João C.M. Barreira

Targeting excessive free radicals with peels and juices of citrus fruits: Grapefruit, lemon, lime and orange

A comparative study between the antioxidant properties of peel (flavedo and albedo) and juice of some commercially grown citrus fruit (Rutaceae), grapefruit (Citrus paradisi), lemon (Citrus limon), lime (Citrusxaurantiifolia) and sweet orange (Citrus sinensis) was performed. Different in vitro assays were applied to the volatile and polar fractions of peels and to crude and polar fraction of juices: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, reducing power and inhibition of lipid peroxidation using beta-carotene-linoleate model system in liposomes and thiobarbituric acid reactive substances (TBARS) assay in brain homogenates. Reducing sugars and phenolics were the main antioxidant compounds found in all the extracts. Peels polar fractions revealed the highest contents in phenolics, flavonoids, ascorbic acid, carotenoids and reducing sugars, which certainly contribute to the highest antioxidant potential found in these fractions. Peels volatile fractions were clearly separated using discriminant analysis, which is in agreement with their lowest antioxidant potential.

Dietary antioxidant supplements: benefits of their combined use.

Several dietary supplements claim medicinal benefits due to their composition in hydrophilic and lipophilic molecules, natural extracts or synthetic compounds with antioxidant properties. In the present work, the antioxidant activity of selected supplements taken in pills, capsules or infusions were studied either individually or combined. Linear discriminant analysis (LDA) was used to categorize the condensed formulations (pills and capsules), infusion bags and combined samples according with their antioxidant activity measured by radical scavenging activity, reducing power and lipid peroxidation inhibition using brain homogenates as models. AAF proved to have the highest antioxidant activity in all the assayed methods, either singly taken or included in mixtures. Furthermore, the mixtures containing this supplement revealed synergistic effects in 92% of the cases. The intake of antioxidant mixtures might provide some additional benefits.

Assessing the effects of gamma irradiation and storage time in energetic value and in major individual nutrients of chestnuts.

Chestnut (Castanea sativa Miller) is an important food resource all over the world. In the present study, it is intended to evaluate if the application of gamma irradiation doses ≤ 3 kGy maintain chestnuts chemical and nutritional profiles unaffected. Furthermore, possible interactions among irradiation dose and storage time were accessed using linear discriminate analysis (LDA). The nutritional composition was evaluated through determination of proteins, fat, ash, carbohydrates and energetic value. The chemical composition was focused in the main nutrients found in chestnuts: sugars - sucrose, fatty acids - palmitic, oleic, linoleic and linolenic acids, tocopherols - γ-tocopherol. The obtained results seem to indicate that the irradiation treatment did not affect the nutritional and chemical quality of chestnut fruits. Otherwise, storage time exerted more evident influence in those parameters. The application of gamma irradiation emerges as a promising technology for chestnuts chemical quality, but food safety issues have to be evaluated in order to recommend its application as a useful conservation alternative.

Analysis of organic acids in electron beam irradiated chestnuts (Castanea sativa Mill.): effects of radiation dose and storage time

Since 2010, methyl bromide, a widely used fumigant was banned from the European Union under the Montreal Protocol guidelines, due to its deleterious effects on health and risk to the environment. Since then, many alternatives for chestnut conservation have been studied (hot water dip treatment being the most common), among them, electron beam irradiation has been proposed as being a safe, clean and cheap alternative. Herein, the effects of this radiation at different doses up to 6kGy and over storage up to 60days in the amounts and profile of nutritionally important organic acids were evaluated. Chestnuts contained important organic acids with quinic and citric acids as main compounds. Storage time, which is traditionally well accepted by consumers, caused a slight decrease on quinic (13-9mg/g), ascorbic (1.2-0.8mg/g), malic (5-4mg/g), fumaric (0.4-0.3mg/g) and total organic (33-26mg/g) acids content. Otherwise, irradiation dose did not cause appreciable changes, either individually or in total (28-27mg/g) organic acid contents. Electron beam irradiation might constitute a valuable alternative for chestnut conservation.

Influence of gamma irradiation in the antioxidant potential of chestnuts (Castanea sativa Mill.) fruits and skins

As seasonal products chestnuts have to be post-harvest treated to increase their shelf-life. The most common preservation method for chestnuts is the chemical fumigation with methyl bromide, a toxic agent that is under strictly Montreal Protocol due to its adverse effects on human health and environment. Food irradiation is a possible feasible alternative to substitute the traditional quarantine chemical fumigation treatment. This preliminary study evaluated the influence of gamma irradiation in the antioxidant potential of chestnut fruits and skins, through several chemical and biochemical parameters. The bioactive compounds (phenolics and flavonoids) and DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging activity, reducing power and inhibition of β-carotene bleaching capacity were determined. The obtained results seem to indicate that the storage favoured chestnuts antioxidant potential. Furthermore, the application of gamma irradiation also seems to be advantageous for antioxidant activity, independently of the dose used (0.27 ± 0.04 kGy or 0.54 ± 0.04 kGy).

Effects of electron-beam radiation on nutritional parameters of Portuguese chestnuts (Castanea sativa Mill.).

Chestnuts are a widely consumed fruit around the world, with Portugal being the fourth biggest producer in Europe. Storage of these nuts is an important step during processing, and the most widely used fumigant was banned in the European Union under the Montreal Protocol because of its toxicity. Recently, radiation has been introduced as a cheap and clean conservation method. Previous studies of our research group proved that γ radiation had no negative effect on the nutritional value of chestnuts; in fact, storage time had a much bigger influence on the chestnut quality. In the present study, we report the effect of a less ionizing radiation, electron beam, with doses of 0, 0.5, 1, 3, and 6 kGy in the nutritional value of chestnuts (ash, energy, fatty acids, sugars, and tocopherols), previously stored at 4 °C for 0, 30, and 60 days. The storage time seemed to reduce fat and energetic values but reported a tendency for higher values of dry matter. With regard to fatty acids, there was a higher detected quantity of C20:2 in non-irradiated samples and four fatty acids were only detected in trace quantities (C6:0, C8:0, C10:0, and C12:0). γ-Tocopherol decreased during storage time but did not alter its quantity for all of the radiation doses (as like α-, β-, and δ-tocopherol); in fact, these compounds were present in higher concentrations in the irradiated samples. Sucrose and total sugars were lower in non-irradiated samples, and raffinose was only detected in irradiated samples. Electron-beam irradiation seems to be a suitable methodology, because the effects on chemical and nutritional composition are very low, while storage time seems to be quite important in chestnut deterioration.

Antioxidant Potential of Chestnut (Castanea sativa L.) and Almond (Prunus dulcis L.) By-products

The antioxidant properties of almond green husks (Cvs. Duro Italiano, Ferraduel, Ferranhês, Ferrastar and Orelha de Mula), chestnut skins and chestnut leaves (Cvs. Aveleira, Boa Ventura, Judia and Longal) were evaluated through several chemical and biochemical assays in order to provide a novel strategy to stimulate the application of waste products as new suppliers of useful bioactive compounds, namely antioxidants. All the assayed by-products revealed good antioxidant properties, with very low EC50 values (lower than 380 μg/mL), particularly for lipid peroxidation inhibition (lower than 140 μg/mL). The total phenols and flavonoids contents were also determined. The correlation between these bioactive compounds and DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, reducing power, inhibition of β-carotene bleaching and inhibition of lipid peroxidation in pig brain tissue through formation of thiobarbituric acid reactive substances, was also obtained. Although, all the assayed by-products proved to have a high potential of application in new antioxidants formulations, chestnut skins and leaves demonstrated better results.

Low dose γ-irradiation as a suitable solution for chestnut (Castanea sativa Miller) conservation: effects on sugars, fatty acids, and tocopherols.

Along with dehydration, the development of insects and microorganisms is the major drawback in chestnut conservation. Irradiation has been regaining interest as an alternative technology to increase food product shelf life. In the present work, the effects of low dose gamma irradiation on the sugar, fatty acid, and tocopherol composition of chestnuts stored at 4 °C for different storage periods (0, 30, and 60 days) was evaluated. The irradiations were performed in a 60Co experimental equipment, for 1 h (0.27±0.04 kGy) and 2 h (0.54±0.04 kGy). Changes in sugars and tocopherols were determined by high performance liquid chromatography coupled to refraction index and fluorescence detections, respectively, while changes in fatty acids were analyzed by gas-chromatography coupled to flame ionization detection. Regarding sugar composition, storage time proved to have a higher effect than irradiation treatment. Fructose and glucose increased after storage, with the corresponding decrease of sucrose. Otherwise, the tocopherol content was lower in nonirradiated samples, without a significant influence of storage. Saturated, monounsaturated, and polyunsaturated fatty acids levels were not affected, either by storage or irradiation. Nevertheless, some individual fatty acid concentrations were influenced by one of two factors, such as the increase of palmitic acid in irradiated samples or the decrease of oleic acid after 60 days of storage. Overall, the assayed irradiation doses seem to be a promising alternative treatment to increase chestnut shelf life, without affecting the profile and composition in important nutrients.

In search of synergistic effects in antioxidant capacity of combined edible mushrooms

The antioxidant activity of different edible mushrooms was evaluated considering the different contribution of individual and combined extracts. The radical scavenging capacity was evaluated through hydrogen atom transfer and single electron transfer reaction-based assays: DPPH radical scavenging activity and reducing power, respectively. The inhibition of lipid peroxidation was studied in lipossomes solutions by the β-carotene-linoleate system. Three types of interactions (synergistic, additive and negative synergistic effects) were observed, synergism being the most abundant effect. Marasmius oreades is present in the mixtures with higher antioxidant properties and synergistic effects, while Cantharellus cibarius is present in the mixtures with lowest antioxidant properties and negative synergist effects. Two discriminant analyses were performed considering individual species in one case and mushroom mixtures in the other. The five mushroom species were clustered in five individual groups, but a similar result could not be obtained for the combined mushrooms, for which only the cases containing C. cibarius were separated in individual clusters.

Edible flowers of Viola tricolor L. as a new functional food: Antioxidant activity, individual phenolics and effects of gamma and electron-beam irradiation

Edible flowers are used in food preparations, being also recognized for their beneficial effects on human health. Nevertheless, these species are highly perishable, and irradiation treatment might be applied to ensure food quality and increase their shelf life. Viola tricolor L. is a typical edible flower, with multiple applications and biological properties, mainly provided by the flavonoid content. In the present work, the phenolic compounds were analyzed by HPLC-DAD-ESI/MS, and the antioxidant activity was evaluated using biochemical assays. Linear discriminant analyses (LDA) were performed in order to compare the results obtained with flowers submitted to different irradiation doses and technologies (cobalt-60 and electron-beam). In general, irradiated samples (mostly with 1 kGy) showed the highest phenolic content and antioxidant activity. Furthermore, the significant differences observed in the LDA allow determination of which dose and/or technology is suitable to obtain flowers with higher antioxidant potential.