Determination of coumarins from pterocaulon balansae by an ultra-fast liquid chromatography method in topical applications

Abstract

Coumarins from Pterocaulon balansae Chodat, Asteraceae, have received increasing attention due to their biological activities, especially for the treatment of skin disorders. This study aimed to develop and validate high-performance and ultra-fast liquid chromatography methods for the analytical determination of coumarins in extracts, in topical formulations, as well as in porcine skin after in vitro permeation/retention studies. The chromatographic conditions consisted of a mobile phase with 0.1% formic acid (A) and acetonitrile (B) using a gradient elution; the flow rate and a column oven temperature: HPLC-UV, 1 ml min−1; 30 °C and UFLC-DAD, 0.55 ml min−1; 55 °C for 45 min and 8 min, respectively. The chemical marker 5-methoxy-6,7- methylenedioxycoumarin was used for the construction of the linearity curve in a concentration range from 0.1 to 7.5 μg ml−1. A low matrix effect was observed. The method robustness was confirmed using the Plackett-Burman experimental design. These methodologies proved to be linear, precise, and accurate for the determination of seven coumarins in analytical and bioanalytical samples in both approaches. However, UFLC analysis time was shortened to about one-fourth the time and consumes significantly less eluent, being more eco-friendly in comparison with HPLC.