Squalene concentration from physically refined olive oil deodorizer distillates

Abstract: The health benefits of olive oil are assigned to the unsaponifiable fraction of the oil, rich in squalene. The unsaponifiable matter is partially removed during the conventional refining process and concentrated in the deodorizer distillate fraction (DOD), obtained in the deodorization stage as a by-product. These distillates of physically refined olive oil (PH-O) were used as a feedstock to obtain squalene concentrates using supercritical carbon dioxide (Sc-CO₂) counter current fractionation technology. Several routs of producing concentrated squalene fraction were investigated: 1) direct concentration from PH- O DOD - squalene was concentrated approximately 3 fold (to 22.7%); 2) after saponification of PH-O DOD –squalene was concentrated 11.5 fold (to 95.2%); 3) after enzymatic esterification – squalene was concentrated approximately 8 fold (to 49%); and 4) after high temperature chemical esterification – squalene was concentrated approximately 8 fold as well (to 44.4%) in a single Sc-CO₂ counter current fractionation step. Experimental results showed that producing high concentrated squalene fraction is possible and looks promising if the DOD is properly pre-treated. The most important factor to conduct successful squalene purification using Sc-CO₂ counter current fractionation is a pre-treatment step that influences matrix composition of the input material.