6540 uhd qtof mass spectrometer

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.

IBG (99.9%) was obtained from Guangzhou Insighter Biotechnology (Guangzhou, China). Sprague–Dawley rat liver microsomes and a Phase I metabolic stability kit was purchased from IPHASE Co., Ltd. (Beijing, China, Lot: 111011). Acetonitrile and formic acid of HPLC-grade were bought from Sigma-Aldrich (Oakville, ON, Canada). Deionized water was prepared using a Milli-Q purification system (Millipore Corp., Bedford, MA, USA). An LC-QTOF-MS/MS system equipped with an Agilent 1290 system (Agilent Technologies, Santa Clara, CA, USA) and a 6540 UHD QTOF mass spectrometer (Agilent Technologies, USA) was employed.