Carboxylesterase and Cytochrome P450 Confer Metabolic Resistance Simultaneously to Azoxystrobin and Some Other Fungicides in Botrytis cinerea.

Plant pathogens have frequently shown multidrug resistance (MDR) in the field, often linked to efflux and sometimes metabolism of fungicides. To investigate the potential role of metabolic resistance in B. cinerea strains showing MDR, the azoxystrobin-sensitive strain B05.10 and -resistant strain Bc242 were treated with azoxystrobin. The degradation half-life of azoxystrobin in Bc242 (9.63 days) was shorter than that in B05.10 (28.88 days). Azoxystrobin acid, identified as a metabolite, exhibited significantly lower inhibition rates on colony and conidia (9.34 and 11.98%, respectively) than azoxystrobin. Bc242 exhibited higher expression levels of 34 cytochrome P450s (P450s) and 11 carboxylesterase genes (CarEs) compared to B05.10 according to RNA-seq analysis. The expression of P450 genes Bcin_02g01260 and Bcin_12g06380, along with the CarEs Bcin_12g06360 in Saccharomyces cerevisiae, resulted in reduced sensitivity to various fungicides, including azoxystrobin, kresoxim-methyl, pyraclostrobin, trifloxystrobin, iprodione, and carbendazim. Thus, the mechanism of B. cinerea MDR is linked to metabolism mediated by the CarE and P450 genes.

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.

A powerful helper of azoxystrobin degradation-the unique mechanism of UGT72E2 promoting environmental degradation of azoxystrobin.

In recent years, environmental pollutants such as pesticide residues have become one of the severe public problems that endanger the ecological environment and affect human health. The development of biotechnology to rapidly and efficiently degrade pesticides is essential to reduce their environmental risks. Azoxystrobin (AZ) is representative of the most widely used agricultural fungicide in the world. A large number of studies have shown that AZ has toxic effects on non-target organisms such as fish, algae, earthworms, etc., which may pose a potential threat to the environmental ecosystem. Therefore, it is particularly important to develop new AZ phytoremediation methods. Based on the constructed Arabidopsis UGT72E2 knockout (KO) and overexpression (OE) lines, this study found that overexpression of UGT72E2 in Arabidopsis can enhance resistance to exogenous AZ stress and maintain a relatively stable physiological state while enhancing the metabolic degradation of AZ. Correspondingly, knockout mutants showed the opposite results. The results showed that the AZ glycosylation and malonyl glycosylation products produced by UGT72E2 overexpression lines increased by 10%~20% compared with normal lines, and increased by 7%~47% compared with gene knockout plants, and exhibited lower phytotoxicity. In summary, our findings highlight the critical role of UGT72E2 overexpression in constructing new varieties of phytoremediation and may provide new ideas for reducing the indirect or direct risks of pesticides or other environmental pollutants to non-target organisms and improving biological and environmental resilience.

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.

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.

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.

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.

An improved solvent evaporation method to produce poly (lactic acid) microspheres via foam-transfer.

The aim of this work was to study an improved solvent evaporation method to prepare poly (lactic acid) (PLA) microspheres via foam-transfer. Since the foaming process and its transfer were critical to the improved method, they have been studied. Additionally, the delivery capability of foams was studied as a function of the oil/water ratio, the stirring rate, the concentration of polyvinyl alcohol (PVA) and ethanol (EtOH) in the aqueous phase (ωPVA, ωEtOH). It was found that foaming varied during the preparation process and it influenced the properties of PLA microspheres. When the oil/water ratio (w/w) ≥ 3:10, stirring rate ≥ 600 r/min, ωPVA ≥ 1 wt%, and ωEtOH = 0 wt%, solvent evaporation was able to produce enough foams for foam-transfer, which helped to deliver more than 89 wt% PLA microspheres to the receiving vessel. However, ωPVA ≤ 0.3 wt% and ωEtOH = 20 wt% were unfavorable for maintaining the spherical shape of PLA microspheres and caused the aggregation. The methodology was further used to prepare azoxystrobin-loaded PLA microspheres successfully with a high encapsulation efficiency of 86.54%. This work is meaningful since it enables an efficient and continuous route to prepare functional biodegradable polymer microspheres.

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.

Hazard assessment using an in-silico toxicity assessment of the transformation products of boscalid, pyraclostrobin, fenbuconazole and glyphosate generated by exposure to an advanced oxidative process.

Agricultural pesticide use is ongoing and consumer concern regarding the safety of pesticide residues on produce has generated interest in techniques that can safely reduce residues post-harvest. Recently an advanced oxidative process has shown promise in substantial residue reduction on the surface of produce. Given the potential for oxidative transformation of pesticides to generate transformation products with greater toxicity than the parent residue, take for example the oxon products of the organophosphorus insecticides, it is important to consider what transformation products are generated by pesticide exposure to an oxidative process and their potential toxicity. In this study, previously published transformation products of boscalid, pyraclostrobin, fenbuconazole and glyphosate were identified after exposure to 3% hydrogen peroxide, UV-C irradiation or their combination in an advanced oxidative process on glass, their oral toxicity, carcinogenicity and developmental toxicity were identified in-silico and an initial tier hazard assessment was conducted. Of the 87 total structures that were searched for, 53 were detected by UPLC-QTOF-MS and identified by mass spectra: 15, 13, 22 and 3 structures for boscalid, pyraclostrobin, fenbuconazole and glyphosate respectively, including the parent residues. Oral toxicity of the transformation products of pyraclostrobin and glyphosate was similar to or lower than the parent residue. Several transformation products of boscalid and fenbuconazole were estimated to be significantly more orally toxic than their parent residues. While the majority of the transformation products of boscalid, pyraclostrobin and fenbuconazole were predicted to be carcinogenic there were 11 that were consistently identified to have carcinogenic potential by several assessments. 29 of the 53 molecules were predicted to be probable developmental toxicants. An initial tier hazard assessment was conducted for Cramer rules classification and mutagenicity using the threshold of toxicological concern approach and predicted rat oral LD50. Two exposure scenarios were considered, one highly protective considering each transformation product to be at the highest maximum residue limit (MRL) for the pesticide and whole produce consumption at the highest consumption rate from the USEPA Exposures Handbook, the other considering only apple consumption with the relevant MRL. As indicated by the hazard assessment, several transformation products of boscalid, pyraclostrobin and fenbuconazole should be strongly considered for further testing, either by quantifying their production or in-vivo and in-vitro toxicity tests due to their predicted toxicity and associated hazard.

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.

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.

Evaluation of Pyraclostrobin as an Ingredient for Soybean Seed Treatment by Analyzing its Accumulation-Dissipation Kinetics, Plant-Growth Activation, and Protection Against Phytophthora sojae.

Seed treatment with fungicides has been regarded as a principal, effective, and economic technique for soybean [Glycine max (L.) Merr.] against pathogenic microorganisms during seed germination and seedling growth. Investigation of the characteristics of seed-treatment reagents is an indispensable basis for their application. The aim of the present work is to evaluate the use of pyraclostrobin as an ingredient for soybean seed treatment by investigating its accumulation-dissipation kinetics in plants, plant-growth activation, and protection against Phytophthora sojae. The results showed that the pyraclostrobin stimulated the visible growth (root and shoot length) of soybean plants, increased the chlorophyll level and root activity, and lowered the malonaldehyde (MDA) level. The peak level and bioavailability of pyraclostrobin in soybean roots were 19.9- and 33.2-fold those in leaves, respectively, indicating that pyraclostrobin was mainly accumulated in roots. Pyraclostrobin had a continuous positive effect on the flavonoid levels and the phenylalanine ammonialyase (PAL) activity in roots and leaves, which could enhance the plant defense system. Pyraclostrobin showed in vitro toxicity to P. sojae with a half-inhibition concentration (EC50) of 1.59 and 1.24 µg/mL for pyraclostrobin and pyraclostrobin plus salicylhydroxamic acid (SHAM, an inhibitor of the alternative pathway of respiration), respectively. Seed treatment with pyraclostrobin significantly reduced the severity of Phytophthora root rot, with a control efficacy of 60.7%. To the best of our knowledge, this is the first report on the characteristics of pyraclostrobin used in soybean seed treatment and its efficacy against Phytophthora root rot.

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.

Fungicidal Effect of Pyraclostrobin against Botrytis cinerea in Relation to Its Crystal Structure.

Pyraclostrobin (PYR) is a commonly used strobilurin fungicide, which inhibits mitochondrial respiration at the ubiquinol oxidation center site of the cytochrome bc1 complex. Little information is available regarding the crystal structure of PYR on its fungicidal effect. In this study, the crystal structures of eight PYRs (PYR-A to H) from different sources are determined by using high-resolution X-ray powder diffraction (XRPD) and model construction with the Pawley refinement module. The effects of PYRs on mycelium growth, the kinetics of mycelial growth, conidial germination, and tube elongation of conidia of Botrytis cinerea from tomato are compared. The level of organic acids in the mitochondrial tricarboxylic acid cycle of PYR-treated B. cinerea is analyzed. The results show that PYR-A to PYR-H have their own unique character of XRPD patterns, but the crystal morphology of eight PYRs presents in the triclinic crystal system and space group P1̅. PYR-D with the eclipsed conformation and rational edge angles α (72.599°) and ß (98.612°) in the crystal cell shows the highest inhibitory effect against mycelium growth with EC50 as 3.383 µg mL-1, the best time-dependent effects on the mycelium growth kinetics, and the strongest inhibition on tube elongation of conidia, whereas PYR-E with anticonformation is the worst. Moreover, a significant accumulation of fumarate, malate, and oxalate in the PYR-D-treated mycelium is observed. These findings reinforce the need for a definite crystal structure of PYR to limit usage and mitigate future selection pressure for gray mold management.

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.

Preparation and characterization of a novel controlled-release nano-delivery system loaded with pyraclostrobin via high-pressure homogenization.

BACKGROUND: The development of efficient and safe green pesticides is a scientific strategy to alleviate current pesticide residues, environmental pollution, and threats to non-target organisms. Pesticide controlled-release formulations (CRFs) have attracted wide attention because they can control the rate of release of active ingredients and prolong the effective duration. In particular, nanoscale pesticide sustained-release systems have excellent biological activity and distribution performance because of their small particle size. Some technical difficulties remain in obtaining nanoscale CRFs. RESULTS: We successfully fabricated pyraclostrobin nanosphere CRF by combining high-pressure homogenization technology and emulsion-solvent evaporation methods. The pyraclostrobin nanospheres had a uniform spherical shape with a mean particle size of 450 nm and polydispersity index of less than 0.3. The pyraclostrobin loading capacity reached 53.6%, with excellent storage stability. The contact angle of nanospheres on cucumber leaf surfaces demonstrated that it had good wettability. Compared with pyraclostrobin technical and commercial formulations, the nanosphere systems showed a significantly sustained release of pyraclostrobin for longer (up to 250 h). A preliminary bioassay against Penicillium ochrochloron showed that the bioactivity and long-term efficiency of pyraclostrobin nanospheres were superior to those of the commercial formulation. CONCLUSION: This research introduced a simple, fast, expandable method for preparing pyraclostrobin nanospheres. The results showed that pyraclostrobin nanospheres could prolong the duration of pesticide efficacy and enhance bioactivity. Furthermore, this technology provides a platform for scale-up production of nano-scale pesticide CRFs. © 2020 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.

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.

An effervescence tablet-assisted microextraction based on the solidification of deep eutectic solvents for the determination of strobilurin fungicides in water, juice, wine, and vinegar samples by HPLC.

In this study, a simple, efficient, and green effervescence tablet-assisted microextraction method based on the solidification of deep eutectic solvent (ETA-ME-SDES) was developed to determine picoxystrobin, pyraclostrobin, and trifloxystrobin in water, juice, wine, and vinegar samples by HPLC. An eco-friendly, hydrophobic, deep eutectic solvent (DES, acting as the extraction solvent) was synthesized by thymol and octanoic acid in the molar ratio of 1:5. The extraction solvent dispersed in sample solutions with the assistance of pH adjustment and effervescence reaction, and was collected after solidification in an ice bath. Several essential conditions, including the type and the volume of DESs, the amount of ammonia hydroxide, and the components of effervescence tablets were optimized. The limits of detection ranged from 0.15 to 0.38 µg L-1. Extraction recovery ranged from 77.4 to 106.9%. The proposed method was successful in determining the amount of strobilurin fungicides in water, juice, wine, and vinegar samples.