Synthesis and fungicidal activity of methyl (E)-1-(2-((E)-2-methoxy-1-(methoxyimino)-2-oxoethyl)benzyl)-2-(1-arylidene)hydrazine-1-carboxylates †‡.

To discover novel strobilurin fungicides, a series of methyl (E)-1-(2-((E)-2-methoxy-1-(methoxy-imino)-2-oxoethyl)benzyl)-2-(1-arylidene)hydrazine-1-carboxylates were designed based on the principle of biologically active splicing and the receptor target structure. The fungicidal activity results show that this class of compounds has excellent fungicidal activity, especially against S. sclerotiorum (Lib.) deBary, wheat white powder and puccinia polysora. The result of structure-activity relationship implied that the introduction of t-butyl in the side chain facilitates the hydrophobic interaction between the compound and the active site. The electrostatic effect of the substituents on the benzene ring is also a key factor affecting such activities. Among them, the compound I-1 not only showed a fungicidal effect comparable to that of kresoxim-methyl in vivo, but also had an excellent inhibitory effect on spore germination of P. oryzae Cav in vitro, which indicated that it could be used as a potential commercial fungicide for plant disease control.

Long-term exposure to azoxystrobin induces immunodeficiency in fish that are vulnerable to subsequent rhabdovirus infection.

Azoxystrobin (AZ) is one of the most widely used strobilurin fungicides in the world, and its residue has seriously endangered aquatic ecological security. Our previous data showed that AZ exposure may reduce the resistance of fish to rhabdovirus infection in aquatic environment. Here, we further reported a potential long-term adverse effect of AZ exposure on the antiviral and immunosuppressive recovery in fish, and observed that mitochondrial dynamic balance was disturbed by AZ in which excessive mitochondrial fission occurred in response to decreased ATP levels. When a recovery operation was performed in AZ-exposed cells and fish, infectivity of our model virus, spring viraemia of carp virus (SVCV), was significantly decreased in vitro (using the epithelioma papulosum cyprini [EPC] fish cell line) and in vivo (using zebrafish) in a time-dependent manner. Also, the expression of eight innate antiviral immune genes (IFNs, ISG15, MX1, RIG-I, IRF3, Nrf2 and HO-1) showed a similar change to SVCV replication between the longer exposure period and the expression recovery. Additionally, AZ facilitated horizontal transmission of SVCV in a static cohabitation challenge model, predicting the increase of the potential for the viral outbreak. Therefore, our data suggest that long-term effect of AZ on irreparable impairment in fish made AZ residue potentially greater for ecological risks.

Comprehensive study of the effects of strobilurin-based fungicide formulations on Enchytraeus albidus.

Excessive application of fungicides in crop fields can cause adverse effects on soil organisms and consequently affect soil properties. Existing knowledge on the effects of strobilurin fungicides has been primarily based on toxicity tests with active ingredients, while the effects of fungicide formulations remain unclear. Therefore, this work aims to provide new data on the effects of three commercial formulations of strobilurin fungicides on the soil organism Enchytraeus albidus. The tested fungicide formulations were Retengo® (pyraclostrobin-PYR), Zato WG 50® (trifloxystrobin-TRI) and Stroby WG® (kresoxim-methyl-KM). In laboratory experiments, multiple endpoints were considered at different time points. The results showed that PYR had the greatest impact on survival and reproduction (LC50 = 7.57 mga.i.kgsoil-1, EC50 = 0.98 mga.i.kgsoil-1), followed by TRI (LC50 = 72.98 mga.i.kgsoil-1, EC50 = 16.93 mga.i.kgsoil-1) and KM (LC50 = 73.12 mga.i.kgsoil-1, EC50 ≥ 30 mga.i.kgsoil-1). After 7 days of exposure, MXR activity was inhibited at the highest concentration of all fungicides tested (6 mgPYRkgsoil-1, 15 mgTRIkgsoil-1 and 30 mgKMkgsoil-1). Furthermore, oxidative stress (induction of SOD, CAT and GST) and lipid peroxidation (increase in MDA) were also observed. In addition, there was a decrease in total available energy after exposure to PYR and KM. Exposure to fungicides resulted in a shift in the proportions of carbohydrates, lipids, and proteins affecting the amount of available energy. In addition to the initial findings on the effects of strobilurin formulations on enchytraeids, the observed results suggest that multiple and long-term exposure to strobilurin formulations in the field could have negative consequences on enchytraeid populations.

Detoxification Esterase StrH Initiates Strobilurin Fungicide Degradation in Hyphomicrobium sp. Strain DY-1.

Strobilurin fungicides are widely used in agricultural production due to their broad-spectrum and fungal mitochondrial inhibitory activities. However, their massive application has restrained the growth of eukaryotic algae and increased collateral damage in freshwater systems, notably harmful cyanobacterial blooms (HCBs). In this study, a strobilurin fungicide-degrading strain, Hyphomicrobium sp. strain DY-1, was isolated and characterized successfully. Moreover, a novel esterase gene, strH, responsible for the de-esterification of strobilurin fungicides, was cloned, and the enzymatic properties of StrH were studied. For trifloxystrobin, StrH displayed maximum activity at 50°C and pH 7.0. The catalytic efficiencies (kcat/Km ) of StrH for different strobilurin fungicides were 196.32 ± 2.30 µM-1 · s-1 (trifloxystrobin), 4.64 ± 0.05 µM-1 · s-1 (picoxystrobin), 2.94 ± 0.02 µM-1 · s-1 (pyraclostrobin), and (2.41 ± 0.19)×10-2 µM-1 · s-1 (azoxystrobin). StrH catalyzed the de-esterification of a variety of strobilurin fungicides, generating the corresponding parent acid to achieve the detoxification of strobilurin fungicides and relieve strobilurin fungicide growth inhibition of Chlorella This research will provide insight into the microbial remediation of strobilurin fungicide-contaminated environments.IMPORTANCE Strobilurin fungicides have been widely acknowledged as an essential group of pesticides worldwide. So far, their residues and toxic effects on aquatic organisms have been reported in different parts of the world. Microbial degradation can eliminate xenobiotics from the environment. Therefore, the degradation of strobilurin fungicides by microorganisms has also been reported. However, little is known about the involvement of enzymes or genes in strobilurin fungicide degradation. In this study, a novel esterase gene responsible for the detoxification of strobilurin fungicides, strH, was cloned in the newly isolated strain Hyphomicrobium sp. DY-1. This degradation process detoxifies the strobilurin fungicides and relieves their growth inhibition of Chlorella.

Pyraclostrobin induced AMPK/mTOR pathways mediated autophagy in RAW264.7 macrophages.

Pyraclostrobin(PCT) is a highly effective and broad-spectrum strobilurin fungicide. The mode action of PCT is inhibiting mitochondrial respiration. With the widespread use of PCT in preventing and controlling crop diseases, its potential safety risks to mammals have gradually attracted attention. This paper focuses on the cytotoxicity of PCT and its molecular mechanism, RAW264.7 macrophages were selected as a research model and conducted systematic toxicology studies in vitro, including MTT assay, colony formation assay, alkaline comet assay, fluorescent staining, ATP assay and Western blotting. The results revealed that PCT decreased viability and inhibited the proliferation of RAW264.7 cells in a concentration- dependent manner. Interestingly, PCT induced DNA damage, the resulting autophagosome, the accumulation of Beclin-1, the reduction of p62, the translocation and the formation of LC3-II. Furthermore, the results showed that PCT-induced the production of excessive ROS, leading to mitochondrial permeability transition pore (mPTP) opening, ATP depletion, and the elimination of mitochondria by autophagy. Furthermore, PCT treatment group significantly enhanced the phosphorylation level of AMPK, decreased the mTOR and p70s6k phosphorylation levels and activated the AMPK/mTOR signaling pathway in RAW264.7 cells. In conclusion, these results showed that PCT induced autophagy in the RAW264.7 cells might potentially have risks to mammal safety.

Synthesis of molecularly imprinted polymer adsorbents for solid-phase extraction of strobilurin fungicides from agricultural products.

Molecularly imprinted polymers for strobilurin fungicides were prepared by precipitation polymerization employing azoxystrobin as template molecular together with methacrylic acid monomer and trimethylolpropane triacrylate cross-linker. Morphological characterization showed molecularly imprinted polymers were uniform spherical particles with about 0.2 µm in diameter, while the morphologies of nonimprinted polymers were irregular bulk. The equilibrium binding and selective experiments proved that molecularly imprinted polymers possessed a higher affinity toward four fungicides compared to nonimprinted polymers and heterogeneous binding sites were found in the molecularly imprinted polymers. Molecularly imprinted solid-phase extraction conditions, including sample loading solvents, selective washing, and elution solvents, were carefully optimized. The developed method showed good recoveries (70.0-114.0%) with relative standard deviations in range of 1.0-9.8% (n  = â€¯3) for samples (cucumber and peach) spiked at three different levels (10, 50, and 100 µg/ kg). The detection limit (signal/noise = 3) ranged from 0.01 to 0.08 µg/kg. The results demonstrated good potential use of this convenient and highly efficient method for determining trace strobilurin fungicides in agricultural products.

Respiratory Toxicity of Azoxystrobin, Pyraclostrobin and Coumoxystrobin on Chlorella vulgaris.

Azoxystrobin (AZ), pyraclostrobin (PYR) and coumoxystrobin (COU) exert negative impacts on Chlorella vulgaris. Thus, in this study, C. vulgaris was used to assess the respiratory toxicity of AZ, PYR and COU by determining the acute toxicity, complex III activity and ATP viability. The 96 h-EC50 values of AZ, PYR and COU for C. vulgaris were 1.85, 2.21 and 1.62 mg/L, respectively. AZ, PYR and COU exerted significant effects on complex III activity and ATP viability after exposure to 0.71, 1.01 and 1.08 mg/L of the fungicides. The binding potentials of AZ, PYR and COU toward ubiquinone were - 10.44, - 9.31 and - 12.98 kcal/mol, respectively, which had adverse effects on amino acids. These results provided new insight into the potential acute respiratory toxicity mechanisms of these strobilurin fungicides in algae.

Perceptions of Midwestern Crop Advisors and Growers on Foliar Fungicide Adoption and Use in Maize.

Foliar fungicide use in hybrid maize in the United States was rare before 2000. The decade from 2000 to 2010 saw foliar fungicides increasingly applied to maize in the absence of appreciable disease pressure, a practice seemingly at odds with integrated pest management philosophy. Yet, it is commonly believed that growers do not employ management strategies unless there are perceived benefits. Maize (corn) growers (CGs) and certified crop advisors (CCAs) across four Midwestern states (Iowa, Illinois, Ohio, and Wisconsin) were surveyed to better understand their practices, values and perceptions concerning the use of foliar fungicides during 2005 to 2009. The survey results demonstrated the rapid rise in maize foliar fungicide applications from 2000 through 2008, with 84% of CGs who sprayed having used a foliar fungicide in maize production for the very first time during 2005 to 2009. During 2005 to 2009, 73% of CCAs had recommended using a foliar fungicide, but only 35% of CGs sprayed. Perceived yield gains, conditional on having sprayed, were above the break-even point on average. However, negative yield responses were also observed by almost half of CCAs and a quarter of CGs. Hybrid disease resistance was a more important factor to economically successful maize production than foliar fungicides. Diseases as a yield-limiting factor were more important to CGs than CCAs. As a group, CGs were not as embracing of foliar fungicide as were CCAs, and remained more conservative about the perceived benefits to yield.

Aquatic risk assessment of a novel strobilurin fungicide: A microcosm study compared with the species sensitivity distribution approach.

The ecotoxicological effects of pyraoxystrobin, a novel strobilurin fungicide, were studied using outdoor freshwater microcosms and the species sensitivity distribution approach. The microcosms were treated with pyraoxystrobin at concentrations of 0, 1.0, 3.0, 10, 30 and 100µg/L. Species sensitivity distribution (SSD) curves were constructed by means of acute toxicity data using the BurrliOZ model for fourteen representatives of sensitive invertebrates, algae and fish and eleven taxa of invertebrates and algae, respectively. The responses of zooplankton, phytoplankton and physical and chemical endpoints in microcosms were studied. Zooplankton, especially Sinodiaptomus sarsi was the most sensitive to pyraoxystrobin exposure in the microcosms. Short-term toxic effects (<8 weeks) on zooplankton occurred in 1µg/L treatment group. The duration of toxic effects on S. sarsi could not be evaluated within the initial 56 days. Significant long-term toxic effects were observed at 10, 30 and 100µg/L (>281 days) for S. sarsi and the zooplankton community. Based on the results obtained from the organisms in the microcosm system, 1µg/L was recommended as the NOEAEC (no observed ecologically adverse effect concentration). Also, 0.33µg/L was derived as the Regulatory Acceptable Concentration based on the ecological recovery option (ERO-RAC) of pyraoxystrobin. For all fourteen tested species, the median HC5 (hazardous concentration affecting 5% of species) was 0.86µg/L, and the lower limit HC5 (LL-HC5) was 0.39µg/L. For the eleven taxa of invertebrates and algae tested, the median HC5 was 1.1µg/L, and the LL-HC5 was 0.26µg/L. The present study positively contributes to the suggestion of adequately using acute L(E)C50-based HC5/ LL-HC5 for deriving protective concentrations for strobilurin fungicides, and it should be valuable for full comprehension of the potential toxicity of pyraoxystrobin in aquatic ecosystems.

Determination of strobilurin fungicides in cotton seed by combination of acetonitrile extraction and dispersive liquid-liquid microextraction coupled with gas chromatography.

The simultaneous determination of four strobilurin fungicides (picoxystrobin, kresoxim-methyl, trifloxystrobin, and azoxystrobin) in cotton seed by combining acetonitrile extraction and dispersive liquid-liquid microextraction was developed prior to GC with electron capture detection. Several factors, including the type and volume of the extraction and dispersive solvents, extraction condition and time, and salt addition, were optimized. The analytes were extracted with acetonitrile from cotton seed and the clean-up was carried out by primary secondary amine. Afterwards, 60 µL of n-hexane/toluene (1:1, v/v) with a lower density than water was mixed with 1 mL of the acetonitrile extract, then the mixture was injected into 7 mL of distilled water. A 0.1 mL pipette was used to collect a few microliters of n-hexane/toluene from the top of the aqueous solution. The enrichment factors of the analytes ranged from 36 to 67. The LODs were in the range of 0.1 × 10(-3) -2 × 10(-3) mg/kg. The relative recoveries varied from 87.7 to 95.2% with RSDs of 4.1-8.5% for the four fungicides. The good performance of the method, compared with the conventional pretreatments, has demonstrated it is suitable for determining low concentrations of strobilurin fungicide residues in cotton seed.

Pyraclostrobin induces developmental toxicity and cardiotoxicity through oxidative stress and inflammation in zebrafish embryos.

Pyraclostrobin, a typical representative of strobilurin fungicides, is extensively used in agriculture to control fungi and is often detected in water bodies and food. However, the comprehensive toxicological molecular mechanism of pyraclostrobin requires further study. To assess the toxic effects and underlying mechanisms of pyraclostrobin on aquatic organisms, zebrafish embryos were exposed to pyraclostrobin (20, 40, and 60 µg/L) until 96 h post fertilization (hpf). These results indicated that exposure to pyraclostrobin induces morphological alterations, including spinal curvature, shortened body length, and smaller eyes. Furthermore, heart developmental malformations, such as pericardial edema and bradycardia, were observed. This indicated severe cardiotoxicity induced by pyraclostrobin in zebrafish embryos, which was confirmed by the dysregulation of genes related to heart development. Besides, our findings also demonstrated that pyraclostrobin enhanced the contents of reactive oxygen species (ROS) and malondialdehyde (MDA), up-regulated catalase (CAT) activity, but inhibited superoxide dismutase (SOD) activity. Subsequently, the NF-κb signaling pathway was further studied, and the results indicated that the up-regulation of tnf-α, tlr-4, and myd88 activated the NF-κb signaling pathway and up-regulated the relative expression level of pro-inflammatory cytokines, such as cc-chemokine, ifn-γ, and cxcl-clc. Collectively, this study revealed that pyraclostrobin exposure induces developmental toxicity and cardiotoxicity, which may result from a combination of oxidative stress and inflammatory responses. These findings provide a basis for continued evaluation of the effects and ecological risks of pyraclostrobin on the early development of aquatic organisms.

Uptake, Translocation, and Subcellular Distribution of Strobilurin Fungicides in Cucumber (Cucumis sativa L.).

The absorption, transport, and subcellular distribution of strobilurin fungicides (azoxystrobin, pyraclostrobin, and trifloxystrobin) have been studied in cucumbers. Under hydroponic laboratory conditions, pyraclostrobin and trifloxystrobin mainly accumulated in cucumber roots whereas azoxystrobin accumulated in cucumber leaves. In the subcellular distribution experiment, azoxystrobin mainly accumulated as a soluble component. Pyraclostrobin and trifloxystrobin accumulated more in the organelles and cell walls. Azoxystrobin and pyraclostrobin enter the root primarily through the apoplast pathway, whereas trifloxystrobin enters the root through the symplastic pathway. Azoxystrobin can be transported in cucumber through anion and cation channels, whereas pyraclostrobin and trifloxystrobin can be transported only through anion channels. This study has great significance in evaluating environmental risks and food safety.

Multigenerational and transgenerational effects of azoxystrobin on Folsomia candida.

Soil organisms are exposed to various pollutants during several generations. However standard toxicity tests are often based on exposure in only one generation. Research of multigenerational (MG) and transgenerational (TG) effects are still quite scarce, however evidence accumulates that effects observed in one generation can be significantly different in some of the following generations, with different effects observed. Some studies suggest adaptation to pollutants, while others report severe effects in following generations. Azoxystrobin is commonly used in the prevention and treatment of fungal diseases in a wide range of economically important crops. The main aim of this study was to assess the toxic effects of azoxystrobin (AZO) on F. candida over 3 generations through the application of biochemical and population level biomarkers. Results of reproduction tests showed a significant decrease in estimated EC50 values, with EC50 for F0 being estimated at 104.44 mga.i./kgD.W.soil and only 15.4 mga.i./kgD.W.soil for F1. In F1 a significant reduction in the number of juveniles was observed, and at AZO concentration of 50 mga.i./kgD.W and higher, F1 did not reproduce. Significant oxidative stress was observed in all generations, with increased SOD and lipid damage that slowly decreased in subsequent generations. Transgenerational effects were also observed, with a significantly reduced number of juveniles in F1 and significant oxidative stress and lipid damage in all generations. IBRv2 showed that F1 was most affected, followed by F0, and least affected was F3. When considering the whole body energy budget, F1 to F3 had significantly higher WBEB compared to F0, and a shift in proportion of energy reserves occurred in F1, where the proportion of lipids increased while protein decreased. Results of this research show that considering standard toxicity tests, risks for populations of soil organisms are possibly severely underestimated. Therefore, standard toxicity guidelines should be supplemented by multigenerational tests, when possible.

Analysis of the toxic mechanisms of fluoxastrobin on the earthworm (Eisenia fetida) using transcriptomics.

Fluoxastrobin (FLUO), one of the best-selling strobilurin fungicides, could prevent fungal diseases from oilseed crops, fruits, grains, and vegetables. The widespread use of FLUO leads to the continuous accumulation of FLUO in soil. Our previous studies have demonstrated that FLUO exhibited different toxicity in artificial soil and three natural soils (fluvo-aquic soils, black soils, and red clay). The toxicity of FLUO was greater in natural soil than the artificial soil, specifically, showed the highest toxicity in fluvo-aquic soils. To better investigate the mechanism of FLUO toxicity to earthworms (Eisenia fetida), we selected fluvo-aquic soils as representative soil and used transcriptomics to study the gene expression in earthworms after FLUO exposure. The results demonstrated that the differentially expressed genes in earthworms after FLUO exposure mainly presented in pathways involving protein folding, immunity, signal transduction, and cell growth. It may be the reason why FLUO exposure stressed the earthworms and affected their normal growth activities. The present study fills gaps in the literature regarding the soil bio-toxicity of strobilurin fungicides. It also sounds the alarm for the application of such fungicides even at the low concentration (0.1 mg kg-1).

Magnetic deep eutectic solvent-based dispersive liquid-liquid microextraction for determination of strobilurin fungicides in water, juice, and vinegar by high-performance liquid chromatography.

In this study, a magnetic deep eutectic solvent coupled with dispersive liquid-liquid microextraction using high-performance liquid chromatography (MDES-DLLME-HPLC) was developed to detect strobilurin fungicides. The green hydrophobic MDES synthesized by methyltrioctylammonium chloride, ferric chloride, and heptanoic acid was used as an extraction solvent, which was dispersed by vortex and separated by an external magnetic field. The use of toxic solvents was avoided, and the separation time was reduced. The best experimental results were obtained through single factor and response surface optimization. The method had a good linear relationship with R2 > 0.996. The limit of detection (LOD) ranged from 0.001 to 0.002 mg L-1. The extraction recoveries were 81.9-108.9%. The proposed method was rapid and green, and it has been successfully applied to detection of strobilurin fungicides in water, juice, and vinegar.

Carboxylesterases from bacterial enrichment culture degrade strobilurin fungicides.

Strobilurin fungicides are a class of persistent fungicides frequently detected in the environment. Microbes can effectively degrade strobilurins, but the mechanisms are complex and diverse. Compared with isolated strains, bacterial consortia are more robust in terms of the degradation of multiple pollutants. The enrichment culture XS19 is a group of bacterial strains enriched from soil and degrades six strobilurins at 50 mg/L within 8 d, including azoxystrobin, picoxystrobin, trifloxystrobin, kresoxim-methyl, pyraclostrobin and enestroburin. LC-Q-TOF-MS analysis confirmed that XS19 can demethylate these strobilurins via hydrolysis of the methyl ester group. Analysis of the bacterial communities suggested that Pseudomonas (69.8%), Sphingobacterium (21.2%), Delftia (6.3%), and Achromobacter (1.6%) spp. were highly associated with the removal of strobilurins in the system. Metagenomics-based comprehensive analysis of XS19 suggested that carboxylesterases in Pseudomonas and Sphingobacterium play a central role in the catabolism of strobilurins. Moreover, the carboxylesterase inhibitor bis-p-nitrophenyl phosphate inhibited the degradation activity of strobilurins in XS19. This work proved that XS19 or carboxylesterases can effectively hydrolyze strobilurins, providing a reliable bioremediation paradigm.

Cyclodextrin-based dispersive liquid-liquid microextraction for the determination of fungicides in water, juice, and vinegar samples via HPLC.

Cyclodextrin-based dispersive liquid-liquid microextraction (CD-DLLME) was developed for the determination of triazole and strobilurin fungicides in water, juice, and vinegar samples using high-performance liquid chromatography-diode-array detection (HPLC-DAD). Undecanol, which is a green solvent, was selected as the extraction solvent. A cyclodextrin aqueous solution was chosen as the dispersion solvent and demulsifier to avoid the use of a toxic dispersion solvent and eliminate the centrifugation step. Dispersion and phase separation were completed within 1 and 60 s, respectively. The linear range of this method was 1 to 100 µg L-1. The limits of detection were 0.3 µg L-1 along with the preconcentration factor of 133 and enrichment factor of 124. The recovery was 83.2% to 103.2%. This pretreatment method was fast, simple, and environmentally friendly and was successfully applied to the analysis of triazole and strobilurin fungicide residues in water, juice, and vinegar samples.

Preparation of metal-organic framework @molecularly imprinted polymers for extracting strobilurin fungicides from agricultural products.

The core-shell metal-organic framework coated with molecularly imprinted polymers (ZIF-8@MIPs) were successfully synthesized by surface imprinting technique, and applied as adsorbents for solid-phase extraction of strobilurin fungicides. The obtained hybrid complex was characterized in detail, and their adsorbing and recognition performance were evaluated. The results showed that ZIF-8@MIPs presented typically core-shell structure with MIP shell (about 20 nm), and exhibited larger adsorption capacity (102.5 mg g-1) and fast adsorption ability (only 5 min). Under the optimized conditions, a sensitive, efficient and reliable method for determining six strobilurin fungicides in different agricultural products based on ZIF-8@MIPs coupling with high performance liquid chromatography-tandem mass spectrometry was established. This method showed good linearity with correlation coefficients higher than 0.9990. With spiked at three different concentration levels in agricultural products (apple, pear, banana, Chinese cabbage, cabbage, cucumber), the good recoveries (83.5-129.0%) with relative standard deviations from 0.5 to 10.2% were obtained. The limit of detections and the limit of quantifications were 0.01-1.12 ng g-1 and 0.03-3.73 ng g-1, respectively. Those results demonstrated good potential application of ZIF-8@MIPs for enriching and separating trace strobilurin fungicides in agricultural samples.

A magnetic covalent organic framework as selective adsorbent for preconcentration of multi strobilurin fungicides in foods.

A new method that utilizes a magnetic covalent organic framework (Fe3O4@BTA-TAPM) as a selective solid-phase extraction adsorbent has been developed for the simultaneous preconcentration of trace eleven kinds of strobilurin fungicides (SFs) in foods combined with ultra-high performance liquid chromatography-tandem mass spectrometry. The preconcentration conditions were optimized, including the adsorbent amount, extraction time, pH value, shaking time, elution solvent, desorption time and the interference ions. The maximum adsorption capacity of the adsorbent was ranging from 42.84 mg g-1 to 54.18 mg g-1 for eleven SFs. The low limits of detection were 0.003-0.072 µg kg-1. The recoveries of eleven SFs spiked at levels of 0.25-1000 µg kg-1 ranged from 80.2% to 109.3%, and the relative standard deviations were <6.8% (n = 3). The utility of the adsorbent was demonstrated to determine traces SFs in four different samples.

Extraction and determination of strobilurin fungicides residues in apple samples using ultrasound-assisted dispersive liquid-liquid microextraction based on a novel hydrophobic deep eutectic solvent followed by H.P.L.C-U.V.

In this study, several novel and natural hydrophobic deep eutectic solvents (D.E.Ss) were prepared using methyl trioctylammonium chloride (M.T.O.A.C) as H.B.A and different types of straight chain alcohols as H.B.Ds. One of the D.E.Ss composed of M.T.O.A.C and n-butanol was advantageously used to develop an ultrasound-assisted dispersive liquid-liquid microextraction (U.A-D.L.L.M.E) method combined with high-performance liquid chromatography-ultraviolet detection (H.P.L.C-U.V) for the determination of some strobilurin fungicides in apple samples. Several important parameters influencing extraction efficiency were investigated and optimised, including the type and volume of extractant in ultrasound stage, sonication time, the type and volume of D.E.S, sample solution pH and effect of salt addition. Under optimal experimental conditions, the method showed good linearity with correlation coefficients (R2) of 0.9985 - 0.9991 in the linear range of 4-1500 µg kg-1, low limits of detection of 1.5-2 µg kg-1 and acceptable extraction recoveries in the range of 76-92%. Enrichment factor was in the range of 95-115. The proposed method was successfully applied for the extraction and preconcentration of trace fungicides in apple samples, and the results demonstrated the potential of the synthesised D.E.S for the extraction and determination of contaminants in aqueous samples.