A high performance liquid chromatographic method for estimation of Transitmycin (Tr) in human plasma was developed. The analyte was extracted using solid phase cartridges and the analysis of the eluent was carried out using Atlantis T3 column (150 cm × 4.6 mm ID) and photo diode array detector set at wavelength of 214 nm.  The assay was specific for Tr and linear from 0.5 µg/mL to 20.0 µg/mL. The relative standard deviations for intra- and inter-day assays were less than 10%. The method yielded a recovery for Tr that ranged from 94% to 107% and could be used in toxicology and pharmacokinetic studies.

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