The content of trans resveratrol and quercetin was determined in Fr 2 SySFV, the fraction with higher phenolic content and antioxidant activity. A reversed-phase chromatographic method to determine trans-resveratrol and quercetin has been previously described and validated [4 (link)]. The HPLC analyses were conducted on a Shimadzu liquid chromatograph system (Shimadzu Corp, Kyoto, Japan) equipped with a LC-10 ATvp pump, variable wavelength detector SPD 10AVvp, controller module SCL 10A vp, a LC-10AD vp pump, a vacuum degasser DGU-14A, and an autosampler. The analytes were separated on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm), using a gradient system of two eluents: acetonitrile and water containing 0.1% formic acid (35:65) at a flow rate of 1 mL/min. The detection wavelength was 307 nm for trans-resveratrol and 370 for quercetin. The injection volume was 20 μL. The concentration of each component of interest was calculated based on a calibration curve created from solutions of the trans-resveratrol standard at concentrations of 0.1, 0.3, 0.5, 1.0, 1.5 µg/mL and quercetin at concentrations of 0.5, 0.7, 1.0, 1.5, 2.0 µg/mL.
Liquid chromatograph system
Manufactured by Shimadzu
Sourced in Japan
The Liquid chromatograph system from Shimadzu is a device used for separating and analyzing complex mixtures. It employs liquid solvents to carry sample compounds through a stationary phase packed into a column, allowing different compounds to elute at different times.
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2 protocols using liquid chromatograph system
1
HPLC Analysis of trans-Resveratrol and Quercetin
2
HPLC Analysis of DNP and LRD
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HPLC analysis were performed on Shimadzu liquid chromatograph system (Kyoto, Japan) equipped with a binary pump (LC-10AT), Photo Diode Array (PDA) detector (SPD-M20A), column oven (CTO-10AS) and auto sampler (SIL-HT, Shimadzu, Japan) was used to inject the samples. The HPLC system was equilibrated for approximately 30 min before starting of analysis and chromatography was carried out at 40 ± 0.5 °C. Eluents were monitored at a wavelength of 268 nm using a PDA detector. DNP and LRD were separated on Waters Nova-Pak C18 column (3.9 × 150 mm, 4 μm) with a mobile phase consisting of acetonitrile and ammonium formate (pH 6.4; 0.01 M) (62 : 38% v/v) in an isocratic mode at a flow rate of 1 mL min−1. The injection volume was 50 μL and retention time for DNP and LRD were 3.31 and 3.99 min respectively. Control of hardware, data acquisition and elaboration were performed using LC solution software version 1.24 SP1.
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