Dissipation of penconazole formulation in horticultural crops by ultrahigh performance liquid chromatography-high resolution mass spectrometry: From the active substance to metabolites.

The present study describes the dissipation and metabolism of penconazole in horticultural products by a method based on ultra-high performance liquid chromatography-quadrupole-orbitrap (UHPLC-Q-Orbitrap). Targeted and suspected analysis were carried out. Two independent trials were performed under laboratory conditions (on courgette samples), and under greenhouse conditions (on tomatoes) during 43 and 55 days, respectively. In both studies, a pesticide formulation (TOPAS® EW) containing penconazole was used. The results showed that penconazole was relatively short-lived (<30 days) in horticultural products. The proposed method allowed for the tentative identification and semi-quantification of nine metabolites. In addition, the potential toxicity of these metabolites was evaluated, observing that some of them are even more toxic than penconazole, as triazole lactic acid. This research may provide a starting point for understanding the dissipation process of penconazole, the formation pathways of its main metabolites, their concentrations and toxicity to ensure food safety and the environmental protection.

Understanding the Metabolism and Dissipation Kinetics of Flutriafol in Vegetables under Laboratory and Greenhouse Scenarios.

Flutriafol is a systemic triazole fungicide that is used to control diseases in various crops. A study was developed to evaluate the metabolism and dissipation of flutriafol in two different scenarios: laboratory and greenhouse conditions. Courgette and tomato samples treated with a commercial product (IMPACT® EVO) at the manufacturer recommended dose were analyzed, and courgette samples were also treated at double dose. Ultra-high-performance liquid chromatography coupled with Q-Orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS), performing targeted and non-targeted approaches (suspect screening and unknown analysis), were used to analyze the samples. The dissipation of flutriafol was fitted to a biphasic kinetic model, with a persistence, expressed as half-life (t1/2), lower than 17 days. During suspect screening, three metabolites (triazole alanine, triazole lactic acid and triazole acetic acid) were tentatively identified. Unknown analysis led to the identification of four additional metabolites (C16H14F2N4, C16H14F2N4, C19H17F2N5O2 and C22H23F2N3O6). The results revealed that the proposed methodology is reliable for the determination of flutriafol and its metabolites in courgette and tomato, and seven metabolites could be detected at low concentration levels. The highest concentration of metabolites was found in the laboratory conditions at 34.5 µg/kg (triazole alanine). The toxicity of flutriafol metabolites was also evaluated, and some of them could be more toxic than the parent compound.

Non-targeted analysis of co-formulants in antifungal pesticide formulations by gas chromatography-tandem high resolution mass spectrometry.

In the present study, six commercial pesticide formulations with antifungal activity were characterized. Thus, two complementary injection methods based on gas chromatography were employed: direct injection (DI) and headspace (HS), both coupled to high resolution mass spectrometry (GC-Q-Orbitrap-MS). The combination of both injection modes allowed the tentatively identification of potential co-formulants. Available analytical standards were acquired for their confirmation, and 21 compounds were successfully confirmed. Finally, the concentration of these co-formulants was calculated, finding the highest value in one of the pesticide formulation, at 218.22 g L-1 for cyclohexanone. Results clearly show that this methodology is suitable for the reliable identification of co-formulants in pesticide formulations, offering high sensitivity, and highlighting that five co-formulants were detected by both injection techniques (DI and HS). Moreover, one of the main advantages of the proposed methods was the great capacity for the elucidation of compounds with similar molecular formula, bearing in mind that up to 8 co-formulants with the same molecular formula C10H14, were well differentiated by retention times between 8.46 (1-methyl-3-propylbenzene) and 10.98 min (1,2,3,4- tetramethylbenzene) in one of the pesticide formulation. Toxicity to human health and the environment has been evidenced for the co-formulants detected, finding compounds with relatively high toxicity, as naphthalene and cyclohexanone.

Characterization of the composition of plant protection products in different formulation types employing suspect screening and unknown approaches.

BACKGROUND: Plant protection products (PPPs) are used extensively in agriculture to control crops. These PPPs, which may be found in different types of formulations, are composed of a designated pesticide (active principle) and other inactive ingredients as co-formulants. They perform specific functions in the formulation, as solvents, preservatives or antifreeze agents, among others. RESULTS: A research technique based on ultra-high-performance liquid chromatography (UHPLC) coupled to a Quadrupole-Orbitrap mass analyzer was successfully applied to characterize the composition of six different PPPs in terms of the presence of co-formulants and types of formulations: emulsifiable concentrate (EC), emulsion in water (EW), suspension concentrate and water-dispersible granule. These PPPs (FLINT MAX, MASSOCUR 12.5 EC, IMPACT EVO, TOPAS, LATINO and IMPALA STAR) had antifungal activity, containing one triazole compound as active principle (tebuconazole, penconazole, myclobutanil, flutriafol or fenbuconazole, respectively). Non-targeted approaches, applying suspect and unknown analysis, were carried out and ten compounds were identified as potential co-formulants. Six (glyceryl monostearate, 1-monopalmitin, dimethyl sulfoxide, N,N-dimethyldecanamide, hexaethylene glycol and 1,2-benzisothiazol-3(2H)-one) were confirmed by injecting analytical standards. Finally, these compounds were quantified in the PPPs. CONCLUSION: The current study allowed for detecting co-formulants in a wide range of concentrations, between 0.04 (dimethyl sulfoxide) and 19.00 g L-1 (glyceryl monostearate), highlighting the feasibility of the proposed analytical methodology. Moreover, notable differences among the types of formulations of PPPs were achieved, revealing that EC and EW were the formulations that contained the largest number of co-formulants (four out of six detected compounds). © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Targeted and untargeted analysis of triazole fungicides and their metabolites in fruits and vegetables by UHPLC-orbitrap-MS2.

Two extraction methods based on solid liquid extraction and Quick, Easy, Cheap, Effective, Rugged and Safe procedure were developed for the determination of 21 triazole compounds and 5 metabolites, including triazole derivative metabolites as 1,2,4-triazole and 1,2,4-triazol 1-yl-acetic, in courgette, orange, grape and strawberry. The analysis was performed in 10.5 min, using ultra-high performance liquid chromatography coupled to Q-Orbitrap mass analyser. The proposed method was validated according to SANTE 12682/2019. Limits of quantification were ≤10 µg kg-1 for all the compounds, except for 1,2,4-triazol, 1,2,4-triazol 1-yl-acetic, difenoconazole-alcohol and prothioconazole that were 50 µg kg-1. Finally, the method was successfully applied to the analysis of 30 samples. More than 30% of these samples contained residues of triazole compounds. The fungicide most frequently found was myclobutanil. Furthermore, a suspect screening analysis was carried out to search pesticides present in the samples, detecting some of them at concentrations higher than Maximum Residue Limits.