RESUMEN
This manuscript describes the use of a portable and low cost fluorescence setup to quantify the concentration of ochratoxin A (OTA) in beverage samples using an in-house developed system and different color models. It is reported that OTA is naturally fluorescent, for that reason an ultraviolet light at 365 nm was used to excite the samples and a Complementary Metal Oxide Semiconductor (CMOS) sensor was used to get a photograph of the OTA under excitation conditions, which is controlled by an executable interface designed in MATLAB. For each concentration of OTA, the coordinates with respect to each model color were obtained and plotted to quantify the mycotoxin present in the sample. It was possible to observe that despite the fact no extraction column was employed, the Red, Green, Blue (RGB) model shows a proportional relation to the evaluated concentrations. Despite the fact more analysis and other methods are required to quantify the OTA concentration, the brightness and a,b for the color-opponent dimensions (L*a*b) and Hue, Saturation, Value (HSV) tests provide results whereby it is possible to identify the concentration of OTA in beverage samples such as beer and wine.
Asunto(s)
Bebidas/análisis , Colorimetría/métodos , Ocratoxinas/análisis , Cerveza/análisis , Vino/análisisRESUMEN
The present article describes a portable and low cost fluorescence set-up designed and characterized for in-situ screening of Ochratoxin A (OTA) in cocoa samples at field settings. The sensing module (the set up) consists of a LED with the wavelength of 370-380nm and a color complementary metal oxide semiconductor (CMOS) micro-camera inbuilt at upright position of a black box to obtain an image of the sensing molecule. It allows the user to get an image of the sensing analytes under excitation conditions and process the image in order to predict the toxicity of the samples. The image capturing and processing of the system was based on the OTA concentration in the sample and analyzed data can be presented as RGB values. For each concentration of the OTA, the R, G, B co-ordinates were obtained and plotted to quantify actual OTA presents in the sample. Moreover, the system was tested for real sample analysis using cocoa contaminated with OTA. The system could detect OTA as low as 1.25ng/ml with the maximum recovery of 87.5% in cocoa samples. The OTA was extracted in 1% NaHCO3 and cleaned up using molecular imprinted polymer column (MIP). The method demonstrated a good linear range between 1.25 and 10ng/ml. The obtained results were cross validated using chromatographic method HPLC and also compared with commercially available fluorescence instrument. The developed fluorescence setup is simple, economical, and portable with added advantages of digital image processing. The system could be deployable to cocoa fields for monitoring of OTA in quick successions. It is noteworthy to mention that this is the first report of such portable fluorescence setup where, OTA sensing was explored.
Asunto(s)
Cacao/química , Análisis de los Alimentos/métodos , Contaminación de Alimentos/análisis , Ocratoxinas/análisis , Fluorescencia , Análisis de los Alimentos/economía , Límite de DetecciónRESUMEN
An approach to an inhibition bioelectronic tongue is presented. The work is focused on development of an automated flow system to carry out experimental assays, a custom potentiostat to measure the response from an enzymatic biosensor, and an inhibition protocol which allows on-line detections. A Multi-commuted Flow Analysis system (MCFA) was selected and developed to carry out assays with an improved inhibition method to detect the insecticides chlorpyrifos oxon (CPO), chlorfenvinfos (CFV) and azinphos methyl-oxon (AZMO). The system manifold comprised a peristaltic pump, a set of seven electronic valves controlled by a personal computer electronic interface and software based on LabView® to control the sample dilutions into the cell. The inhibition method consists in the injection of the insecticide when the enzyme activity has reached the plateau of the current; with this method the incubation time is avoided. A potentiostat was developed to measure the response from the enzymatic biosensor. Low limits of detection of 10 nM for CPO, CFV, and AZMO were achieved.
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Acetilcolinesterasa/química , Técnicas Biosensibles/instrumentación , Drosophila/química , Inhibidores Enzimáticos , Insecticidas/análisis , Animales , Azinfosmetilo/análogos & derivados , Azinfosmetilo/análisis , Bencenosulfonatos/análisis , Cloropirifos/análogos & derivados , Cloropirifos/análisis , Potenciometría/métodos , Programas InformáticosRESUMEN
An amperometric biosensor based on laccase, from Trametes versicolor (LTV), was developed and optimized for monitoring the phenolic compounds content in tea infusions. The fungal enzyme was immobilized by entrapment within polyvinyl alcohol photopolymer PVA-AWP (azide-unit pendant water-soluble photopolymer) onto disposable graphite screen-printed electrodes (SPE). Sensitivity optimization in terms of pH, temperature and applied potential was carried out. The linear range, detection limit, operational and storage stabilities were also determined. The laccase biosensor (LTV-SPE) was calibrated for o-, m- and p-diphenol as well as caffeic acid. The highest response was found at 0.1M acetate buffer pH 4.7, though it must be added the good reproducibility and operational stability were also obtained. The useful lifetime of the biosensor is estimated to be greater than 6 months. LTV-SPE was used for the determination of the equivalent phenol content (EPC) in tea infusions by the direct addition into the electrochemical cell: the results were compared with those from the Folin-Ciocalteu spectrophotometric method. The amperometric detection exhibits some interesting advantages such as high simplicity, minimal sample preparation and shorter response time. A stable and sensitive amperometric response was obtained toward standard diphenolic compounds and herbal infusions. These biosensors are useful for easy and fast monitoring of EPC that can be related to the antioxidant capacity of natural extracts.
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Técnicas Biosensibles , Electroquímica/métodos , Análisis de los Alimentos/métodos , Fenol/química , Té , Antioxidantes/química , Calibración , Electrodos , Concentración de Iones de Hidrógeno , Polímeros/química , Sensibilidad y Especificidad , Espectrofotometría/métodos , Temperatura , Agua/químicaRESUMEN
Amperometric acetylcholinesterase biosensors have been developed for quantification of the pesticides carbofuran, carbaryl, methylparaoxon, and dichlorvos in phosphate buffer containing 5% acetonitrile. Three different biosensors were built using three different acetylcholinesterase (AChE) enzymes-AChE from electric eel, and genetically engineered (B394) and wild-type (B1) AChE from Drosophila melanogaster. Enzymes were immobilized on cobalt(II) phthalocyanine-modified electrodes by entrapment in a photocrosslinkable polymer (PVA-AWP). Each biosensor was tested against the four pesticides. Good operational stability, immobilisation reproducibility, and storage stability were obtained for each biosensor. The best detection limits were obtained with the B394 enzyme for dichlorvos and methylparaoxon (9.6 x 10(-11) and 2.7 x 10(-9) mol L(-1), respectively), the B1 enzyme for carbofuran (4.5 x 10(-9) mol L(-1)), and both the B1 enzyme and the AChE from electric eel for carbaryl (1.6 x 10(-7) mol L(-1)). Finally, the biosensors were used for the direct detection of the pesticides in spiked apple samples.
Asunto(s)
Acetonitrilos , Acetilcolinesterasa/metabolismo , Técnicas Biosensibles/métodos , Carbaril/análisis , Carbofurano/análisis , Diclorvos/análisis , Paraoxon/análogos & derivados , Animales , Carbaril/metabolismo , Carbofurano/metabolismo , Diclorvos/metabolismo , Drosophila melanogaster/enzimología , Electrophorus/metabolismo , Malus/química , Estructura Molecular , Paraoxon/análisis , Paraoxon/metabolismo , Plaguicidas/análisis , Plaguicidas/metabolismoRESUMEN
Genetically modified acetylcholinesterase (AChE) from Drosophila melanogaster (dm) and from commercial sources, Electric eel (ee), Bovine erythrocites (be) and Human erythrocites (he), were investigated as biological receptors for the detection of methamidophos pesticide based on inhibition studies. Most engineered variant of AChE from dm showed enhanced sensitivity toward methamidophos pesticide. Among 24 dmAChE variants tested, 12 presented a sensitivity comparable to the commercially available eeAChE, but higher than AChEs from be and he. Four were found more sensitive and six others were insensitive to methamidophos insecticide. The D375G,Y370F,Y374A,F376L mutant was the most sensitive, with a ki value of 2.2 X 10(6) mol(-1) L min(-1), three orders of magnitude higher than eeAChE (1.1 X 10(3) mol(-1) L min(-1)). The sensor constructed with genetically modified enzyme showed better characteristics with respect to detection limit and sensitivity compared with those using commercial eeAChE. Differential pulse polarography and chronoamperometry were used as electrochemical techniques to characterize the AChE biosensors. The lower detection limit of 1 ppb was obtained with D375G,Y370F,Y374A,F376L mutant of dmAChE, compared to 90 ppb for the commercial eeAChE. This study may stimulate scientists to develop more sensitive and selective procedures for organophosphorus insecticides detection by using engineered variant of dmAChE.