Agilent 1290 infinity system

Liquid chromatography (LC) analysis was carried out by ultra-performance liquid chromatography (UPLC) Agilent 1290 Infinity system coupled to an Agilent 6120 Quadrupole mass spectrometer equipped with an electrospray ionization source (positive ESI mode) (Agilent Technologies, Santa Clara, CA, USA). The chromatographic separation was performed on a reversed phase column Zorbax Eclipse + C18 Agilent (50 mm × 2.1 mm; 1.8 µm). The LC method and mass spectrometry analysis parameters were the same as described previously [14 (link)].

Anthocyanins and related compounds were extracted with a solvent mixture of methanol, acetic acid, and water (4:1:5, v:v:v), and the extracts were filtered through a ZORBAX Eclipse Plus C18 column (Agilent Technologies, Tokyo, Japan). The extract solution of each sample was then analyzed using HPLC with an Agilent 1290 infinity system (Agilent Technologies). A linear gradient of 10-60 % of solvent B (1.5 % H₃PO₄, 20 % acetic acid, and 25 % acetonitrile) in solvent A (1.5 % H₃PO₄) was run over a 9-min period. The anthocyanins and CADs were identified on the basis of absorption spectra obtained using a photodiode array detector, and pigment quantity was determined on the basis of absorption values at wavelengths of 340 nm (CADs) and 530 nm (anthocyanins).

Lipid profiling of CVF was conducted using Agilent 1290 Infinity system equipped with Agilent 6546 quadrupole-time-of-flight mass spectrometry from Agilent Technologies (Santa Clara, CA, USA). The ACQUITY UPLC BEH C18 Column (2.1 × 100 mm, 1.7 μm, Waters, Milford, MA, USA) assisted lipid separation under a binary gradient elution with a flow rate of 0.15 mL/min at 40 °C. Mobile phase A was acetonitrile/water (2:8, v/v) and mobile phase B was isopropanol/acetonitrile (6:4, v/v), both including 10 mM ammonium acetate and 0.1% acetic acid. The solvent gradient condition was programmed as follows: 0.0–3.0 min, 40% B; 3.0–10.0 min 85% B; 10.0–23.0 min 90% B; 23.0–28.0 min 100% B; 28.0–38.0 min 100% B; 38.0–40.0 min 40% B. The injection volume was 5 µL. Separated lipids were detected by electrospray ionization and Q-TOF detector with parameters as follows: sheath gas temperature 350 °C; sheath gas flow, 11 L/min; nebulizer, 17 psi, gas temperature 200 °C; dry gas flow, 8 L/min; capillary voltage, 3500 V; nozzle voltage, 1000 V; fragmentor, 175 V. After obtaining chromatograms using untargeted LC-MS analysis with scan mode, targeted MS/MS with the same parameters confirmed a spectral match with the fragment pattern in database.