. T. M. C.,
Bronzati, V., Fonseca, M. J. V. Evaluation of
the antioxidant activity as an additional
parameter to attain the functional quality of
natural extracts. Latin American Journal of
Pharmacy, v.27, n.3, p. 325–332, 2008.
Mendonça, L.S., Mendonça, F.M.R., Maia-
Araújo, Y.L.F., Araújo, E.D., Ramalho, S.A.,
Narain, N., Padilha, F.F., Jain, S., Orellana,
S.C., Cardoso, J.C. (Accepted) Chemical
markers and antifungal activity of red propolis
from Sergipe, Brazil. Food Science and
Technology (Campinas).
Miguel, M.G., Nunes, S., Dandlen, S. A.,
Cavaco, A.M. and Antunes, M.D. Phenols and
antioxidant activity of hydro-alcoholic extracts
of propolis from Algarve, South of Portugal.
Food and Chemical Toxicology, v.48, p.3418–
3423, 2010.
Moraes, C.S. Isolamento e identificação de
formononetina da própolis de João Pessoa-PB,
estudo de sua sazonalidade e avaliação de suas
atividades biológicas. 2009. Tese de doutorado,
Universidade Estadual de Campinas. Faculdade
de Engenharia de Alimentos. São Paulo, SP,
Brasil.
Park, Y.K., Ikegaki, M., Alencar, S.M.
Classification of Brazilian propolis by both
physicochemical methods and biological
activity. Mensagem Doce, v.58, p.2-7. 2000.
Pascual C., Gonzales R., Torricella R. G.
Scavenging action of propolis extract against
oxygen radicals. Journal Ethnopharmacology,
v.4, p.9-13, 1994.
Pedersen P.L., Greenawalt J.W., Reynafarje B.,
Hullihen J., Decker G.L., Soper J.W.
Bustamente E. Preparation and characterization
of mitochondria and submitochondrial particles
of rat liver and liver-derived
tissues. Methods in Cell Biology, v..20, p.411–
481, 1978.
Pierini, G. D., Granero, A.M., Di Nezio, M.S.,
Centurión, M.E., Zon, M.A., Fernández, H.
Development of an electroanalytical method for
the determination of lead in Argentina raw
propolis based on bismuth electrodes.
Microchemical Journal, v.106, p.102–106,
2013.
Pinheiro, M.S. Avaliação da atividade
antimicrobiana e citoprotetora gástrica dos
extratos de mangaba, caju e própolis vermelha.
2009. Dissertação de mestrado, Universidade
Tiradentes – UNIT. Aracaju, SE, Brasil, 2009.
Pinheiro, K. S., Ribeiro,D.R, Alves, A.V.F.,
Pereira-Filho, R.N., Oliveira, C.R., Lima, S.O.
Modulatory activity of brazilian red propolis on
chemically induced dermal carcinogenesis.
Acta Cirúrgica Brasileira, v.29, n.2, p.111-
117, 2014.
Ramanauskiene, K., Kalveniene, Z.,
Kasparaviciene, G., Briedisis, V. Assay of
correlation between antioxidant activity of
propolis extracts and their chemical
components. European Journal of
Pharmaceutical Sciences, v.32, p. S45, 2007.
Rodgers, E.H., Grant, M.H. The effect of the
flavonoids, quercetin, myricetin and
epicathechin on the growth and enzyme
activities of MCF7 human breast cancer cells.
Chemical Biologic Interactions, v.116, n.1-2,
p.213-228, 1998.
Silva, F.A.S.E., Azevedo, C.A. A New Version
of The Assistat-Statistical Assistance Software.
In: World Congress on Computers in
Agriculture, 4, Orlando-FLUSA: Anais.
Orlando: American Society of Agricultural
Engineers, p.393-396, 2006.
Tominaga, Y., Kobayachi, Y., Goto,
T., Kasemura, K., Nomura, M., Yasshi, Y. DPPH
Radical–scavenging effect of
several phenylpropanoid compounds and their
glycoside derivatives. Journal of the
Pharmaceutical Society of Japan, v. 125, n.4,
p.371–375, 2005.
Torel, J., Cillard, J., and Cillard, P. Antioxidant
activity of flavonoids and reactivity with peroxy
radical. Phytochemistry, v.25, n.2, p. 383–385,
1986.
10. ______________________________________
1. Departamento de Biologia,
Universidade Federal de Sergipe, São Cristóvão,
Sergipe – Brasil (ylmaia@yahoo.com.br –
Maia-Araújo, Y.L.F),
(lucyana_biologia@yahoo.com.br – Mendonça,
L.S.) e (edaraujo@yahoo.com.br – Araújo,
E.D.);
2. Departamento de Engenharia de
Alimentos, Universidade Federal de Sergipe,
São Cristóvão, Sergipe – Brasil
(nayjaracarvalho@hotmail.com – Gualberto,
N.C.) e (narendra.narain@gmail.com – Narain,
N.);
3. Fundação Oswaldo Cruz, Centro de
Pesquisa René Rachou, Belo Horizonte, Minas
Gerais, Brasil (sara_cuadros@yahoo.com –
Cuadros-Orellana, S.);
4. Instituto de Tecnologia e Pesquisa
(ITP), Aracaju, Sergipe – Brasil
(juaracaju@yahoo.com.br – Cardoso, J.C.);
5. Departamento de Morfologia,
Universidade Federal de Sergipe, São Cristóvão,
Sergipe – Brasil (sonajain24@yahoo.com –
Jain, S.) *.
Corresponding author*
Sona Jain, Av. Marechal Rondon, S/N, Bairro
Rosa Elze. Universidade Federal de Sergipe,
Departamento de Morfologia, São Cristóvão –
SE. CEP 49100-000. e-mail:
sonajain24@yahoo.com
82