The chromatographic assay was performed in the Pharmacognosy Laboratory of the Biotechnology and Prototypes Unit of the Facultad de Estudios Superiores Iztacala, UNAM.
HPLC was used to characterize 30 μL of the propolis extract chemically. The extract was injected with a concentration of 3 mg/mL into a Hewlett-Packard HP model 1100 series (Hewlett-Packard, Wilmington DE, USA) HPLC equipped with a diode array detector (DAD) 1100 operated with ChemStation A0903 under the following parameters: isocratic separation using a mobile phase, methanol: acetronile: water (25 :25 :50) acidified with formic acid (1%) for 60 minutes; column, Discovery C-18 (250 x 4.6 mm), at 269 bar pressure and a temperature range of 22°C-23°C; flow rate, 1mL/min; detector array of diodes with detector setting at 260 nm; and full scanning of 200-400 nm. The constituents were identified based on a comparison of the retention time and UV spectrum with those of the standards.
HPLC was used to characterize 30 μL of the propolis extract chemically. The extract was injected with a concentration of 3 mg/mL into a Hewlett-Packard HP model 1100 series (Hewlett-Packard, Wilmington DE, USA) HPLC equipped with a diode array detector (DAD) 1100 operated with ChemStation A0903 under the following parameters: isocratic separation using a mobile phase, methanol: acetronile: water (25 :25 :50) acidified with formic acid (1%) for 60 minutes; column, Discovery C-18 (250 x 4.6 mm), at 269 bar pressure and a temperature range of 22°C-23°C; flow rate, 1mL/min; detector array of diodes with detector setting at 260 nm; and full scanning of 200-400 nm. The constituents were identified based on a comparison of the retention time and UV spectrum with those of the standards.