Inhibition of melanoma cell proliferation by strobilurins isolated from mushrooms and their synthetic analogues.

Strobilurins A and X, isolated from Mucidula venosolamellata culture extracts, demonstrated potent inhibition of human melanoma G-361 cell proliferation. Strobilurin X exhibited milder inhibitory effects on human fibroblast cells (NB1RGB) compared to strobilurin A. Additional strobilurin-related compounds were isolated from the other mushroom species. Oudemansins A and B displayed weaker activities on G-361 cells than strobilurins A and B, respectively, emphasizing the importance of a conjugated double-bond structure. Among isolated compounds, strobilurin G showed the lowest IC50 value for G-361 cells. Additional strobilurins bearing various substituents on the benzene ring were synthesized. Synthetic intermediates lacking the methyl ß-methoxyacrylate group and a strobilurin analogue bearing modified ß-methoxyacrylate moiety showed almost no inhibitory activity against G-361 cells. The introduction of long or bulky substituents at the 4' position of the benzene ring of strobilurins enhanced the activity and selectivity, suggesting differential recognition of the benzene ring by G-361 and NB1RGB cells.

Evaluation of Anticholinesterase Activity of the Fungicides Mefentrifluconazole and Pyraclostrobin.

Triazoles are compounds with various biological activities, including fungicidal action. They became popular through cholinesterase studies after the successful synthesis of the dual binding femtomolar triazole inhibitor of acetylcholinesterase (AChE, EC 3.1.1.7) by Sharpless et al. via in situ click chemistry. Here, we evaluate the anticholinesterase effect of the first isopropanol triazole fungicide mefentrifluconazole (Ravystar®), developed to overcome fungus resistance in plant disease management. Mefentrifluconazole is commercially available individually or in a binary fungicidal mixture, i.e., with pyraclostrobin (Ravycare®). Pyraclostrobin is a carbamate that contains a pyrazole ring. Carbamates are known inhibitors of cholinesterases and the carbamate rivastigmine is already in use for the treatment of Alzheimer's disease. We tested the type and potency of anticholinesterase activity of mefentrifluconazole and pyraclostrobin. Mefentrifluconazole reversibly inhibited human AChE and BChE with a seven-fold higher potency toward AChE (Ki = 101 ± 19 µM). Pyraclostrobin (50 µM) inhibited AChE and BChE progressively with rate constants of (t1/2 = 2.1 min; ki = 6.6 × 103 M-1 min-1) and (t1/2 = 1.5 min; ki = 9.2 × 103 M-1 min-1), respectively. A molecular docking study indicated key interactions between the tested fungicides and residues of the lipophilic active site of AChE and BChE. Additionally, the physicochemical properties of the tested fungicides were compared to values for CNS-active drugs to estimate the blood-brain barrier permeability. Our results can be applied in the design of new molecules with a lesser impact on humans and the environment.

Efficient synthesis of carbon-14 labeled metabolites of the strobilurin fungicide mandestrobin using biomimetic iron-porphyrin catalyzed oxidation.

Biomimetic oxidation using synthetic iron-porphyrin (F20 TPPFeCl) as a catalyst eliminated a xylene moiety of the fungicide mandestrobin, uniformly labeled with carbon-14 at the benzyl ring, to produce the corresponding radiolabeled metabolite 1. This reaction mechanism was investigated by identifying chemical structures of intermediate 5 and p-xyloquinone derivatives 6 and 7, as by-products. Optimization of reaction factors based on the mechanism improved the yield of 1 from mandestrobin up to 87%. Finally, various carbon-14 labeled metabolites of mandestrobin were prepared from 1.

Structural Investigation and Molecular Modeling Studies of Strobilurin-Based Fungicides Active against the Rice Blast Pathogen Pyricularia oryzae.

The increasing emergence of fungicide-resistant pathogens requires urgent solutions for crop disease management. Here, we describe a structural investigation of new fungicides obtained by combining strobilurin and succinate dehydrogenase inhibitor pharmacophores. We identified compounds endowed with very good activity against wild-type Pyricularia oryzae, combined in some cases with promising activity against strobilurin-resistant strains. The first three-dimensional model of P. oryzae cytochrome bc1 complex containing azoxystrobin as a ligand was developed. The model was validated with a set of commercially available strobilurins, and it well explains both the resistance mechanism to strobilurins mediated by the mutation G143A and the activity of metyltetraprole against strobilurin-resistant strains. The obtained results shed light on the key recognition determinants of strobilurin-like derivatives in the cytochrome bc1 active site and will guide the further rational design of new fungicides able to overcome resistance caused by G143A mutation in the rice blast pathogen.

Enhanced Fungicidal Efficacy by Co-Delivery of Azoxystrobin and Diniconazole with Cauliflower-Like Metal-Organic Frameworks NH2-Al-MIL-101.

Long-term use of a single fungicide increases the resistance risk and causes adverse effects on natural ecosystems. Controlled release formulations of dual fungicides with different modes of action can afford a new dimension for addressing the current issues. Based on adjustable aperture and superhigh surface area, metal-organic frameworks (MOFs) are ideal candidates as pesticide release carriers. This study used Al3+ as the metal node and 2-aminoterephthalic acid as the organic chain to prepare aluminum-based metal-organic framework material (NH2-Al-MIL-101) with "cauliflower-like" structure and high surface area of 2359.0 m2/g. Fungicides of azoxystrobin (AZOX) and diniconazole (Dini) were simultaneously encapsulated into NH2-Al-MIL-101 with the loading content of 6.71% and 29.72%, respectively. Dual fungicide delivery system of AZOX@Dini@NH2-Al-MIL-101 demonstrated sustained and pH responsive release profiles. When the maximum cumulative release rate of AZOX and Dini both reached about 90%, the release time was 46 and 136 h, respectively. Furthermore, EC50 values as well as the percentage of inhibition revealed that AZOX@Dini@NH2-Al-MIL-101 had enhanced germicidal efficacy against rice sheath blight (Rhizoctonia solani), evidenced by the synergistic ratio of 1.83. The present study demonstrates a potential application prospect in sustainable plant protection through co-delivery fungicides with MOFs as a platform.

Residue dissipation, transfer and safety evaluation of picoxystrobin during tea growing and brewing.

BACKGROUND: Picoxystrobin is a new osmotic and systemic broad-spectrum methoxyacrylate fungicide with a good control effect on tea anthracnose, so it has been proposed to spray picoxystrobin before the occurrence and onset of tea anthracnose during tea bud growth in order to protect them. However, there are few reports about the residue analysis method, field dissipation, terminal residue and risk assessment of picoxystrobin in tea. And there is no scientific and reasonable maximum residue limit of picoxystrobin in green tea. RESULTS: A rapid and sensitive analysis method for picoxystrobin residue in fresh tea leaf, green tea, tea infusion and soil was established by UPLC-MS/MS. The spiked recoveries of picoxystrobin ranged from 73.1% to 111.0%, with relative standard deviations from 1.8% to 9.2%. The limits of quantitation were 20 µg kg-1 in green tea, 8 µg kg-1 in fresh tea leaves and soil and 0.16 µg kg-1 in tea infusion. The dissipation half-lives of picoxystrobin in fresh tea leaf and soil were 2.7-6.8 and 2.5-14.4 days, respectively. And the maximum residue of picoxystrobin in green tea was 15.28 mg kg-1 with PHI at 10 days for terminal test. The total leaching rate of picoxystrobin during green tea brewing was lower than 35.8%. CONCLUSIONS: According to safety evaluation, the RQc and RQa values of picoxystrobin in tea after 5 to 14 days for the last application were significantly lower than 1. Therefore, the maximum residue limit value of picoxystrobin in tea that we suggest to set at 20 mg kg-1 can ensure the safety of tea for human drinking. © 2020 Society of Chemical Industry.

Bagging and non-bagging treatment on the dissipation and residue of four mixed application pesticides on banana fruit.

BACKGROUND: Bananas are vulnerable to disease and insect pests after producing fruit. In order to increase the yield and produce high-quality fruit, the insecticides and fungicides are mixed and applied 2-3 times on banana, then the fruit is bagged. Buprofezin, imidacloprid, difenoconazole, and pyraclostrobin are widely used on banana. However, there is a lack of research on the effect of fruit bagging on pesticide dissipation and residues on bananas. RESULTS: A versatile liquid chromatography-tandem mass spectrometry method with modified QuEChERS sample preparation has been developed for the determination of buprofezin, imidacloprid, difenoconazole, and pyraclostrobin in bananas. The recovery of four pesticides was satisfactory (74.96-98.63%) with reasonable relative standard deviation (≤ 8.78%). In Hainan and Guangzhou, the half-lives of the four pesticides were 4.68-13.9 and 5.63-20.4 days in non-bagged and bagged bananas, respectively. The significance analysis of the half-lives in the two sites showed that the dissipation rates of the three pesticides (imidacloprid, difenoconazole, pyraclostrobin) on whole bananas were significantly decreased by the effect of bagging (P < 0.05). However, there was no significant difference in the degradation of half-life of buprofezin under bagging and without bagging (P > 0.05). CONCLUSION: The high vapor pressure and the non-systemic property cause buprofezin to evaporate and dissipate the fastest among the four studied pesticides. The ultimate residues of four pesticides in bananas are lower than the maximum residue limits in China after three times of mixed applications under bagging or non-bagging. The results provide scientific data for evaluating the safety of four pesticides in banana bagging. © 2020 Society of Chemical Industry.

Fabrication a controlled-release pesticide for improving UV-shielding properties and reducing toxicity via coating polydopamine.

Controlled-release formulations (CRFs) have potential applications in modern agriculture, for it can not only prolong the duration of agrochemicals but also alleviate the adverse effect on non-target organism. In this study, we constructed pyraclostrobin@SiO2@polydopamine microcapsule (Pyr@SiO2@PDA MC). The resulting microcapsule is a near-rod shape (about 1.15 µm), which has a drug-loading efficiency of 55%. Fourier transform infrared (FTIR) and thermogravimetric analysis (TG) revealed the successful entrapment of the pesticide. The coating of polydopamine (PDA) endowing the microcapsule with superior UV-shielding properties than pyraclostrobin@SiO2 microcapsule (Pyr@SiO2 MC). Compared with pyraclostrobin emulsifiable concentrate (EC), the Pyr@SiO2@PDA MC exhibited 9.07-, 5.50-, 4.93- and 4.16-fold higher fungicidal activity against Rice blast fungus (Pyricularia oryzae) at concentrations of 0.5, 1, 2 and 4 mg/L. Moreover, acute toxicity tests demonstrated that the sample on zebrafish with lower toxicity on the first day. These results showed that the obtained microcapsule may process broader application potential in agriculture.

Crystal structure of bacterial cytochrome bc1 in complex with azoxystrobin reveals a conformational switch of the Rieske iron-sulfur protein subunit.

Cytochrome bc1 complexes (cyt bc1), also known as complex III in mitochondria, are components of the cellular respiratory chain and of the photosynthetic apparatus of non-oxygenic photosynthetic bacteria. They catalyze electron transfer (ET) from ubiquinol to cytochrome c and concomitantly translocate protons across the membrane, contributing to the cross-membrane potential essential for a myriad of cellular activities. This ET-coupled proton translocation reaction requires a gating mechanism that ensures bifurcated electron flow. Here, we report the observation of the Rieske iron-sulfur protein (ISP) in a mobile state, as revealed by the crystal structure of cyt bc1 from the photosynthetic bacterium Rhodobacter sphaeroides in complex with the fungicide azoxystrobin. Unlike cyt bc1 inhibitors stigmatellin and famoxadone that immobilize the ISP, azoxystrobin causes the ISP-ED to separate from the cyt b subunit and to remain in a mobile state. Analysis of anomalous scattering signals from the iron-sulfur cluster of the ISP suggests the existence of a trajectory for electron delivery. This work supports and solidifies the hypothesis that the bimodal conformation switch of the ISP provides a gating mechanism for bifurcated ET, which is essential to the Q-cycle mechanism of cyt bc1 function.

Discovery of metyltetraprole: Identification of tetrazolinone pharmacophore to overcome QoI resistance.

Quinone outside inhibitors (QoIs) are one of the major agricultural fungicide groups used worldwide. However, the development of resistance by different pathogenic species associated with specific mutation at the target gene site is becoming a critical issue for the sustainable use of QoIs. The authors aimed to design a novel QoI molecule to overcome the aforementioned issue. A rational approach to avoid steric hindrance between the QoI molecule and the mutated target site was successfully employed. The resulting compound, metyltetraprole, is characterized by 3-substituted central ring with a tetrazolinone moiety, the key structure to retain potent activity against QoI-resistant mutants. Metyltetraprole is a promising new fungicide under commercial development, and its development in this study has paved the way to overcoming resistance to QoI fungicides.

Antimalarial 9-Methoxystrobilurins, Oudemansins, and Related Polyketides from Cultures of Basidiomycete Favolaschia Species.

Fourteen new compounds, oudemansins 1-4, oudemansinols 5-7, favolasins 8-10, favolasinin (12), polyketides 13-15, and (R,E)-2,4-dimethyl-5-phenyl-4-pentene-2,3-diol (16), together with nine known compounds were isolated from the basidiomycete fungus Favolaschia sp. BCC 18686. Two new compounds, favolasin E (11) and 9-oxostrobilurin E (17), were isolated from the closely related organism Favolaschia calocera BCC 36684 along with nine ß-methoxyacrylate-type derivatives. Compounds in the class of oudemansins and strobilurins exhibited moderate to strong antimalarial activity with relatively low cytotoxicity against Vero cells (African green monkey kidney fibroblasts). Potent antimalarial activity was demonstrated for 9-methoxystrobilurins G, K, and E (IC50 values 0.061, 0.089, and 0.14 µM, respectively). The structure-activity relationships (SAR) for antimalarial activity is proposed on the basis of the activity of the new and several known ß-methoxyacrylate derivatives in combination with the data from previously isolated compounds. Furthermore, several compounds showed specific cytotoxicity against NCI-187 cells (human small-cell lung cancer), although the SAR was different from that for antimalarial activity.

Adsorption Properties of Comb-Shaped Polycarboxylate Dispersant onto Different Crystal Pyraclostrobin Particle Surfaces.

The stability of the suspension system of the two crystal forms of pyraclostrobin is evaluated using multiple light technology, and the adsorption performance of polycarboxylate dispersant on the surface of two different crystalline pyraclostrobin particles is compared in combination with XRD, FTIR, XPS, and SEM from the microscopic view. The adsorption kinetics and thermodynamics studies of 2700 on the surfaces of different crystalline forms of pyraclostrobin particles show that the adsorption process of 2700 on the surfaces of pyraclostrobin crystal forms II and IV conform to pseudo-second-order kinetic adsorption model. The Ea values for crystal forms II and IV are 12.93 and 14.39 kJ∙mol-1, respectively, which indicates that both adsorption processes are physical adsorption. The adsorption models of 2700 on the surfaces of pyraclostrobin crystal forms II and IV are in accordance with Langmuir adsorption isotherms. The ∆Gad values of crystal forms II and IV are negative and the ∆Sad values are positive at different temperatures. Therefore, the adsorption processes are spontaneous and accompanied by entropy increase. The results of this study provide an important theoretical basis for the selection of polycarboxylate dispersants in the suspension of pyraclostrobin. This study also provides a reference for the research of polycrystalline pesticide suspension concentrate.

Natural Compound-derived Cytochrome bc1 Complex Inhibitors as Antifungal Agents.

The high incidence of fungal pathogens has become a global issue for crop protection. A promising strategy to control fungal plant infections is based on the use of nature-inspired compounds. The cytochrome bc1 complex is an essential component of the cellular respiratory chain and is one of the most important fungicidal targets. Natural products have played a crucial role in the discovery of cytochrome bc1 inhibitors, as proven by the development of strobilurins, one of the most important classes of crop-protection agents, over the past two decades. In this review, we summarize advances in the exploration of natural product scaffolds for the design and development of new bc1 complex inhibitors. Particular emphasis is given to molecular modeling-based approaches and structure-activity relationship (SAR) studies performed to improve the stability and increase the potency of natural precursors. The collected results highlight the versatility of natural compounds and provide an insight into the potential development of nature-inspired derivatives as antifungal agents.

Dissipation, residues and risk assessment of pyraclostrobin and picoxystrobin in cucumber under field conditions.

BACKGROUND: Pyraclostrobin and picoxystrobin are two representative pesticides of strobilurins used to treat cucumber downy mildew, which have raised issues of food safety and human health. A new formulation containing these two compounds is being prepared for marketing in China. RESULTS: The dissipation and residual levels of pyraclostrobin and picoxystrobin in cucumbers under field conditions were determined simultaneously by a validated method via liquid chromatography-tandem mass spectrometry (LC-MS/MS). The dissipation rules were described by first-order kinetics and the half-lives of pyraclostrobin and picoxystrobin were less than 8.2 days and 3.4 days. The highest terminal residue of pyraclostrobin was 0.014 mg kg-1 which was lower than maximum residue limit (MRL) in China (0.5 mg kg-1 ) and of picoxystrobin was 0.029 mg kg-1 , respectively. In the long-term intake risk assessment of pyraclostrobin and picoxystrobin for general population (18-79 years), the chronic risk quotient (RQc ) varied from 5.64% to 21.97%. The assessment of short-term risks included children (1-6 years) and adults (18-79 years) and in which the RQa values were 0.38% and 2.85%. Both results showed the intake risks of cucumber were acceptable. CONCLUSION: Pyraclostrobin and picoxystrobin degraded easily in cucumbers under open field conditions. The long-term and short-term risks caused by final residues of pyraclostrobin and picoxystrobin were insignificant. The recommended pre-harvest interval of 3 days was safe. The article will be helpful in rational use of these pesticides and MRL formulation of picoxystrobin on cucumber. © 2020 Society of Chemical Industry.

Meptyldinocap and azoxystrobin residue behaviors in different ecosystems under open field conditions and distribution on processed cucumber.

BACKGROUND: Several diseases and insects may cause damage to the normal growth of cucumber. Azoxystrobin and meptyldinocap, because of their novel mode of action, are effective against pathogens that have developed reduced sensitivity to other fungicides. Azoxystrobin is persistent in various crops and environments. However, there is a lack of research on the dissipation of these two pesticides, especially meptyldinocap. RESULTS: Analytes could be quantified with decent recoveries of 90-101%, with relative standard deviations (RSDs) of 3.0-10.1%. The terminal residues of meptyldinocap and azoxystrobin in cucumber were all < limit of quantification (LOQ) (0.02 and 0.05 mg kg-1 ). The half-lives of meptyldinocap and azoxystrobin were 0.8-1.1 and 1.2-2.8 days, respectively. The processing factors (PFs) for washing were all < 1, but the removal rate for washing was < 29.0%. Peeling had a significant effect on the removal of pesticide. The largest residue reductions were noticed through the pickling process, but special care should be taken regarding residues in the pickling solution as pesticides could transfer to them from cucumber. A more interesting finding was that the degradation of two pesticides was accelerated by the addition of calcium oxide. CONCLUSION: Pesticide residues on cucumber decreased after these processes. These results enable the health-risks from dietary exposures to pesticide residues to be characterized. They enable maximum residue limits (MRLs) to be established for pesticide residues in food products. They also assist the optimization of food processing with regard to pesticide residue dissipation. © 2019 Society of Chemical Industry.

Dissipation behavior, residue distribution and risk assessment of three fungicides in pears.

BACKGROUND: Fungicides are often applied to pears before they are kept in storage facilities. The scientific application of pesticides can reduce unnecessary exposure, which in turn could benefit both humans and the environment. RESULTS: We investigated dissipation behavior and residue distribution, and conducted risk assessments for prochloraz, pyraclostrobin, and tebuconazole in pears stored under different conditions using ultra-performance liquid chromatography (UPLC). The recoveries of the three fungicides ranged from 76.5% to 114.5%, and the coefficients of variation were 1.0%-8.5%. The half-life (t1/2 ) ranges for degradation of the three fungicides in Dangshan Su pear peel were 8.8-13.9 days after storage at 25 °C and 99.0-346.6 days after storage at 2 °C. Among the three fungicides, tebuconazole had the lowest residue concentration in pear pulp (maximum of 0.226 mg·kg-1 ) and the longest half-life (≥ 231.0 days). Accordingly, among these fungicides, tebuconazole is the most suitable for the preservation of Dangshan Su pears during storage. Finally, we analyzed samples of six pear varieties from markets in China and found that the residue concentrations of the three fungicides in pear pulp and fruit met Chinese standards. CONCLUSION: The results provide a scientific basis for rationalizing the use of prochloraz, pyraclostrobin, and tebuconazole, and improving the safety of pears for eating. © 2019 Society of Chemical Industry.

Determination of azoxystrobin and its impurity in pesticide formulations by liquid chromatography.

A method was developed for the simultaneous qualitative and quantitative determination of azoxystrobin and its relevant impurity (Z)-azoxystrobin using high performance liquid chromatography with diode array detector (HPLC-DAD) in suspension concentrate (SC) pesticide formulations, with the aim of the product quality control. Method validation was realized according to SANCO/3030/99 rev. 5. The proposed method was characterized by acceptable accuracy and precision. The repeatability expressed as ratio standard deviation (%RSD) to relative standard deviation (%RSDr) was lower than 1, whereas individual recoveries were in the range of 97-103% and 90-110% for azoxystrobin and (Z)-azoxystrobin, respectively. The limit of quantification (LOQ) for the impurity ((Z)-azoxystrobin) amounted to 0.3 µg mL-1 and was acceptable because it was lower than the maximum permitted level according to Commission Implementing Regulation (EU) No 703/2011 of 20 July 2011 for the active substance (azoxystrobin) being 25 g kg-1 of the azoxystrobin content found. The method described in this paper is simple, precise, accurate and selective as well as represents a new and reliable way of simultaneous determination of azoxystrobin and its relevant impurity in formulated products.

Self-Assembling Micelles Based on an Intrinsically Disordered Protein Domain.

The self-assembly of micellar structures from diblock polymers that contain hydrophilic and hydrophobic domains has been of great interest for the encapsulation of drugs and other hydrophobic molecules. While most commercially used surfactants are derived from hydrocarbon sources, there have been recent efforts to replace these with biodegradable, nontoxic, biologically synthesized alternatives. Previous examples have primarily examined naturally occurring self-assembling proteins, such as silk and elastin-like sequences. Herein, we describe a new series of fusion proteins that have been developed to self-assemble spontaneously into stable micelles that are 27 nm in diameter after enzymatic cleavage of a solubilizing protein tag. The sequences of the proteins are based on a human intrinsically disordered protein, which has been appended with a hydrophobic segment. The micelles were found to form across a broad range of pH, ionic strength, and temperature conditions, with critical micelle concentration (CMC) values in the low micromolar range, 3 orders of magnitude lower than the CMC of commonly used surfactant sodium dodecyl sulfate (SDS). The reported micelles were found to solubilize hydrophobic metal complexes and organic molecules, suggesting their potential suitability for catalysis and drug delivery applications. Furthermore, the inherent flexibility in the design of these protein sequences enables the encoding of additional functionalities for many future applications. Overall, this work represents a new biomolecular alternative to traditional surfactants that are based on nonrenewable and poorly biodegradable hydrocarbon sources.

One-step microencapsulation and spraying of pesticide formulations for improved adhesion and sustained release.

Aim: To reduce the contamination arising from abuse of commercial pesticide formulations, the coaxial electrospray (CES) method was used for one-step microencapsulation and spraying of pesticides. Methods: After optimisation of process parameters, polymeric microcapsules with different structures were fabricated as the carriers of azoxystrobin (AZS). For the resultant microcapsules, the sustained pesticide release was verified in vitro and the adhesion properties were investigated through a normalised rinsing test. Results: The maximum encapsulation efficiency of the fabricated AZS-loaded microcapsules was 99.14%. Compared to commercial AZS aqueous suspension, the microcapsules fabricated by the CES method exhibited improved sustained release performance of AZS, which could be readily controlled by adjusting the shell thicknesses. Moreover, highly enhanced adhesion performance was observed for the AZS-loaded microcapsules directly sprayed in CES process. Conclusions: The CES process is promising to be applied as a one-step microencapsulation and spraying technology for improving pesticide utilisation and reducing environmental pollution.

Identification of Azoxystrobin Glutathione Conjugate Metabolites in Maize Roots by LC-MS.

Xenobiotic detoxification in plant as well as in animals has mostly been studied in relationship to the deactivation of the toxic residues of the compound that, surely for azoxystrobin, is represented by its ß-methoxyacrylate portion. In maize roots treated for 96 h with azoxystrobin, the fungicide accumulated over time and detoxification compounds or conjugates appeared timewise. The main detoxified compound was the methyl ester hydrolysis product (azoxystrobin free acid, 390.14 m/z) thought to be inactive followed by the glutathione conjugated compounds identified as glutathione conjugate (711.21 m/z) and its derivative lacking the glycine residue from the GSH (654.19 m/z). The glycosylated form of azoxystrobin was also found (552.19 m/z) in a minor amount. The identification of these analytes was done by differential untargeted metabolomics analysis using Progenesis QI for label free spectral counting quantification and MS/MS confirmation of the compounds was carried out by either Data Independent Acquisition (DIA) and Data Dependent Acquisition (DDA) using high resolution LC-MS methods. Neutral loss scanning and comparison with MS/MS spectra of azoxystrobin by DDA and MSe confirmed the structures of these new azoxystrobin GSH conjugates.