Accela 1250 uplc system

Mycotoxins were determined on an Accela 1250 UPLC system (Thermo Fisher Scientific, San Jose, CA, USA) coupled to a TSQ VantageTM (Thermo Fisher Scientific, San Jose, CA, USA) triple stage quadrupole mass spectrometer. Separation was performed on an Agilent Extend-C18 column (100 mm × 4.6 mm, 3.5 μm) at 30°C with a flow rate of 350 μL/min. The mobile phase consisted of water containing 5 mM ammonium acetate (A) and methanol (B). The gradient was as follows: 0 min 20% B, 1 min 20% B, 2 min 50% B, 8 min 100% B, 10 min 100% B, 13 min 20% B, 15 min 20% B. The injection volume was 10 μL.
For MS/MS analysis, the parameters were set as follows: interface voltage of 2.5 kV (ESI⁻); desolvation temperature of 270°C; nebulizing gas (N₂) pressure of 50 psi and drying gas (N₂) pressure of 25 psi; heat block temperature of 300°C. The quantitation and identification of target mycotoxins were performed in selected reaction monitoring (SRM) mode. The optimized MS/MS parameters for each analyte are listed in Table 1. Xcalibur™ software (Thermo Fisher Scientific, San Jose, CA, USA, 2011) was used for data processing.

Samples were extracted by mixing flour with 10 mL of acetonitrile: water: formic acid (840:159:1 v/v). Then, they were centrifuged at 4000 rpm/min for 10 min and filtered through a 0.22 μm nylon filter. From the filtered solution, 1 mL was taken and stored in sampler vials at −20 °C until LC–MS/MS analysis.
Mycotoxins were determined using an Accela 1250 UPLC system (Thermo Fisher Scientific, San Jose, CA, USA) coupled to a TSQ VantageTM (Thermo Fisher Scientific, San Jose, CA, USA) triple-stage quadrupole mass spectrometer. The chromatographic column used in this method was an AgilentExtend-C18 (100 mm × 4.6 mm, 3.5 μm) column with a flow rate of 0.35 mL/min at 30 °C. The injection volume was 10 μL. For FBs, the mobile phase consisted of water containing 0.1% formic acid (A) and methanol containing 0.1% formic acid (B). The gradient was as follows: 0 min 29% B, 3 min 74% B, 5 min 74% B, 5.2 min 100% B, 5.7 min 100% B, 5.8 min 29% B, and 10 min 24% B. Mass spectrometry analysis was performed in positive ionization mode (ESI + 3.5 kV) using selected reaction monitoring (SRM). For MS/MS analysis, the optimized conditions were set as follows: vaporizer temperature at 300 °C; capillary temperature at 300 °C; sheath gas pressure at 10 psi; aux gas pressure at 15 psi. Data analysis was performed with XcaliburTM software 4.1 (Thermo Fisher Scientific, San Jose, CA, USA, 2011).