1290 lc system

The LC–MS analysis was performed as described previously by Hanhineva et al.^{43 (link)}. In brief, the samples were analyzed using liquid chromatography quadrupole time-of-flight mass spectrometry UHPLC–QTOF–MS system (Agilent Technologies), which consisted of a 1290 LC system, a Jetstream electrospray ionization (ESI) source, and a 6540 UHD QTOF mass spectrometer. The samples were separated using reversed-phase (RP) chromatography (Zorbax Eclipse XDB-C18, particle size 1.8 µm, 2.1 × 100 mm; Agilent Technologies). The elution solvents were water (A) and HPLC grade methanol (B), both containing formic acid 0.1% v/v. The gradient was as follows for the ratio of solvent B: 0–10 min: 2 → 100% B; 10–14.5 min: 100% B; 14.5–14.51 min: 100 → 2% B; 14.51–16.5 min: 2% B. Data was acquired with both positive and negative polarity. Quality controls were injected at the beginning and at the end of the MS run and after every ten injections. Automatic data-dependent MS/MS analysis was performed on one sample representing each sample type. The sample tray was kept at +4 °C during the analysis. Three replicates for malted grain and rootlet and three technical replicates for whole grains, water extract, spent grain, and wort, were analyzed in a completely randomized order.

LC separation was carried out on a 1290 LC system (Agilent technologies). Separation was performed with a reversed-phase Zorbax 300 SB-C18 column maintained at 60°C. The mobile phases consisted in (A) 0.1% FA in water and (B) 0.1% FA in MeOH. After an isocratic step of 2 min at 2% B, a linear gradient from 10 to 70% B was run over the next 13 min with a flow rate of 50 μL/min. The column was then washed for 1 min with 90% B and re-equilibrated during 5 min with 2% B. Eluent flow before 2 min and after 15 min was discarded with a divert valve to reduce contamination of the mass spectrometer.