Cto 10acvp column oven

A LC-VP series (Shimadzu, Kyoto, Japan) consisting of an SIL-HTc autosampler, LC-10ADvp Pump, CTO-10ACvp column oven, and DGU-14AM degasser was used to set the reverse-phase liquid chromatographic conditions. The column was a Cadenza CD-C18 column (100 × 2 mm i.d., 3 μm, Imtakt Corp., Kyoto, Japan) used at 45°C. The mobile phase included water/acetonitrile/formic acid 95/5/0.05 (v/v/v, Solvent A) and water/acetonitrile/formic acid 35/65/0.05 (v/v/v, Solvent B). The gradient elution programs were 0% B (0-1 min with an isocratic gradient), 0–40% B (1-2 min with a linear gradient), 40% B (2–7 min with an isocratic gradient), 40–100% B (7–12 min with a linear gradient), 100% B (12–14 min with an isocratic gradient), 100–0% B (14-15 min with a linear gradient), and 0% B (15-16 min with an isocratic gradient) at a flow rate of 0.3 mL/min. The autosampler tray was cooled to 45°C and the injection volume was 5 μL. HPLC grade acetonitrile and formic acid were purchased from WAKO.