High-performance liquid chromatography was used to study the stability of folate vitamers in two types of rye breads after baking and 16 weeks of frozen storage. Bread made using sourdough seeds contained less total folate (74.6μg/100 g dry basis, expressed as folic acid) than the whole rye flour (79.8μg/100 g dry basis) and bread leavened only with baker's yeast (82.8μg/100 g dry basis). Most importantly, it was generated by a significant decrease in 5-CH3-H4folate form. The baking process caused some changes in folate distribution. Storage of breads at −18°C for 2 weeks did not have a significant effect (p<0.05) on total folates compared to the content directly after baking. After a 5-weeks storage period, a significant decrease (p<0.05) in the content of total folates was recorded and it dropped on average by 14% for both type of breads. After a longer period of storage (16 weeks), a 25% loss of folates in the bread made with baker's yeast and a 38% loss in the bread fermented with sourdough seeds was found. Retention of 5-CH3-H4folate and 10-HCO-H2folate forms were much lower in the bread made with a sourdough addition than with baker's yeast only.