Chemical Profile of Brazilians Propolis Samples

Author(s)

Adne Abbud Righi

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Instituto de Biociências (IBIOC/SB)
• Defense date: 2008-11-24

Examining board members

Antonio Salatino; Marize Terezinha Lopes Pereira Peres; Deborah Yara Alves Cursino dos Santos

• Advisor: Antonio Salatino

Abstract

Propolis is a resinous substance comprising exsudates collected from plants and substances produced by bees, with texture and colour varying according to the plant source of resin and respective phenology. This product is used in the hive with various purposes: to seal holes and line the hive entrance and to embalm the bodies of killed invaders. The interest in propolis has increased recently, mainly due to experimental evidences of its biological activities, such as cytotoxic, anti-herpes, anti-HIV, antitumoral, anti-microbial and antioxidant. The chemical composition of propolis is complex, including, in addition to beeswax, flavonoids, phenolic acids, terpenoids and volatile substances, among other constituents. The composition depends on many factors, mainly the region, where it was produced. In the present work the chemical profile of the Brazilian propolis was established based on the content of total phenolic substances, total flavonoids and waxes of samples from distinct regions of the Brazilian territory: Ponta Grossa (PR), Bauru and Pariquera-Açu (SP), Lavras and Mira Bela (MG), Pirenópolis (GO), Cabo Verde (BA), Maceio (AL) and Picos (PI). The contents of total phenols were obtained by the Folin-Ciocalteau method, using p-coumaric acid as reference. Determination of total flavonoids was based on two spectrophotometric complementary methods: alluminum chloride and dinitrophenyl-hydrazine, using quercetin and pinocembrin, respectively, as references. Determination of wax contents was carried out by extraction with chloroform, treatment with methanol and weight of the residue. The results revealed a wide chemical variety among the samples, even between samples from the same state, as are the cases of samples from Lavras and Mira Bela and from Bauru and Pariquera-Açu. Total phenol contents varied in the range 0,91%-41,63%, highest figures corresponding to red propolis from Maceio. The contents determined are higher than those obtained with propolis from Uruguai, Argentina, Paraguai, China and Europe, and also with Brazilian green propolis. Regarding values of total flavonoids, they ranged from 0,31% to 4,43%, similar to results reported by other researchers with Brazilian propolis, but lower than contents of European propolis, which has varied in the range 1,1%-22,3%. Wax contents varied from 5,37% to 45,88%, such figures being higher than values of Brazilian propolis reported by other authors, but comparable with Chinese propolis, which content is close to 30%. Based on the results obtained a Principal Components Analysis (PCA) was carried out. Samples from Maceio, Cabo Verde, Pirenópolis and Bauru emerged isolated. Pariquera-Açu and Mira Bela standed out as a group in the other end of the graphic, due to low contents of phenolic compounds and high levels of waxes. The samples from Picos, Lavras and Ponta Grossa comprised another group, based on salient contents of total phenols and relatively high contents of waxes. Chloroform and methanolic fractions from each sample were analyzed by HPLC-MS and GC-MS. These analyses showed that samples from Bauru, Lavras and Cabo Verde have composition typical of green propolis, with many prenylated phenylpropanoids and cafeoylquinic acid derivatives. The composition of the Maceio propolis differed very much, standing out by having exclusive constituents, such as chalcones, a class of flavonoids never reported for propolis. Propolis from Pirenopolis and Picos are alike due to many glycosilated flavonoids, substances also never reported for propolis. Samples from Ponta Grossa, Pariquera-Açu and Mira Bela have chemical profiles intermediate between green propolis and those with glycosilated flavonoids. The commented chemical affinities were evident in clustering analysis carried out with the identified substances as variables and UPGMA method. These results constitute evidence that Brazilian propolis cannot be grouped in only twelve types, as has been suggested. The present chemical analyses revealed three additional types: Maceió, Pirenópolis/Picos e Ponta Grossa/Pariquera-Açu/Mira Bela. The huge diversity of the composition of propolis produced in Brazil highlight the importance of studies for determination of chemical profiles and resin botanical sources. Such findings are essential for adequate use of propolis products both in the national and international market. (AU)