The present study aimed to evaluate in vivo and in vitro the antitumoural activity of a propolis extract obtained with edible vegetable oil and its fractions and also to investigate its chemical composition by LC-MS and LC-MS/MS. To evaluate the toxicological aspects related to the propolis extract treatment, hematological, biochemical, histopathological and morphological analyses of treated animals were performed. All propolis extracts showed an in vivo antitumour activity in the experimental model with a moderate toxicity effect at experimental exposure levels. The oil extract was as effective as the ethanolic extract at inhibiting tumour growth. In vitro assays showed that the whole oil extract produced better inhibition of tumour cells than its fractions. LC-MS and LC-MS/MS identified four phenolic acids and three flavonoids. The anticancer potential of the oil extract of propolis has been demonstrated and the edible vegetable oil was shown as an attractive alternative solvent to extract bioactive natural propolis components.

Recently there has been a growing number of patents dealing with new solvents for preparing propolis extracts. This study aimed to prepare edible oil propolis extracts and compare their chemical composition and biological activity with ethanolic propolis extracts. ESI-MS and spectrophotometric methods were used for qualitative and quantitative analyses, respectively. Antibacterial activity was evaluated by diffusion in agar. Cytotoxicity was tested by MTT assay using tumor cell lines. The oil is able to extract bioactive compounds from propolis. Further studies are needed to improve extraction efficiency and to characterize the active components.Keywords: oil propolis extract; phenolic compounds; antibacterial and cytotoxic activities. introduçãoA própolis é uma resina de ampla utilidade para as abelhas e para o homem.1,2 A composição química da própolis é complexa e está relacionada com a flora da região em que foi originada e a época da coleta.3-6 Sua atividade farmacológica tem sido atribuída aos compostos fenólicos, entre eles flavonóides e ácidos fenólicos, cujos teores têm sido propostos como parâmetros para o controle da qualidade. 7,8 Devido às inúmeras propriedades benéficas da própolis, o seu uso comercial em produtos farmacêuticos, cosméticos e de higiene pessoal na forma de extratos líquidos é amplo. Para a elaboração desses produtos, é comumente utilizado o extrato obtido com álcool de cereais 70% v/v e tempos de extração que variam de 1 dia até 6 meses.9 Entretanto, a presença de álcool na formulação confere um sabor não agradável para alguns consumidores.Buscando superar esses inconvenientes, têm surgido muitas patentes que propõem novos métodos de extração da própolis com baixo teor alcoólico ou isento de álcool. [10][11][12][13][14][15] Dentre as alternativas propostas, destaca-se o extrato de própolis obtido com óleo vegetal, o qual conserva bem as características organolépticas da própolis e possibilita a apresentação do produto em cápsulas gelatinosas. Existe uma dúvida, porém, quanto à eficiência do óleo comestível em extrair os principais compostos bioativos da própolis e se os produtos elaborados com o extrato oleoso possuem ou não as propriedades farmacológicas adequadas para a finalidade à qual são destinados.Neste trabalho avaliamos o extrato oleoso de própolis quanto a sua composição química qualitativa e quantitativa de compostos fenólicos extraídos e comparamos os resultados com aqueles obtidos de diferentes extratos etanólicos de própolis. Este estudo empregou as técnicas de espectrometria de massas com ionização por electrospray (ESI-MS) e espectrofotometria no UV-VIS. Como as atividades antibacteriana e citotóxica de extratos hidroalcoólicos e de alguns compostos isolados de própolis são bem conhecidas, 2 procurou-se avaliar se o extrato oleoso também possuía essas atividades. Neste trabalho o extrato oleoso da própolis foi, então, avaliado in vitro em diferentes cepas de organismos patogênicos e em linhagens de célu-las tumorais humanas para estimar suas atividades antibacteriana...

Objective. As propolis is a highly valued bee product, we aimed to verify the quality of aged propolis, investigating their phenolic and flavonoid composition, levels of toxic metals, radical scavenging and antimicrobial activities. Material and Methods. Samples of fresh and aged propolis of six different beekeepers, from the same geographical location, were investigated in terms of their phenolic and flavonoid composition and levels of Pb, Cd, and Cr, as well as radical scavenging and antimicrobial activities. Results. The two groups of propolis had similar qualitative composition by HPLC-PDA and ESI(-)-MS. Fresh propolis and aged propolis show no differences when average values of extraction yield, flavonoids, EC50, or MIC were compared and both types of propolis showed good antimicrobial activity at low concentrations. Only levels of phenolic compounds were higher in fresh propolis. Conclusion. The propolis samples considered in this study, aged or fresh, had similar qualitative composition, although they were collected in different periods. Samples only differed in their levels of total phenolic content. Moreover, aged propolis conserves significant radical scavenging and antimicrobial properties. We suggest that aged propolis should not be discarded but explored for alternative applications.

Concentrations of eleven representative metals (Al, Ca, Cd, Cr, Cu, K, Mg, Mn, Na, Pb and Zn) in forty-two propolis samples were measured by electrothermal atomization and flame atomic absorption spectrometry after calcination in a muffle furnace. Samples were collected from different regions from Paraná State - Brazil where apiculture is an important economic activity. Results showed that the average content of Al, Ca, K, Mg, Mn, Na and Zn in propolis was 0.68, 1.66, 7.59, 1.27, 0.08, 0.58 and 0.02 mg g(-1), respectively. Levels of Al, Ca, and Mg were statistically different in some regions of Paraná and could be used to assign the geographical origin of the propolis. The average concentration of the Cd, Cr, and Pb in raw propolis was 0.13, 5.53 and 9.85 μg g(-1), respectively, and allowed for identification of specific areas with environmental contamination.

The study aimed to evaluate the antifungal potential of an extract of propolis obtained with edible vegetable oil (ODEP) and to identify antifungal compounds in this extract. Propolis sample was extracted with canola oil. After filtration, the oily liquid extract was submitted to solvent partition and dried to obtain ODEP, which was tested in vitro against Candida albicans strains. ODEP was fractioned on Sephadex and by high-performance liquid chromatography (HPLC). Active fractions and/or a pure compound were analyzed by nuclear magnetic resonance, liquid chromatographic mass spectrometry (LC-MS) and liquid chromatographytandem mass spectrometry (LC-MS/MS) to characterize their chemical composition. Bioassay-guided fractionation allowed the isolation of dihydrokaempferide, which was also quantified in ODEP by HPLC together with the total flavonoid content. Kaempferide and isosakuranetin were identified by LC-MS and LC-MS/MS in fractions with a potential activity against C. albicans. In vitro assays showed that some fractions from ODEP produced better inhibition of C. albicans than the propolis extract itself. PRACTICAL APPLICATIONSAqueous or ethanolic extracts of propolis (EEP) are widely used in alternative homemade medicine products mainly because of their antimicrobial and healing properties. Edible vegetal oils are known to extract bioactive compounds from propolis. The resulting extract (ODEP) has several advantages over the common EEP, such as the possibility of new pharmaceutical presentations for topical or internal applications. Because of the absence of ethanol in its formulation, edible ODEP are expected to be more tolerated. In this study, ODEP was active in vitro against Candida albicans strains. This supports the use of ODEP as an alternative to EEP and also its potential for topical application in combination with antifungal drugs. In addition, the ingestion of ODEP (e.g., contained in gelatin capsules), which have a high percentage of flavonoids, could be of nutritional value, helping to reach the recommended daily amounts of flavonoids and phenolic compounds. bs_bs_banner Journal of Food Quality

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Total levels of caffeoylquinic, p-coumaric, and benzoic acid derivatives, flavonols, and dihydroflavonols were investigated in 44 brown propolis samples from the state of Paraná(Brazil) together with their cytotoxicity and antibacterial activities. The aim was to discover chemical and biological activity-based patterns. Great quantitative variation was found within individual samples; however, a typical extract might contain 11.14−21.45 mg of caffeoylquinic acid derivatives/g, 6.27−12.17 mg of p-coumaric acid derivatives/g, 3.18−7.45 mg of benzoic acid derivatives/g, 9.35−23.55 mg of flavonols/g, and 0.17−4.25 mg of dihydroflavonols/g. MIC values poorly correlated to the levels of p-coumaric acids, flavonols, and dihydroflavonols. Most propolis samples showed medium-to-high cytotoxic activity against at least one tumoral cell. The most susceptible cell was OVCAR-8, followed by SF-295, but no correlation was found between the inhibition of OVCAR-8 and SF-295 and levels of any chemical class investigated. The concentration of bioactive compounds in brown propolis samples together with biological activities distinguished samples produced in northern, southern, eastern, and western Parana, being characteristics associated with the geographical origin, and may help to establish a pattern for the propolis produced in the state of Parana.

Total phenolic, flavonoid and mineral contents and antiradical activity are usually used as a measure of propolis extract quality. The purpose of this study was to compare the quality of propolis extracts obtained with different solvents such as water, ethanolic solutions (30, 70, 95%, v/v in water) and edible vegetable oils (canola and soybean). LC-MS was used to identify constituents of the extracts. Antiradical activity was assayed using chemiluminescence from the luminol/hemin/H 2 O 2 /CTAB system and the DPPH test. Both antiradical assays were strongly correlated (r = -0.97). Principal Component Analysis indicated positive correlations for phenolics, flavonoid levels and antiradical activity showing that all phenolic compounds are important for the scavenging activity of Brazilian propolis. The extract obtained with ethanol 30% v/v had the highest antiradical activity mainly due to its high content of dicaffeoylquinic acids. Efecto del solvente de extracción en la actividad antirradical de los extractos obtenidos de propóleos ResumenEl total de compuestos fenólicos, flavonoides, minerales y la actividad antirradical son parámetros usados comúnmente como medida de la calidad de extractos de propóleo. El objetivo de este estudio fue comparar la calidad del extracto de propóleo obtenido con diferentes disolventes como agua, disoluciones etanólicas (30, 70, 95%, v/v en agua) y aceites vegetales comestibles (Canola and Soya). LC-MS fue utilizada para identificar constituyentes de los extractos. La actividad antirradical fue evaluada a partir de la quimioluminiscencia del sistema luminol/hemina/H 2 O 2 /CTAB y por el ensayo del DPPH. Estos dos ensayos se mostraron fuertemente correlacionados (r = -0.97). El analice de los componentes principales indicó correlaciones positivas entre las cantidades de fenólicos, flavonoides y la actividad antirradical mostrando que todos los compuestos fenólicos son importantes para esta actividad del propóleo brasileño. El extracto obtenido con 30% de etanol presentó la mayor actividad antirradical, principalmente debido al contenido elevado de ácidos dicafeoilquínicos.