Mixtures of sodium nitrite, amino acids and ascorbic acid at pH 6.3 or 7.2 were filter sterilized or heated for various times at 121°C. Samples were analyzed for nitrite concentration immediately after treatment and after storage at 37 or 43°C. Heating the complete medium for 15 min at 121°C reduced the nitrite concentration by 30–50% independent of the initial nitrite concentration. Storage of complete filter sterilized medium at 43°C for 18–24 hr resulted in 50–65% loss of nitrite while in heat sterilized medium the loss was as great as 90%. None of the 19 amino acids stored individually with sodium nitrite at 37°C for up to 4 hr affected the nitrite concentration, whereas ascorbic acid resulted in total disappearance of nitrite. During heating at 121°C of the 19 amino acids only cystine resulted in any significant nitrite loss after 15 min of treatment while ascorbic acid effected total nitrite disappearance after 5 min. The defined medium containing nitrite demonstrated inhibitory activity against seven strains ofClostridium