Development and validation of a UPLC‐UV method for the determination of daptomycin in rabbit plasma

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Abstract
Daptomycin (DPT) is a lipopeptide antibiotic with potent bactericidal activity in vitro against Gram‐positive bacteria, which has attracted the attention of the scientific community due to its unique mechanism of action and due to the immediate need for new antibiotics in the era of multidrug resistance. In order to assess its pharmacokinetics in rabbits a new analytical method has been developed and validated using ultra performance liquid chromatography in conjugation with ultraviolet detection for the quantitation of the antibiotic in rabbit plasma, using the internal standard methodology. The separation was achieved employing a C18 column with gradient elution using 0.1% aq. trifluoroacetic acid and methanol. The total analysis time was 2.5 min. The sample pretreatment employed protein precipitation with acetonitrile–methanol mixture and centrifugation. The method was validated in terms of linearity, precision, accuracy, sensitivity, robustness, short‐term and freeze–thaw stability and was applied to the quantification of DPT in plasma samples obtained from rabbits treated with 25 mg kg−1 DPT. Copyright © 2009 John Wiley & Sons, Ltd.

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