Differentiating between Monofloral Portuguese Bee Pollens Using Phenolic and Volatile Profiles and Their Impact on Bioactive Properties.

Nowadays, bee products are commended by consumers for their medicinal and dietary properties. This study aimed to differentiate between monofloral bee pollens originating from Portugal using phenolic and volatile profiles and investigate their antioxidant and cytotoxic activity. Total phenolic and flavonoid compounds were recorded between 2.9-35.8 mg GAE/g and 0.7-4.8 mg QE/g, respectively. The LC/DAD/ESI-MSn analytical results allowed us to identify and quantify a total of 72 compounds, including phenolic and phenylamide compounds, whereas GC-MS results revealed the presence of 49 different compounds, mostly ketones, aldehydes, esters, hydrocarbons, and terpenes. The highest DPPH• radical scavenging activity, EC50: 0.07 mg/mL, was recorded in the sample dominated by Castanae sp. pollen, whereas the Rubus sp. (1.59 mM Trolox/mg) and Cistaceae sp. (0.09 mg GAE/g) pollen species exhibited the highest antioxidant activity in ABTS•+ and reducing power assays, respectively. Regarding the anti-carcinogenic activity, only Carduus sp. showed remarkable cytotoxic potential against MCF-7.

Bee pollen and bee bread nutritional potential: Chemical composition and macronutrient digestibility under in vitro gastrointestinal system.

Bee pollen (BP) and bee bread (BB) have been often investigated as potential functional foods. Both bee products are generally characterized by their high nutritional content, with BB being referred as more digestible than BP, however, there is a lack of scientific studies proving this claim. Here, we present a comparative evaluation of the macronutrient digestibility of BP and BB after applying a simulated in vitro gastrointestinal digestive system, together with the evaluation of its nutritional value and chemical composition. The digestibility scores for protein content were calculated on average as 69% and 76% for BP and BB, respectively, whereas digestibility scores for soluble sugars varied depending on bee product and sugar type. The results demonstrated that the nutritional values of both bee products changed depending on their botanical origin but BB is more accessible in the intestinal lumen, especially regarding protein.

Evaluation of Antioxidant and Anticancer Activity of Mono- and Polyfloral Moroccan Bee Pollen by Characterizing Phenolic and Volatile Compounds.

Bee pollen is frequently characterized as a natural source of bioactive components, such as phenolic compounds, which are responsible for its pharmaceutical potential and nutritional properties. In this study, we evaluated the bioactive compound contents of mono- and polyfloral bee pollen samples using spectroscopic and chromatographic methods and established links with their antioxidant and antitumor activity. The findings demonstrated that the botanical origin of bee pollen has a remarkable impact on its phenolic (3-17 mg GAE/g) and flavonoid (0.5-3.2 mg QE/g) contents. Liquid chromatography-mass spectrometry analysis revealed the presence of 35 phenolic and 13 phenylamide compounds in bee pollen, while gas chromatography-mass spectrometry showed its richness in volatiles, such as hydrocarbons, fatty acids, alcohols, ketones, etc. The concentration of bioactive compounds in each sample resulted in a substantial distinction in their antioxidant activity, DPPH (EC50: 0.3-0.7 mg/mL), ABTS (0.8-1.3 mM Trolox/mg), and reducing power (0.03-0.05 mg GAE/g), with the most bioactive pollens being the monofloral samples from Olea europaea and Ononis spinosa. Complementarily, some samples revealed a moderate effect on cervical carcinoma (GI50: 495 µg/mL) and breast adenocarcinoma (GI50: 734 µg/mL) cell lines. This may be associated with compounds such as quercetin-O-diglucoside and kaempferol-3-O-rhamnoside, which are present in pollens from Olea europaea and Coriandrum, respectively. Overall, the results highlighted the potentiality of bee pollen to serve health-promoting formulations in the future.

Quality assessment of Portuguese monofloral honeys. Physicochemical parameters as tools in botanical source differentiation.

The quality evaluation and physicochemical parameters assessment of Portuguese monofloral honeys were performed. Fifty-one monofloral honeys were collected from several regions from mainland Portugal, and from the Azores islands, producer labelled as carob tree (n = 5), chestnut (n = 2), eucalyptus (n = 5), bell heather (n = 6), incense (n = 4), lavender (n = 8), orange (n = 9), rape (n = 2), raspberry (n = 2), rosemary (n = 1), sunflower (n = 3), and strawberry tree (n = 4). Pollen analysis and microbiological safety were evaluated, and the parameters such as colour index, moisture content, electrical conductivity, hydroxymethylfurfural, pH, free and total acidity, diastase activity, proline, and sugar profile were assessed for physicochemical characterization, in all 51 monofloral honeys. After melissopalynological examination, the honeys were either confirmed as monofloral, or classified as multifloral with predominance of a specific pollen type or multifloral. Microbiological analysis showed that honeys were safe for human consumption. Pairwise comparisons of physicochemical parameters, using only honey types with n ≥ 3, revealed significant differences between honey types. Despite some homogeneity in sugar profile among honeys, eucalyptus honey was significantly different in glucose, maltose and maltulose content compared to incense, orange and sunflower honeys, and also exhibited a higher isomaltose amount compared to all analyzed honeys. Electrical conductivity, colour index, free and total acidity, and diastase activity showed significant differences between the analyzed honeys, indicating that these parameters may provide an additional tool in monofloral honey identification.

Production of chitosan-based biodegradable active films using bio-waste enriched with polyphenol propolis extract envisaging food packaging applications.

Developing biodegradable active films has been a promising green approach to overcoming global concerns over the environmental pollution and human health caused by plastic utilization. This study aimed to develop active films based on chitosan (CS), produced from waste crayfish (Procambarus clarkii) shells enriched with bioactive extract (5-20%) of propolis (PS) and to characterize its properties, envisaging food packaging applications. The chromatographic profile of PS extract confirmed its richness, with 41 phenolic compounds. With increasing extract addition to the chitosan, the thickness of the films increased from 61.7 to 71.7 µm, causing a reduction in the light transmission rate, along with a greenish colour shift. The interactions between PS extract and CS was confirmed by infrared spectroscopy, at the same time that the microstructural integrity of the films was checked on the scanning electron microscopy micrographs. The findings also showed that addition of PS enhanced the films thermal stability and mechanical properties e.g., tensile modulus, yield strength, and stress at break. Besides, it improved the antioxidant and antimicrobial activities. Overall, CS-based composite films seem a promising green alternative to petroleum-based synthetic plastics allowing to extend the shelf life of food products due to their eco-friendly nature.

Description of the volatile fraction of Erica honey from the northwest of the Iberian Peninsula.

Heather honey is highly appreciated by consumers for its sensorial profile, which varies depending on the flora used by the honeybees. Volatile compounds contribute to these qualities. Characterisation of the volatile profile related to the botanical origin is of great interest for the standardization of unifloral honey. For this reason, 33 heather honey samples from northwest of the Iberian Peninsula were analysed by headspace solid-phase microextraction (HS-SPME) to identify the key volatile compounds in this type of honey. The aim of this research was to provide a descriptive analysis of these compounds, and to find whether there is any relationship with the main Erica species. A total of 58 volatile organic compounds were found, with hotrienol, phenylacetaldehyde, and cis-linalool being the most abundant. A principal component analysis and Spearman's rank correlation showed the homogeneity of the volatile profile in the samples, and their close relationship with the main pollen types.

Chemical characterization, antioxidant, anti-inflammatory and cytotoxic properties of bee venom collected in Northeast Portugal.

Bee venom (BV) or apitoxin is a complex mixture of substances with reported biological activity. In the present work, five bee venom samples obtained from Apis mellifera iberiensis from the Northeast Portugal (two different apiaries) were chemically characterized and evaluated for their antioxidant, anti-inflammatory and cytotoxic properties. The LC/DAD/ESI-MS(n) analysis of the samples showed that melittin was the most abundant compound, followed by phospholipase A2 and apamin. All the samples revealed antioxidant and anti-inflammatory activity but without a direct relation with any of the individual chemical components identified. The results highlight that there are specific concentrations (present in BV5) in which these compounds are more active. The BV samples showed similar cytotoxicity for all the tested tumour cell lines (MCF-7, NCI-H460, HeLa and HepG2), being MCF-7 and HeLa the most susceptible ones. Nevertheless, the studied samples seem to be suitable to treat breast, hepatocellular and cervical carcinoma because at the active concentrations, the samples were not toxic for non-tumour cells (PLP2). Regarding the non-small cell lung carcinoma, BV should be used under the toxic concentration for non-tumour cells. Overall, the present study corroborates the enormous bioactive potential of BV being the first report on samples from Portugal.

Is honey able to potentiate the antioxidant and cytotoxic properties of medicinal plants consumed as infusions for hepatoprotective effects?

Due to the enormous variety of phytochemicals present in plants, their extracts have been used for centuries in the treatment of innumerable diseases, being perceived as an invaluable source of medicines for humans. Furthermore, the combination of different plants was reported as inducing an improved effect (synergism) in comparison with the additive activity of the plants present in those mixtures. Nevertheless, information regarding the effects of plant infusions added with honey is still rather scarce. Accordingly, the aim of this study was to evaluate the interaction between chestnut honey, a natural product with well-reported beneficial properties, and three medicinal plants (either as a single plant or as combinations of two and three plants), with regard to their antioxidant activity and hepatotoxicity. Antioxidant activity was evaluated by comparing the results from four different assays; hepatotoxicity was assessed in two different cell lines. Results were compared by analysis of variance and linear discriminant analysis. The addition of honey to the infusions had a beneficial result in both cases, producing a synergistic effect in all samples, except ß-carotene bleaching inhibition for artichoke + milk thistle + honey preparation and also preparations with lower hepatotoxicity, except in the case of artichoke + honey. Moreover, from the discriminant linear analysis output, it became obvious that the effect of honey addition overcame that resulting from using single plant or mixed plant based infusions. Also, the enhanced antioxidant activity of infusions containing honey was confirmed by lower hepatotoxicity.

In vitro evaluation of Portuguese propolis and floral sources for antiprotozoal, antibacterial and antifungal activity.

Propolis is a beehive product with a very complex chemical composition, used since ancient times in several therapeutic treatments. As a contribution to the improvement of drugs against several tropical diseases caused by protozoa, we screened Portuguese propolis and its potential floral sources Populus x Canadensis and Cistus ladanifer against Plasmodium falciparum, Leishmania infantum, Trypanosoma brucei and Trypanosoma cruzi. The toxicity against MRC-5 fibroblast cells was evaluated to assess selectivity. The in vitro assays were performed following the recommendations of WHO Special Programme for Research and Training in Tropical Diseases (TDR) and revealed moderate activity, with the propolis extracts presenting the relatively highest inhibitory effect against T. brucei. Additionally, the antimicrobial activity against Staphylococcus aureus, Candida albicans, Trichophyton rubrum and Aspergillus fumigatus was also verified with the better results observed against T. rubrum. The quality of the extracts was controlled by evaluating the phenolic content and antioxidant activity. The observed biological activity variations are associated with the variable chemical composition of the propolis and the potential floral sources under study.

Potentiating effects of honey on antioxidant properties of lemon-flavoured black tea.

Health benefits including antioxidant potential of black tea (Camellia sinensis), lemon (Citrus limon) and honey bees (Apis mellifera) have been extensively reported. Nevertheless, nothing is reported about the effects of their concomitant use. Herein, those effects were evaluated in infusions of lemon-flavoured black tea with three different kinds of honey (light amber, amber and dark amber) from Lavandula stoechas, Erica sp. pl. and other indigenous floral species from north-east Portugal, a region with high amounts of this food product. Data obtained showed that the use of honey (dark amber>amber>light amber) potentiates the antioxidant activity of lemon-flavoured black tea, increasing the reducing power and lipid peroxidation inhibition properties, as also the antioxidant contents such as phenolics, flavonoids and organic acids including ascorbic acid.

Phenolic characterization of Northeast Portuguese propolis: usual and unusual compounds.

In this study, an ethanolic extract from Portuguese propolis was prepared, fractionated by high-performance liquid chromatography, and the identification of the phenolic compounds was done by electrospray mass spectrometry in the negative mode. This technical approach allowed the identification of 37 phenolic compounds, which included not only the typical phenolic acids and flavonoids found in propolis from temperate zones but also several compounds in which its occurrence have never been referred to in the literature. Four of the novel phenolic compounds were methylated and/or esterified or hydroxylated derivatives of common poplar flavonoids, although six peculiar derivatives of pinocembrin/pinobanksin, containing a phenylpropanoic acid derivative moiety in their structure, were also identified. Furthermore, the Portuguese propolis sample was shown to contain a p-coumaric ester derivative dimer.

Effect of microwave heating with different exposure times on physical and chemical parameters of olive oil.

This study reflects the effect of different microwave heating times (1, 3, 5, 10 and 15 min) on physical and chemical characteristics of three Portuguese olive oils from three protected designation of origin (PDO), "Azeite de Trás-os-Montes PDO", "Azeites da Beira Interior PDO", and "Azeite de Moura PDO". The parameters evaluated were free acidity, peroxide value, specific extinction coefficients (K232 and K270), color and chlorophylls and carotenoids content. A differential pulse voltammeter was also used to monitor the changes in alpha-tocopherol content. The results showed that microwave heating produce losses in the quality of the different analysed olive oils. The heating time did not promote the occurrence of hydrolysis in the samples since no changes in free acidity values were found. All other parameters were affected by exposure time in a similar way: in the first 3 min no marked changes were observed, after that the quality of the oil decrease significantly. The microwave heating time also affects the total chlorophylls, carotenoids and alpha-tocopherol contents which clearly decreased as long as the exposure time increases. After 15 min of heating the electrochemical signal, due to the alpha-tocopherol, disappear completely in the voltamogram.