Prostar 230 pump

The high-performance liquid chromatography system composed of the autosampler, Varian (Les Ulis, France) Prostar 230 pump, as well as the Varian Prostar 335 photodiode array detector was used to analyze and controlled with the Galaxie software (Varian v1.9.3.2). The separation was then carried out on the Purospher RP-18 column (250 × 4.0 mm internal diameter; 5 µm) (Merck Chemicals, Molsheim, France) at 40 °C. The mobile phase consisted of the mixture of methanol (solvent A) and HPLC grade water (solvent B), which were acidified with 0.05% formic acid. Then, the linear gradient, from 0 to 60 min was applied to this mobile phase variation, ranging from the 5:95 (v/v) to 100:0 (v/v) mixture of solvents A and B, respectively, using a flow rate of 1.30 mL/min. The injection volume was 3 µL, and the maximum back pressure was 110 bar. Detection was performed at 320 nm. The flavonoid compounds were identified by comparison with authentic standards (Sigma Aldrich).

The high-performance liquid chromatography system, consisting of an autosampler, a Varian (Les Ulis, France) Prostar 230 pump and a Varian Prostar 335 photodiode array detector, was used; this was controlled using Galaxie software (Varian v1.9.3.2). The separation was performed at 35 °C using a C18 core-shell column with iso-butyl side chains with TMS endcapping (Kinetex 5 µm XB-C18, 100 Å, LC Column 150 × 4.6 mm, core-shell silica, Phenomenex Le Pecq France). The mobile phase composed of a methanol (solvent A) and HPLC-grade water (solvent B) mixture, both being acidified with 0.05% formic acid. A linear gradient was applied for the mobile phase variation from a 5:95 (v/v) to a 100:0 (v/v) mixture of solvents A and B, respectively; the flow rate was 1.30 mL/min and the injection volume was 3 µL. The maximum back pressure was 110 bar. Detection was conducted using PDA, and the quantification was performed at 320 nm (the maximum absorbance wavelength of the studied flavonoids). Flavonoid compounds were identified by means of comparison with the authentic standards (Extrasynthese, Genay, France). The limits of detection (LOD) and limits of quantification (LOQ) were investigated based on the signal-to-noise ratios (S:N) of 3:1 and 10:1, respectively, following the previous study [20 (link)].

In this study, the HPLC system consisting of an autosampler, Varian (Les Ulis, France) Prostar 230 pump, as well as a Varian Prostar 335 photodiode array detector was used, and controlled by Galaxie software (Varian v1.9.3.2). The separation was performed at 35 °C using C18 core-shell column with iso-butyl side chains with TMS endcapping (Kinetex 5 µm XB-C18, 100 Å, LC Column 150 × 4.6 mm, core-shell silica, Phenomenex Le Pecq France). The mobile phase was composed of a methanol (solvent A) and HPLC grade water (solvent B) mixture, both being acidified with 0.05% formic acid. The linear gradient was applied for the mobile phase variation from a 5:95 (v/v) to a 100:0 (v/v) mixture of solvents A and B, respectively. Flow rate was 1.30 mL/min, and injection volume was 3 µL. Maximum back pressure was 110 bar. The detection was conducted using a PDA (photodiode array detector), and the quantification was performed at 320 nm (the maximum absorbance wavelength of the studied flavonoids). Flavonoid phytochemicals were then identified by means of comparison with the authentic standards (Extrasynthese, Genay, France), as described previously [36 (link)].