Methanol solvent a

The phenolic profiles of OA extract consisting of cyanidin-3-glucoside (Sigma-Aldrich, USA), gallic acid (Sigma-Aldrich, USA), and quercetin-3-O-rutinoside (Sigma-Aldrich, USA) were determined by high-performance liquid chromatography (HPLC). Chromatography was performed by using a Waters® system equipped with a Waters® 2998 photodiode array detector. Chromatographic separation was performed using Purospher® STAR, C-18 encapped (5 μm), LiChroCART® 250-4.6, and HPLC-Cartridge, Sorbet Lot no. HX255346 (Merck, Germany). The mobile phase (HPLC grade) consisted of 100% methanol (solvent A) (Fisher Scientific, USA) and 2.5% acetic acid (solvent B) (Fisher Scientific, USA) in deionized (DI) water was used to induce gradient elution. The gradient elution was carried out at a flow rate of 1.0 ml/min with the following gradient: 0-17 min, 70% A; 18-20 min, 100% A; and 20.5-25 min, 10% A. The sample was filtered (0.45 μm, Millipore), and a direct injection of the tested sample at the volume of 20 μl on the column was performed. The chromatograms were recorded at 280 nm using the UV detector, and data analysis was performed using Empower™ 3.

High-performance liquid chromatography (HPLC) analysis was used to determine the fingerprint chromatogram. Chromatography was performed by using a Waters® system equipped with a Waters® 2998 photodiode array detector. Chromatographic separation was performed using a Purospher® STAR, C-18 encapped column (5 μm) and LiChroCART® 250-4.6 HPLC cartridge, Sorbet Lot No. HX255346 (Merck, Germany). The mobile phase (HPLC-grade) consisted of 100% methanol (solvent A) (Fisher Scientific, USA), and 2.5% acetic acid (solvent B) (Fisher Scientific, USA) in deionized (DI) water was used to induce gradient elution. The gradient elution was carried out at a flow rate of 1.0 ml/min with the following gradient: 0-17 min, 70% A, 18-20 min, 100% A; 20.5-25 min, 10% A. The sample was filtered (0.45 μm, Millipore), and a direct injection of the tested sample at the volume of 20 μl on the column was performed. Chromatogram detection was performed at 280 nm using a UV detector, and data analysis was performed using EmpowerTM3.

The phenolic profiles of mulberry extract and encapsulated mulberry extract consisting of cyanidin-3-glucoside (Sigma-Aldrich, USA), gallic acid (Sigma-Aldrich, USA), and quercetin-3-O-rutinoside (Sigma-Aldrich, USA) were determined by high-performance liquid chromatography (HPLC). Chromatography was performed by using a Waters® system equipped with a Waters 2998 photodiode array detector. Chromatographic separation was performed using Purospher® STAR, C-18 encapped (5 μm), LiChroCART® 250-4.6, and HPLC-Cartridge, Sorbet Lot number HX255346 (Merk, Germany). The mobile phase (HPLC grade) consisted of 100% methanol (solvent A) (Fisher Scientific, USA) and 2.5% acetic acid (solvent B) (Fisher Scientific, USA) in deionized (DI) water was used to induce gradient elution. The gradient elution was carried out at a flow rate of 1.0 ml/min with the following gradient: 0–17 min, 70% A; 18–20 min, 100% A; and 20.5–25 min, 10% A. The sample was filtered (0.45 μm, Millipore), and a direct injection of tested sample at the volume of 20 μl on the column was performed. The chromatograms were recorded at 280 nm using UV detector, and data analysis was performed using EmpowerTM3.