Determination Of Pesticides Research Articles

On-line immobilized acetylcholinesterase microreactor based on capillary electrophoresis for the determination of organophosphorus pesticide residues

• Acetylcholinesterase is simply immobilized on the capillary via polydopamine coating. • The CE-based acetylcholinesterase microreactor is developed for the rapid detection of acephate. • The proposed method is capable to achieve rapid analysis of acephate with simple operation. In this study, a capillary electrophoresis-based online immobilized acetylcholinesterase (AChE) microreactor was developed for the determination of organophosphorus pesticide (OPs) residues in six fruit and vegetable juice samples. AChE was immobilized on the inner wall at the inlet of the capillary via polydopamine (PDA) coating. The immobilized AChE activity and other kinetic parameters were evaluated by measuring the peak area of the product (thiocholine). The value of Michaelis-Menten constant ( K m ) was determined to be 3.11 mM, and the half-maximal inhibitory concentration (IC 50 ) and inhibition constant ( K i ) of tacrine on immobilized AChE were measured to be 0.46 μM and 0.27 μM, respectively. The activity of the immobilized AChE remained more than 85.0% of its initial immobilized enzyme activity after 30 runs. Then, the developed strategy was successfully applied to detect the acephate in six fruit and vegetable juice samples (apple, pear, peach, tomato, cucumber and lettuce juice) with a limit of detection (LOD) of 11.0 ng/mL. Results show that the recovery of spiked samples are in the range of 86.0-96.0% with RSD of less than 5.7%. Therefore, the proposed method is capable of achieving rapid analysis of acephate with simple operation when there are no other types of OPs in the real samples.

Determination of new generation amide insecticide residues in complex matrix agricultural food by ultrahigh performance liquid chromatography tandem mass spectrometry

Eight new generation amide insecticide residues analysis by multiwalled carbon nanotubes (MWCNs) cleanup, combined with QuEChERS and ultrahigh performance liquid chromatography tandem mass spectrometry has been developed and successfully applied in complex matrix such as orange, celery, onion, litchi, mango, shallot, chives, avocado, garlic. The matric effect of MWCNs is optimized and compared with ordinary cleanup materials. The results show that the performance of MWCNs is fine and effectively reduce matrix interference. Through chemical structure skeletons analyzed, chlorantraniliprole, bromoantraniliprole, and cyantraniliprole can cause same product ions of m/z 286.0 or 177.1 in the ESI+ mode, then tetrachlorantraniliprole and cyclaniliprole can produce collective ions of m/z 146.9 in the ESI− mode. The coefficients (R2) were greater than 0.9990, the limit of quantification ranges from 0.03 to 0.80 μg/kg, the recovery rate ranges from 71.2 to 120%, and the relative standard deviation (RSD) ranges from 3.8 to 9.4%. The method is fast, simple, sensitive, and suitable for the rapid determination of amide pesticides in complex matrix agricultural food.

Ringer Tablet-Based Micelle-Mediated Extraction-Solvent Back Extraction Coupled with High-Performance Liquid Chromatography for Preconcentration and Determination of Neonicotinoid Pesticides

Ringer Tablet-Based Micelle-Mediated Extraction-Solvent Back Extraction Coupled with High-Performance Liquid Chromatography for Preconcentration and Determination of Neonicotinoid Pesticides

Facile Detection and Quantification of Acetamiprid Using a Portable Raman Spectrometer Combined with Self-Assembled Gold Nanoparticle Array

Rapid and facile determination of pesticides is critically important in food and environmental monitoring. This study developed a self-assembled gold nanoparticle array based SERS method for highly specific and sensitive detection of acetamiprid, a neonicotinoid pesticide that used to be difficult in SERS analysis due to its low affinity with SERS substrates. SERS detection and quantification of acetamiprid was conducted with self-assembled gold nanoparticle arrays at the interface of chloroform and water as the enhancing substrate. Since targets dissolved in chloroform (organic phase) also have access to the hot-spots of Au NP array, the developed method exhibited good sensitivity and specificity for acetamiprid determination. Under the optimal conditions, SERS intensities at Raman shifts of 631 cm−1 and 1109 cm−1 displayed a good linear relationship with the logarithm concentration of acetamiprid in the range of 5.0 × 10−7 to 1.0 × 10−4 mol/L (0.11335 ppm to 22.67 ppm), with correlation coefficients of 0.97972 and 0.97552, respectively. The calculated LOD and LOQ of this method were 1.19 × 10−7 mol/L (0.265 ppb) and 2.63 × 10−7 mol/L (0.586 ppb), respectively, using SERS signal at 631 cm−1, and 2.95 × 10−7 mol/L (0.657 ppb) and 3.86 × 10−7 mol/L (0.860 ppb) using SERS signal at 1109 cm−1, respectively. Furthermore, the developed SERS method was successfully applied in determining acetamiprid on the surface of apple and spinach. This method offers an exciting opportunity for rapid detection of acetamiprid and other organic pesticides considering its advantages of simple preparation process, good specificity and sensitivity, and short detection time (within 1 h).

Multivariate Assessment of Procedures for Molecularly Imprinted Polymer Synthesis for Pesticides Determination in Environmental and Agricultural Samples.

In the case of quantitative and qualitative analysis of pesticides in environmental and food samples, it is required to perform a sample pre-treatment process. It allows to minimalize the impact of interferences on the final results, as well as increase the recovery rate. Nowadays, apart from routinely employed sample preparation techniques such as solid-phase extraction (SPE) or solid-phase microextraction (SPME), the application of molecularly imprinted polymers (MIPs) is gaining greater popularity. It is mainly related to their physicochemical properties, sorption capacity and selectivity, thermo-mechanical resistance, as well as a wide range of polymerization techniques allowing to obtain the desired type of sorption materials, adequate to a specific type of pesticide. This paper targets to summarize the most popular and innovative strategies since 2010, associated with the MIPs synthesis and analytical procedures for pesticides determination in environmental and food samples. Application of multi-criteria decision analysis (MCDA) allows for visualization of the most beneficial analytical procedures in case of changing the priority of each step of analysis (MIPs synthesis, sample preparation process—pesticides extraction, chromatographic analysis) bearing in mind metrological and environmental issues.

Use of Poly(vinyl alcohol)-Malic Acid (CLHPMA) Hydrogels and Chitosan Coated Calcium Alginate (CCCA) Microparticles as Potential Sorbent Phases for the Extraction and Quantitative Determination of Pesticides from Aqueous Solutions.

Pesticides are used worldwide to increase crop yields in agriculture. However, their toxicity and accumulation capacity can make them toxic to the environment, animals and humans. In the case of workers chronically exposed to these substances, they must be sampled continuously, so urine is an excellent option. In this sense, this study proposes to use poly(vinyl alcohol)-malic acid hydrogels, and chitosan-coated calcium alginate as new sorbent phases to be used in pesticide determination processes in urine. To better understand the behavior of these materials in the capture and desorption process, molecular dynamics simulations (MDS) were used, and desorption experiments were performed, using mechanical agitation, ultrasound, and pH variation in the desorption process, in order to optimize the parameters to obtain better recoveries. Under the optimal experimental conditions, the maximum recoveries were of the order of 11% (CFN), 3% (KCF), 53% (DMT), 18% (MTD) and 35% (MTL). Although the recoveries were not exhaustive, they are a first approximation for the use of these new sorbent phases in the determination of this type of compound in aqueous solutions and urine.

Determination of 197 pesticide residues in edible vegetable oil by gas chromatography-time-of-flight mass spectrometry

建立了气相色谱-飞行时间质谱(GC-TOF-MS)同时测定食用植物油中197种农药残留的方法。样品经乙腈超声提取,冷冻除脂,C18和PSA吸附剂共同净化;目标物经HP-5MS UI毛细管柱(30 m×0.25 mm×0.25 μm)分离,电子轰击源电离,全扫描模式采集质谱信息;MassHunter软件对数据进行定性与定量分析,设置精确质量数偏差为±5×10-5,保留时间偏差为±0.1 min。实验考察了基质效应情况和方法学性能。结果表明,大多数农药表现出基质增强效应,需采用基质标准工作溶液进行定量。在优化的条件下,174种农药定量限可以达到0.01 mg/kg,占全部被测农药的88%,另外23种农药的定量限为0.025~0.1 mg/kg。除联苯的线性范围为2~100 μg/L外,其余农药的线性范围均为定量限~200 μg/L,相关系数(R2)均大于0.99。在3个添加水平(0.1、0.25和0.5 mg/kg)下,有156种农药的回收率为70%~120%,占全部被测农药的79%,有185种农药的相对标准偏差<10%,占全部被测农药的94%。应用该方法对23份市售植物油样品进行了检测,结果在12个样品中检出13种农药。该方法操作简便,一次进样即可实现近200种农药的同时检测,且检测结果的准确度和灵敏度良好,适用于食用植物油中197种农药残留的快速检测。

Adsorptive stripping voltammetric determination of carbofuran in herbs on chromatographic sorbent modified electrode

• Novel carbon-containing modified electrode was developed for carbofuran determination. • Carbon ink containing chromatographic sorbent, polyethylene glycol, and iron acetylacetonate was used. • The practical applicability of the novel electrode was verified on determination of carbofuran in herbs. The determination of carbofuran (CBF) in herbs (wild chamomile and Saint-John's-wort) was developed using a carbon-containing electrode (CCE) modified with carbon ink (CI) containing chromatographic sorbent (CS) based on chromaton (Ch), polyethylene glycol (PEG), and iron acetylacetonate (Fe(AA) 3 ). The surface morphology of the Ch/PEG/Fe(AA) 3 /CI/CCE was investigated by scanning electron microscopy (SEM) in backscattered-electron (BSE) mode and by electrochemical impedance spectroscopy (EIS). Electrochemically inactive CBF was converted to an electrochemically active phenolic analogue CBFP by an alkaline hydrolysis. The electrochemical behavior of CBFP at Ch/PEG/Fe(AA) 3 /CI/CCE was studied by cyclic voltammetry (CV) in a phosphate buffer solution (PBS) pH 6.86 in the potential range from −0.4 to 1.2 V. Under the optimal experimental conditions, linear concentration dependences in the concentration range from 0.25 to 10 µmol·L −1 were obtained by linear scan voltammetry in the first derivative mode (LSVFD) with limit of detection ( LOD ) 0.08 µmol·L −1 . The proposed Ch/PEG/Fe(AA) 3 /CI/CCE demonstrated good sensitivity and selectivity for the determination of CBF after its conversion to CBFP. The use of sorbents as modifying agents of electrodes for the determination of carbamate pesticides opens up the possibilities for “green” electroanalytical chemistry.

Liquid–liquid extraction procedure for nonvolatile pesticides determination in acacia honey as environmental biomonitor

Liquid–liquid extraction procedure for nonvolatile pesticides determination in acacia honey as environmental biomonitor

Usefulness of Dilution for Rapid Simultaneous Determination of Residual Pesticides in Agricultural Products by LC-MS/MS

A rapid, easy and versatile, simultaneous analytical method by LC-MS/MS based on extraction of QuEChERS method (EN 15662:2008) was developed for the determination of residual pesticides in agricultural products. In this method, it allowed to prepare a test solution only dilution without purification using solid phase extraction column. The method was assessed for 210 pesticide residues in 21 kinds of vegetables and fruits at the fortification levels of 0.01 and 0.1 μg/g according to the method validation guideline (Ministry of Health, Labour and Welfare of Japan). As a result, 194 to 210 analytes met the management criteria of the guideline. Thus, the present method could be useful for a rapid simultaneous determination of residual pesticides in agricultural products.

Determination of Antibiotics, Pesticides, Herbicides, Fungicides and Hormones in Water Bodies in Italy in Occurrence with European Watch List Mechanism by Using an UHPLC-MS/MS System: Method Validation, Quantification and Evaluations

In recent years, the quality of aquatic ecosystems has received increasing attention from European institutions. The Commission Implementing Decision (EU) 2018/840 drafted a Watch List (WL) of compounds to be monitored in Europe. In this study, we report a method based on solid phase extraction with ultra-high-performance liquid chromatography, coupled with a triple-quadrupole mass spectrometer (UHPLC-MS/MS) to analyze the whole water sample. The method was developed and validated for the determination of 12 listed compounds. The employment of solid-phase extraction by a horizon system ensures the analysis of the entire body of samples and minimizes sample manipulation. Different ng L−1 detection limits (from 2 to 50 ng L−1), linearities (from 2 to 500 ng L−1), accuracy (from 70 to 130%) and levels of precision (RSD less 20% at LOQs levels) were assessed to be satisfactory for quantification and confirmation at the levels of interest. The developed method was applied for quantitative analysis for Watch List compounds (with the exception of hormones) in surface water samples from different Italian sites during monitoring activities by the Regional Environmental Protection Agencies in the years 2019 and 2020.

Fluorometric determination of ziram using CsPbBr3 quantum dots.

Astrategy based on CsPbBr3 quantum dots (QDs) isdescribed for the determination of ziram pesticide. A facile and inert gas-free method was used for the synthesis of CsPbBr3 QDs. The obtained CsPbBr3 QDs displayed turn-off fluorescence behavior for ziram. The fluorescence intensity of the CsPbBr3 QDs (Ex/Em = 365/516nm) was inversely proportional to the concentration of ziram (0.10 to 50.0ppm) with a detection limit of 0.086ppm. Notably, satisfactory recoveries (100 ± 0.25 to 107 ± 5.72%) were obtained in spiked fruit samples, which demonstrated that this method iscapable of detecting ziram in real samples. In addition, the mechanism for the detection of ziram was investigated in detail. According to the results, this mechanism can be tentatively explained by fluorescence quenching originating from the increased surface defects and the structural changes of the CsPbBr3 QDs. The detection ability of this strategy shows promising applicability in food safety.

On-line coupling of supercritical fluid extraction with enantioselective supercritical fluid chromatography-triple quadrupole mass spectrometry for the determination of chiral pesticides in hemp seeds: A proof-of-principle study

The present research can be considered as a proof-of-principle study focused on the determination of chiral pesticides using a supercritical fluid extraction instrument coupled on-line with an enantioselective supercritical fluid chromatography-triple quadrupole mass spectrometry. To the best of Authors’ knowledge, this is the first description of an on-line approach for the extraction and determination of chiral pesticides. Metalaxyl, benalaxyl and dimethenamid were investigated in nine hemp seed samples belonging to four varieties of Cannabis sativa; only in one case a pesticide was found at levels above the method limit of quantification (LoQ), though within the EU maximum residue level value. The figures-of-merit determined were linearity, precision, limit of detection (LoD), and LoQ. Regression coefficients were between 0.9856 and 0.9973, the LoDs were in the 0.04–0.41 μg kg−1 range, the LoQs were in the 0.12–1.38 μg kg−1 range, while coefficients of variation were between 1 and 3% (10 μg kg−1 level).

Detection of trizole contaminated waste water using biocatalyst and effective biodegradation potential of flubendiamide

Flubendiamide is a new class of chemical pesticide with broad spectrum activity against lepidopteran pests. Due to limited approach and high specificity towards various non targeted organisms, the unrestricted application of this pesticide as a prominent alternate for organochlorine and organophosphate pesticides, causing serious environmental pollution. In this study, wastewater was used for the determination of microbial strains and pesticide degrading fungi. Microbial population and flubendiamide resistant fungal strains were characterized using enriched medium. Aerobic bacteria (6.38 ± 0.23 log CFU/mL), nitrifying bacteria (2.73 ± 0.31 CFU/mL), Lactobaillus (0.72 ± 0.03 log CFU/mL), actinomycetes (5.36 ± 0.27 log CFU/mL) and fungi (4.79 ± 0.22 log CFU/mL) were detected. The prominent fungi genera were, Fusarium, Trichoderma, Cladophialophora, Paecilomyces, Talaromyces, Penicillium, Aspergillus, Candida, Phyllosticta, Mycosphaerella, Ochroconis, and Mucor. Minimum inhibitory concentration of the rapidly growing organism (FR04) revealed its ability to tolerate up to 1250 mg/L flubendiamide concentration. Morphological, biochemical and molecular analysis revealed that the strain was Aspergillus terreus FR04. The residual pesticide was detected using a High Performance Liquid Chromatography (HPLC). High performance liquid chromatography analysis revealed that 89 ± 1.9% pesticide removal efficiency was observed in strain FR04 at optimized culture conditions (96 h, pH 6.5, 30 °C and 300 mg/L pesticide concentration). The strain FR04 degraded pollutants from the wastewater and improved water quality. A. terreu sFR04 is an indigenous fungus and has the ability to degrade trizole pesticides from the wastewater significantly.

A Lab-in-a-Syringe Device Integrated with a Smartphone Platform: Colorimetric and Fluorescent Dual-Mode Signals for On-Site Detection of Organophosphorus Pesticides

Herein, a portable lab-in-a-syringe device integrated with a smartphone sensing platform was designed for rapid, visual quantitative determination of organophosphorus pesticides (OPs) via colorimetric and fluorescent signals. The device was chiefly made up of a conjugate pad labeled with cetyltrimethylammonium bromide-coated gold nanoparticles (CTAB-Au NPs) and a sensing pad modified by ratiometric probes (red-emission quantum dots@SiO2 nanoparticles@green-emission quantum dots, rQDs@SiO2@gQDs probe), which was assembled through a disposable syringe and reusable plastic filter. In the detection system, thiocholine (Tch), the hydrolysis product of thioacetylcholine (ATch) by acetylcholinesterase (AchE), could trigger the aggregation of CTAB-Au NPs, resulting in a significant color change from red to purple. Then, CTAB-Au NPs flowed vertically upward and bound to the rQDs@SiO2@gQDs probe on the sensing pad, reducing the fluorescence resonance energy transfer effect between CTAB-Au NPs and gQDs. Meanwhile, rQDs embedded in SiO2 NPs remained stable as internal reference fluorescence, achieving a color transition from red to green. Thus, based on the inhibition of AChE activity by OPs, a colorimetric and fluorescent dual-mode platform was constructed for on-site detection of OPs. Using glyphosate as a model, with the support of a color recognizer application (APP) on a smartphone, the ratio of red and green channel values could be utilized for accurate OP quantitative analysis ranging from 0 to 10 μM with a detection limit of 2.81 nM (recoveries, 90.8-122.4%; CV, 1.2-3.4%). Overall, the portable lab-in-a-syringe device based on a smartphone sensing platform integrated sample monitoring and result analysis in the field, implying great potential for on-site detection of OPs.

Needle-trap device packed with the MIL-100(Fe) metal–organic framework for the extraction of the airborne organochlorine pesticides

This study was presented the development of the needle trap device (NTD) packed with the MIL-100(Fe) metal–organic framework (MOF) for extraction of organochlorine pesticides (Hexachlorobenzene, Aldrin, Alpha-Chlordane, Dildrin and DDT isomers) from the air environment for the first time. Specifications of synthesized MIL-100(Fe) were identified by Fourier-transfer infrared spectroscopy (FT-IR), X-ray Diffraction (XRD), field emission scanning electron microscopy (FE-SEM), energy dispersive spectroscopy (EDS) and elemental mapping techniques. The optimization of sampling and desorption parameters such as humidity, time, and temperature were performed by response surface methodology (RSM). Sampling and analysis of organochlorine pesticides were accomplished under different conditions in the laboratory scale and then was employed in the field study for the real air samples. The highest response was obtained at desorption temperature of 280 °C and desorption time of 5 min. The temperature and humidity of the sampling site were set to 30 °C and 25%. Furthermore, the Limit of Detection (LOD) and Limit of Quantification (LOQ) for the targeted analytes were determined in the range of 0.04–0.41 µg/m3 and 0.21–1.82 µg/m3, respectively. Moreover, from the recovery standpoint, any significant change was not detected on the analytes after 6 days of storage at 4 °C. Also, the results of this study showed a high correlation (R2 = 0.9882) between the results of the proposed method with the standard NIOSH method. This study exhibited that the proposed MIL-100(Fe)@NTD method can be employed as an eco-friendly, reliable, sensitive, and efficient procedure for sampling and determination of the organochlorine pesticides compared to the standard NIOSH method.

Development of Paraquat Pesticide Determination Methodology in Urine Samples by UHPLC-MS/MS

In technical-scientific development, mankind has developed different ways of managing pests, diseases and weeds, increasing agricultural production impacting social, economic and environmental aspects. As major grains producer (such as corn, soybeans and beans), the Brazilian State, through the Brazilian agencies as Institute for the Environment and Renewable Natural Resources (IBAMA), the National Health Surveillance Agency (ANVISA), and the Ministry of Agriculture, Livestock and Supply (MAPA) authorize and regulate the use of different pesticides, including Paraquat, to improve and guarantee the production. However, the direct contact and long-term exposure to these substances offer environmental and occupational risks, impacting negatively on the workers, population of neighbor cultures, and environmental health. Considering the potential damages, it is of utmost importance to develop effective analytical methodologies for the biomonitoring and assessment of the level of exposure of farmers and ranchers in direct contact with pesticides. Thus, this work proposed to develop a sensitive and selective analytical method, using the ultra-high efficiency liquid chromatography technique, coupled to mass spectrometry seeking a fast and simple sample preparation for the Paraquat quantification in urine samples. To achieve this, two sample preparations were compared in terms of speed and practicality. The first method (A) used C18 silica as adsorbent to remove the non-polar interferences. The second preparation (B) consisted in direct sample dilution in acetonitrile. Both methodologies used centrifugation under refrigeration to precipitate suspended artifacts. The linearity of the Paraquat detection by the analytical methodology, developed in a HILIC column, was evaluated between 10 to 70 µg L-¹ (r²=0.9911; y=19427x+479868), enabling the matrix evaluation after applying the best sample preparation. The method developed was simple, fast, which makes it useful and efficient for toxicology laboratories routine to monitor the exposure levels of farmers dealing with Paraquat daily.

Utilization of montmorillonite nanocomposite incorporated with natural biopolymers and benzyl functionalized dicationic imidazolium based ionic liquid coated fiber for solid-phase microextraction of organochlorine pesticides prior to GC/MS and GC/ECD

A novel montmorillonite clay (MMT) bionanocomposite modified with chitosan (CH), carboxymethyl cellulose (CMC), and benzylimidazolium based dicationic ionic liquid with tetraethylene glycol linker (DIL) was fabricated on stainless steel wire by in situ process. The MMT–CH–CMC-DIL coated solid-phase microextraction (SPME) fiber was examined for the determination of organochlorine pesticides (OCPs) in real samples by HS-SPME-GC method using mass spectrometry (MS) and electron capture detector (ECD). Under optimized conditions, the proposed method exhibited low limits of detection (0.5 ng L−1 with MS and 0.1 ng L−1 with ECD detection), good linearities (R2 = 0.9972–0.9993 with MS and 0.9987–0.9998 with ECD detection), favorable single-fiber repeatability, and fiber-to-fiber reproducibility (less than 8.2% and 9.9% for both types of detection) and high reusability around 125 cycles. Recovery studies were carried out for OCPs in tap water, green tea, and milk samples to verify the applicability of the developed SPME-GC method.

Contamination of Water, Sediment and Fish with Residues of Pesticides Used in Cotton Production in Northern Benin.

In Northern Benin, insecticides are used for cotton production. These insecticides can be easily transferred to water ponds close to cotton fields. To monitor insecticides levels in water, sediments and fish samples from water ponds, a GC-MS analytical method was developed to detect residues of endosulfan, DDT and its parent compounds, isomers of HCH, pyrethroids and chlorpyrifos. In addition, the influence of storage conditions of water sample on pesticides determination performance has been studied. The limits of quantification were between 0.16 and 0.32µg/L in water, 0.5 and 1μg/kg in sediment and 1 and 2μg/kg in fish. Twenty samples of water, twenty of sediments and forty of fish were taken in four different water reservoirs at five different times. Alpha-endosulfan, lambda-cyhalothrin and permethrin were identified in sediment while p,p'-DDE, α- and β-HCH, chlorpyrifos, lambda-cyhalothrin and permethrin were detected in fish. Only organochlorines were determined in water because of the lack of recovery of pyrethroids from water stored in glass. Concentrations of insecticide residues in sediment for all water ponds ranged from non-detected to 101µg/kg and from non-detected to 36µg/kg in fish. Preliminary risk assessment for consumers of the North of Benin showed that the Estimated Daily Intakes were lower than the Acceptable Daily Intakes and Acute Reference Doses for all consumers. However, as one fish can be contaminated by five pesticide residues at the same time, it is not possible to exclude a risk for the consumer due to his exposure to mixtures of pesticides.

Enhanced Bio-Barcode Immunoassay Using Droplet Digital PCR for Multiplex Detection of Organophosphate Pesticides.

A bio-barcode immunoassay based on droplet digital polymerase chain reaction (ddPCR) was developed to simultaneously quantify triazophos, parathion, and chlorpyrifos in apple, cucumber, cabbage, and pear. Three gold nanoparticle (AuNP) probes and magnetic nanoparticle (MNP) probes were prepared, binding through their antibodies with the three pesticides in the same tube. Three groups of primers, probes, templates, and three antibodies were designed to ensure the specificity of the method. Under the optimal conditions, the detection limits (expressed as IC10) of triazophos, parathion, and chlorpyrifos were 0.22, 0.45, and 4.49 ng mL-1, respectively. The linear ranges were 0.01-20, 0.1-100, and 0.1-500 ng mL-1, and the correlation coefficients (R2) were 0.9661, 0.9834, and 0.9612, respectively. The recoveries and relative standard deviations (RSDs) were in the ranges of 75.5-98.9 and 8.3-16.7%. This study provides the first insights into the ddPCR for the determination of organophosphate pesticides. It also laid the foundation for high-throughput detection of other small molecules.