Development and Validation of an UPLC Method for Quality Control of Plantago major

Plantago major L. and Plantago australis Lam., Plantaginaceae, have been used in folk medicine mainly as anti-inflammatory and healing agents. The two species, known by the same popular names, are morphologically similar and frequently P. australis appears as adulterant or substitute of P. major. In this study, an efficient and reproducible extractive method was developed for quality control of P. major samples, capable of identifying differences in the chromatographic profile of both plants. A reflux extraction has been proposed and optimized by fractional factorial design and subsequent Box-Behnken design. The following conditions were defined to evaluate the verbascoside content: particle size 180 µm, without solvent renewal, mass solvent ratio in 1:60, percentage of solvent in 89