Abstract
Acacetin (1), 4′,7-dimethyl naringenin (2), and 4′,7-dimethyl apigenin (3)—three of the main markers active components of propolis—were chosen for the development and validation of a suitable HPLC method for quality control of the crude drug. A 23 factorial design was employed to study the main interactions of independent variables, and the efficient separation was achieved using an XBridge C18 column with an isocratic binary phase consisting of deionized water (TFA; 0.1 % v/v) (eluent A) in methanol (eluent B) (45:65, v/v) at a flow rate of 1.0 mL min−1. The method allows the quantification of 1–3; for each compound, a linear response was evaluated within the range of 100–1,000 μg mL−1 for 1, 25–200 μg mL−1 for 2, and 8–60 μg mL−1 for 3. All the calibration curves showed good linearity within the test ranges (r2 ≥ 0.999). Percentage recoveries of the selected flavonoids 1–3 ranged between 98.3 and 101.2 % (RSD ≤2.0 %). The intra- and interday precision RSDs were no more than 2.0 %, while the repeatability variation was no more than 2.0 %. Finally, the method was applied to establish some seasonal and geographical variations of the markers in propolis obtained from several regions of Mexico. Thus, 4′,7-dimethyl apigenin (3) was selected as a marker compound from Mexican propolis, and, along with 2, could be useful for quality control procedures focused in to establish some geographical variation of Mexican propolis.
Published Version
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