Improved microbiological procedure for determining bacitracin in premixes and mixed feeds

An improved microbiological procedure is presented for determining bacitracin in premixes and mixed feeds. Premixes are extracted with acidic 50% aqueous methanol, and mixed feeds are extracted with 50% dimethylformamide after washing with acetone to remove fats and pigments. Extracts are diluted with pH 6.0 phosphate buffer and assayed by using a 2-point assay system with Micrococcus flavus as the test organism. The simpler extraction coupled with the 2-point assay system measured bacitracin levels as low as 10 g/ton. Accuracy and repeatability of the 2-point system were at least equivalent to those for official AOAC methods.