Development and Validation of a Simple Reversed-Phase HPLC-UV Method for Determination of Malondialdehyde in Olive Oil

A simple, precise, accurate and selective method was developed and validated for determination of malondialdehyde (MDA) in olive oil. Separation was achieved on a reversed-phase C-8 column using a mobile phase consisting of methanol/0.8 % phosphoric acid (10:90, v/v), at a flow rate of 1.0 ml/min and UV detection at 220 nm. This method was validated according to the requirements for new methods, which include accuracy, precision, selectivity, robustness, a limit of detection, limit of quantitation (LOQ), linearity and range. The current method demonstrated good linearity over the range of 0.5-1000 ppm of MDA with r (2) greater than 0.999. The recovery of MDA in olive oil ranged from 97.1 to 99.1 %. The method was selective where MDA was distinctly separated from other compounds of the oil with good resolution. The method was also precise where the RSD of the peak areas of replicate injections of MDA standard solution were less than 1 %. The degree of reproducibility of the results obtained as a result of small deliberate variations in the method parameters and by changing the analytical operators proved that the method is robust and rugged. The low LOQ of MDA (0.5 ppm) using this method enables quantitation of MDA at low concentration.