Browsing by Author "Angulo, I."
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- Active Packaging Produced by Extrusion with Shrimp Waste: Migration of Astaxanthin into Food SimulantsPublication . Sanches-Silva, A.; Ribeiro, T.; Albuquerque, T.G.; Paseiro, P.; Sendón, R.; Bernaldo de Quirós, A.; López-Cervantes, J.; Sánchez-Machado, D.; Soto Valdez, H.; Angulo, I.; Pardo Aurrekoetxea, G.; Costa, H.S.Introduction: Astaxanthin (3,3’-dihydroxy-β-β´-carotene-4-4´-dione), a potent antioxidant, is one of the major carotenoids in crustaceans. In the frame of the project ‘Preparation of active packaging with antioxidant and antimicrobial activity based on astaxanthin and chitosan’, a methodology for the incorporation of compounds obtained from shrimp waste in plastic matrices was developed to produce an active packaging with antioxidant properties. The aim of the present work was to develop and optimize a method to determine astaxanthin by ultra-high pressure liquid chromatography in fermented shrimp waste. Moreover, the method was also applied to determine the migration of astaxanthin from plastic films containing different amounts of shrimp waste to food simulants. Material and Methods: The method was optimized to determine astaxanthin by ultra-high pressure liquid chromatography (UHPLC) with diode array detection (DAD). The chromatographic separation was achieved using a vanguard pre-column (UPLCÒ BEH, 1.7 µm particle size) and a column (UPLCÒ BEH, 2.1 x 50 mm, 1.7 µm particle size) at 20 °C. The mobile phase was a gradient of A (dichloromethane/methanol with ammonium acetate/acetonitrile 5:20:75 (v/v)) and B (ultrapure water) with a flow rate of 0.5 mL/min. The optimized UPLC method allowed an excellent resolution of astaxanthin. The method was also evaluated in what concerns to validation parameters such as linearity, precision, limit of detection, limit of quantification and recovery. Low density polyethylene plastic films produced by extrusion with different amounts of the lipid fraction of shrimp waste were prepared and tested regarding migration into fatty food stimulants (isooctane and ethanol 95%, v/v). Results and conclusion: The proposed method to determine astaxanthin in shrimp waste is simple and has a low detection level (0.054 μg/mL). The concentration of astaxanthin found in the lipid fraction of fermented shrimp waste was 453.8 μg/g. The films produced by extrusion with the lipid fraction of the fermented shrimp waste did not originate astaxanthin migration into the tested fatty food simulants. Further studies could be made in order to evaluate the capacity of these films in protecting packed food from oxidation.
- Activity of chitosan films against different microorganismsPublication . Sanches-Silva, A.; Maia, C.; Furtado, R.; Ribeiro, T.; Paseiro, P.; Sendón, R.; Rodríguez-Bernaldo de Quirós, A.; López-Cervantes, J.; Sánchez-Machado, D.I.; Bueno, C.; Soto Valdez, H.; Angulo, I.; Aurrekoetxea, G.P.; Bilbao, A.; Costa, H.S.Chitosan is a hydrophilic polysaccharide which derives from chitin by deacetylation. It has several applications, namely as a film that can be applied to preserve the quality and increase the shelf-life of food. Chitosan is insoluble in most solvents but it is soluble in dilute organic acids such as formic acid and acetic acid[1]. The properties of chitosan depend on the degree of deacetylation (DA) and molecular weight (MW). A broad antimicrobial activity has been attributed to chitosan, either for gram-negative, gram-positive bacteria and fungi. The aim of the present study is to evaluate the antimicrobial activity of a chitosan film prepared by casting. The chitosan was obtained from shrimp waste collected from shrimp processing factories of South Sonora (Mexico). Four bacteria (Bacillus cereus; Escherichia coli; Staphylococcus aureus and Listeria monocytogenes) and one fungus (Botrytis cinerea) were evaluated. Although L. monocytogenes and B. cinerea growth was not inhibited by the chitosan film, results showed a clear growth-inhibitory effect, at the two bacteria concentration levels tested, for B. Cereus, E. coli and S. aureus. Different antibacterial mechanisms have been proposed to explain chitosan antimicrobial activity[2-3]: i) chitosan may form an external barrier which inhibits essential nutrients adsorption; ii) chitosan can also penetrate the microbial cell, disturbing the metabolism of the cell by inhibiting the mRNA and protein synthesis; iii) chitosan may have an ionic surface interaction with the bacteria originating wall cell leakage. Although these mechanisms may take place simultaneously, the antimicrobial activity may also depend on the properties of chitosan (DA and MW).
- Astaxanthin from shrimp by-products for active packagingPublication . Sanches-Silva, A.; Ribeiro, T.; Albuquerque, T.G.; Paseiro, P.; Sendón, R.; Bernaldo de Quirós, A.; López-Cervantes, J.; Sánchez-Machado, D.; Soto Valdez, H.; Angulo, I.; Pardo Aurrekoetxea, G.; Costa, H.S.
- Characterization of chitosan intended to develop antimicrobial films: Microscopical StudiesPublication . Lago, M.A.; Sendón, R.; Rodríguez-Bernaldo de Quirós, A.; Sanches-Silva, A.; Costa, H.S.; Sánchez-Machado, D.I.; Soto Valdez, H.; Angulo, I.; Aurrekoetxea, G.P.; López-Cervantes, J.; Paseiro, P.Accumulation of organic wastes in intensive crustaceans culture ponds and nearby coastal waters has become a serious environmental and economical problem. For this reason, new ecofriendly and economically feasible products from agricultural wastes or byproducts for shrimp farms have been developed. This biowaste could be used as an important source of the useful biopolymer chitin and others components such as proteins or carotenoids like asthaxanthin [1]. Chitin is the most abundant polysaccharide after cellulose and the main source is the shell of crustaceans. Chitosan, derived from chitin, has proven useful for a wide range of applications due to its biodegradability, biocompatibility, antimicrobial activity, non-toxicity and versatile physicochemical properties. These properties make the chitosan an excellent candidate to use in food packaging [2]. The development of active packaging with antimicrobial and antioxidant activity based on chitosan and asthaxanthin obtained from shrimp waste is the main goal of the project: “Preparation of active packaging with antioxidant and antimicrobial activity based on asthaxanthin and chitosan” funded by FONCYCIT. The characterization of chitosan in the development of active materials is a key issue since their properties play an important role in its effectiveness as an antimicrobial agent. These properties are mainly molecular weight (Mw), acetylation degree (DA) and polymerization degree (PA). In addition, in mediums of low pH, the antimicrobial activity of chitosan increases [3]. The objective of the present study was characterized three different samples of chitosan obtained from shrimp waste by using two microscopy techniques, Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Additionally, the films developed after the chitosan incorporation in the polyamide matrix were also characterized. Shrimp waste (heads and cephalotorax) samples were collected from local shrimp processing factories in South Sonora, Mexico. The waste was minced, fermented and centrifuged. After the treatment, three fractions were obtained: chitin-rich fraction, protein rich liquor and lipid fraction. Scanning Electron Microscopy was used to investigate the structure properties relationships of chitosan. Samples were spread on a carbon conducting adhesive tape pasted on a metallic stub, subjected to gold covering and observed. The samples for TEM observation were embedded in EPON resin and polymerized at 60 ºC; then were cut at (-120 ºC) using a Leica Ultracut crio-ultramicrotome. The images obtained showed the particle of chitosan embedded into the polyamide matrix.
- Characterization of chitosan meant for antimicrobial food packagingPublication . Sendón, R.; Rodríguez Bernaldo de Quirós, A.; Bueno, C.; Pereira, D.; Sanches-Silva, A.; Costa, H.S.; Sánchez-Machado, D.I.; Soto Valdez, H.; Angulo, I.; Aurrekoetxea, G.P.; López-Cervantes, J.; Paseiro, P.Chitosan (CAS nº 9012-76-4) is a natural polysaccharide obtained by the partial deacetylation of chitin. It is a linear polymer of β (1-4) 2-acetamido-2-deoxy-D-glucose and 2-amino-2-deoxy-D-glucose in different proportions. Chitin is the most abundant polysaccharide after cellulose and the main source is the shells of crustaceans. It has been demonstrated that some important properties are directly related to the antimicrobial activity of chitosan. Some of these properties are the molecular weight (Mw), the degree of polymerisation (DP) and the degree of deacetylation (DD). In this work several analytical techniques (FTIR (Fourier-transform Infrared Spectroscopy), NMR (Nuclear Magnetic Resonance Spectroscopy) and SEM (Scanning Electron Microscopy)) were attempted to characterize two different samples obtained from shrimp waste. It can be concluded that sample 1 should be more suitable to be added as an active agent to a film.
- Compilation of analytical methods to characterize and determine chitosan, and main applications of the polymer in food active packagingPublication . Lago, M.; Rodríguez-Bernaldo de Quirós, A.; Sendon, R.; Sanches-Silva, A.; Costa, H.S.; Sánchez-Machado, D.; López-Cervantes, J.; Soto-Valdez, H.; Aurrekoetxea, G.P.; Angulo, I.; Paseiro, P.Antimicrobial films for food packaging applications have received increasing attention from the industry in recent years. Due to their exceptional properties, such as non-toxicity, biodegradability, antimicrobial characteristics, and biocompatibility, chitosan has proven useful for the development of active materials. This review aims to provide an overview of the main techniques used for the characterization of chitin and chitosan, including Fourier transform infrared spectroscopy (FTIR), 1H and 13C nuclear magnetic resonance (NMR) spectroscopy, UV spectrophotometry, viscosimetry, elemental analysis, X-ray diffraction (XRD), thermogravimetric analysis (TGA), titrations, scanning electron microscopy SEM) and size exclusion chromatography (SEC) among others. In addition, the main applications of the polymer in food packaging are also reported.
- Determination of Glucosamine by Ultra-high Pressure LC in Shrimp By-ProductsPublication . Sanches-Silva, A.; Ribeiro, T.; Albuquerque, T.G.; Paseiro, P.; Sendón, R.; López-Cervantes, J.; Sánchez-Machado, D.I.; Soto Valdez, H.; Angulo, I.; Aurrekoetxea, G.P.; Costa, H.S.Glucosamine is one of the most abundant monosacharides and is part of the structure of the chitosan and chitin, which compose the exoskeletons of crustaceans, other arthropods and the fungi cell walls. Shrimp by-products are a good source of glucosamine because about 45% of the animal is composed of inedible cephalothorax and exoskeleton. The project ‘Preparation of active packaging with antioxidant and antimicrobial activity based on astaxanthin and chitosan’ aims to develop a methodology for the incorporation of compounds obtained from shrimp waste in plastic matrices for the development of an active packaging with antimicrobial and antioxidant properties. In the frame of this project, shrimp by-products were fermented and insoluble chitin was purified by a sequence of steps: depigmentation, deproteinization, demineralization and blanching. The aim of the present work was to optimize a method to determine glucosamine by Ultra-high Pressure Liquid chromatography (Ultra Performance Liquid Chromatography, UPLC) with diode array detection (DAD) from shrimp by-products. First, an acid hydrolysis of the chitin of shrimp by-products is carried out. Afterwards, a derivatization is done because glucosamine does not contain a chromophore. The selected derivatization reagent was 9-fluorenylmethyl-chloroformate (Fmoc-Cl). The chromatographic separation is achieved using a vanguard pre-column (Acquity UPLCÒ HSS T3, 1.8 µm particle size) and a column (Acquity UPLCÒ HSS T3, 2.1 x 50 mm, 1.7 µm particle size) at 38 °C. Quantification of glucosamine was carried out at 265 nm. The mobile phase is a gradient of phase A [water (pH 6.5)/ methanol, 85:15 (v/v)] and phase B (acetonitrile) with a flow rate of 0.3 mL/min. The optimized UPLC method allows an excellent separation in just 10 min of the two anomers of glucosamine, of the Fmoc-Cl and of its corresponding alcohol, which is the hydrolysis product of the spontaneous reaction between Fmoc-Cl with water. Several samples have been analysed, some are commercial samples (chitosan with low molecular weight; chitosan with medium molecular weight; chitosan from shrimp shells; chitosan from crab shells), and three other samples produced from shrimp by-products in the frame of the project. Commercial samples presented a higher amount of glucosamine content than in-house produced samples.
- Determination of α-tocopherol in shrimp waste to evaluate its potential to produce active packagingPublication . Sanches-Silva, A.; Costa, H.S.; Bueno-Solano, C.; Sendón, R.; Sánchez-Machado, D.I.; Soto Valdez, H.; Colín-Chávez, C.; Angulo, I.; Aurrekoetxea, G.P.; Paseiro, P.; López-Cervantes, J.The possibility of using the lipid fraction from fermented shrimp waste as a source of natural α-tocopherol to produce packaging with antioxidant properties was evaluated. A fast reverse-phase ultra-performance liquid chromatographic (RP-UPLC) method coupled with a diode array detector was developed to determine α-tocopherol in the lipid fraction of shrimp waste. The α-tocopherol level found using acetonitrile as extraction solvent was 50.5 mg/100 g sample.
- Evaluación físico-química de aceite pigmentado obtenido de la cabeza de camarónPublication . Núñez-Gastélum, J.A.; Sánchez-Machado, D.I.; López-Cervantes, J.; Paseiro-Losada, P.; Sendón, R.; Sanches-Silva, A.T.; Costa, H.S.; Aurrekoetxea, G.P.; Angulo, I.; Soto-Valdez, H.En este trabajo se presenta el análisis proximal, caracterización físico-química, perfil de ácidos grasos y contenido de astaxantina en aceite pigmentado aislado por fermentación láctica de los residuos de camarón. Los lípidos son los componentes mayoritarios (95%). El índice de saponificación es 178.62 mg KOH/g, el de yodo 139.8 cg yodo/g, y los peróxidos no fueron detectados. La densidad y la viscosidad fueron de 0.92 mg/ml y 64 centipoises, respectivamente. Los ácidos grasos en mayor cantidad fueron el linoleico (C18:2n6), oleico (C18:1n9) y palmítico (C16:0). El ácido eicosapentaenoico (C20:5n3, EPA) y el docosahexaenoico (C22:6n3, DHA) suman el 9% del total. El contenido promedio de astaxantina fue de 2.72 mg/g base seca. El aceite pigmentado es una fuente dietética de nutrientes con alto valor como la astaxantina.
- Migration of chitosan films prepared by solvent evaporation and extrusionPublication . Sanches-Silva, A.; Ribeiro, T.; Albuquerque, T.G.; Paseiro, P.; Sendón, R.; Bernaldo de Quirós, A.; López-Cervantes, J.; Sánchez-Machado, D.; Soto Valdez, H.; Angulo, I.; Pardo Aurrekoetxea, G.; Costa, H.S.Introduction: Chitosan has multiple applications and as inhibitor of microbial growth, there is interest in developing new methodologies to its incorporation into plastics, which would avoid microbial growth in foods. In the frame of the project “Preparation of active packaging with antioxidant and antimicrobial activity based on astaxanthin and chitosan” (PAPAAABAC), chitosan was extracted from shrimp by-products and used to prepare plastic films by solvent evaporation (casting) and extrusion. Material and Methods: Chitosan films were prepared by extrusion of polyamide and by casting. By casting, films were prepared at different concentrations (1%, 2%, and 3%) by dissolving chitosan in acetic acid aqueous solution 1% (w/v), with and without plasticizer (1% glycerol). By extrusion, chitosan pellets were made of polyamide 6 at 2%, 5%, 6%, 8%, and 10% with two different particle sizes (180 and 300 µm). Then, they were rolled in an extruder using a specific screw for polyamide. In order to determine chitosan in food simulants (ultrapure water and ethanol 95% (v/v)), it was degraded into the glucosamine units by hydrolysis and quantified by ultra high pressure liquid chromatography coupled with diode array detection after its derivatization with 9-fluorenylmethyl chloroformate. Results and conclusion: From the plastic films prepared by casting and by extrusion, only those prepared by casting without plasticizer presented chitosan migration that increases with the amount of chitosan added. These films could not be used to pack aqueous foodstuffs. However, the addition of a plasticizer (glycerol) has avoided the migration of chitosan. Therefore, the use of casting films with chitosan shall include a plasticizer in the formulation. Films prepared by extrusion presented no migration into both simulants indicating suitability to pack both aqueous and fatty foodstuffs.