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Development and validation of a simultaneous extraction procedure for HPLC-MS quantification of daptomycin, amikacin, gentamicin, and rifampicin in human plasma

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Abstract

A simultaneous extraction method to measure daptomycin, amikacin, gentamicin, and rifampicin in human plasma, by high-performance liquid chromatography, was developed and validated. The method involved a rapid sample preparation by protein precipitation with acetonitrile followed by direct injection into a high-performance liquid chromatography system coupled with mass detection. Drug retention times were 10.00 ± 0.25, 2.00 ± 0.25, 3.50 ± 0.25, 11.50 ± 0.25, and 12.50 ± 0.25 min for daptomycin, amikacin, gentamicin, rifampicin, and quinoxaline, respectively. Good linearity (mean r 2 = 0.998) was obtained for all drugs quantified over the range of clinically relevant concentrations in human plasma and the use of the internal standard quinoxaline improves accuracy (RSD% <14.9%) and intra-day (RSD% <11.56) and inter-day (RSD% <12.10) precision for the analytical procedure. The limits of quantification for daptomycin, amikacin, gentamicin, and rifampicin were 1.56, 2.34, 0.63, 0.63 μg/ml, respectively. Moreover, the addition of ion pair trifluoroacetic acid in the sample allowed the majority of gentamicin and amikacin separation. A rapid, specific, sensitive, accurate, and reproducible HPLC method was developed and validated to measure daptomycin, amikacin, gentamicin, and rifampicin in human plasma. This method is suitable for clinical pharmacokinetic studies.

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Authors and Affiliations

  1. Laboratory of Clinical Pharmacology and Pharmacogenetics, Department of Infectious Diseases, University of Torino, Amedeo di Savoia Hospital, Corso Svizzera 164, Turin, 10149, Italy

    Lorena Baietto, Antonio D’Avolio, Francesco Giuseppe De Rosa, Silvia Garazzino, Marianna Michelazzo, Giusi Ventimiglia, Marco Siccardi, Marco Simiele, Mauro Sciandra & Giovanni Di Perri

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  1. Lorena Baietto
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  2. Antonio D’Avolio
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Correspondence to Lorena Baietto.

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Baietto, L., D’Avolio, A., De Rosa, F.G. et al. Development and validation of a simultaneous extraction procedure for HPLC-MS quantification of daptomycin, amikacin, gentamicin, and rifampicin in human plasma. Anal Bioanal Chem 396, 791–798 (2010). https://doi.org/10.1007/s00216-009-3263-1

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  • DOI: https://doi.org/10.1007/s00216-009-3263-1

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