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Suarez-Pantaleon C.,IATA CSIC | Wichers J.,Wageningen UR Food and Biobased Research | Abad-Somovilla A.,University of Valencia | Van Amerongen A.,Wageningen UR Food and Biobased Research | Abad-Fuentes A.,IATA CSIC
Biosensors and Bioelectronics | Year: 2013

Rapid analytical methods enabling the determination of diverse targets are essential in a number of research areas, from clinical diagnostics to feed and food quality and safety. Herein, the development of a quantitative immunochromatographic assay for the detection of the synthetic phytoregulator forchlorfenuron (CPPU) is described. The competitive lateral flow immunoassay (LFIA) was based on the immobilization onto a nitrocellulose membrane of an ovalbumin-CPPU conjugate (test line) and on the use of an immunodetection ligand consisting of carbon nanoparticles labeled with an anti-CPPU monoclonal antibody through interaction with a secondary antibody. The presence of CPPU in horticultural samples was visually interpreted by the decrease in the black signal intensity of the test line, according to the competitive character of the format. The quantitative determination of the analyte was easily performed by a two-step procedure consisting of flatbed scanning of the strips followed by computer-based image analysis of the pixel gray volumes of the test lines. Under optimized conditions, the immunochromatographic test afforded a limit of quantification in buffer of 89. ng/L. The accuracy of the strip test was assessed by the analysis of fruit samples with incurred residues, and the obtained results were compared with those derived from two reference methods, ELISA and HPLC. The LOQ of the CPPU-specific LFIA in kiwifruits and grapes was established at 33.4 μg/kg. The excellent analytical performance of the developed strip test demonstrates the potential of immunochromatographic assays for the quantitative monitoring of small organic molecules in complex matrices. © 2012 Elsevier B.V.

Gomez-Mascaraque L.G.,IATA CSIC | Soler C.,University of Valencia | Lopez-Rubio A.,IATA CSIC
Food Hydrocolloids | Year: 2016

Micro-hydrogels are very promising systems for the protection and controlled delivery of sensitive bioactives, but limited knowledge exists regarding the impact of this encapsulation on their bioaccessibility. In this work, two different hydrogel-forming biopolymers (gelatin and chitosan) were compared as wall materials for the microencapsulation of a model flavonoid, (-)-epigallocatechin gallate (EGCG). Results showed that gelatin was more adequate as wall material for the encapsulation of EGCG than chitosan, achieving higher encapsulation efficiencies (95% ± 6%), being more effective in delaying EGCG release and degradation in aqueous solution and exhibiting a 7 times higher bioaccessibility of the bioactive compound (in terms of antioxidant activity) after in-vitro gastrointestinal digestion. A very low bioaccessibility of EGCG in chitosan was observed, due to the neutralization of the carbohydrate in the basic simulating salivary conditions, thus precluding subsequent flavonoid release. Moreover, gelatin micro-hydrogels also hindered dimer formation during in-vitro digestion, thus suggesting greater bioavailability when compared with free EGCG. © 2016 .

Esteve-Turrillas F.A.,IATA CSIC | Abad-Fuentes A.,IATA CSIC
Biosensors and Bioelectronics | Year: 2013

Quantum dots (QDs) are semiconductor nanoparticles with very interesting optical properties, like high quantum yield or narrow and size-tuneable fluorescence spectra. Current applications of QDs are widespread, their use as fluorescence labels in bioassays being one of the most promising. These nanoparticles are usually conjugated to highly specific biomolecules like antibodies, oligonucleotides, enzymes or aptamers to improve assay selectivity. In this review, QD surface passivation, conjugation to biomolecules, and purification strategies are discussed with special emphasis to the development of QD-based immunoassays for the detection of low molecular weight compounds given the relevance of this sort of analytes in health, food safety, pharmaceutical, or environmental monitoring areas. The aim of this review is to summarise the main achievements attained so far and to initialise researchers in the field of antibody-based assays employing QDs as labels, such as fluorescence-linked immunosorbent assay (FLISA), fluorescence (or Förster) resonance energy transfer (FRET), immunochromatographic methods, and immunosensors. © 2012 Elsevier B.V.

Tomas-Navarro M.,CSIC - Center of Edafology and Applied Biology of the Segura | Vallejo F.,CSIC - Center of Edafology and Applied Biology of the Segura | Sentandreu E.,IATA CSIC | Navarro J.L.,IATA CSIC | Tomas-Barberan F.A.,CSIC - Center of Edafology and Applied Biology of the Segura
Journal of Agricultural and Food Chemistry | Year: 2014

The effect of two technological treatments on orange juice flavanone bioavailability in humans was assessed. Processing affected flavanone solubility and particle size of the cloud. Volunteers were stratified in high, medium, and low urinary excretion capabilities. Flavanones from high-pressure homogenized juice showed better absorption than those of conventional pasteurized juice in high excretors. These differences were not observed in medium and low excretors. High flavanone excretors took advantage of the high-pressure homogenization juice attributes (smaller cloud particle size) and showed an improved absorption/excretion. Stratification of the individuals by their excretion capability is more relevant than technological treatments in terms of flavanone bioavailability. This stratification should be considered in clinical studies with citrus juices and extracts as it could explain the large interindividual variability that is often observed. © 2013 American Chemical Society.

Mercader J.V.,University of Valencia | Agullo C.,IATA CSIC | Abad-Somovilla A.,IATA CSIC | Abad-Fuentes A.,University of Valencia
Organic and Biomolecular Chemistry | Year: 2011

The design and synthesis of functional chemical derivatives of small organic molecules is usually a key step for the intricate production of a variety of bioconjugates. In this respect, the derivatization site at which the spacer arm is introduced in immunizing conjugates constitutes a highly critical parameter for the generation of high-affinity and selective antibodies. However, due to the usual complexity of the required synthetic procedures, the appropriate comparison of alternative tethering positions has often been neglected. In the present study, meticulous strategies were followed to prepare synthetic derivatives of pyraclostrobin with the same linkers located at diverse rationally-chosen sites. Activity appraisal of antibodies and bioconjugates was carried out by bidimensional competitive direct and indirect immunoassays, and a superior performance of two of the three synthesized haptens was found. Finally, a detailed analysis of the conformations of the target molecule and the synthesized haptens in aqueous solution was done using computer assisted molecular modeling techniques. This study suggested that the lower titers and affinities of one set of antibodies are most probably due to conformational effects of the spacer arm in the immunizing bioconjugate. © The Royal Society of Chemistry 2011.

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