Uv vis detector

APIs were quantified using a Dionex Ultimate 3000 U-HPLC (Thermo Fischer, Dreieich) equipped with a Synchronis C18 50 × 2.1 mm, 1.7 µm column (Thermo Fischer, Germany) and a UV–VIS detector (Thermo Fischer, Germany). (i) In case of LVX, 10 mg of dry powder was dissolved in 1 mL PBS containing 0.1% Triton-X and eluted with a 18% of mobile phase A (acetonitrile (ACN)) and 72% of mobile phase B (0.5% trimethylamine buffer at pH 2.5) at a flow of 0.3 mL/min (see Supporting Information, Fig. 7). (ii) In case of BTZ043 (Selleckchem; Houston, TX, USA), powder was dissolved in a 60:40 mixture of ACN:PBS at a concentration of 10 mg/mL of dry powder. A binary solvent gradient was applied at a flow rate of 0.3 mL/min and programmed as follows: 80% H₂O and 20% ACN at 0 min to 0.5 min, progressing linearly at 5% H₂O and 95% ACN at 2 min, followed by the hold in H₂O 5% ACN to 95% for 1 min, and finally returning to the initial gradient until 5 min (cf. Supporting Information, Figs. 8 and 9). Data analysis was performed with Chromeleon 7 software (Thermo Fischer, Germany).

Pesticides were analyzed according to Hafeez et al. (2015) using the HPLC-DAD technique with some modifications [50 (link)]. For the qualitative–quantitative determination of pesticides, HPLC analysis was performed using a UHPLC+ Dionex UltiMate 3000 system (Thermo Fisher Scientific Inc., Waltham, MA, USA) equipped with a UV–Vis detector (Thermo Fisher Scientific Inc., Waltham, MA, USA) and NUCLEOSIL C18 column (4.6 × 250 mm, 5 µm particle size; Macherey-Nagel, Germany). Isocratic elution was carried out with 70/30 (v/v) acetonitrile/water as the mobile phase. The flow rate was set to 1.0 mL/min, and the column temperature was 25 °C. The injection volume was 20 µL. Detection and quantification were performed with two wavelengths: 219 nm for chlorpyrifos and coumaphos and 270 nm for imidacloprid. Quantification was performed using an external standard method. All measurements were made twice.

AFB₁ in the control groups and samples was extracted three times using chloroform with an equal volume before the solvent evaporated under N₂ at room temperature [23 (link),26 (link),57 (link),58 (link)]. Dimethyl sulfoxide (DMSO) (50 µL) was used to dissolve the dried extracts, and 20 µL of the mixture was injected in UltiMateTM3000 HPLC (Thermo Scientific, Bremen, Germany). HPLC analysis was conducted with a C18 Polaris column (250 mm × 4.6 mm i.d., 5 µm) in a mobile phase of water and methanol in a 1:1 ratio (v/v). The flow rate was set as 1 mL min⁻¹, and a UV/VIS detector (Thermo Scientific, Germany) was used for absorbance measurements at a wavelength of 360 nm. The column temperature was set to 35 °C for detection. The software Chromeleon v6.8 was used for data analysis. The rate of AFB₁ degradation was determined and calculated with (1 − AFB₁ peak area in treatment/AFB₁ peak area in control) × 100%.