A simple micellar liquid chromatography (MLC) method was developed and validated according to ICH Guidelines for the determination of sesquiterpenic acids (valerenic, hydroxyvalerenic, and acetoxyvalerenic acids) in root and rhizome extract from Valeriana officinalis L. and valerian dry hydroalcoholic extract. Samples were analyzed on Nucleosil C18 column (, 5 μm) using an isocratic mobile phase which consisted of Brij 35 (5% (w/v) aqueous solution; pH by phosphoric acid) and 1-butanol (6%

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... ); UV detection was at 220 nm. Micellar mobile phase using allows to fully separate valerenic acids within 25 minutes. Linearity for hydroxyvalerenic, acetoxyvalerenic, and valerenic acids was 1.9–27.9, 4.2–63.0, and 6.1–91·3 μg.mL−1, and limit of detection was 0.14, 0.037, and 0.09 μg·mL−1, respectively. Intraday and interday precisions were not less than 2% for all investigated compounds. The proposed method was found to be reproducible and convenient for quantitative analysis of sesquiterpenic acids in valerian root and related preparations.