310 ms tq mass spectrometer

All analyses were carried out by using a 1260 Agilent Technologies system consisting of a binary pump and a vacuum degasser, connected to a Varian autosampler Model 410 Prostar (Hansen Way, CA, USA) equipped with a 20 μL loop coupled with a Varian 310-MS TQ Mass Spectrometer. The separation of mycotoxins was performed using a Kinetex PFP (100 × 2.10 mm 2.6 μm, Phenomenex, Torrance, CA, USA) under a flow of 200 μL/min. The column temperature was set at 35 °C. Solvent A was H₂O with 2 mM NH₄HCO₃, solvent B was CH₃OH with 2 mM NH₄HCO₃. HPLC analysis was performed using a linear gradient from 40% to 100% of solvent B in 12 min.
Samples were ionized using an electrospray (ESI) ion source operating in negative ion mode. For the MRM experiments two transitions were selected for each compound: for TeA, m/z 196 > 112 CE 24V (monitoring) and m/z 196 > 139 CE 20V (quantification); for AME, m/z 257 > 147 CE 34V (monitoring) and m/z 257 > 213 CE 22V(quantification); for AOH, m/z 271>228 CE 28V (monitoring) and m/z 271 > 256 CE 22V (quantification); for ALT, m/z 291 > 247 CE 20V (monitoring) and m/z 291 > 229 CE 12V (quantification); for TEN, m/z 413 > 141 CE 18V (monitoring) and m/z 413 > 271 CE 16V (quantification). The collision gas (Ar) pressure was set at 2 mbar for all experiments.

Stilbenes and ABA were quantified starting from 200 mg of frozen leaves. Reversed-phase high-performance liquid chromatography analysis was performed on a 1,260 Agilent Technologies system. Chromatographic separation was performed with a Phenomenex (Torrance, CA) Luna C18 column (150 × 2.1 mm, 3 μm particle size), with a C18 SecurityGuard column (4.0 × 3.0 mm ID), operated at room temperature. Elution was carried out using aqueous formic acid (0.1% v/v; mobile phase A) and acetonitrile (mobile phase B). HPLC analysis was performed using a gradient from 20 to 60% of mobile phase B in 15 min, then from 60 to 100% of B in 4 min; after washing for 5 min with solvent B, the column was re-equilibrated. The flow rate was 0.2 mL/min and injection volume was 10 μL. A triple quadrupole mass spectrometer (Varian 310-MS TQ Mass Spectrometer) equipped with an ESI interface operated in negative ion mode was used. The detection of analytes was carried out in multiple reaction monitoring (MRM) mode by monitoring two transitions for each compound. The concentration of analytes was quantified using an external calibration method. Original standards of resveratrol (purity ≥99%), polydatin (purity ≥95%), and viniferin (purity ≥95%), purchased from Sigma-Aldrich, were used to prepare the calibration curves. Similarly, the ABA content was quantified as described by Siciliano et al. (2015 (link)).

Analyses were performed using a 1260 Agilent Technologies system consisting of a binary pump and a vacuum degasser, connected to a Varian autosampler, Model 410 Prostar (Hansen Way, CA, USA), equipped with a 20 μL loop coupled to a Varian 310-MS TQ Mass Spectrometer. The separation of mycotoxins was performed using a Gemini-NX C18 (150 × 2 mm, 3 μm, Phenomenex, Torrance, CA, USA) under a flow of 200 μL/min. Solvent A was H₂O acidified with formic acid 0.05%, while solvent B was CH₃CN. Elution gradient started with 30% of solvent B for 5 min, increased to 50% in 10 min and remained at 50% for 10 min, then increased to 100% in 20 min. During the next 6 min, the column was washed and readjusted to the initial conditions and equilibrated for 10 min. The volume of the injection was 10 μL. Samples were ionized using an electrospray (ESI) ion source operating in positive and negative ion modes in different segments. For the multiple reaction monitoring (MRM) experiments, two transitions were selected for each compound (Table 1) and the collision gas (Ar) pressure was set at 2 mbar for all the experiments.