The extracts of the chilto samples (peel, pulp, and seeds) were analyzed by HPLC-DAD to obtain a phenolic fingerprint. The HPLC system used was a Shimadzu equipment (Shimadzu Corporation, Kyoto, Japan) consisting of an LC-20AT pump, an SPD-M20A diode array detector, CTO-20AC column oven and a LabSolution software. An Inertsil ® ODS-3 RP-C18 (5μ, 250 × 4.6 mm) column (GL-Sciences Inc, Tokio, Japan) maintained at 25 °C. The analyses were performed using a linear gradient solvent system consisting of 1% formic acid in water (A) and acetonitrile (B) as follow: 90%–85% A over 15 min, maintained to 20 min, then 85% A for 20–25 min, 85% to 70% A for 25–50 min, maintained to 70% A for 50–80 min, and returning to 90% A for 80.01–90 min. The flow rate was 0.5 mL/min and the volume injected was 20 μL. The compounds were monitored at 254, 280, 320 and 350 nm and spectra from 200 to 600 nm were recorded.
The main phenolics were quantified. The reference compounds rosmarinic acid (98%, Sigma Aldrich, St. Louis, MO, USA) and 3-caffeoylquinic acid (98%, Phytolab GmbH, Verstenbergsgreuth, Germany) were used in concentrations ranging from 5-500 mg/L. Results were expressed as mg of the corresponding compound/100g extract.
The main phenolics were quantified. The reference compounds rosmarinic acid (98%, Sigma Aldrich, St. Louis, MO, USA) and 3-caffeoylquinic acid (98%, Phytolab GmbH, Verstenbergsgreuth, Germany) were used in concentrations ranging from 5-500 mg/L. Results were expressed as mg of the corresponding compound/100g extract.