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Open Access Research article

Bioassay guided purification of the antimicrobial fraction of a Brazilian propolis from Bahia state

Myrella Lessio Castro1, Walquíria Ribeiro Vilela2, Rogéria Cristina Zauli2, Masaharu Ikegaki3, Vera Lúcia Garcia Rehder4, Mary Ann Foglio4, Severino Matias de Alencar2 and Pedro Luiz Rosalen1*

Author Affiliations

1 Department of Physiological Sciences, Piracicaba Dental School, University of Campinas (UNICAMP); Avenida Limeira, 901; Piracicaba, SP, 13414-903, Brazil

2 Department of Agri-Food Industry, Food and Nutrition, Escola Superior de Agricultura "Luiz de Queiroz", University of São Paulo (USP); Avenida Pádua Dias, 11; Piracicaba, SP, 13418900, Brazil

3 Department of Pharmacy, Federal University of Alfenas, Alfenas, MG, Brazil; Avenida Gabriel Monteiro da Silva, 700; Alfenas, MG, 37130-000, Brazil

4 Chemical, Biological and Agricultural Pluridisciplinary Research Center (CPQBA), University of Campinas (UNICAMP); C.P. 6171; Campinas, SP, 13081-970, Brazil

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BMC Complementary and Alternative Medicine 2009, 9:25  doi:10.1186/1472-6882-9-25

Published: 30 July 2009

Abstract

Background

Brazilian propolis type 6 (Atlantic forest, Bahia) is distinct from the other types of propolis especially due to absence of flavonoids and presence of other non-polar, long chain compounds, but presenting good in vitro and in vivo antimicrobial activity. Several authors have suggested that fatty acids found in this propolis might be responsible for its antimicrobial activity; however, so far no evidence concerning this finding has been reported in the literature. The goals of this study were to evaluate the antibacterial activity of the main pure fatty acids in the ethanolic extract and fractions and elucidate the chemical nature of the bioactive compounds isolated from Brazilian propolis type 6.

Methods

Brazilian propolis type 6 ethanolic extract (EEP), hexane fraction (H-Fr), major fatty acids, and isolated sub-fractions were analyzed using high performance liquid chromatography (HPLC), high resolution gas chromatography with flame ionization detection (HRGC-FID), and gas chromatography-mass spectrometry (GC-MS). Three sub-fractions of H-Fr were obtained through preparative HPLC. Antimicrobial activity of EEP, H-Fr, sub-fractions, and fatty acids were tested against Staphyloccus aureus ATCC 25923 and Streptococcus mutans Ingbritt 1600 using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC).

Results

EEP and H-Fr inhibited the growth of the microorganisms tested; nevertheless, no antimicrobial activity was found for the major fatty acids. The three sub-fractions (1, 2, and 3) were isolated from H-Fr by preparative HPLC and only sub-fraction 1 showed antimicrobial activity.

Conclusion

a) The major fatty acids tested were not responsible for the antimicrobial activity of propolis type 6; b) Sub-fraction 1, belonging to the benzophenone class, was responsible for the antimicrobial activity observed in the present study. The identification of the bioactive compound will improve the development of more efficient uses of this natural product.