Chemical analysis of the bioactive compounds of red propolis

Author(s)

Begoña Gimenez Cassina Lopez

• Document type: Doctoral Thesis
• Press: Campinas, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
• Defense date: 2017-04-24

Examining board members

Alexandra Christine Helena Frankland Sawaya; Elfriede Marianne Bacchi; Paulo César Pires Rosa; Humberto Márcio Santos Milagre; Rosana Maria Alberici

• Advisor: Marcos Nogueira Eberlin; Alexandra Christine Helena Frankland Sawaya

Abstract

Red propolis is a resinous bee product typical from the northeastern states of Brazil. Its composition varies widely depending on the botanical and geographical origin, and on the type of bee involved in the collecting process. Red propolis from Alagoas, received the mention of geographical origin indication, however this resin is also produced in other regions and it displays promising biological activities such as antimicrobial, antioxidant and cytotoxic among others. An experimental design was assessed aiming to identify the potential bioactive compounds. Then, an isolation method was developed by counter current chromatography (CCC) followed by the scale-up of the method, reaching fractions and isolated compounds. The purification of the fractions was made by preparative HPLC and the identification was performed by high-resolution mass spectrometry (QTOF). There are many variables that influence the composition of red propolis and it is necessary to standardize the composition due to the popular use of the resin. Thus, an analytical method was developed and validated to quantify nine of the substances present in red propolis (formononetin, pinocembrin, liquiritigenin, isoliquiritigenin, daidzein, biochanin A, retusapurpurin A, 7-O-methylvesitol and C38H50O6) including the ones isolated in this study. The identification and quantification method was applied to twenty samples of red propolis from Alagoas, Bahia, Paraiba and Sergipe, together with samples of Dalbergia ecastophyllym (the botanical source) from Paraiba and Brasilia, as well as ten samples collected throughout the year in Sergipe to verify the seasonal variation of the composition. The antioxidant and antimicrobial activity of the isolated compounds was evaluated as well as the synergistic action of these compounds. In this work, it was possible to develop a method for isolation of bioactive compounds by CCC and it was possible to isolate three compounds (retusapurpurin A, 7-O-methylvesitol and C38H50O6). An analytical method was developed and validated and it was applied to red propolis samples from different states of the northeastern region of Brazil, and a variation of the composition was observed in samples from the same region as well as samples from different geographical origins. The seasonal variation of red propolis from Sergipe showed that samples collected in March presented less amount of the compounds evaluated. Finally, it was demonstrated that all the tested compounds are bioactive (formononetin, pinocembrin, liquiritigenin, isoliquiritigenin, daidzein, biochanin A, retusapurpurin A, 7-O-methylvesitol and C38H50O6) and a synergic action was observed in the antimicrobial and antioxidant activities (AU)

• FAPESP's process: 13/20313-3 - Identification of the substances present in red Brazilian propolis by UPLC-ESI-MS and NMR and evaluation of the antioxidant and antimicrobial activity of the isolated compounds
• Grantee: Begona Gimenez-Cassina Lopez
• Support Opportunities: Scholarships in Brazil - Doctorate