An improved RP-HPLC method for simultaneous analyses of squalene and cholesterol especially in aquatic foods.

A simple, rapid and simultaneous method for the quantitative evaluation of squalene and cholesterol in oil recovered from different body parts of fresh water and marine fish species was developed using reverse phase High performance liquid chromatography (RP-HPLC). A modified fractional crystallization method was developed using absolute ethanol as a solvent to obtain a fraction of squalene and cholesterol in the oil extracted from different body parts of fish samples. Elution of squalene and cholesterol was carried out isocratically with 100 % acetonitrile at 195 nm by diode array detector. Excellent linearity of the calibration curve was observed. The limit of detection for squalene and cholesterol were found to be 5.0 ng ml(-1) and 1.0 μg ml(-1) respectively. A possible correlation was established by determining the ratio between cholesterol and squalene which was found to be ranging from 2 to 10. This method was successfully employed for the determination of squalene and cholesterol in the above mentioned samples.