In developing the microbiological assay for nystatin1 in feeds, reported by Pagano, studies were made of the following factors: effect of heat and of pH on nystatin stability; effect of solvents on nystatin recovery; effect of feed extractives on Candida tropicalis; recovery of nystatin at different concentrations in feed; and preparation of a nystatin-free feed. Heat and pH must be rigidly controlled to keep nystatin loss at a minimum. Best recoveries were obtained with a hexane wash and methanol extraction.

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