The Analysis of Docosahexaenoic Acid (DHA) in Dried Dog Food Enriched with an Aurantiochytrium limacinum Biomass: Matrix Extension Validation and Verification of AOAC Method 996.06.

BACKGROUND: Docosahexaenoic acid (DHA) plays an important role in brain and retinal development in dogs. However, supranutritional dietary supplementation can result in health issues, including gastrointestinal bleeding, making the accurate analysis of DHA in dog food important for nutritional and welfare regulatory compliance. OBJECTIVE: The aim of this study was to conduct a validation and verification of the AOAC 996.06 method, and hence establish its fitness for purpose, for the analysis of DHA in dried dog food supplemented with a heterotrophically grown unextracted DHA-rich Aurantiochytrium limacinum biomass. METHODS: The AOAC 996.06 method, which involves the use of gas chromatography coupled to flame ionization detection (GC-FID), was used to conduct a validation of the analysis of DHA in dried dog food and the results were verified in a second laboratory. RESULTS: The method was found to be linear over the ranges analyzed and results were found to be within the acceptance criteria for precision and accuracy, verifying the applicability for this matrix. The selectivity and sensitivity of the method were also determined. CONCLUSIONS: The AOAC 996.06 method is fit for purpose for the analysis of DHA in dry dog food kibble. HIGHLIGHTS: The method can be applied to various dog food samples, supplemented with an unextracted Aurantiochytrium limacinum biomass, using alternative manufacturing methods, i.e. pelleted and extruded with no significant matrix effects being observed.