Determination of squalene in olive oil using fractional crystallization for sample preparation

A simple, rapid method for the determination of squalene in virgin olive oil was developed using RP‐HPLC with detection at 208 nm. Fractional crystallization from methanol/acetone (7∶3, vol/vol) was applied to obtain squalene in the liquid fraction of the oil prior to HPLC. Elution of squalene was then carried out isocratically with acetone/acetonitrile (40∶60 vol/vol) within 11 min. The detection limit was 23 mg/kg, and the limit of quantification 79 mg/kg. The precision of the crystallization procedure (CV%=3.76, n=7) and the mean recovery (92.5 and 81.5% for the 7,000 and 700 mg/kg levels of addition, respectively) were satisfactory. The method is easily applicable to fulfill future needs for nutrition labeling.