In this study, the stability of 5-methyltetrahydrofolate (5-MTHF) in the model system and folate-enriched egg yolk and strategies for 5-MTHF stabilization were investigated. The oxygen, temperature and light affect the stability of 5-MTHF in the model system, among which oxygen is the main factor. In thermal pasteurization and spray-drying with normal air media, 5-MTHF is sensitive to oxidation, with the retention rate of blank group only reaching 74.96% ± 1.28%. The addition of vitamin C or vitamin E can protect 5-MTHF in egg yolk from degradation and the latter has a better protective effect. By adding 0.2% (w/v) vitamin E to egg yolk liquid, the retention rate of 5-MTHF during thermal pasteurization and spray-drying with normal air media were 94.16% ± 0.48% and 84.80% ± 0.82% respectively. Additionally, the spray-drying technique with inert gas media (N) was also an effective method to improve the stability of the 5-MTHF in egg yolk. Our study explored the factors affecting the stability of 5-MTHF in both model systems and egg yolk liquid and provided effective strategies for the protection of 5-MTHF during the processing of egg.
