Development and Validation of a Reversed-Phase HPLC Method for Determination of Elaidic Acid in Oils and Fats

Abstract

A simple, precise, accurate, and selective method with low limit of quantitation (LOQ) was developed and validated for analysis of elaidic acid which is the predominant trans fatty acids in partially hydrogenated vegetable oils. Separation was achieved on a reversed-phase C18 column, using mobile phase consisting of acetonitrile/water (80:20, v/v) containing 0.1% acetic acid, and using UV detection at 205 nm. This method was validated according to the requirements for new methods, which include accuracy, precision, selectivity, robustness, limit of detection (LOD), LOQ, linearity and range. The current method demonstrates good linearity over the range of 3-1000 mg L-1 of elaidic acid with r2 greater than 0.999. The recovery of elaidic acid in oils and fats ranges from 94.5 to 98.7%. The method is selective where elaidic acid is good separated from oleic acid and other components of fats and oils with good resolution. The method is also precise where the RSD of the peak areas of replicate injections of elaidic acid solution is 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 analytical operators has proven that the method is robust and rugged.