A novel, sensitive and robust reverse phase liquid chromatographic method was developed for assay of mupirocin lithium in ointment or cream as its dosage form. For chromatographic study, water symmetry C8 (150 mm × 4.6 mm, 5 μm) column was used in on isocratic system at ambient temperature. The selected mobile phase made up of buffer and acetonitrile in proportion 74:26% (v/v). The flow rate of 1 ml / min. was used in study. The effluent study was done at wavelength 221 nm. The retention time was observed at 5.3 min. The linearity was observed in concentration range as 50 to 150 μg/ml. The correlation coefficient is less than 1. The ICH guidelines were used for system suitability, linearity, accuracy and precision studies. The accuracy and precision were found to be well within the acceptable limit. The method was successfully applied for assay of mupirocin lithium in cream or ointment with good recoveries.