How To Stabilize ω‑3 Polyunsaturated Fatty Acids (PUFAs) in an Animal Feeding Study?Effects of the Temperature, Oxygen Level, and Antioxidant on Oxidative Stability of ω‑3 PUFAs in a Mouse Diet

Abstract Substantial studies have shown that ω-3 polyunsaturated fatty acids (PUFAs) have various health-promoting effects; however, there are inconsistent results from animal studies that showed that ω-3 PUFAs have no effects or even detrimental effects. Emerging research suggests that oxidized ω-3 PUFAs have different effects compared to unoxidized ω-3 PUFAs; therefore, lipid oxidation of dietary ω-3 PUFAs could contribute to the mixed results of ω-3 PUFAs in animal studies. Here, we prepared an AIN-93G-based, semi-purified, powder diet, which is one of the most commonly used rodent diets in animal studies, to study the oxidative stability of fortified ω-3 PUFAs in animal feed. We found that lowering the storage temperature or the addition of a certain antioxidant, notably tert-butylhydroquinone (TBHQ), helps to stabilize ω-3 PUFAs and suppress ω-3 oxidation in the animal diet, while reducing the level of oxygen in the storage atmosphere is not very effective. The addition of 50 ppm of TBHQ in the diet inhibited 99.5 ± 0.1% formation of primary oxidation products and inhibited 96.1 ± 0.7% formation of secondary oxidation products, after 10 days of storage of the prepared diet at a typical animal-feeding experiment condition. Overall, our results highlight that ω-3 PUFAs are highly prone to lipid oxidation in a typical animal-feeding experiment, emphasizing the critical importance to stabilize ω-3 PUFAs in animal studies.