A Box-Behnken Design for Optimal Green Extraction of Compounds from Olive Leaves That Potentially Activate the AMPK Pathway

Olive leaves contain bioactive compounds that have been shown to activate AMP-activated protein kinase (AMPK), which decreases intracellular lipid accumulation. Microwave-assisted extraction (MAE) is a green extraction technique that is frequently used in the recovery of phenolic compounds from plants. Thus, in this study, a Box-Behnken design was used to optimize MAE conditions such as temperature, percentage of ethanol and extraction time to obtain the maximum content of total compounds and compounds that activate AMPK. To this end, all extracts were characterized by High-Performance Liquid Chromatography Coupled to Electrospray Ionization Time-of-Flight Mass Spectrometry (HPLC-ESI-TOF-MS). The optimum conditions to obtain the highest content of total compounds were 123 °C, 100% of ethanol/water (v/v) and 23 min, whereas the optimum conditions for the highest amount of compounds that activate AMPK were 111 °C, 42% of ethanol/water (v/v) and 23 min. Thus, a multi-analysis by desirability was carried out to establish MAE optimal conditions for both responses. The optimum conditions were 111 °C, 100% EtOH and 23 min with a desirability of 0.97, which means that the responses are close to their individual optimal values. As a result, the olive leaf extract obtained at these optimal MAE conditions has great potential to be effective in the treatment of obesity.