A novel strategy for identification of pesticides in different categories by concentration-independent model based on a nanozyme with multienzyme-like activities.

Conventional rapid detection methods are difficult to identify or distinguish various pesticide residues at the same time. And sensor arrays are also limited by the complexity of preparing multiple receptors and high cost. To address this challenge, a single material with multiple properties is considered. Herein, we first found that different categories of pesticides have diverse regulatory behaviors on the multiple catalytic activities of Asp-Cu nanozyme. Thus, a three-channel sensor array based on the laccase-like, peroxidase-like, and superoxide dismutase-like activities of Asp-Cu nanozyme was constructed and successfully used for the discrimination of eight kinds of pesticides (glyphosate, phosmet, isocarbophos, carbaryl, pentachloronitrobenzene, metsulfuron-methyl, etoxazole, and 2-methyl-4-chlorophenoxyacetic acid). In addition, a concentration-independent model for qualitative identification of pesticides has been established, and 100% correctness was achieved in the recognition of unknown samples. Then, the sensor array also exhibited excellent interference immunity and was reliable for real sample analysis. It provided a reference for pesticide efficient detection and food quality supervision.

Understanding the impact of a non-ionic surfactant alkylphenol ethoxylate on surface-enhanced Raman spectroscopic analysis of pesticides on apple surfaces.

Pesticide active ingredients (AIs) are often applied with adjuvants to facilitate the stability and functionality of AIs in agricultural practice. The objective of this study is to investigate the role of a common non-ionic surfactant, alkylphenol ethoxylate (APEO), on the surface-enhanced Raman spectroscopic (SERS) analysis of pesticides as well as its impact on pesticide persistence on apple surfaces, as a model fresh produce surface. The wetted areas of two AIs (thiabendazole and phosmet) mixed with APEO were determined respectively to correct the unit concentration applied on apple surfaces for a fair comparison. SERS with gold nanoparticle (AuNP) mirror substrates was applied to measure the signal intensity of AIs with and without APEO on apple surfaces after a short-term (45 min) and a long-term (5 days) exposure. The limit of detection (LOD) of thiabendazole and phosmet using this SERS-based method were 0.861 ppm and 2.883 ppm, respectively. The result showed that APEO decreased the SERS signal for non-systemic phosmet, while increased SERS intensity of systemic thiabendazole on apple surfaces after 45 min pesticide exposure. After 5 days, the SERS intensity of thiabendazole with APEO was higher than thiabendazole alone, and there was no significant difference between phosmet with and without APEO. Possible mechanisms were discussed. Furthermore, a 1% sodium bicarbonate (NaHCO3) washing method was applied to test the impact of APEO on the persistence of the residues on apple surfaces after short-term and long-term exposures. The results indicated that APEO significantly enhanced the persistence of thiabendazole on plant surfaces after a 5-day exposure, while there was no significant impact on phosmet. The information obtained facilitates a better understanding of the impact of the non-ionic surfactant on SERS analysis of pesticide behavior on and in plants and helps further develop the SERS method for studying complex pesticide formulations in plant systems.

An insect acetylcholinesterase biosensor utilizing WO3/g-C3N4 nanocomposite modified pencil graphite electrode for phosmet detection in stored grains.

This study was undertaken to assess the potential of Tribolium castaneum (Red flour beetle) acetylcholinesterase (Tc-AChE) based electrochemical biosensor integrating WO3/g-C3N4 nanocomposite modified Pencil graphite electrode to detect an organophosphate insecticide, Phosmet. The WO3/g-C3N4 nanocomposite provides a non-toxic, biocompatible surface for binding the enzyme on the electrode surface, attributed to its large surface area, high conductivity, and low ohmic resistance. The proposed biosensor shows a very good analytical performance with LOD 3.6 nM for Phosmet and effectively determined Phosmet in wheat with a 99% recovery rate. Furthermore, molecular docking deciphers the binding interactions of Phosmet with Tc-AChE using a modified AutoDock LGA algorithm and an AMBER03 force field in YASARA. The kinetic parameters strongly suggest the high potency of inhibitor with the enzyme. This study presents an adaptable, rapid, and straightforward approach that opens ways towards real progress in developing commercial biosensors for pesticide detection.

Facile synthesis of a core-shell structured magnetic covalent organic framework for enrichment of organophosphorus pesticides in fruits.

A facile strategy was developed for the fabrication of a magnetic covalent organic framework (COF) via grafting of the monomers, 2,5-dihydroxyterephthalaldehyde (Dt) and 1,3,5-tris(4-aminophenyl) benzene (Tb) onto surface-modified Fe3O4 nanoparticles. The magnetic COF, named as magnetic COF-DtTb, was readily fabricated without high temperature or harsh reaction conditions. The synthesized magnetic COF-DtTb nanoparticles were fully characterized, presenting a regular core-shell spherical structure, large specific surface area, superparamagnetism, and good thermal stability. Their potential as an enrichment adsorbent was investigated to establish an efficient magnetic solid-phase extraction method for the determination of organophosphorus pesticide residues in fruits. Systematic method validation revealed good linearity in the concentration range of 1-200 µg L-1 (correlation coefficient >0.9957). The method limits of detection were in the range of 0.002-0.063 µg kg-1, the method limit of quantification was 1.00 µg kg-1 and recoveries ranged from 72.8% to 111% with RSDs lower than 12.3%. The results indicated that magnetic COF-DtTb possesses superior trace enrichment properties for organophosphorus pesticides in fruits.

A spectroscopic approach to detect and quantify phosmet residues in Oolong tea by surface-enhanced Raman scattering and silver nanoparticle substrate.

There have been increasing concerns among consumers about pesticide residues in Oolong tea. This study aimed to establish surface-enhanced Raman scattering (SERS) method for rapid measurement of chemical contaminants in Oolong tea. Synthesis of SERS substrate was achieved by synthesizing silver nanoparticles (AgNPs) via a reduction method. AgNPs were spherical and highly monodispersed, which created remarkable electromagnetic fields during SERS activities to measure phosmet in the methanol-water solution and Oolong tea. Partial least squares regression models were established to predict the concentrations of phosmet in the methanol-water solution (r = 0.934; slope = 0.880; RMSEP = 1.001 mg/L) and Oolong tea samples (r = 0.927; slope = 0.938; RMSEP = 1.157 mg/kg) with the detection limit of 0.1 mg/kg. The results indicate that SERS coupled with silver nanoparticles is a fast, sensitive, and reliable method for detection and characterization of pesticide contaminants in Oolong tea products.

Effectiveness of Commercial and Homemade Washing Agents in Removing Pesticide Residues on and in Apples.

Removal of pesticide residues from fresh produce is important to reduce pesticide exposure to humans. This study investigated the effectiveness of commercial and homemade washing agents in the removal of surface and internalized pesticide residues from apples. Surface-enhanced Raman scattering (SERS) mapping and liquid chromatography tandem mass spectrometry (LC-MS/MS) methods were used to determine the effectiveness of different washing agents in removing pesticide residues. Surface pesticide residues were most effectively removed by sodium bicarbonate (baking soda, NaHCO3) solution when compared to either tap water or Clorox bleach. Using a 10 mg/mL NaHCO3 washing solution, it took 12 and 15 min to completely remove thiabendazole or phosmet surface residues, respectively, following a 24 h exposure to these pesticides, which were applied at a concentration of 125 ng/cm2. LC-MS/MS results showed, however, that 20% of applied thiabendazole and 4.4% of applied phosmet had penetrated into the apples following the 24 h exposure. Thiabendazole, a systemic pesticide, penetrated 4-fold deeper into the apple peel than did phosmet, a non-systemic pesticide, which led to more thiabendazole residues inside the apples, which could not be washed away using the NaHCO3 washing solution. This study gives us the information that the standard postharvest washing method using Clorox bleach solution for 2 min is not an effective means to completely remove pesticide residues on the surface of apples. The NaHCO3 method is more effective in removing surface pesticide residues on apples. In the presence of NaHCO3, thiabendazole and phosmet can degrade, which assists the physical removal force of washing. However, the NaHCO3 method was not completely effective in removing residues that have penetrated into the apple peel. The overall effectiveness of the method to remove all pesticide residues diminished as pesticides penetrated deeper into the fruit. In practical application, washing apples with NaHCO3 solution can reduce pesticides mostly from the surface. Peeling is more effective to remove the penetrated pesticides; however, bioactive compounds in the peels will become lost too.

Biosensor-assisted selection of optimal parameters for designing molecularly imprinted polymers selective to phosmet insecticide.

Molecularly imprinted polymers (MIPs) for phosmet insecticide were synthesized by batch polymerization. The affinity of functional monomers to phosmet was tested using an original method involving an electrochemical biosensor based on acetylcholinesterase inhibition. It was demonstrated that association of phosmet with appropriate functional monomers resulted in a decrease of enzyme inhibition. Using this method, it was shown that N,N-methylenebis(acrylamide) displayed the highest interactions with phosmet using DMSO as solvent. These results were in good accordance with those obtained by conventional computational modeling. Molecularly imprinted polymers (MIPs) and non-imprinted polymers (NIPs) were synthesized and adsorption isotherms were studied to describe their interaction with phosmet. Freundlich isotherm was able to fit phosmet adsorption on MIPs with good agreement (R2 = 0.9). The pre-exponential factor KF determined for MIPs was 1.439mg(1-N)g-1LN, more that 10 times higher than for NIPs (0.125mg(1-N)g-1. LN), indicating an increase of binding sites number and average affinity.

Ultra-sensitive biosensor based on genetically engineered acetylcholinesterase immobilized in poly (vinyl alcohol)/Fe-Ni alloy nanocomposite for phosmet detection in olive oil.

An ultra-sensitive screen-printed biosensor was successfully developed for phosmet detection in olive oil, based on a genetically-engineered acetylcholinesterase (AChE) immobilized in a azide-unit water-pendant polyvinyl alcohol (PVA-AWP)/Fe-Ni alloy nanocomposite. Fe-Ni not only allowed amplifying the response current but also lowering the applied potential from 80 mV to 30 mV vs Ag/AgCl. The biosensor showed a very good analytical performance for phosmet detection, with a detection limit of 0.1 nM. This detection limit is lower than the allowable concentrations set by international regulations. In addition to the good reproducibility, operational and storage stability, the developed biosensor was successfully used for the determination of phosmet in olive oil samples without any laborious pre-treatment. The phosmet recovery rate was about 96% after a simple liquid-liquid extraction.

Breaking the take home pesticide exposure pathway for agricultural families: workplace predictors of residential contamination.

BACKGROUND: Pesticides used in agriculture can be taken into worker homes and pose a potential risk for children and other family members. This study focused on identification of potential intervention points at the workplace. METHODS: Workers (N = 46) recruited from two tree fruit orchards in Washington State were administered a 63-item pesticide safety questionnaire. Dust was collected from commuter vehicles and worker homes and analyzed for four organophosphorus (OP) pesticides (azinphosmethyl, phosmet, chlorpyrifos, malathion). RESULTS: Geometric mean azinphosmethyl concentrations in dust for three worker groups (16 pesticide handlers, 15 green fruit thinners, 15 organic orchard workers) ranged from 0.027-1.5 µg/g, with levels in vehicle dust higher than in house dust, and levels in house dust from handlers' homes higher than levels from tree fruit thinners' homes. Vehicle and house dust concentrations of azinphosmethyl were highly associated (R(2) = 0.44, P < 0.001). Significant differences were found across worker groups for availability of laundry facilities, work boot storage, frequency of hand washing, commuter vehicle use, parking location, and safety training. CONCLUSIONS: These findings support a focus on intervention activities to reduce take home pesticide exposure closer to the source of contamination; specifically, the workplace and vehicles used to travel to the workplace.

Surface plasmon resonance sensor for phosmet of agricultural products at the ppt detection level.

A surface plasmon resonance (SPR) immunoassay using a PH-BSA immobilized sensor chip was developed to measure phosmet in food samples. The in-competitive inhibition assay showed highly sensitive and good specificity of the cross-reactivity with analogue structure pesticides. The biosensor exhibited a linear detection range from 8.0 to 60.0 ng/L of phosmet with a lower detection limit of 1.6 ng/L (S/N = 3). The sensitivity obtained with the present SPR affinity biosensor was significantly higher than most of the sensors reported with different measurement methodologies for phosmet. A recovery test of pesticide quantification in peaches, apples, cabbages, and rapes was also studied. Good recoveries (86.4-102.8%) and coefficients of variation (CVs) (5.1-12.6%) were obtained in all cases. The SPR biosensor assay method was compared with cd-ELISA in terms of analysis time, antibody dosage, recoveries, precision, detection limit, pretreatment, and testing costs, and clear advantages could be seen over the traditional ELISA-based detection systems. The developed SPR method was suitable for the rapid quantitative or qualitative determination of phosmet in agricultural products.

Organophosphate pesticide exposure and residential proximity to nearby fields: evidence for the drift pathway.

OBJECTIVES: Residential proximity to pesticide-treated farmland is an important pesticide exposure pathway. METHODS: In-person interviews and biological samples were collected from 100 farmworker and 100 non-farmworker adults and children living in Eastern Washington State. We examined the relationship of residential proximity to farmland to urinary metabolite concentrations of dimethylphosphate (DMTP) and levels of pesticide residues in house dust. RESULTS: DMTP concentrations were higher in farmworkers than non-farmworkers (71 µg/L vs 6 µg/L) and in farmworker children than non-farmworker children (17 µg/L vs 8 µg/L). Compared to non-farmworker households, farmworker households had higher levels of azinphos-methyl (643 ng/g vs 121 ng/g) and phosmet (153 ng/g vs 50 ng/g). Overall, a 20% reduction in DMTP concentration was observed per mile increase in distance from farmland. CONCLUSIONS: Lower OP metabolite concentrations correlated with increasing distance from farmland.

Hapten synthesis and enzyme-linked immunosorbent assay for phosmet residues: assay optimization and investigation of matrix effects from different food samples.

In this study, a panel of haptens was synthesized for immunoconjugate preparation, and several haptens for heterologous tracer conjugates were also prepared. A highly sensitive polyclonal antibody against the organophosphorus insecticide phosmet was obtained and competitive direct enzyme-linked immunosorbent assays (cd-ELISA) for this pesticide were developed. In the cd-ELISA, the limit of detection (IC(15)) was 0.6 microg kg(-1) and the sensitivity (IC(50)) was 20 microg kg(-1). The suitability of the ELISA for pesticide quantification in peach, apple, orange juice, and apple juice was also studied. Good accuracy and precision were obtained with mean recoveries between 78% and 102.3% and mean coefficients of variation below 13.63%. Validation of the ELISA was conducted by high-performance liquid chromatography. The correlation between the data obtained using the microwell assay and the high-performance liquid chromatography was good (R(2) = 0.9849). The developed immunoassay methods were suitable for the rapid quantitative or qualitative determination of phosmet in food samples.

Effects of postharvest preparation on organophosphate insecticide residues in apples.

Apples were sampled directly from orchard trees at 96, 45, and 21 days postapplication with one of three organophosphate insecticides (azinphos methyl, phosalone, or phosmet, respectively). Individual apples were prepared for analysis following one of three postharvest preparations: no preparation, rinsed with deionized water for 10-15 s, or rinsed and peeled. Azinphos methyl, phosalone, and phosmet concentrations ranged from below the level of detection to 5.26 ng/g, 94.7 to 5720 ng/g, and 0.011 to 663 ng/g in the apples that received no postharvest preparation, respectively. Although rinsed apples had lower maximum concentrations than observed in apples with no preparation, levels were not significantly lower. Concentrations of all three OP insecticides in apples that were rinsed followed by peeling, however, were much lower (below detection limits to 0.733 ng/g, azinphos methyl; 0.322-219 ng/g, phosalone; and below detection limits to 44.0 ng/g, phosmet) than observed in apples that had been rinsed alone. Rinsing and peeling of apples resulted in a 74.5-97.9% reduction in OP residues, while rinsing alone lowered mean concentrations by 13.5-28.7% relative to apples that received no postharvest preparation.

Performance of two monoclonal immunoassays in mixtures of cross-reacting dithiophosphorus pesticides.

A statistical approach for the analysis of complex samples by immunoassay is proposed in this article. Two enzyme-linked immunosorbent assays (ELISAs), one of them in the conjugate-coated format and the other in the antibody-coated format, were evaluated for their suitability to the analysis of mixtures of three organodithiophosphorus pesticides: azinphos-methyl, azinphos-ethyl and phosmet. It was found that the apparent affinity of the antibody to each analyte changed in the presence of a cross-reacting compound in the antibody-coated ELISA format, but not when the conjugate-coated ELISA format was used. The assays were thereafter applied to the analysis of mixtures of the three recognized pesticides. With the conjugate-coated ELISA format, accurate and precise determinations of mixtures could be performed if an azinphos-methyl standard curve was employed, with recoveries between 71% and 130% and with coefficients of variation lower than 12.7%. Neither accurate nor precise measurements could be accomplished with the enzyme immunoassay using the antibody-coated ELISA format, independently of the standard curve used. It is thought that the study presented here will have applicability in a variety of cases where the analytical goal is semiquantitative screening based on the total quantity of an unknown mixture of related compounds.

Gas chromatography with pulsed flame photometric detection multiresidue method for organophosphate pesticide and metabolite residues at the parts-per-billion level in representatives commodities of fruits and vegetable crop groups.

A gas chromatographic method with a pulsed flame photometric detector (P-FPD) is presented for the analysis of 28 parent organophosphate (OP) pesticides and their OP metabolites. A total of 57 organophosphates were analyzed in 10 representative fruit and vegetable crop groups. The method is based on a judicious selection of known procedures from FDA sources such as the Pesticide Analytical Manual and Laboratory Information Bulletins, combined in a manner to recover the OPs and their metabolite(s) at the part-per-billion (ppb) level. The method uses an acetone extraction with either miniaturized Hydromatrix column partitioning or alternately a miniaturized methylene dichloride liquid-liquid partitioning, followed by solid-phase extraction (SPE) cleanup with graphitized carbon black (GCB) and PSA cartridges. Determination of residues is by programmed temperature capillary column gas chromatography fitted with a P-FPD set in the phosphorus mode. The method is designed so that a set of samples can be prepared in 1 working day for overnight instrumental analysis. The recovery data indicates that a daily column-cutting procedure used in combination with the SPE extract cleanup effectively reduces matrix enhancement at the ppb level for many organophosphates. The OPs most susceptible to elevated recoveries around or greater than 150%, based on peak area calculations, were trichlorfon, phosmet, and the metabolites of dimethoate, fenamiphos, fenthion, and phorate.

Pesticide residues on fruits and vegetables from Ontario, Canada, 1991-1995.

For the 5-year period 1991 to 1995, 1536 vegetable and 802 fruit samples were analyzed. The purpose of this study was to determine if pesticides were present on Ontario-produced fruits and vegetables, and if so, to determine if residues violated maximum residue limits (MRLs). Overall, 31.5% of the samples had no detectable pesticide residues, whereas 68.5% contained one or more residues. Most of the residues were present at very low concentrations; 48% of the detections were < 0.1 parts per million (ppm), and 86% were < 1 ppm. However, violations of MRL were observed in only 3.2% of the vegetables samples and 3.1% of the fruit samples. In addition, 4.8% of the samples contained a "technical" violation, that is, there was no specified MRL for the pesticide-commodity combination and the residues exceeded 0.1 ppm. Of the detectable residues, 63% were < 10% of the MRL, whereas 89% were < 50% of the MRL. More fruit samples (91.4%) had a detectable residue, compared with vegetable samples (56.6%). Fruit is often treated close to harvest or post harvest to ensure that wholesome produce reaches the consumer. Forty-six percent of the samples contained 2 or more residues, and 2% of all samples had more than 5 different pesticides detected; fruit samples tended to have more multiple residues. The most frequently found pesticides were captan, the dithiocarbamate fungicides, endosulfan, azinphos-methyl, phosmet, parathion, and iprodione. These pesticides were also used in the greatest quantity for crop production. Overall, the data agree fairly closely with those reported for the U.S. Department of Agriculture Pesticide Data Program because the 2 programs have similar analytical goals and objectives.

Exposure of farmers to phosmet, a swine insecticide.

OBJECTIVES: The goal of this study was to measure dermal and inhalation exposures to phosmet during application to animals and to identify what determinants of exposure influence the exposure levels. METHODS: Ten farmers were monitored using dermal patches, gloves, and air sampling media during normal activities of applying phosmet to pigs for insect control. Exposures were measured on the clothing (outer), under the clothing (inner), on the hands, and in the air. Possible exposure determinants were identified, and a questionnaire on work practices was administered. RESULTS: The geometric mean of the outer exposure measurements was 79 microg/h, whereas the geometric mean of the inner exposure measurements was 6 microg/h. The geometric mean for hand exposure was 534 microg/h, and the mean air concentration was 0.2 microg/m3. Glove use was associated with the hand and total dermal exposure levels, but no other determinant was associated with any of the exposure measures. The average penetration through the clothing was 54%, which dropped to 8% when the farmers wearing short sleeves were excluded. The farmers reported an average of 40 hours a year performing insecticide-related tasks. CONCLUSIONS: Farmers who applied phosmet to animals had measurable exposures, but the levels were lower than what has been seen in other pesticide applications. Inhalation exposures were insignificant when compared with dermal exposures, which came primarily from the hands. Clothing, particularly gloves, provided substantial protection from exposures. No other exposure determinant was identified.

Phosmet residues in an orchard and adjacent recreational area.

Two cover sprays of phosmet were applied to an orchard adjoining a camping area and a bird sanctuary with a resident goose population. Insecticide residues were monitored on orchard leaves, orchard ground cover, ground cover in the camp-site and along the adjacent lakeshore. Despite attempts to minimize drift, significant spray residues were found outside the target area. Residues on ground cover and leaves were reduced by sprinkler irrigation subsequent to spray application.