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Optimization and characterization of olive leaf (olea europaea) obtained ultrasonic stimulations and vacuum by response surface methodology
* 1, 2 , 1, 2 , 1, 2 , 1 , 1 , 1 , 2 , 1
1  Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche (IDiBE), Universidad Miguel Hernández de Elche, 03202 Elche, Spain
2  Instituto Universitario de Ingeniería de Alimentos-FoodUPV, Universitat Politècnica de València, 46022 Valencia, Spain
Academic Editor: Antonios Koutelidakis

Abstract:

INTRODUCTION
Olive leaves, a byproduct of olive tree pruning, are considered an industrial residue with significant potential due to their content of bioactive compounds, such as oleuropein and hydroxytyrosol, which exhibit antioxidant, anti-inflammatory, and antimicrobial properties. Traditional extraction methods for these compounds typically require large volumes of solvents, prompting the need for innovative technologies that minimize solvent use and reduce processing times. Conventional methods often involve the use of heat and agitation to enhance the transfer of compounds from olive leaves into the solvent. A new extraction technique has been developed, using high temperature, ultrasound, and vacuum to improve the efficiency of compound extraction.

METHODS
To optimize the process, response surface methodology was employed, evaluating the influence of key variables (% EtOH, temperature, and number of cycles) at three levels (low, medium, high). A total of 27 assays were conducted. The samples were analyzed by HPLC-DAD and Folin-Cocolteau method for the assay of total phenols; DPPH and FRAP methods for antioxidant activity methods.

RESULTS
The highest yield of the extrations of oleuropein and hydroxytyrosol was observed at medium level of temperature, high level of hydroalcoholic solution, and medium level of number of cycles obtaining 43.53 mg/g and 2.73 mg/g for oleuropein and hydroxytyrosol, respectively. In contrast, samples processed without the new technology, yielded 35.08 mg/g and 2.70 mg/g for oleuropein and hydroxytyrosol, respectively. The antioxidant activity, assessed by the FRAP and DPPH methods, showed that untreated samples yielded 1.5 mg TE/g, which increased to 3.31 mg TE/g under conditions of high % EtOH, low temperature, and high flash cycles.

CONCLUSIONS
The optimal combination of variables enhanced the extraction yield by up to 24% for bioactive compounds like oleuropein, compared to unprocessed samples.

Keywords: Optimization; olive leaf; ultrasonic; vacuum
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