Coloring conditions of thiobarbituric acid test for detecting lipid hydroperoxides

Abstract The coloring reaction of the thiobarbituric acid test for hydroperoxides was completely inhibited by the addition of EDTA. Therefore, it was necessary to add a metal salt to the reaction mixture to complete the reaction and also to add an antioxidant to prevent autoxidation when unoxidized unsaturated fatty acids co‐exist. The optimal pH of the reaction was found at 3.6 using glycine‐hydrochloric acid buffer.