Ultrasound-assisted extraction process of phenolic antioxidants from Olive leaves: a nutraceutical study using RSM and LC-ESI-DAD-MS.

In recent years, there has been an ever growing interest in finding new natural sources of food antioxidants. As a main fruit crop, olive is also valued due to its phenolic-containing leaves. Mathematically based optimization methods are used as powerful tools to extract different antioxidant compounds. The present study is aimed to provide an efficient extraction method for total phenol content (TPC), total flavonoid content (TFC) and antioxidant ability (DPPH scavenging assay and FRAP). The effects of ultrasonic temperature (35-65 °C), ultrasonic time (5-15 min), and ethanol to water ratio (Et: W) (25-75%) were evaluated. Second-order polynomial models were used through a rotatable Box-Behnken design (BBD) consisting of 15 experimental runs with three replicates at the center point. Interactional effects of the studied factors were significant in most cases for all responses. The highest extraction efficiency was found to be fifty-one percent of ethanol (65 °C, 15 min) to water ratio. Under optimal conditions, values for TPC, TFC, DPPHsc and FRAP assay were 183.4 (mg GAE. g-1 DW), 696.77 (mg Quercetin. g-1 DW), 78.98 (DPPHsc %) and 1942 µmol Fe+2/g DW, respectively. R2 values (R2 > 0.92) showed that RSM models could efficiently predict the yield of all responses. In the LC-ESI-MS-DAD profiling of the optimized extract, 27 compounds were identified with oleuropein as the main compound. In the present study, olive leaf is introduced as a promising source of natural antioxidant and can be used in food industries following further studies.