Quantification of EA and GA was conducted on a HPLC (Ultimate 3000, Thermo Fisher Scientific®), coupled to a diode array detector (DAD; Thermo Fisher Scientific®) and equipped with a binary pump (HPG-3x00RS, Thermo Fisher Scientific®), a degasser, and an autosampler equipped with a loop of 20 µL (ACC-3000, Thermo Fisher Scientific®). For data analysis and processing, the software Chromeleon 6.8 (Dionex, Thermo Fisher Scientific®, USA) was used. The wavelength was set at 254 nm for detection of the EA and 270 nm for GA, in accordance with the maximum absorption measured by DAD. The chromatographic separations were achieved with a C18 column (250 mm × 4.6 mm i.d., particle size 5 µm) from Dionex® equipped with a guard column (C18, 4 mm × 3.9 µm, Phenomenex®). Separations were carried out at a column oven temperature of 24°C. The mobile phase consisted of purified water (A) and methanol (B), both acidified with 0.05% trifluoracetic acid (TFA), at a flow rate adjusted to 0.8 mL/min. A gradient program was applied as follows: 0–10 min, 12.5–25% B; 10–15 min, 25–40% B; 15–25 min, 40–75% B; 25–30 min, 75–75% B; 30–33 min, 75–12.5% B.
Hpg 3x00rs
Manufactured by Thermo Fisher Scientific
The HPG-3x00RS is a high-performance gradient pump designed for liquid chromatography applications. It features precision flow control, accurate gradient composition, and reliable performance for a wide range of analytical and preparative separations.
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2 protocols using hpg 3x00rs
1
Quantification of Ellagic and Gallic Acids
2
HPLC Analysis of Phenolic Compounds
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A Thermo Scientific system (Ultimate 3000 Thermo Fisher Scientific®) equipped with a DAD (Thermo Fisher Scientific®), binary pump (HPG-3x00RS, Thermo Fisher Scientific®), degasser and an autosampler equipped with a 20 μL loop (ACC-3000, Thermo Fisher Scientific®) was used to perform the HPLC analyses. Chromeleon 6.8 software (Dionex®) was used for data acquisition and data processing.
Chromatographic separation was performed with a C18 column (250 mm × 4.6 mm inner diameter, 5 μm; Dionex®) that was protected by a guard column made of the same material (Phenomenex®). A gradient elution was achieved by varying the proportion of solvent B (methanol with 0.05%, v/v, trifluoracetic acid) to solvent A (water with 0.05%, v/v, trifluoracetic acid) at a flow rate of 0.8 mL/min, according to the following gradient program: 10–25% B (10 min), 25–40% B (5 min), 40–70% B (10 min), 75% B (5 min), and 75–10% B (1 min). Separations were carried out in a column oven at a temperature of 23 ± 2 °C. Wavelengths of 254 nm, 270 nm, and 350 nm were used for detecting ellagic acid, gallic acid, and myricitrin, respectively, according to the maximum absorption measured by DAD.
Chromatographic separation was performed with a C18 column (250 mm × 4.6 mm inner diameter, 5 μm; Dionex®) that was protected by a guard column made of the same material (Phenomenex®). A gradient elution was achieved by varying the proportion of solvent B (methanol with 0.05%, v/v, trifluoracetic acid) to solvent A (water with 0.05%, v/v, trifluoracetic acid) at a flow rate of 0.8 mL/min, according to the following gradient program: 10–25% B (10 min), 25–40% B (5 min), 40–70% B (10 min), 75% B (5 min), and 75–10% B (1 min). Separations were carried out in a column oven at a temperature of 23 ± 2 °C. Wavelengths of 254 nm, 270 nm, and 350 nm were used for detecting ellagic acid, gallic acid, and myricitrin, respectively, according to the maximum absorption measured by DAD.
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