HPLC Analysis of Propolis Extracts

The three extracts obtained, PPF, PSDE, and MPE, were submitted to high-performance liquid chromatography (HPLC) using Shimadzu equipment with a CBM-20A controller, a LC-20AT quaternary pump, an SPD-M 20A diode matrix detector, and Shimadzu LC software, version 1.21 SP1. The mobile phase consisted of methanol and aqueous formic acid solution (0.1% v/v), pH 2.7. The method consisted of a 20–95% gradient for 77 min at a flow rate of 0.8 mL/min in a CLC-ODS column (4.6 mm × 250 mm, particle diameter 5 µm, pore diameter 100 A). Detection was set at 275 nm. The chemical markers: caffeic acid, p-Coumaric acid, 3,5-dicapheoylquinic acid, 4,5-dicapheoylquinic acid, aromadendrin-4′-O-methyl-ether, drupanin, chrysin, galangin, artepillin C, and baccharin were identified and quantified, according to Berretta et al. [6 (link)].