Kinetex biphenyl c18 100 rp column

The changes in major sinapates obtained from the pre-heat-treated and extracted canola meal samples were evaluated by High Performance Liquid Chromatography (HPLC) described by Nandasiri et al. (9 (link)). Phenolic compounds were analyzed by reversed-phase High Performance Liquid Chromatography-Diode Array Detection (HPLC-DAD) (Ultimate 3000, Dionex, Sunnyvale, Torrance, CA, United States). The separation was carried out on a Kinetex® Biphenyl C₁₈ 100 Å RP column (2.6 mm, 150 × 4.6 mm, Phenomenex, Torrance, CA, United States), with flow rate of 0.4 mL/min and 10 μL injection volume. The column oven was maintained at 30°C. Extract of 70% aqueous methanol was used in the analysis. Using the authentic standards (sinapine, sinapic acid, and canolol) phenolic compounds were identified. The separation was conducted using gradient elution with water [0.1% (v/v) formic acid] as solvent A, and methanol [0.1% (v/v) formic acid] as solvent B. The chromatograms were acquired at 270 nm (canolol) and 330 nm (sinapine and sinapic acid) using Chromeleon software Version 7.2 SR4 (Dionex Canada Ltd., Oakville, ON, Canada).

The changes in the phenolic composition of the extracts obtained with MAE were analyzed using high-performance liquid chromatography with diode array detection (HPLC-DAD) (Ultimate 3000; Dionex, Sunnyvale, Torrance, CA, USA) according to the method described by Nandasiri et al. [5 (link)]. The HPLC analysis was done using a reversed phased Kinetex Biphenyl C₁₈ 100 Å RP column (2.6 µm, 150 × 4.6 mm, Phenomenex, Torrance, CA, USA) with a flow rate of 0.4 mL/min and 10 μL injection volume. The column was maintained at 30 °C for improved separation. The gradient consisted of formic acid (0.1%, v/v) in water as solvent A, and formic acid (0.1%, v/v) in methanol as solvent B. The gradient system was operated as follows: 25% B (0–3 min), 25–40% B (3–8 min), 40% B (8–13 min), 40–60% B (13–25 min), 60–70% B (25–38 min), 70–100% B (38–41 min), 100% B (41–44 min), 100–25% B (44–47 min), and 25% B (47–57 min). The chromatograms were acquired at both 320 nm (sinapine and sinapic acid) and 270 nm (canolol) using Chromeleon software Version 7.2 SR4 (Dionex Canada Ltd., Oakville, ON, Canada). Major sinapates were identified using the authentic standards with a detection limit of 0.001 mg/mL. Calibration curves for each standard were obtained from 1.0 to 100 μg/mL (n = 11) concentration range with R² = 0.999 for sinapine, sinapic acid, and canolol.