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4 protocols using kresoxim methyl

1

Chemical Library Acquisition and Verification

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The chemical library was donated by the EPA ToxCast Program. For verification and replication experiments, the following chemicals were purchased from Sigma-Aldrich: DL-α tocopherol acetate (T3376), DL-sulforophane (S4441), vincristine sulfate salt (V8879), oxyfluorfen (35031), rotenone (45656), fenamidone (33965), pyraclostrobin (33696), trifloxystrobin (46447), myxothiazol (T5580), pyridaben (46047), azoxystrobin (31697), fluoxastrobin (33797), fenpyroximate (31684) and kresoxim-methyl (37899). Famoxadone was purchased from Chem Service, Inc (N-11943). Topotecan hydrochloride was purchased from Tocris (4562). Paclitaxel was purchased from Fisher Scientific (AC32842). All chemical stocks were prepared in DMSO unless otherwise noted. Vehicle samples were prepared with an equivalent DMSO concentration of ≤0.5% in feeding medium.
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2

Antifungal Compounds Assessment Protocol

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Antifungal compounds, kresoxim methyl (Kre-Me; strobilurin), octylgallate (octyl 3,4,5-trihydroxybenzoic acid; OG), veratraldehyde (3,4-dimethoxybenzaldehyde; VT), benzohydroxamic acid (BHAM) (Figure 1), were procured from Sigma Co. (St. Louis, MO, USA). Each compound was dissolved in dimethylsulfoxide (DMSO; absolute DMSO amount: <2% in media) before incorporation into culture media (except for those plates used in aflatoxin assays; see below). Throughout this study, control plates (No treatment) contained DMSO at levels equivalent to that of cohorts receiving antifungal agents, within the same set of experiments.
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3

Pesticides Analysis in Samples

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The pesticides standards of deltamethrin, penconazole, kresoxim-methyl, cyproxonazole, epoxiconazole and azoxystrobin were supplied by Sigma-Aldrich (Seelze, Germany). The stock solutions (1000 mg/L) were prepared by dissolving the standards in methanol HPLC grade (99.9%) purchased by Sintorgan (Buenos Aires, Argentina), and stored under freezing condition (-18°C ± 1°C) in dark bottles sealed with PTFE/silicone caps. The working standard solutions (50 mg/L) were prepared in acetonitrile (ACN) of high purity grade for residue analysis, provided by Merck (Darmstadt, Germany).
Deionized water obtained from an E-pure water purification system (Barnstead/Thermolyne, Bedford, MA, United States) was used for sample swelling in the extraction step.
For extraction and partitioning, sodium chloride and anhydrous sodium sulfate were purchased from Biopack (Buenos Aires, Argentina); sodium hydrogencitrate sesquihydrate and sodium citrate dihydrate were obtained from Sigma-Aldrich (Seelze, Germany). For clean-up using d-SPE, C18 and PSA bulk powder were obtained from Agilent Technologies (Santa Clara, United States).
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4

Pesticides Analysis in Samples

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The pesticides standards of deltamethrin, penconazole, kresoxim-methyl, cyproxonazole, epoxiconazole and azoxystrobin were supplied by Sigma-Aldrich (Seelze, Germany). The stock solutions (1000 mg/L) were prepared by dissolving the standards in methanol HPLC grade (99.9%) purchased by Sintorgan (Buenos Aires, Argentina), and stored under freezing condition (-18°C ± 1°C) in dark bottles sealed with PTFE/silicone caps. The working standard solutions (50 mg/L) were prepared in acetonitrile (ACN) of high purity grade for residue analysis, provided by Merck (Darmstadt, Germany).
Deionized water obtained from an E-pure water purification system (Barnstead/Thermolyne, Bedford, MA, United States) was used for sample swelling in the extraction step.
For extraction and partitioning, sodium chloride and anhydrous sodium sulfate were purchased from Biopack (Buenos Aires, Argentina); sodium hydrogencitrate sesquihydrate and sodium citrate dihydrate were obtained from Sigma-Aldrich (Seelze, Germany). For clean-up using d-SPE, C18 and PSA bulk powder were obtained from Agilent Technologies (Santa Clara, United States).
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