Abstract
The predominant majority of bioactive compounds in natural products are polyphenols. Reverse Phase High Performance Liquid Chromatography (HPLC) is the most employed analytical method for determining the polyphenol profiles in natural products. Analyses are conducted based on methods validated according to the number and type of phenolic standards used. In this study, validation experiments were performed using HPLC- photodiode array detector for 26 frequently encountered phenolic compounds in bee products and natural substances. The analysis was carried out utilizing a C18 column (250 mm x 4.6 mm, 5 μm; GL Sciences) with a gradient program. The HPLC method was developed, determining the limit of detection within the range of 0.019-0.072 μg/mL, and the limit of quantification within the range of 0.063-0.239 μg/mL. All calibration curves exhibited linear corelations with R² values exceeding 0.994 across the specified range. The developed method has been optimized and validated by assessing detection and quantification limits, accuracy, repeatability, and recovery data suitable for phenolic analysis. It has been concluded that the optimized method allows for the rapid and reliable evaluation of the phenolic content of natural products and their quantitative determination.
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