Response surface optimised extraction and chromatographic purification of rosmarinic acid from Melissa officinalis leaves

The extraction of lemon balm (Melissa officinalis) leaves with aqueous methanol was optimised using response surface methodology. Fifteen runs were conducted following a Box-Behnken design (BBD) followed by ridge analysis using the concentration of methanol, the extraction temperature and time as the independent variables and taking the extraction yield of RA from lemon balm as the response variable. The optimal extraction conditions were a methanol concentration of 59.0% (v/v), a temperature of 54.8 °C and a time of 64.8 min, which gave a maximal RA yield of 46.1 mg RA/g dry materials. The RA extract was loaded onto a column packed with Sephadex LH-20 and then was eluted with 100% methanol, which resulted in RA with a purity of 38.8% and a yield of 43.8%. The purity of RA increased by 3.1-fold when compared to its initial purity in the extract obtained from extraction.