Ethanolic Extraction of Propolis and Bee Pollen

The procedure described by Dziedzic et al. (2013) [20 (link)] with modifications was used to prepare an ethanolic extract of propolis and ethanolic extract bee pollen. Crushed propolis (40g) or bee pollen (40 g) (apiary, Poland) was mixed with 96.0% ethanol (100ml). The suspensions were stored in the darkness at room temperature (25°C) with shaking (200 rpm, 6 hours per day) for 4 days. Then the infusion was placed at -20°C for precipitating alcohol insoluble compounds of propolis. After 24 hours, the resulting infusion was filtrated through sterile filters (0.45 μm, Millipore). The 40.0% EEP and 40.0% EEBP were stored at room temperature (25°C).