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- New Trends in Food Allergens Detection: Toward Biosensing StrategiesPublication . Alves, Rita C.; Barroso, M. Fátima; González-García, María Begoña; Oliveira, M. Beatriz P.P.; Delerue-Matos, CristinaFood allergens are a real threat to sensitized individuals. Although food labeling is crucial to provide information to consumers with food allergies, accidental exposure to allergenic proteins may result from undeclared allergenic substances by means of food adulteration, fraud or uncontrolled cross-contamination. Allergens detection in foodstuffs can be a very hard task, due to their presence usually in trace amounts, together with the natural interference of the matrix. Methods for allergens analysis can be mainly divided in two large groups: the immunological assays and the DNA-based ones. Mass spectrometry has also been used as a confirmatory tool. Recently, biosensors appeared as innovative, sensitive, selective, environmentally friendly, cheaper and fast techniques (especially when automated and/or miniaturized), able to effectively replace the classical methodologies. In this review, we present the advances in the field of food allergens detection toward the biosensing strategies and discuss the challenges and future perspectives of this technology.
- Detection of the peanut allergen Ara h 6 in foodstuffs using a voltammetric biosensing approachPublication . Alves, Rita C.; Pimentel, Filipa B.; Nouws, Henri P. A.; Correr, Wagner; González-García, María Begoña; Oliveira, M. Beatriz P. P.; Delerue-Matos, CristinaA voltammetric biosensor for Ara h 6 (a peanut allergen) detection in food samples was developed. Gold nanoparticle-modified screen-printed carbon electrodes were used to develop a sandwich-type immunoassay using two-monoclonal antibodies. The antibody-antigen interaction was detected through the electrochemical detection of enzymatically deposited silver. The immunosensor presented a linear range between 1 and 100 ng/ml, as well as high precision (inter-day RSD ≤9.8 %) and accuracy (recoveries ≥96.7 %). The detection and quantification limits were 0.27 and 0.88 ng/ml, respectively. It was possible to detect small levels of Ara h 6 in complex food matrices.
- Improving the extraction of Ara h 6 (a peanut allergen) from a chocolatebased matrix for immunosensing detection: Influence of time, temperature and additivesPublication . Alves, Rita C.; Pimentel, Filipa B.; Nouws, Henri P.A.; Silva, Túlio H.B.; Oliveira, M. Beatriz P.P.; Delerue-Matos, CristinaThe extraction of Ara h 6 (a peanut allergen) from a complex chocolate-based food matrix was optimized by testing different temperatures, extraction times, and the influence of additives (NaCl and skimmed milk powder) in a total of 36 different conditions. Analyses were carried out using an electrochemical immunosensor. Three conditions were selected since they allowed the extraction of the highest levels of Ara h 6. These extractions were performed using 2 g of sample and 20 ml of Tris-HNO3 (pH = 8) containing: a) 0.1 M NaCl and 2 g of skimmed milk powder at 21 C for 60 min; b) 1 M NaCl and 1 g of skimmed milk powder at 21 C for 60 min; and c) 2 g of skimmed milk powder at 60 C for 60 min. Recoveries were similar or higher than 94.7%. This work highlights the importance to adjust extraction procedures regarding the target analyte and food matrix components.
- Total antioxidant capacity of plant infusions: Assessment using electrochemical DNA-based biosensor and spectrophotometric methodsPublication . Barroso, M. Fátima; Ramalhosa, Maria João; Alves, Rita C.; Dias, Alexandra; Soares, Cristina M. D.; Oliva-Teles, M. Teresa; Delerue-Matos, CristinaNine plant infusions were assessed in terms of their polyphenolic compounds and antioxidant capacity properties by performing a “battery” of assays. Bioactive compounds (total phenolic and flavonoid content) were measured using spectrophotometric procedures and the total antioxidant capacity was determined by DPPHradical dot-RSA, FRAP and DNA-based biosensor against the reactive oxygen species: HOradical dot and H2O2. Although the optical methods are based on the donation of hydrogen or electrons between antioxidants and radicals, the DNA-based biosensor intends to simulate an in vivo assay by exposing DNA layer to biological ROS and antioxidants. In general, green and black tea showed higher polyphenolic content and antioxidant capacity evaluated by the described methods. All the analysed plant beverages showed to be a good source of bioactive compounds, particularly the green and black tea, peppermint and senna.
- Targeting specific nutrient deficiencies in proteinrestricted diets: some practical facts in PKU dietary managementPublication . Pimentel, Filipa B.; Alves, Rita C.; Oliva-Teles, M. Teresa; Costa, Anabela S. G.; Fernandes, Telmo J. R.; Almeida, Manuela F.; Torres, Duarte; Delerue-Matos, Cristina; Oliveira, M. Beatriz P. P.Among aminoacidopathies, phenylketonuria (PKU) is the most prevalent one. Early diagnosis in the neonatal period with a prompt nutritional therapy (low natural-protein and phenylalanine diet, supplemented with phenylalanine-free amino acid mixtures and special low-protein foods) remains the mainstay of the treatment. Data considering nutrient contents of cooked dishes is lacking. In this study, fourteen dishes specifically prepared for PKU individuals were analysed, regarding the lipid profile and iron and zinc contents. These dishes are poor sources of essential nutrients like Fe, Zn or n-3 fatty acids, reinforcing the need for adequate supplementation to cover individual patients’ needs. This study can contribute to a more accurate adjustment of PKU diets and supplementation in order to prevent eventual nutritional deficiencies. This study contributes to a better understanding of nutrient intake from PKU patients’ meals, showing the need for dietary supplementation.
- Detection of Arah1 (a major peanut allergen) in food using an electrochemical gold nanoparticle-coated screen-printed immunosensorPublication . Alves, Rita C.; Pimentel, Filipa B.; Nouws, Henri P.A.; Marques, Raquel C.B.; González-García, María Begoña; Oliveira, M. Beatriz P.P.; Delerue-Matos, CristinaA gold nanoparticle-coated screen-printed carbon electrode was used as the transducer in the development of an electrochemical immunosensor for Ara h 1 (a major peanut allergen) detection in food samples. Gold nanoparticles (average diameter=32 nm) were electrochemically generated on the surface of screen-printed carbon electrodes. Two monoclonal antibodies were used in a sandwich-type immunoassay and the antibody–antigen interaction was electrochemically detected through stripping analysis of enzymatically (using alkaline phosphatase) deposited silver. The total time of the optimized immunoassay was 3 h 50 min. The developed immunosensor allowed the quantification of Ara h 1 between 12.6 and 2000 ng/ml, with a limit of detection of 3.8 ng/ml, and provided precise (RSD <8.7%) and accurate (recovery >96.6%) results. The immunosensor was successfully applied to the analysis of complex food matrices (cookies and chocolate), being able to detect Ara h 1 in samples containing 0.1% of peanut.
- Improving the extraction of Ara h 6 (a peanut allergen) from a chocolate-based matrix for immunosensing detection: Influence of time, temperature and additivesPublication . Alves, Rita C.; Pimentel, Filipa B; Nouws, Henri P.A.; Silva, Túlio H B; Oliveira, M Beatriz P P; Delerue-Matos, CristinaThe extraction of Ara h 6 (a peanut allergen) from a complex chocolate-based food matrix was optimized by testing different temperatures, extraction times, and the influence of additives (NaCl and skimmed milk powder) in a total of 36 different conditions. Analyses were carried out using an electrochemical immunosensor. Three conditions were selected since they allowed the extraction of the highest levels of Ara h 6. These extractions were performed using 2g of sample and 20ml of Tris-HNO3 (pH=8) containing: a) 0.1M NaCl and 2g of skimmed milk powder at 21°C for 60min; b) 1M NaCl and 1g of skimmed milk powder at 21°C for 60min; and c) 2g of skimmed milk powder at 60°C for 60min. Recoveries were similar or higher than 94.7%. This work highlights the importance to adjust extraction procedures regarding the target analyte and food matrix components.