Ana M.M. Sousa

Agar extraction from integrated multitrophic aquacultured Gracilaria vermiculophylla: Evaluation of a microwave-assisted process using response surface methodology

Microwave-assisted extraction (MAE) of agar from Gracilaria vermiculophylla, produced in an integrated multitrophic aquaculture (IMTA) system, from Ria de Aveiro (northwestern Portugal), was tested and optimized using response surface methodology. The influence of the MAE operational parameters (extraction time, temperature, solvent volume and stirring speed) on the physical and chemical properties of agar (yield, gel strength, gelling and melting temperatures, as well as, sulphate and 3,6-anhydro-L-galactose contents) was evaluated in a 2(4) orthogonal composite design. The quality of the extracted agar compared favorably with the attained using traditional extraction (2 h at 85 degrees Celsius) while reducing drastically extraction time, solvent consumption and waste disposal requirements. Agar MAE optimum results were: an yield of 14.4 + or - 0.4%, a gel strength of 1331 + or - 51 g/cm(2), 40.7 + or - 0.2 degrees Celsius gelling temperature, 93.1 + or - 0.5 degrees Celsius melting temperature, 1.73 + or - 0.13% sulfate content and 39.4 + or - 0.3% 3,6-anhydro-L-galactose content. Furthermore, this study suggests the feasibility of the exploitation of G. vermiculophylla grew in IMTA systems for agar production.

Electrospinning of agar/PVA aqueous solutions and its relation with rheological properties

In this work, we report the successful fabrication of agar-based nanofibers by electrospinning technique, using water as solvent media. A tubeless spinneret was attached inside the electrospinning chamber, operating at 50°C, to avoid agar gelation. Agar pure solution (1 wt%) showed inadequate spinnability regardless of the used electrospinning conditions. The addition of a co-blending polymer such as PVA (10 wt% starting solution) improved the solutions viscoelasticity and hence, the solutions spinnability. Agar/PVA solutions were prepared with different mass ratios (100/0, 50/50, 40/60, 30/70, 20/80 and 0/100) and electrospun at various sets of electrospinning conditions. Best nanofibers were obtained with 30/70 and 20/80 agar/PVA blends while samples with higher agar contents (50/50 and 40/60 agar/PVA) were harder to process and led to discontinuous fibrous mats. This first set of encouraging results can open a new window of opportunities for agar-based biomaterials in the form of nanofibers.

Influence of the extraction process on the rheological and structural properties of agars.

Agars obtained by traditional hot-water (TWE) and microwave-assisted (MAE) extractions were compared in terms of their rheological and physicochemical properties and molecular self-association in solutions of low (0.05%, w/w) and high (1.5%, w/w) polymer concentrations. At low concentration, thin gelled layers were imaged by AFM. Slow or rapid cooling of the solutions influenced structure formation. In each case, TWE and MAE agar structures were different and apparently larger for MAE. At high concentration, progressive structural reinforcement was seen; while TWE agar showed a more open and irregular 3D network, MAE agar gel imaged by cryoSEM was denser and fairly uniform. The rheological (higher thermal stability and consistency) and mechanical (higher gel strength) behaviors of MAE agar seemed consistent with a positive effect of molecular mass and 3,6-anhydro-α-l-galactose content. MAE produced non-degraded agar comparable with commercial ones and if properly monitored, could be a promising alternative to TWE.

Biodegradable Agar Extracted from Gracilaria Vermiculophylla: Film Properties and Application to Edible Coating

Gracilaria is a red algal genus that biosynthesizes a polymer called agar that is extensively used in the food and pharmaceutical industries as gelling and stabilizing agent. In the last years, many studies have focused on gel properties of this biopolymer; but the agar films and coatings still have few studies reported. The edible film and coating have a protective function, preventing moisture, oxygen and flavour transfers between food and surroundings. The objectives of this work are the production of biodegradable agar films from Gracilaria vermiculophylla, collected in Ria de Aveiro, Portugal, and the study of the effect of glycerol, an hydrophilic plasticizer, on the properties of the films and on subsequent application in edible coating of fresh fruits and vegetables. The agar extraction was carried out at specific optimum parameters determined in previous work (3.5 h pre-treatment duration, 6% NaOH concentration and 2 h extraction time). Agar films were made using the knife coating technique and compared with commercial agar. The physical properties of films such as hygroscopicity, mechanical resistance (Young’s modulus, tensile strength and elongation), and permeability to water vapour and oxygen were characterized. As expected, the plasticizer addition revealed and increase on elongation and decrease on tensile strength. The films were transparent and optically clear, showing good properties similar to the commercial agar films. The potential application of the agar/glycerol solution to fresh vegetable preservation was tested. Model fruits and vegetables were coated with the biopolymer/plasticizer solution and compared with a control sample in terms of colour, firmness, weight loss and shelf life. Considering on one hand the abundance of the raw algal material which is actually an invasive species, and the properties of the agar films and coatings obtained on the other hand, commercial use of G.vermiculophylla from Ria de Aveiro is well justified.

Choline chloride based ionic liquid analogues as tool for the fabrication of agar films with improved mechanical properties.

In the present paper, we test the suitability of ChCl/urea (DES-U) and ChCl/glycerol (DES-G) eutectic mixtures, each one prepared at 1:2 molar ratio, for the production of agar films. A three-step process is proposed: pre-solubilization of polymer in DES followed by compression-molding and subsequent drying. The mechanical properties, water resistance and microstructure of the films were evaluated at different polymer concentrations (i.e. 2-6%, w/w). DES-U showed by far, the best film forming ability. Agreeing with the diffusion and SEM data, films with the best mechanical properties were found at the lowest and highest agar concentrations (tensile strengths of 24.2-42 MPa and elongations of 15.4-38.9%). The water sorption and contact angle studies suggested increased hydrophilicity for the film containing the lowest concentration of agar. The use of choline chloride based ionic liquid analogues as solvent and plasticizer might be a promising tool for the development of new non-aqueous materials based on seaweed polysaccharides.

Shaping the molecular assemblies of native and alkali-modified agars in dilute and concentrated aqueous media via microwave-assisted extraction

The use of agar-based biomaterials for the development of emerging areas, such as tissue engineering or ‘smart materials’ production has recently gained great interest. Understanding how these gel-forming polysaccharides self-organise in aqueous media and how these associations can be tuned to meet the specific needs of each application is thus of great relevance. As an extension of previous pioneering research concerning the application of the microwave-assisted extraction (MAE) technique in the recovery of native (NA) and alkali-modified (AA) agars, this article focuses on the different molecular assemblies assumed by these novel NA and AA when using different MAE routes. The molecular architectures in dilute (5, 10, 50 and 100 μg mL−1) and concentrated (1.5% (w/w)) aqueous media were imaged by AFM and cryoSEM, respectively. Relevant structural and physicochemical properties were investigated to support the microscopic data. Different extraction routes led to polysaccharides with unique properties, which in turn resulted in different molecular assemblies. Even at 5 μg mL−1, AFM images included individual fibers, cyclic segments, aggregates and local networks. At higher polymer concentrations, the structures further aggregated forming multilayer polymeric networks for AA. The more compact and denser 3D networks of AA, imaged by cryoSEM, and their higher resistance to large deformations matched the 2D-shapes observed by AFM. Depending on the nature of the AA chains, homogeneous or heterogeneous growth of assemblies was seen during network formation. The obtained results support well the view of double helix formation followed by intensive double helix association proposed for agar gelation.

Thermo-compression molding of chitosan with a deep eutectic mixture for biofilms development

A eutectic mixture of choline chloride (ChCl) and citric acid (CA) was successfully used for the preparation of chitosan (Chit) bio-films by thermo-compression molding. Optimization of the film preparation conditions was carried out using response surface methodology and a Box-Behnken design based on the best mechanical properties and lowest energetic requirements (lower compression load and time). The optimum film, made with chitosan and ChCl–CA (eutectic mixture), was compared with films prepared using only CA in their formulation. Chit–ChCl–CA films presented higher elasticity, opacity and total color difference, and a lower tensile strength in relation to the Chit–CA films. Films prepared with the eutectic mixture presented higher water vapor permeability values. These results were associated with the films microstructures. FTIR analysis confirms the occurrence of chemical changes in the processed films. The thermo-compression process significantly affects the crystallinity of the pristine chitosan. The films thermal stability is depressed when compared to the stability of each component.

Electrospun ultra-fine cellulose acetate fibrous mats containing tannic acid-Fe3+ complexes

Cellulose acetate (CA) fibrous mats with improved mechanical and antioxidant properties were produced by a simple, scalable and cost-effective electrospinning method. Fibers loaded with small amounts of TA-Fe3+ complexes showed an increase in tensile strength of ∼117% when compared to that of neat CA and were more resistant than those loaded with TA alone. The water uptake of the fibers increased upon TA or TA-Fe3+ incorporation while their thermal behavior was only slightly affected. Fibrous mats loaded with TA-Fe3+ showed comparable antioxidant activity with that of CA/TA mats, and a much slower TA release. These results suggest that TA-Fe3+ complexes can be incorporated into electrospun CA fibers to improve their mechanical properties and antioxidant activity which may be of interest for the development of active packaging that can extend the shelf life of perishable foods.

Influence of Prior Exercise on VO2 Kinetics Subsequent Exhaustive Rowing Performance

Prior exercise has the potential to enhance subsequent performance by accelerating the oxygen uptake (VO2) kinetics. The present study investigated the effects of two different intensities of prior exercise on pulmonary VO2 kinetics and exercise time during subsequent exhaustive rowing exercise. It was hypothesized that in prior heavy, but not prior moderate exercise condition, overall VO2 kinetics would be faster and the VO2 primary amplitude would be higher, leading to longer exercise time at VO2max. Six subjects (mean ± SD; age: 22.9±4.5 yr; height: 181.2±7.1 cm and body mass: 75.5±3.4 kg) completed square-wave transitions to 100% of VO2max from three different conditions: without prior exercise, with prior moderate and heavy exercise. VO2 was measured using a telemetric portable gas analyser (K4b2, Cosmed, Rome, Italy) and the data were modelled using either mono or double exponential fittings. The use of prior moderate exercise resulted in a faster VO2 pulmonary kinetics response (τ1 = 13.41±3.96 s), an improved performance in the time to exhaustion (238.8±50.2 s) and similar blood lactate concentrations ([La−]) values (11.8±1.7 mmol.L−1) compared to the condition without prior exercise (16.0±5.56 s, 215.3±60.1 s and 10.7±1.2 mmol.L−1, for τ1, time sustained at VO2max and [La−], respectively). Performance of prior heavy exercise, although useful in accelerating the VO2 pulmonary kinetics response during a subsequent time to exhaustion exercise (τ1 = 9.18±1.60 s), resulted in a shorter time sustained at VO2max (155.5±46.0 s), while [La−] was similar (13.5±1.7 mmol.L−1) compared to the other two conditions. Although both prior moderate and heavy exercise resulted in a faster pulmonary VO2 kinetics response, only prior moderate exercise lead to improved rowing performance.

Complexation of WPI and microwave-assisted extracted agars with different physicochemical properties

The complex formation between whey protein isolate (WPI) and agar has been investigated and their interactions were monitored as a function of the physicochemical properties of agar, the pH and the ionic strength of the medium. Agars from Gracilaria vermiculophylla were extracted under different MAE conditions and characterized according to their physicochemical properties. By using microwave irradiation a wide variety of agars was obtained, as different MAE conditions results in polyelectrolytes with distinct properties. UV-vis (in optical dispersion (O.D.) model) spectroscopy and isothermal titration calorimetry (ITC) have been used to study the formation of insoluble (coacervate) complexes. MAE agars revealed excellent properties for complex formation with WPI. The binding of WPI to MAE agar samples has been shown to be the result of different contributions. O.D. and ITC results showed that the molecular mass and the sulfate content of different agars had a determinant effect on coacervate formation.