A bispecific nanobody with high sensitivity/efficiency for simultaneous determination of carbaryl and its metabolite 1-naphthol in the soil and rice samples.

The simultaneous determination of carbaryl and its metabolite 1-naphthol is essential for risk assessment of pesticide exposure in agricultural and environmental samples. Herein, several bispecific nanobodies (BsNbs) with different lengths of hydrophilic linkers and junction sites were prepared and characterized for the simultaneous recognition of carbaryl and its metabolite 1-naphthol. It was found that the affinity of BsNbs to the analytes could be regulated by controlling linker length and linking terminal. Additionally, molecular simulation revealed that linker lengths affected the conformation of BsNbs, leading to alteration in sensitivity. The BsNb with G4S linker, named G4S-C-N-VHH, showing good thermal stability and sensitivity was used to develop a bispecific indirect competitive enzyme-linked immunosorbent assay (Bic-ELISA). The assay demonstrated a limit of detection of 0.8 ng/mL for carbaryl and 0.4 ng/mL for 1-naphthol in buffer system. Good recoveries from soil and rice samples were obtained, ranging from 80.0% to 112.7% (carbaryl) and 76.5%-110.8% (1-naphthol), respectively. Taken together, this study firstly provided a BsNb with high sensitivity and efficiency against environmental pesticide and its metabolite, and firstly used molecular dynamics simulation to explore the influence of linker on recognition. The results are valuable for the application of immunoassay with high efficiency in the fields of environment and agriculture.

Au nanogap SERS substrate for the carbaryl pesticide determination in juice and milk using chemomterics.

Conventional spectroscopic methods like IR, and Raman are not very effective at detecting low levels of pesticides or harmful chemicals in food matrices. A quick, highly accurate approach that can identify pesticides present in different food products at lower levels must be developed in order to address this problem and ensure food safety. In this study, a highly sensitive and uniform wafer-scale Au nanogap surface-enhanced Raman spectroscopy (SERS) substrate was used for the quantitative analysis of carbaryl pesticide levels in standard solution, mango juice, and milk samples using chemometrics. Carbaryl was detected up to 3 ppb concentration levels for all three group of samples. However, due to the higher sensitivity, uniformity, and enhancement factors of the SERS substrate used in this investigation, the limit of detection (LOD) values for the standard solution, mango juice, and milk were 0.37 ppb, 0.57 ppb, and 0.15 ppb at 1380 cm-1, 1380 cm-1, and 1364 cm-1 wavenumber ranges. In order to predict different carbaryl concentrations (1, 2, 3, 4, and 5 ppb), the variable importance in projection (VIP) method combined with partial least squares regression (PLSR) and attained the coefficient of determination (R2) values of 0.994, 0.989, and 0.978 along with minimum root mean square error (RMSE) values of 0.112, 0.190, and 0.278 ppb for the prediction datasets. Furthermore, PLS-DA was able to distinguish between pure and adulterated samples with the highest classification accuracy of 100 % for a standard solution, and mango juice and 94.4 % for milk samples. Considering this, we can conclude that the SERS Au Nanogap substrate can rapidly and effectively detect carbaryl pesticides quantitatively and qualitatively in mango juice and milk.

Ovalbumin-coated gold nanoparticles with interesting colloidal stability for colorimetric detection of carbaryl in complex media.

Pesticide carbaryl can cause serious environmental pollution and its sensitive detection is of increasing interest. Gold nanoparticles (AuNPs) are classically colorimetric probes for detection of many analytes, but the instability in complex media limits their application. Here, Au@Ova NPs have been developed as a stable, effective, sensitive, and selective sensing system for colorimetric detection of carbaryl. Au@Ova NPs present unique and proper colloidal stability in various medias containing salt, small molecules, organic solvent (DMSO), and seawater, which are distinct from previous ones including citrate (or rhodamine B) capped AuNPs. Compared with Au@BSA NPs, Au@Ova NPs showed efficient responses to carbaryl by inhibiting acetylcholinesterase, with a linear concentration range between 0 and 25 µg/L and a detection limit of 0.25 µg/L. In addition, this nanoprobe also has good selectivity and can be applied in different real samples analysis, including fruit juice (tomato and apple) and real water samples (artificial urine and seawater).

Digital image colorimetry in combination with chemometrics for the detection of carbaryl based on the peroxidase-like activity of nanoporphyrins and the etching process of gold nanoparticles.

Carbaryl is a typical carbamate pesticide that plays an essential role in agricultural production, but its residues cause serious harm to the environment and human health. Here, we developed a polychromatic colorimetric sensor based on ZnTPyP-DTAB peroxidase activity and gold nano-bipyramids (Au NBPs) etching to detect carbaryl. ZnTPyP-DTAB catalyzes the decomposition of H2O2 to hydroxyl radicals, and Au NBPs are etched. The coordination of zinc and nitrogen in nanometer porphyrins was affected by the steric effects of carbaryl, which resulted in decreased activity of ZnTPyP-DTAB peroxidase. The detection limit of carbaryl was 0.26 mg/kg. The recoveries of carbaryl in reaal sample ranged from 91 % to 107% (RSD ≤ 0.7%). The sensor platform displayed a series of high-resolution multicolor variations of rainbow colors within the above concentration range. The rich color variation facilitates the acquisition of digital images. RGB value transformation combined with partial least squares regression model can accurately and quantitatively detect carbaryl in vegetables, fruits and Chinese medicinal materials.

Highly porous seeding-free boron-doped ultrananocrystalline diamond used as high-performance anode for electrochemical removal of carbaryl from water.

Boron-doped diamond (BDD) electrodes are regarded as the most promising catalytic materials that are highly efficient and suitable for application in advanced electrochemical oxidation processes targeted at the removal of recalcitrant contaminants in different water matrices. Improving the synthesis of these electrodes through the enhancement of their morphology, structure and stability has become the goal of the material scientists. The present work reports the use of an ultranano-diamond electrode with a highly porous structure (B-UNCDWS/TDNT/Ti) for the treatment of water containing carbaryl. The application of the proposed electrode at current density of 75 mA cm-2 led to the complete removal of the pollutant (carbaryl) from the synthetic medium in 30 min of electrolysis with an electric energy per order of 4.01 kWh m-3 order-1. The results obtained from the time-course analysis of the carboxylic acids and nitrogen-based ions present in the solution showed that the concentrations of nitrogen-based ions were within the established maximum levels for human consumption. Under optimal operating conditions, the proposed electrode was successfully employed for the complete removal of carbaryl in real water. Thus, the findings of this study show that the unique, easy-to-prepare BDD-based electrode proposed in this study is a highly efficient tool which has excellent application potential for the removal of recalcitrant pollutants in water.

Passivated graphene quantum dots for carbaryl determination in juices.

This paper reports a simple method for the preparation of suitable graphene quantum dots after surface passivation, to be used for the determination of carbaryl in juice samples. A comparison of synthetic conditions for the preparation of graphene quantum dots following the top-down approach is described. In the one-step route selected, evaluation of diverse reaction time for cutting and modulating the oxidizing sites in the broken pieces of the initial graphene layer is conducted with a mixture of concentrated acids. Exploring the passivation effect on the purified graphene quantum dots, we demonstrated the suitability of the selected graphene quantum dots for practical application in the detection of carbaryl using fluorometric detection. Higher sensitivity was achieved after 8 min of contact, in which graphene quantum dots promotes the degradation of carbaryl into naphthol, being the latter responsible for the analytical signal. The detection and quantification limits were 0.36 and 1.21 µg/L, respectively, being the response linear up to 26 µg/L with excellent precision (better than 3.2% at the limit of detection). The recovery of the analyte from commercial juice samples (91.4-96.7%) testifies to the applicability of the proposal for the analytical problem selected.

Core-shell magnetic Ag-molecularly imprinted composite for surface enhanced Raman scattering detection of carbaryl.

Surface-enhanced Raman scattering (SERS) is a promising technique for rapid detection of pesticide residues. However, conventional SERS substrates require extraction processes which are time consuming and they also lack selectivity, stability and reproducibility. Herein, we present a multifunctional stable zero-valent iron based core-shell substrate. It combines magnetic separation, selective adsorption by molecular imprinting technique and sensitive detection of carbaryl by SERS. The core-shell substrate was successfully prepared by immobilizing silver on the surface of zero-valent iron microspheres. Subsequent molecular imprinting on the bimetallic magnetic silver microspheres ensured selective removal and detection. The substrate exhibited magnetization saturation of 8.89 emu/g providing efficient analyte separation. It showed high sensitivity and selectivity toward carbaryl detection to nanomolar concentration level. Linear regression models for peaks at Raman shift 1599 cm-1 and 2233 cm-1 demonstrated a good linear fit with R2=0.9738 and R2=0.8952 respectively. The composite was successfully applied on spiked water samples resulting in average recovery rate of 89%. The findings of this study demonstrate great substrate potential for application in separation and detection of trace quantities of chemical contaminants for environment safety and protection.

A ratiometric fluorescence nanoprobe using CdTe QDs for fast detection of carbaryl insecticide in apple.

In this study, a novel, simple and sensitive ratiometric fluorescence method is presented for the detection of very low quantities of the carbaryl in Iranian apple using cadmium telluride quantum dots (CdTe QDs) nanoprobe. The principle of the proposed strategy relies on the rapid hydrolysis of the carbaryl under an alkaline condition and production of the 1-naphthol with a blue emission at 470 nm. Besides, using the CdTe QDs with a yellow emission at 580 nm, as a reference, improves the visual tracking of carbaryl through changes in color tonality. The herein described methodology is applied for enzyme-free visual detection of carbaryl with satisfactory results in the presence of other common pesticides in Iranian apple sample. Additionally, the calculated limit of detection (LOD) of 0.12 ng mL-1 for carbaryl is much lower than the maximum residue limits of carbaryl warned value reported by the European Union and United States pesticides database, which is so promising for carbaryl sensing in the monitoring of fruits. Furthermore, the ability of the proposed method for the detection of carbaryl residues in Iranian apple was confirmed by the high-performance liquid chromatography (HPLC) method as a standard method through statistical analysis. This fast and highly sensitive naked-eye ratiometric sensor may hold great promise to provide the technical support for the rapid and valid detection of other targets in food safety fields.

Digital image colorimetry detection of carbaryl in food samples based on liquid phase microextraction coupled with a microfluidic thread-based analytical device.

This research used a digital image colorimetry (DIC) method to detect carbaryl in food samples using effervescence-assisted liquid phase microextraction based on solidification of switchable hydrophilicity solvent combined with a microfluidic thread-based analytical device (EA-LPME-SSHS-µTAD). 1-naphthol, the hydrolysate of carbaryl, was extracted into octanoic acid by the adjustment of pH values of the sample solution and separated through solidification in an ice bath. Then 1-naphthol contained in the extracted solution was coupled with 4-methoxybenzenediazonlum tetrafluoroborate (MBDF) fixed on the µTAD to produce tangerine compounds. The inherent colour variation was captured by a smartphone and processed to calculate the intensity (I). Under the optimal conditions, the limit of quantification was within 0.020-0.027 mg kg-1. The recovery was varied in the range from 92.3% to 105.9% with a relative standard deviation (RSD) below 5%. The developed method provides an alternative strategy to extract and detect pesticides for food samples.

The effect of ozonation on the degradation of carbaryl in aqueous solution.

The aim of this study is to investigate the synergetic effect of ozonation on ultraviolet irradiation on the degradation of carbaryl (CBL) (1-naphthyl-N-methylcarbamate) in aqueous solutions (40 mg L-1). The degradation of CBL and the intermediates formed during ozonation were determined by gas chromatography-mass spectrometer. The change in formaldehyde, dissolved oxygen, pH, and total acidity were determined as a function of ozonation time. It was found that the dissolved oxygen and total acidity were increased with the increasing ozonation time but the pH was decreased with the increasing ozonation time. The aliphatic acids formed during ozonation were followed by ion chromatography as a function of ozonation time. It was found that the effect of ozonation is highly effective on the degradation of CBL. The complete degradation of CBL has occurred at 75-second ozonation time. As a consequence of the obtained results the possible degradation pathway was proposed. The results confirmed that ozonation is an effective method for the degradation of CBL.

Ratiometric Strategy for Electrochemical Sensing of Carbaryl Residue in Water and Vegetable Samples.

Accurate analysis of pesticide residue in real samples is essential for food safety and environmental protection. However, a traditional electrochemical sensor based on single-signal output is easily affected by background noise, environmental conditions, electrode diversity, and a complex matrix of samples, leading to extremely low accuracy. Hence, in this paper, a ratiometric strategy based on dual-signal output was adopted to build inner correction for sensing of widely-used carbaryl (CBL) for the first time. By comparison, Nile blue A (NB) was selected as reference probe, due to its well-defined peak, few effects on the target peak of CBL, and excellent stability. The effects of a derivatization method, technique mode, and pH were also investigated. Then the performance of the proposed ratiometric sensor was assessed in terms of three aspects including the elimination of system noise, electrode deviation and matrix effect. Compared with traditional single-signal sensor, the ratiometric sensor showed a much better linear correlation coefficient (r > 0.99), reproducibility (RSD < 10%), and limit of detection (LOD = 1.0 µM). The results indicated the introduction of proper reference probe could ensure the interdependence of target and reference signal on the same sensing environment, thus inner correction was fulfilled, which provided a promising tool for accurate analysis.

Detection of DDT and carbaryl pesticides in honey by means of immunosensors based on high fundamental frequency quartz crystal microbalance (HFF-QCM).

BACKGROUND: In recent years there has been a concern about the presence of pesticides in honey because residues of DDT and carbaryl were found in honey samples. Traditional techniques, such as chromatography, reach the required limits of detection (LOD) but are not suitable for in situ implementation in the honey-packaging industry due to their high cost and the need for highly qualified staff for routine operation. Biosensors offer simplicity, low cost, and easy handling for analytical purposes in food applications. RESULTS: Piezoelectric immunosensors based on high fundamental frequency quartz crystal microbalance (HFF-QCM) have been developed for the detection of carbaryl and DDT in honey. Biorecognition was based on competitive immunoassays in the conjugate-coated format, using monoclonal antibodies as specific immunoreagents. The assay LODs attained by the HFF-QCM immunosensors were 0.05 µg L-1 for carbaryl and 0.24 µg L-1 for DDT, reaching a similar level of detectability to that of the usual reference techniques. The practical LODs in honey samples were 8 µg kg-1 for carbaryl and 24 µg kg-1 for DDT. The immunosensors' analytical performance allow the detection of these pesticides in honey at EU regulatory levels with good accuracy (recovery percentages ranging from 94% to 130% within the working range of each pesticide standard curve) and precision (coefficients of variation in the 9-36% range). CONCLUSION: The proposed immunosensor is a promising analytical tool that could be implemented for quality control in the honey packaging industry, to simplify and to reduce the cost of the routine pesticide analysis in this appreciated natural food. © 2020 Society of Chemical Industry.

The effectiveness of chemical solutions on the removal of carbaryl residues from cucumber and chili presoaked in carbaryl using the HPLC technique.

This research aimed to compare the efficiency of five washing solutions (0.9% NaCl, 0.1% NaHCO3, DI water, 0.001% KMnO4, and 0.1% acetic acid) for removing carbaryl residues from cucumber and chili. The vegetables were soaked in 10 mg/L of carbaryl solution for 30 min and then washed for 30 min in one of the five washing solutions and the results compared. Each experiment was performed in triplicate and the amounts of carbaryl residues remaining were determined using high-performance liquid chromatography (HPLC). The results showed that 0.001% KMnO4 was the most effective at removing carbaryl from both vegetables. Washing with 0.001% KMnO4 reduced carbaryl residues to 64% and 28%, respectively, of the original concentrations. Washing with DI water was the least effective method of removing carbaryl residues. Hopefully, the results will encourage further research, into reducing carbaryl contamination by washing with chemical solutions, which will enable producers to reduce pesticide residues.

Leaching of insecticides used in blueberry production and their toxicity to red worm.

Soil columns were collected from a blueberry field, and insecticide solutions were allowed to leach through these columns. Insecticides from four different chemical classes were applied at two different rates: the concentration at which the insecticides wash off blueberries under rainfall conditions and the labeled field rate at which they are sprayed. The soil columns were divided into thirds; top, middle and bottom. Soil bioassays using Eisenia foetida Savigny, as an indicator species, were set up to determine the toxicity of the insecticides at a top, middle and bottom layer of the soil column. The mass of E. foetida was also measured after the bioassay experiment was completed. The concentrations at which insecticides wash-off of blueberries from rainfall were not lethal to E. foetida. In order to support mortality data, insecticide residues were quantified in the soil layers for each insecticide. Under field rate leaching conditions, carbaryl showed the high levels of toxicity in the top and middle layers of soil suggesting that it has the highest risk to organisms from leaching. This study will help blueberry growers make informed decisions about insecticide use, which can help minimize contamination of the environment.

Reduced graphene oxide decorated on Cu/CuO-Ag nanocomposite as a high-performance material for the construction of a non-enzymatic sensor: Application to the determination of carbaryl and fenamiphos pesticides.

A novel electrochemical sensor based on the reduced graphene oxide-Cu/CuO-Ag nanocomposite modified glassy carbon electrode (rGO/Cu/CuO-Ag/GCE) has been applied for the simultaneous analysis of carbaryl and fenamiphos as two important pesticides. The electrochemical behavior of carbaryl and fenamiphos at rGO/Cu/CuO-Ag/GCE was studied by cyclic voltammetry and differential pulse voltammetry. The modified electrode exhibited two separated oxidation signals for the simultaneous determination of both carbaryl and fenamiphos with excellent sensitivity. The characteristics of the modified electrode were studied with transmission electron microscopy, X-ray diffraction and Fourier transform-infrared spectroscopy techniques. Under optimized conditions, the rGO/Cu/CuO-Ag/GCE detected carbaryl and fenamiphos with the wide linear ranges of 0.05-20 and 0.01-30 µM, and the detection limits were 0.005 and 0.003 µM, respectively. This developed electrochemical platform applied as a simple and cost-effective sensor for the detection of low levels of carbaryl and fenamiphos in fruit and vegetable samples successfully.

One-step immunoassay for the insecticide carbaryl using a chicken single-chain variable fragment (scFv) fused to alkaline phosphatase.

Immunoassays provide a high-throughput method for monitoring pesticides in foods and the environment. Due to easy generation and capable of being manipulated, chicken single-chain variable fragment (scFv) is attractive in the development of immunoassays for pesticides. Two scFvs (X1 and X2) against the insecticide carbaryl were generated from a chicken immunized with hapten C1 conjugated to keyhole limpet hemocyanin and fused with alkaline phosphatase (AP) to develop a rapid one-step enzyme-linked immunosorbent assay for this pesticide. X2-AP showed higher binding affinity to carbaryl than X1-AP. The X2-AP-based ELISA had a half-maximum signal inhibition concentration of 15 ng mL-1 and a limit of detection of 1.6 ng mL-1. This assay showed negligible cross-reactivity with other carbamate pesticides (<0.1%) and low cross-reactivity with 1-naphthol (5%). The average recoveries of carbaryl spiked in soil, apple and pear samples by the one-step assay ranged from 90% to 114% and agreed well with those of high-performance liquid chromatography. The chicken scFv-based assay showed promise as a high-throughput screening tool for carbaryl in environmental and food matrices.

Development of an immunoassay for the detection of carbaryl in cereals based on a camelid variable heavy-chain antibody domain.

BACKGROUND: The variable domain of camelid heavy-chain antibodies (VHH) is increasingly being adapted to detect small molecules in various matrices. The insecticide carbaryl is widely used in agriculture while its residues have posed a threat to food safety and human health. RESULTS: VHHs specific for carbaryl were generated from an alpaca immunized with the hapten CBR1 coupled to keyhole limpet hemocyanin. An enzyme-linked immunosorbent assay (ELISA) based on the VHH C1 and the coating antigen CBR2-BSA was developed for the detection of carbaryl in cereals. This assay, using an optimized assay buffer (pH 6.5) containing 10% methanol and 0.8% NaCl, has a half-maximum signal inhibition concentration of 5.4 ng mL-1 and a limit of detection (LOD) of 0.3 ng mL-1 for carbaryl, and shows low cross reactivity (≤0.8%) with other tested carbamates. The LOD of carbaryl using the VHH-based ELISA was 36 ng g-1 in rice and maize and 72 ng g-1 in wheat. Recoveries of carbaryl in spiked rice, maize and wheat samples were in the range of 81-106%, 96-106% and 83-113%, respectively. Relative standard deviations of repeatability and intra-laboratory reproducibility were in the range of 0.8-9.2% and 2.9-9.7%, respectively. CONCLUSION: The VHH-based ELISA was highly effective in detecting carbaryl in cereal samples after simple sample extraction and dilution. © 2019 Society of Chemical Industry.

Engineering the effector specificity of regulatory proteins for the in vitro detection of biomarkers and pesticide residues.

Transcriptional regulatory proteins (TRPs)-based whole-cell biosensors are promising owing to their specificity and sensitivity, but their applications are currently limited. Herein, TRPs were adapted for the extracellular detection of a disease biomarker, uric acid, and a typical pesticide residue, carbaryl. A mutant regulatory protein that specifically recognizes carbaryl as its non-natural effector and activates transcription upon carbaryl binding was developed by engineering the regulatory protein TtgR from Pseudomonas putida. The TtgR mutant responsive to carbaryl and a regulatory protein responsive to uric acid were used for in vitro detection, based on their allosteric binding of operator DNA and inducer molecules. Based on the quantitative polymerase chain reactions (qPCRs) output, the minimum detectable concentration was between 1 nM-1 µM and 1-10 nM for uric acid and carbaryl, respectively. Our results demonstrated that engineering the effector specificity of regulatory proteins is a potential technique for generating molecular recognition elements for not only in vivo but also in vitro applications.

Design of an ultrasonic piezoelectric injection port for analysis of thermally unstable compounds using corona discharge ion mobility spectrometry.

This paper reports on a novel ultrasonic injection port designed and constructed to analyze thermally unstable chemical compounds using corona discharge ion mobility spectrometry (CD-IMS). In order to achieve the highest possible efficiency with the device, some parameters such as the solvent type, carrier gas flow rate and sample volume were exhaustively investigated. Through a comparative study conducted, it was revealed that unlike the thermal desorption system, the proposed ultrasonic injection port could easily be used for the analysis of some thermally unstable compounds such as carbaryl, propoxur and vitamin B1, by means of CD-IMS. To evaluate the potential of the device, carbaryl, extracted from different samples by dispersive liquid-liquid microextraction technique, was quantitatively analyzed. The CD-IMS-based results brought forth the detection limit of 0.03 mg L-1, and dynamic range of 0.1-10.0 mg L-1 with the determination coefficient of 0.9981. The relative standard deviations for one day and three consecutive days were 4 and 6%, respectively. Further, the spiked samples of agricultural wastewater, underground water, and tomato analyzed culminated in the recovery values of 83%, 98% and 82%, respectively. The satisfactory results proved an acceptable capability of the sample introduction system, to be conveniently used for routine analysis of thermally unstable compound, without any tedious derivatization.

Nano carbon black-based screen printed sensor for carbofuran, isoprocarb, carbaryl and fenobucarb detection: application to grain samples.

An electrochemical screening assay for the detection of phenyl carbamates (i.e. carbaryl, carbofuran, isoprocarb and fenobucarb) was developed and applied to grains samples (i.e. durum wheat, soft wheat and maize). Nano carbon black (CB) was strategically employed to realize an effective, reproducible, fouling resistant, low cost, delocalisable screen printed sensor (CB-SPE). CB-SPEs morphology (SEM and FEM) and electrochemical property (CV and EIS) were studied. The final pesticides analysis protocol consist of: (i) extraction of the analyte (just by mixing), (ii) alkaline hydrolysis (10 min R.T.), (iii) DPV detection directly of 100 µL of extract on the CB-SPE surface. Linear range between 1.0 × 10-7 and 1.0 × 10-4 mol L-1, good determination coefficients (R2 ≥ 0.9971) and satisfactory sensitivity (≥ 3.90 × 10-1 A M-1 cm-2) and LODs (≤ 8.0 × 10-8 mol L-1) were obtained for all the analytes. Excellent recoveries (78-102%) and accuracy (relative error vs. HPLC-MS/MS between 9.0% and -7.8%) resulted from the analysis of grains samples. The proposed CB-SPE based approach has demonstrated to be able to detect carbaryl at Maximum residue limits levels (MRLs), allowing class selective detection of commonly employed phenyl carbamates in food samples.