Person: Benito Peña, María Elena
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First Name
María Elena
Last Name
Benito Peña
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Química Analítica
Area
Química Analítica
Identifiers
4 results
Search Results
Now showing 1 - 4 of 4
Item Molecular super-gluing: a straightforward tool for antibody labelling and its application to mycotoxin biosensing(Analytical and Bioanalytical Chemistry, 2022) Prádanas González, Fernando; Glahn Martínez, Ana Bettina; Benito Peña, María Elena; Arola, Henri O.; Nevanen, Tarja K.; Moreno Bondi, María CruzMycotoxins are low molecular weight toxic compounds, which can cause severe health problems in animals and humans. Immunoassays allow rapid, simple and cost-efective screening of mycotoxins. Sandwich assays with a direct readout provide great improvement in terms of selectivity and sensitivity, compared to the widely used competitive assay formats, for the analysis of low molecular weight molecules. In this work, we report a non-competitive fuorescence anti-immune complex (IC) immunoassay, based on the specifc recognition of HT-2 toxin with a pair of recombinant antibody fragments, namely antigen-binding fragment (Fab) (anti-HT-2 (10) Fab) and single-chain variable fragment (scFv) (anti-IC HT-2 (10) scFv). The SpyTag and SpyCatcher glue proteins were applied for the frst time as a bioconjugation tool for the analysis of mycotoxins. To this aim, a SpyTag-mScarlet-I (fuorescent protein) and scFv-SpyCatcher fusion proteins were constructed, produced and fused in situ during the assay by spontaneous Tag-Catcher binding. The assay showed an excellent sensitivity with an EC50 of 4.8±0.4 ng mL−1 and a dynamic range from 1.7±0.3 to 13±2 ng mL−1, an inter-day reproducibility of 8.5% and a high selectivity towards HT-2 toxin without cross-reactivity with other Fusarium toxins. The bioassay was applied to the analysis of the toxin in an oat reference material and in oat samples, with a LOD of 0.6 µg kg−1, and the results were validated by analysing a certifcate reference material and by HPLC–MS/MS.Item Extracting mycotoxins from edible vegetable oils by using green, ecofriendly deep eutectic solvents(Food Chemistry, 2023) Pradanas-González, Fernando; Aragoneses-Cazorla, Rubén; Ángel Merino-Sierra, Miguel; Andrade-Bartolomé, Elena; Navarro-Villoslada, Fernando; Benito-Peña, Elena; Moreno-Bondi, María Cruz; Moreno Bondi, María Cruz; Benito Peña, María Elena; Navarro Villoslada, Fernando; Prádanas González, FernandoIn this work, we developed an environmentally friendly liquid–liquid microextraction method using a natural deep eutectic solvent in combination with liquid chromatography for the simultaneous determination of four mycotoxins (deoxynivalenol, alternariol, ochratoxin A and zearalenone) in edible vegetable oils. A chemometric approach assessed the effect of the operational parameters on the mycotoxin extraction efficiency. The extracts were analyzed by HPLC coupled with a diode array and fluorescence detector. The optimum NADES composition resulted in the highest extraction recoveries, and it was applied to coextract the target mycotoxins in several types of edible vegetable oils without using hazardous solvents or requiring further clean-up. The limits of detection ranged from 0.07 to 300μg/kg, and recoveries were close to 100%, except for zearalenone(viz. 35%), with relative standard deviations below 9% in all cases. The proposed method was validated following the European Commission 2002/657/EC and 2006/401/EC.Item Homogeneous immunoassay for cyclopiazonic acid based upon mimotopes and upconversion-resonance energy transfer(Biosensors and Bioelectronics, 2023) Prádanas González, Fernando; Peltomaa, Riikka Johanna; Satu Lahtinen; Luque Uria, Álvaro; Vicente Más; Rodrigo Barderas; Chris M. Maragos; Canales Mayordomo, María Ángeles; Tero Soukka; Benito Peña, María Elena; Moreno Bondi, María CruzStrains of Penicillium spp. are used for fungi-ripened cheeses and Aspergillus spp. routinely contaminate maize and other crops. Some of these strains can produce toxic secondary metabolites (mycotoxins), including the neurotoxin α-cyclopiazonic acid (CPA). In this work, we developed a homogeneous upconversion-resonance energy transfer (UC-RET) immunoassay for the detection of CPA using a novel epitope mimicking peptide, or mimotope, selected by phage display. CPA-specific antibody was used to isolate mimotopes from a cyclic 7-mer peptide library in consecutive selection rounds. Enrichment of antibody binding phages was achieved, and the analysis of individual phage clones revealed four different mimotope peptide sequences. The mimotope sequence, ACNWWDLTLC, performed best in phage-based immunoassays, surface plasmon resonance binding analyses, and UC-RET-based immunoassays. To develop a homogeneous assay, upconversion nanoparticles (UCNP, type NaYF4:Yb3+, Er3+) were used as energy donors and coated with streptavidin to anchor the synthetic biotinylated mimotope. Alexa Fluor 555, used as an energy acceptor, was conjugated to the anti-CPA antibody fragment. The homogeneous single-step immunoassay could detect CPA in just 5 min and enabled a limit of detection (LOD) of 30 pg/mL (1.5 μg/kg) and an IC50 value of 0.36 ng/mL. No significant cross-reactivity was observed with other co-produced mycotoxins. Finally, we applied the novel method for the detection of CPA in spiked maize samples using high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) as a reference method.Item Mycotoxin extraction from edible insects with natural deep eutectic solvents: a green alternative to conventional methods(Journal of Chromatography A, 2021) Prádanas González, Fernando; Álvarez-Rivera, Gerardo; Benito Peña, María Elena; Navarro Villoslada, Fernando; Cifuentes, Alejandro; Herrero, Miguel; Moreno Bondi, María CruzEdible insects are widely consumed in Africa, Asia, Oceania and Latin America, but less commonly so in Western countries. Since the turn of the millennium, however, entomophagy has aroused growing interest worldwide in response to the increasing scarcity of food resources. In fact, edible insects can be a source of high-quality protein, and also of fat, energy, minerals and vitamins. However, the lack of regulatory guidelines for microbiologically or chemically hazardous agents potentially present in these new foods (e.g., mycotoxins) may make their consumption unsafe. In this work, we developed an environmentally friendly analytical method using natural deep eutectic solvents (NADES or natural DES) in combination with ultra-high performance liquid chromatography/tandem mass spectrometry (UHPLC-MS/MS) for the simultaneous determination of six mycotoxins of great concern owing to their toxic effects on humans and animals (namely, fumonisin B 1 , fumonisin B 2 , T-2 toxin, HT-2 toxin, ochratoxin A and mycophenolic acid) in insect-based food products. The target mycotoxins were co-extracted from cricket flour by using the optimum DES composition (namely, a mixture of choline chloride and urea, in a 1:2 mole ratio, containing 15% water which resulted in the highest extraction recoveries for all toxins). An experimental design method (Fractional Factorial Design (FFD) was used to examine the influence of the operational variables DES volume and water content, amount of sample, extraction time and extraction temperature on the extraction efficiency for each mycotoxin. Under optimum conditions, extraction recoveries were close to 100% except for fumonisin B 2 (70%) and T-2 toxin (50%), with relative standard deviations (RSDs) below 13% in all cases. The proposed NADES-UHPLC–MS/MS method was validated in accordance with the European Commission 2002/657/EC and 2006/401/EC decisions, and used to determine the target compounds in cricket flour, silkworm pupae powder and black cricket powder.