A rapid and simple method based on ultra performance liquid chromatography (UPLC) with ultra violet detection has been developed for the determination of daptomycin (DPT) and rifampicin (RFM) in rabbit plasma using 4-nitrophenol as internal standard (IS). Sample preparation involved protein precipitation with an acetonitrile:methanol mixture and centrifugation. Chromatographic separation was achieved on an Acquity BEH C18 column (100mmx2.1mm, 1.7microm) using gradient elution with methanol and 0.1% aqueous TFA. The total analysis time was 4.5min with DPT and RFM eluting at 1.9 and 2.1min, respectively. The method was fully validated with a lower limit of quantitation (LLOQ) of 2microgmL(-1) for both DPT and RFM. The intra- and inter-day precision, measured as % relative standard deviation, were less than 12.1 for DPT and 10.7 for RFM, respectively. This validated method was successfully applied to a pharmacokinetic study involving intravenous administration of 14mgkg(-1) DPT and 30mgkg(-1) RFM to rabbits.
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