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Abstract
Propolis obtained from honeybee hives has been widely used in medicine, cosmetics, and industry due to its versatile biological activities (antioxidant, antimicrobial, fungicidal, antiviral, antiulcer, immunostimulating, and cytostatic). These activities are mainly attributed to the presence of flavonoids in propolis, which points out the interest in quantifying these constituents in propolis preparations, as well as validation of analytical methodologies. High‐performance liquid chromatography (HPLC) methods have been reported to quantify isolated flavonoids or these compounds in complex biological matrices, such as herbal raw materials and extractive preparations. An efficient, precise, and reliable method was developed for quantification of propolis extractive solution using HPLC with UV detection. The chromatograms were obtained from various gradient elution systems (GES) tested in order to establish the ideal conditions for the analysis of propolis extractive solution, using methanol and water:acetonitrile (97.5:2.5, v/v) as mobile phase. Gradient reversed phase chromatography was performed using a stainless steel column (250 × 4.6 mm i.d., 5 µm) filled with Chromsep RP 18 (Varian), column temperature at 30.0 ± 0.1°C and detection at 310 nm. The main validation parameters of the method were also determined. The method showed linearity for chrysin in the range 0.24–2.4 µg mL−1 with good correlation coefficients (0.9975). Precision and accuracy were determined. The obtained results demonstrate the efficiency of the proposed method. The analytical procedure is reliable and offers advantages in terms of speed and cost of reagents.
Acknowledgments
The authors thank the CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) for their financial support.