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- Response surface methodology in the optimization of extraction conditions for Gracilaria gracilis extracts for use in thermoplastic food coatingsPublication . Reboleira, João; Andrade, Mariana A.; Sanches-Silva, Ana; Ganhão, Rui; Mendes, Susana; Vilarinho, Fernanda; Félix, Rafael; Pinheiro, Joaquina; Bernardino, Susana M.Incorporation of antioxidant agents in edible films and packages often relies in the usage of essential oils and other concentrated hydrophobic liquids, with reliable increases in antimicrobial and antioxidant activities of the overall composite, and consequent improvements in product shelf life. These oils are often obtained from plant sources and the usage of organic solvents in their extraction has been criticized as to whether it constitutes a health hazard for consumers [1, 2, 3]. The use of water-soluble antioxidant agents is less common, due to lower activities and extraction yields. As such, using commonly available seaweeds as the source for both the main polymers and the supplementing antioxidant/antimicrobial agents in bioactive films can potentially reduce production costs and create a safer, more sustainable product [1]. Hydroethanolic extracts of commercially available red macroalgae Gracilaria gracilis were evaluated for their antioxidant potential and phenolic content, as part of the preliminary assays for the selection of algal biomass for the enrichment of thermoplastic films. The extracts were obtained through use of solid-liquid extractions, over which yield, DPPH radical reduction capacity, total phenolic content, and FRAP activity assays were measured [4, 5]. Solid to liquid ratio (SLR), extraction time, and ethanol to water ratio were selected as independent variables with experimental ranges and configurations obtained using a Box-Behnken design with three factors, resulting in 15 experimental conditions. Extraction duplicates were used [4]. Response surface methodology was then used to estimate the effect of each extraction condition on the tested bioactivities. Bioactivities were significantly (p<0.05) and positively affected by the presence of ethanol in the extraction solvent, while extraction yield was reduced. Lower solid to liquid ratios (higher solvent proportions) resulted in higher yields, but had no significant (p<0.05) impact on bioactivities. Time was not a relevant factor in any of the measured variables, and as such will be minimized in any further work. 100% ethanol with minimum extraction time (10 minutes) and 1g to 5mL SLR maximize antioxidant activities per gram of dry extract. Maximum ethanolic yields were obtained with a theoretical extraction of 100 minutes and a 1g to 25mL SLR. Aqueous extractions displayed similar results in terms of optimum conditions, but with overall lower bioactivities and higher yields, very likely due to high amounts of soluble polysaccharides. While the high antioxidant activities from the ethanolic extracts encourage the selection of this solvent for extraction of bioactives, low yields may make them later unfeasible. Future studies over the compatibility of these extracts in the polymeric matrix of the films, as well as further optimization of the extraction process will be necessary before the definitive choice for bioactive origin and processing is made.
- Novel active food packaging films based on whey protein incorporated with seaweed extract: development, characterization, and application in fresh poultry meatPublication . Andrade, Mariana A.; Barbosa, Cássia H.; Souza, Victor G. L.; Coelhoso, Isabel M.; Reboleira, João; Bernardino, Susana; Ganhão, Rui; Mendes, Susana; Fernando, Ana Luisa; Vilarinho, Fernanda; Silva, Ana Sanches; Ramos, FernandoAlgae and seaweeds are used in cookery since the beginnings of human civilization, particularly in several Asian cultures. Phenolic compounds are secondary metabolites produced by aquatic and terrestrial plants for their natural defense against external stimuli, which possess powerful antimicrobial and antioxidant properties that can be very important for the food industry. The main objective of this study was to develop a whey protein concentrate active coating, incorporated with a Fucus vesiculosus extract in order to delay the lipid oxidation of chicken breasts. Ten hydroethanolic extracts from F. vesiculosus were obtained and their antioxidant capacity was evaluated through two antioxidant activity assays: the DPPH radical scavenging activity and -carotene leaching assay. The total content in phenolics compounds was also determined by Folin-Ciocalteu method. The chosen extract was the one obtained from the freeze-dried F. vesiculosus using 75% (v/v) ethanol as extraction solvent. The extract was successfully incorporated into a whey protein film and successfully strengthened the thickness, tensile strength, and elastic modulus. The active film also was able to inhibit the chicken breasts lipid oxidation for 25 days of storage.
- Enhancement of the antioxidant and antimicrobial activities of porphyran through chemical modification with tyrosine derivativesPublication . Adão, Pedro; Reboleira, João; Teles, Marco; Santos, Beatriz; Ribeiro, Nádia; Teixeira, Carlos M.; Guedes, Mafalda; Pessoa, João Costa; Bernardino, Susana M.The chemical modification of porphyran hydrocolloid is attempted, with the objective of enhancing its antioxidant and antimicrobial activities. Sulfated galactan porphyran is obtained from commercial samples of the red algae Porphyra dioica using Soxhlet extraction with water at 100ºC and precipitation with isopropyl alcohol. The extracted porphyran is then treated with modified L-tyrosines in aqueous medium in the presence of NaOH, at ca. 70ºC. The modified tyrosines L1 and L2 are prepared through a Mannich reaction with either thymol or 2,4-di-tert-butylphenol, respectively. While the reaction with 2,4-di-tert-butylphenol yields the expected tyrosine derivative, a mixture of products is obtained with thymol. The resulting polysaccharides are structurally characterized and the respective antioxidant and antimicrobial activities are determined. Porphyran treated with the N-(2-hydroxy-3,5-di-tert-butyl-benzyl)-L-tyrosine derivative, POR-L2, presents a noticeable superior radical scavenging and antioxidant activity compared to native porphyran, POR. Furthermore, it exhibited some antimicrobial activity against S. aureus. The surface morphology of films prepared by casting with native and modified porphyrans is studied by SEM/EDS. Both POR and POR-L2 present potential applicability in the production of films and washable coatings for food packaging with improved protecting characteristics.
- Optimization of extraction conditions for Gracilaria gracilis extracts and their antioxidative stability as mart of microfiber food coating additivesPublication . Reboleira, João; Ganhão, Rui; Mendes, Susana; Adão, Pedro; Andrade, Mariana; Vilarinho, Fernanda; Sanches-Silva, Ana; Sousa, Dora; Mateus, Artur; Bernardino, SusanaIncorporation of antioxidant agents in edible films and packages often relies in the usage of essential oils and other concentrated hydrophobic liquids, with reliable increases in antimicrobial and antioxidant activities of the overall composite, but with less desirable synthetic sources and extraction methods. Hydroethanolic extracts of commercially-available red macroalgae Gracilaria gracilis were evaluated for their antioxidant potential and phenolic content, as part of the selection of algal biomass for the enrichment of thermoplastic film coatings. The extracts were obtained through use of solid-liquid extractions, over which yield, DPPH radical reduction capacity, total phenolic content, and FRAP activity assays were measured. Solid-to-liquid ratio, extraction time, and ethanol percentages were selected as independent variables, and response surface methodology (RSM) was then used to estimate the e ect of each extraction condition on the tested bioactivities. These extracts were electrospun into polypropylene films and the antioxidant activity of these coatings was measured. Similar bioactivities were measured for both 100% ethanolic and aqueous extracts, revealing high viability in the application of both for antioxidant coating purposes, though activity losses as a result of the electrospinning process were above 60% in all cases.
- Poultry shelf-life enhancing potential of nanofibers and nanoparticles containing Porphyra dioica extractsPublication . Reboleira, João; Adão, Pedro; Guerreiro, Sara F.C.; Dias, Juliana R.; Ganhão, Rui; Mendes, Susana; Andrade, Mariana; Vilarinho, Fernanda; Sanches-Silva, Ana; Mateus, Artur; Alves, Nuno; Bernardino, Susana M.Aqueous extracts of commercially available red macroalgae Porphyra dioica were integrated as inner coatings of food-grade polypropylene (PP) films through use of electrospinning and electrospraying technologies. Two coating formulations (A = 5 wt% P. dioica extract and 7.5 wt% polyvinyl alcohol (PVA); B = 1 wt% P. dioica extract, 1 wt% PVA, and 17% gelatine) were evaluated as to their capacity to delay spoilage of minced chicken breasts, through monitoring of microbial growth (total mesophile aerobic colony counts), colour stability, lipid oxidation (thiobarbituric acid reactive substances (TBARS)), and sensory analysis over a 4-day refrigerated storage. Scanning electron microscopy (SEM) imaging revealed an increased nanofiber and nanoparticle density on extract-enriched fibers, without compromise to their morphology or the homogeneity of the coatings. Total microbial counts on coating B samples was significantly (p < 0.001) reduced compared to uncoated plastic wraps. The coated samples also exhibited fewer colour degradation, though the coatings did not di er substantially from uncoated plastic wrap. Sensory analysis test subjects successfully distinguished the raw samples based on their treatment and gave a positive approval rating (66.7%) to the extract-enriched coatings when asked about edibility post storage.
- Red seaweed-derived compounds as a potential new approach for acne vulgaris carePublication . Januário, Adriana P.; Félix, Rafael; Félix, Carina; Reboleira, João; Valentão, Patrícia; Lemos, Marco F.L.Acne vulgaris (AV) is a chronic skin disease of the pilosebaceous unit affecting both adolescents and adults. Its pathophysiology includes processes of inflammation, increased keratinization, sebum production, hormonal dysregulation, and bacterial Cutibacterium acnes proliferation. Common AV has been treated with antibiotics since the 1960s, but strain resistance has emerged and is of paramount concern. Macroalgae are known producers of substances with bioactive properties, including anti-viral, antibacterial, antioxidant, and anti-inflammatory properties, among several others. In particular, red algae are rich in bioactive compounds such as polysaccharides, phenolic compounds, lipids, sterols, alkaloids, and terpenoids, conferring them antioxidant, antimicrobial, and anti-inflammatory activities, among others. Thus, the exploration of compounds from marine resources can be an appealing approach to discover new treatment options against AV. The aim of this work is to provide an overview of the current knowledge of the potentialities of red macroalgae in the treatment of AV by reviewing the main therapeutic targets of this disease, and then the existence of compounds or extracts with bioactive properties against them.
- Uncovering the bioactivity of Aurantiochytrium sp.: a comparison of extraction methodologiesPublication . Reboleira, João; Félix, Rafael; Vicente, Tânia F. L.; Januário, Adriana P.; Félix, Carina; Melo, Marcelo M.R. de; Silva, Carlos M.; Ribeiro, Ana C.; Saraiva, Jorge A.; Bandarra, Narcisa M.; Sapatinha, Maria; Paulo, Maria C.; Coutinho, Joana; Lemos, Marco F.L.Aurantiochytrium sp. is an emerging alternative source of polyunsaturated fatty acids (PUFAs), docosahexaenoic acid (DHA), and squalene, playing an important role in the phasing out of traditional fish sources for these compounds. Novel lipid extraction techniques with a focus on sustainability and low environmental footprint are being developed for this organism, but the exploration of other added-value compounds within it is still very limited. In this work, a combination of novel green extraction techniques (high hydrostatic pressure extraction (HPE) and supercritical fluid extraction (SFE)) and traditional techniques (organic solvent Soxhlet extraction and hydrodistillation (HD)) was used to obtain lipophilic extracts of Aurantiochytrium sp., which were then screened for antioxidant (DPPH radical reduction capacity and ferric-reducing antioxidant potential (FRAP) assays), lipid oxidation protection, antimicrobial, anti-aging enzyme inhibition (collagenase, elastase and hyaluronidase), and anti-inflammatory (inhibition of NO production) activities. The screening revealed promising extracts in nearly all categories of biological activity tested, with only the enzymatic inhibition being low in all extracts. Powerful lipid oxidation protection and anti-inflammatory activity were observed in most SFE samples. Ethanolic HPEs inhibited both lipid oxidation reactions and microbial growth. The HD extract demonstrated high antioxidant, antimicrobial, and antiinflammatory activities making, it a major contender for further studies aiming at the valorization of Aurantiochytrium sp. Taken together, this study presents compelling evidence of the bioactive potential of Aurantiochytrium sp. and encourages further exploration of its composition and application.
- Seaweed fermentation within the fields of food and natural productsPublication . Reboleira, João; Silva, Susana; Chatzifragkou, Afroditi; Niranjan, Keshavan; Lemos, Marco F.L.Background: Seaweeds are promising substrates for biotransformation via fermentation, something that has been primarily utilized by the field of biofuels but focused less attention from other fields of research. Considering that the fermentation of abundant land resources has become an important means by which new added-value compounds can be obtained, exploring the same process for seaweeds can contribute to an effective and sustainable exploitation of marine resources. Scope and approach: In this review, recent advances demonstrating the potential behind the fermentation of seaweeds are evaluated. A breakdown of the most relevant seaweed compounds and their effect on potential bioprocesses is presented, along with pre-processing techniques that have become popular in biofuel fermentations. The applications of seaweed fermentation products in the fields of natural product research, functional foods and nutraceuticals, as well as the limitations and opportunities of seaweed fermentation are also highlighted. Key findings and conclusions: Research revealing that seaweed fermentation can be used to create novel food and nutraceutical products that demonstrate high bioactivity and sensory quality was presented. The studies included demonstrate the use of this process in algal tissues and extracts as an enhancer of antioxidant, antimicrobial, antiinflammatory and antidiabetic activities, among others. Many of the difficulties related to fermenting seaweed have been addressed by research within the field of biofuels, providing insight on the conditions and pretreatments necessary to improve seaweed fermentability. Food applications for seaweed fermentation products are still underdeveloped, but the nutritional, sensory and bioactive profiles collected so far highly encourage further developments.