Safety assessment of citrus and olive by-products using a sustainable methodology based on natural deep eutectic...

In this work, the application of betaine-based hydrophilic natural deep eutectic solvents (NADESs) as green extraction solvents was proposed for the first time for the evaluation of twelve pesticides in citrus and olive by-products intended to be applied as potential sources of compounds with neuroprotective activity against Alzheimer Disease. Ultrasound-assisted extraction of selected pesticides was followed by separation and determination using gas chromatography coupled to single quadrupole mass spectrometry. Eight NADESs were tested using different hydrogen bond donors (i.e. citric and lactic acid, fructose, glucose, glycerol, propylene glycol, propionic and butanoic acid). Other factors affecting extraction efficiency were also evaluated using a step-by-step approach. Eight mL of a mixture composed of 60% betaine:propylene glycol NADES at a molar ratio 1:4 and 40% of water, as well as 30 min of ultrasound-assisted extraction were selected as the most adequate conditions. The methodology was validated prior to its application in citrus and olive by-products. Recovery values were between 73 and 115% (RSD% < 20%), while limits of quantification of the method were in the range 8.5-128.8 µg/kg, which demonstrates the suitability of the procedure to determine the selected group of pesticides, usually applied in citrus and olive crops, at the legislated levels. The greenness of the procedure was also evaluated using AGREE calculator. Finally, the whole method was applied for the safety assessment of seven olive leaf samples and seven citrus by-products produced in Spain, finding the presence of several of the evaluated compounds at concentrations higher than the established limits for similar products.