HPLC columns with different length, particle size, and chemical properties of sorbent were tested and compared for the application in the development of the universal HPLC assay for determination of phenolic compounds, which could be present in some medicinal plants from the Lamiaceae family (Melissa officinalis, Rosmarinus officinalis, Salvia officinalis, Thymus serpyllum, and Origanum vulgare). More RP-18 columns have been chosen for the simultaneous separation of all phenolic compounds in the study. The basic chromatographic characteristics were evaluated and the HPLC method using one of the suitable columns was validated for all determined analytes. The recommended separation conditions were applied for phenolic compound monitoring in extracts of plant material. Yields of analytes present in all plant samples were evaluated.

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