Determination of pesticide residues in coconut water by liquid-liquid extraction and gas chromatography with electron-capture plus thermionic specific detection and solid-phase extraction and high-performance liquid chromatography with ultraviolet detection

Two simple methods were developed to determine, 11 pesticides in coconut water, a natural isotonic drink rich in salts, sugars and vitamins consumed by the people and athletes. The first procedure involves solid-phase extraction using Sep-Pak Vac C-18 disposable cartridges with methanol for elution. Isocratic analysis was carried out by means of high-performance liquid chromatography with ultraviolet detection at 254 nm to analyse captan, chlorothalonil, carbendazim, lufenuron and diafenthiuron. The other procedure is based on liquid-liquid extraction with hexane-dichloromethane (1:1, v/v), followed by gas chromatographic analysis with effluent splitting to electron-capture detection for determination of endosulfan, captan, tetradifon and trichlorfon and thermionic specific detection for determination of malathion, parathion-methyl and monocrotophos. The methods were validated with fortified samples at different concentration levels (0.01-12.0 mg/kg). Average recoveries ranged from 75 to 104% with relative standard deviations between 1.4 and 11.5%. Each recovery analysis was repeated at least five times. Limits of detection ranged from 0.002 to 2.0 mg/kg. The analytical procedures were applied to 15 samples and no detectable amounts of the pesticides were found in any samples under the conditions described. (C) 2002 Elsevier B.V. B.V. All rights reserved.

Specific detection of Lettuce mosaic virus isolates belonging to the Most type

Lettuce mosaic virus (LMV)-Most isolates can infect and are seed-borne in cultivars containing the mol gene. A reverse transcription and polymerase chain reaction (RT-PCR)-based test was developed for the specific detection of LMV-Most isolates. Based on the complete genome sequences of three LMV isolates belonging respectively to the Most type, the Common type and neither of these two types, three different assays were compared: (i) presence of a diagnostic restriction site in the region of the genome encoding the variable N-terminus of the capsid protein, in the 3' end of the genome, (ii) RT-PCR using primers designed to amplify a cDNA corresponding to a portion of the P1 coding region, in the 5' end of the genome and (iii) RT-PCR using primers designed to amplify a central region of the genome. The assays were performed against a collection of 21 isolates from different geographical origins and representing the molecular variability of LMV. RT-PCR of the central region of the genome was preferred because its results are expected to be less affected by natural recombination between LMV isolates, and it allows sensitive detection of LMV-Most in situations of single as well as mixed contamination. (C) 2004 Elsevier B.V. All rights reserved.

Determination of herbicides residues in soil by small scale extraction

Herbicidas tais como trifluralina, simazina, atrazina, metribuzina e metolaclor são usados na agricultura brasileira. A eficiência de um método em pequena escala para a determinação destes herbicidas e dois produtos de degradação em amostras de solo foi avaliada. Os compostos foram extraídos das amostras de solo (5 g) com 20 ml de acetato de etila em agitador mecânico por 50 min. Após a extração, o sobrenadante foi filtrado em sulfato de sódio anidro, concentrado e analisado por cromatografia a gás de alta resolução com detector termiônico específico. Valores médios de recuperação obtidos de amostras de solo fortificadas em três níveis diferentes variaram de 81 a 115 % com valores de desvio padrão relativo entre 1,2 e 12,7 %. Os limites de detecção do método variaram de 0,01 a 0,06 mg kg-1. A metodologia foi aplicada a amostras de solo de fazendas localizadas próximas à cidade de Araraquara, estado de São Paulo, Brasil.

A simple and rapid extraction for gas chromatographic determination of thiabendazole and imazalil residues in lemons

A rapid and efficient method is described for the determination of thiabendazole and imazalil residues in lemons (peel and pulp). The procedure is based on the extraction with an hexane:ethyl acetate mixture (1:1, v/v) and gas chromatographic analysis using thermionic specific detection (TSD). The possibility of matrix effect was also studied. Mean recoveries from 8 replicates of fortified samples ranged from 79% to 109%, with relative standard deviation values between 2.4% to 12.8%. The detection and quantification limits of the method were 0.2 mg kg-1 and 0.5 mg kg-1, respectively.

Desenvolvimento e aplicação de métodos para análise de resíduos dos fungicidas Pirimetanil, Procloraz e Fluquinconazol e dos acaricidas Clofentezina e Cihexatina em frutas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detection of Babesia bigemina in cattle of different genetic groups and in Rhipicephalus (Boophilus) microplus tick

Babesia bigemina infections were investigated in four genetic groups of beef cattle and in Rhipicephalus (Boophilus) microplus engorged female ticks. Blood samples and engorged female ticks were collected from 15 cows and 15 calves from each of the following genetic groups: Nelore, Angus x Nelore, Canchim x Nelore, and Simmental x Nelore. Microscopic examination of blood smears and tick hemolymph revealed that merozoites of B. bigemina (6/60) as well as kinetes of Babesia spp. (9/549) were only detected in samples (blood and ticks, respectively) originated from calves. PCR-based methods using primers for specific detection of B. bigemina revealed 100% infection in both calves and cows, regardless the genetic group. Tick infection was detected by nested-PCR amplifications showing that the frequency of B. bigemina was higher (P 0.01) in female ticks collected from calves (134/549) than in those collected from cows (52/553). The frequency of B. bigemina was similar in ticks collected from animals, either cows or calves, of the four genetic groups (P > 0.05). (C) 2008 Elsevier B.V. All rights reserved.

Tethered DNA scaffolds on optical sensor platforms for detection of hipO gene from Campylobacter jejuni

DNA biosensors have gained increased attention over traditional diagnostic methods due to their fast and responsive operation and cost-effective design. The specificity of DNA biosensors relies on single-stranded oligonucleotide probes immobilized to a transduction platform. Here, we report the development of biosensors to detect the hippuricase gene (hipO) from Campylobacter jejuni using direct covalent coupling of thiol- and biotin-labeled single-stranded DNA (ssDNA) on both surface plasmon resonance (SPR) and diffraction optics technology (DOT, dotLab) transduction platforms. This is the first known report of the dotLab to detect targeted DNA. Application of 6-mercapto-1-hexanol as a spacer thiol for SPR gold surface created a self-assembled monolayer that removed unbound ssDNA and minimized non-specific detection. The detection limit of SPR sensors was shown to be 2.5 nM DNA while dotLab sensors demonstrated a slightly decreased detection limit of 5.0 nM (0.005 μM). It was possible to reuse the SPR sensor due to the negligible changes in sensor sensitivity (∼9.7 × 10 -7 ΔRU) and minimal damage to immobilized probes following use, whereas dotLab sensors could not be reused. Results indicated feasibility of optical biosensors for rapid and sensitive detection of the hipO gene of Campylobacter jejuni using specific ssDNA as a probe. © 2011 Elsevier B.V.

Graphene-based protein biomarker detection

An ability to detect and quantify protein molecules, harbingers of specific pathologies, potentially underpins both early disease diagnosis and an assessment of treatment efficacy. However, the specific detection of a particular protein biomarker in a complex environment is by no means an easy task and requires a progressive improvement in sensor technology. The high surface area, volume, electrical conductance, atomic level thickness and apparent biocompatibility of graphene makes it potentially an exceedingly powerful transducer of biorecognition events; the demands of its application in biosensing, and progress to date are reviewed herein.

Microscopia de força atômica aplicada em imunoensaios

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detecção parasitológica, molecular e sorológica de Erlichia canis e babesia canis em cães da região Centro-Oriental da Colômbia

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Pesticidas mais empregados na cultura de soja no município de Dourados (MS): determinação em água para consumo humano

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detection of infectious bronchitis virus and specific anti-viral antibodies using a concanavalin A-Sandwich-ELISA

Concanavalin A-Sandwich ELISA (Con A-S-ELISA) was developed for the detection of infectious bronchitis virus (IBV) or chicken specific anti-viral antibodies. The antigen detection limit for the Con A-S-ELISA was 10(5,1) EID50/mL. Three homologous and four heterologous IBV strains were similarly detected. This assay was highly effective in detecting the virus after infected tissue homogenates were passed once in embryonated chicken eggs, showing a good agreement with virus isolation technique. The Con A-S-ELISA was also used to measure anti-IBV chicken antibodies and showed a high coefficient of correlation (r = 0.85) and an agreement of k = 0.80 with the commercially available Indirect-ELISA. The relative sensitivity and specificity between these two tests were, respectively, 92.86% and 95.65% with an accuracy of 93.39%. Thus, the Con A-S-ELISA proved to be able to detect alternatively homologous and heterologous IBV strains or specific chicken anti-IBV antibodies, using the Con A as capture reagent of this assay.

A virtual experiment for the detection of specific features in the frequency response of a coupled nonlinear and linear oscillator

This paper presents an investigation into some practical issues that may be present in a real experiment, when trying to validate the theoretical frequency response curve of a two degree-of-freedom nonlinear system consisting of coupled linear and nonlinear oscillators. Some specific features, such as detached resonance curves, have been theoretically predicted in multi degree-of-freedom nonlinear oscillators, when subject to harmonic excitation, and the system parameters have been shown to be fundamental in achieving such features. When based on a simplified model, approximate analytical expression for the frequency response curves may be derived, which may be validated by the numerical solutions. In a real experiment, however, the practical achievability of such features was previously shown to be greatly affected by small disturbances induced by gravity and inertia, which led to some solutions becoming unstable which had been predicted to be stable. In this work a practical system configuration is proposed where such effects are reduced so that the previous limitations are overcome. A virtual experiment is carried out where a detailed multi-body model of the oscillator is assembled and the effects on the system response are investigated.

Development of Y-chromosome-Specific SCAR Markers Conserved in Taurine, Zebu and Bubaline Cattle

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development and application of a Saccharomyces cerevisiae-expressed nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibodies against infectious bronchitis virus

A Saccharomyces cerevisiae-expressed nucleocapsid (N) polypeptide of the M41 strain of infectious bronchitis virus (IBV) was used as antigen in a recombinant yeast-expressed N protein-based enzyme-linked immunosorbent assay (Y-N-ELISA). The Y-N-ELISA was rapid, sensitive, and specific for detecting chicken serum antibodies to IBV, and it compared favorably with a commercial ELISA.