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      Brazilian Red Propolis: Extracts Production, Physicochemical Characterization, and Cytotoxicity Profile for Antitumor Activity

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          Abstract

          Brazilian red propolis has been proposed as a new source of compounds with cytotoxic activity. Red propolis is a resinous material of vegetal origin, synthesized from the bees of the Appis mellifera family, with recognized biological properties. To obtain actives of low polarity and high cytotoxic profile from red propolis, in this work, we proposed a new solvent accelerated extraction method. A complete 2 3 factorial design was carried out to evaluate the influence of the independent variables or factors (e.g., temperature, number of cycles, and extraction time) on the dependent variable or response (i.e., yield of production). The extracts were analyzed by gas chromatography coupled with mass spectrometry for the identification of chemical compounds. Gas chromatography analysis revealed the presence of hydrocarbons, alcohols, ketones, ethers, and terpenes, such as lupeol, lupenone, and lupeol acetate, in most of the obtained extracts. To evaluate the cytotoxicity profile of the obtained bioactives, the 3-(4,5-dimethyl-2-thiazole)-2,5-diphenyl-2- H-tetrazolium bromide colorimetric assay was performed in different tumor cell lines (HCT116 and PC3). The results show that the extract obtained from 70 °C and one cycle of extraction of 10 min exhibited the highest cytotoxic activity against the tested cell lines. The highest yield, however, did not indicate the highest cytotoxic activity, but the optimal extraction conditions were indeed dependent on the temperature (i.e., 70 °C).

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          Recent Global Patterns in Prostate Cancer Incidence and Mortality Rates

          Previous studies have reported significant variation in prostate cancer rates and trends mainly due to differences in detection practices, availability of treatment, and underlying genetic susceptibility.
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            Chemical characterization and biological activity of six different extracts of propolis through conventional methods and supercritical extraction

            Propolis is a natural product with many demonstrated biological activities and propolis extract has been used in the food, pharmaceutical and cosmetics industries. Different works have showed the variations in the chemical composition, and consequently, on the biological activity of the propolis that are associated with its type and geographic origin. Due to this study evaluated propolis extracts obtained through supercritical extraction and ethanolic extraction (conventional) in three samples of different types of propolis (red, green and brown), collected from different regions in Brazil (state of Bahia). Analyses were performed to determine the humidity, water activity, the content of total ash, proteins, lipids and fiber in raw propolis samples. The content of phenolic compounds, flavonoids, in vitro antioxidant activity (DPPH), catechin, ferulic acid and luteolin and antimicrobial activity against two bacteria (Staphylococcus aureus and Escherichia coli) were determined for all extracts. For the green and red ethanolic extracts the anti-leishmanicidal potential was also evaluated. The physicochemical profiles showed agreement in relation to the literature. The results identified significant differences among the extracts (p>0.05), which are in conformity with their extraction method, as well as with type and botanical origin of the samples. The extraction with supercritical fluid was not efficient to obtain extracts with the highest contents of antioxidants compounds, when compared with the ethanolic extracts. The best results were shown for the extracts obtained through the conventional extraction method (ethanolic) indicating a higher selectivity for the extraction of antioxidants compounds. The red variety showed the largest biological potential, which included the content of antioxidants compounds. The results found in this study confirm the influence of the type of the raw material on the composition and characteristics of the extracts. The parameters analysis were important to characterize and evaluate the quality of the different Brazilian propolis extracts based on the increased use of propolis by the natural products industry.
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              Chemical Composition and Biological Activity of Extracts Obtained by Supercritical Extraction and Ethanolic Extraction of Brown, Green and Red Propolis Derived from Different Geographic Regions in Brazil

              The variations in the chemical composition, and consequently, on the biological activity of the propolis, are associated with its type and geographic origin. Considering this fact, this study evaluated propolis extracts obtained by supercritical extraction (SCO2) and ethanolic extraction (EtOH), in eight samples of different types of propolis (red, green and brown), collected from different regions in Brazil. The content of phenolic compounds, flavonoids, in vitro antioxidant activity (DPPH and ABTS), Artepillin C, p-coumaric acid and antimicrobial activity against two bacteria were determined for all extracts. For the EtOH extracts, the anti-proliferative activity regarding the cell lines of B16F10, were also evaluated. Amongst the samples evaluated, the red propolis from the Brazilian Northeast (states of Sergipe and Alagoas) showed the higher biological potential, as well as the larger content of antioxidant compounds. The best results were shown for the extracts obtained through the conventional extraction method (EtOH). However, the highest concentrations of Artepillin C and p-coumaric acid were identified in the extracts from SCO2, indicating a higher selectivity for the extraction of these compounds. It was verified that the composition and biological activity of the Brazilian propolis vary significantly, depending on the type of sample and geographical area of collection.
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                Author and article information

                Journal
                Biomolecules
                Biomolecules
                biomolecules
                Biomolecules
                MDPI
                2218-273X
                06 May 2020
                May 2020
                : 10
                : 5
                : 726
                Affiliations
                [1 ]Tiradentes University (UNIT), Av. Murilo Dantas, 300, Aracaju 49010-390, Brazil; felipemdadc@ 123456gmail.com (F.M.d.A.d.C.); jadersonqmc@ 123456gmail.com (J.K.S.); carlavfj@ 123456gmail.com (C.V.F.d.J.); laiza_canielas@ 123456hotmail.com (L.C.K.); pattypharma@ 123456gmail.com (P.S.); cleide18@ 123456yahoo.com.br (C.M.F.S.); elina@ 123456ufrgs.br (E.C.B.); fpadilha@ 123456yahoo.com (F.F.P.); svfloresta@ 123456hotmail.com (S.V.F.G.)
                [2 ]Institute of Technology and Research (ITP), Av. Murilo Dantas, 300, Aracaju 49032-490, Brazil
                [3 ]Federal University of Sergipe (UFS), Avenida Marechal Rondon, São Cristovão 49100-000, Brazil; luciana.nalone@ 123456hotmail.com (L.N.d.A.); ricardoamaral23@ 123456hotmail.com (R.G.A.)
                [4 ]Federal University of Bahia (UFBA), Salvador/BA 40170-110, Brazil; jmdavid@ 123456ufba.br
                [5 ]Tiradentes Institute, 150 Mt Vernon St, Dorchester, MA 02125, USA
                [6 ]Department of Agricultural Sciences, University of Napoli Federico II, Via Università 100, 80055 Portici, Italy; rafcapas@ 123456unina.it
                [7 ]Department of Pharmacy, University of Napoli Federico II, Via D. Montesano 49, 80131 Napoli, Italy; asantini@ 123456unina.it
                [8 ]Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal
                [9 ]CEB-Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal
                Author notes
                [* ]Correspondence: ebsouto@ 123456ff.uc.pt (E.B.S.); ricardo.patologia@ 123456uol.com.br (R.L.C.d.A.-J.)
                Author information
                https://orcid.org/0000-0001-6527-6612
                https://orcid.org/0000-0002-5932-9842
                https://orcid.org/0000-0002-5892-4252
                https://orcid.org/0000-0002-3335-1822
                https://orcid.org/0000-0001-5505-3327
                https://orcid.org/0000-0002-9737-6017
                https://orcid.org/0000-0003-4444-7632
                Article
                biomolecules-10-00726
                10.3390/biom10050726
                7277404
                32384801
                5a0d0347-8c6c-4698-99ff-38d80be65f31
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 20 March 2020
                : 01 May 2020
                Categories
                Article

                red propolis,extraction,supercritical liquids,antitumor activity

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