Quantitative Determination of Usnic Acid in Usnea lichen and Its Products by Reversed-Phase Liquid Chromatography with Photodiode Array Detector

Abstract Usnic acid, a lichen substance, has a wide range of pharmaceutical applications, including antibiotic, antimycotic, antifeedant, antitubercular, antitumor, and analgesic activities. Some products containing usnic acid are marketed as weight control supplements; however, hepatotoxicity and acute liver failures were reported as severe side effects. The usnic acid content present in the plant materials and market products was analyzed by reversed-phase high-pressure liquid chromatography with a photodiode array detector at 233 nm. A Waters XTerra RP18 (150 × 4.6 mm; 5 μm particle size) column was the stationary phase; mobile phase was aqueous 0.1% acetic acid and acetonitrile gradient at flow rate of 1.0 mL/min. The temperature was held constant at 30°C. The retention time of usnic acid was approximately 13.3 min. Acetone extraction of the samples took place with sonication. The precision of the method was confirmed by a standard deviation below 3.0% (n = 3) and usnic acid recovery was 99.0%. Limit of detection was 0.4 μg/mL and the response was linear from 1.4 to 570.0 μg/mL with a correlation coefficient (R2) of 0.9991. The content of usnic acid in 4 raw materials and 22 finished products was analyzed.