Sequant zic philic lc column

The LC-MS conditions were identical to previously established methods [62 ]. For the chromatographic metabolite separation, the Vanquish UPLC systems were coupled to a Q Exactive HF (QE-HF) mass spectrometer equipped with HESI (Thermo Fisher Scientific, Waltham, MA). The column was a SeQuant ZIC-pHILIC LC column, 5 μm, 150 × 4.6 mm (MilliporeSigma, Burlington, MA) with a SeQuant ZIC-pHILIC guard column, 20 × 4.6 mm (MilliporeSigma, Burlington, MA). Mobile phase A was 10 mM (NH₄)₂CO₃ and 0.05% NH₄OH in H₂O while mobile phase B was 100% ACN. The column chamber temperature was set to 30 °C. The mobile phase condition was set according to the following gradient: 0–13 min: 80%–20% of mobile phase B, 13–15 min: 20% of mobile phase B. The ESI ionization mode was negative. The MS scan range (m/z) was set to 60–900. The mass resolution was 120,000 and the AGC target was 3 × 10⁶. The capillary voltage and capillary temperature were set to 3.5 KV and 320 °C, respectively. 5 μL of sample was loaded. For the non-targeted metabolomics approach, the LC-MS peaks were automatically extracted and aligned using the Automated Feature Detection function of EL-Maven. After the normalization with the median value of the intensities of LC-MS peaks, the statistical analysis was conducted. For the isotope tracing experiment, the natural abundance isotope correction was performed using EL-Maven.