Development of a Gas Chromatography-Flame Ionization Method for the Detection and Quantification of 12 Flavoring Agents in Supplementary Feed

A flavoring agent is a compound that serves to add flavor with a pleasant scent and is used as a feed additive. Current flavor analysis methods include reflux pretreatment, titration, neutralization titration, and inversion; these are all analytical methods in which deviations and errors between experiments are generated. Titration methods are characterized by difficult selectivity analysis both for mixtures containing two or more types of flavoring agents and also for very low content samples. Current analysis methods are therefore particularly unsuitable for these sample types. Thus, more precise and accurate analysis of flavor agents is needed. This study intends to develop and verify a multi-component simultaneous analysis method that can accurately confirm the guaranteed content of 12 flavor agents of supplementary feeds distributed in the market, the goal being to establish a universally trusted method. Method validation was performed according to the International Conference on Harmonization (ICH) and International Union of Pure and Applied Chemistry (IUPAC) guidelines. Method validation was performed in terms of linearity, sensitivity, selectivity, accuracy, and precision. The limits of detection (LOD) for the instrument employed in these experiments ranged from 0.44–4.77 mg/kg, and the limits of quantification (LOQ) ranged from 1.32–14.31 mg/kg. Average recoveries of the 12 flavoring agents ranged from 75.1–111.4%. Maximum %RSD values for intraday and interday peak area variation are 13.09% and 13.08%, respectively. A novel and simple method for detecting 12 flavoring agents in animal feed supplements using a gas chromatography-flame ionization detector (GC-FID) was developed.