Rapid quantification of the aminoglycoside arbekacin in serum using high performance liquid chromatography–tandem mass spectrometry

This project entails the development and validation of a method for quantification of the aminoglycoside antibiotic arbekacin in serum using liquid chromatography tandem mass spectrometry (LC–MS/MS) for therapeutic drug monitoring in future clinical trials. Following a protein precipitation with 0.3...

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Bibliographic Details
Published in:Clinica chimica acta 2013-03, Vol.418, p.102-106
Main Authors: Breaud, Autumn R., Henemyre-Harris, Claudia L., Schools, Sabitha, Emezienna, Nkechinyere, Clarke, William
Format: Article
Language:English
Subjects:
Aminoglycosides
Arbekacin
Chromatography, High Pressure Liquid
Dibekacin - analogs & derivatives
Dibekacin - blood
High performance liquid chromatography–tandem mass spectrometry
Humans
Tandem Mass Spectrometry
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Summary:This project entails the development and validation of a method for quantification of the aminoglycoside antibiotic arbekacin in serum using liquid chromatography tandem mass spectrometry (LC–MS/MS) for therapeutic drug monitoring in future clinical trials. Following a protein precipitation with 0.3mol/l perchloric acid containing internal standard dibekacin at a concentration of 0.6μg/ml, human serum samples containing arbekacin were analyzed using a Hypersil Gold PFP column and a liquid chromatography system. Elution occurred with a gradient of water and acetonitrile, each containing 0.05% (v/v) trifluoroacetic acid and 0.1% (v/v) formic acid. Analytes were detected over a 3.25minute run time using a tandem mass spectrometer with a heated electrospray-ionization (HESI) source in positive ionization mode with selected reaction monitoring (SRM). Matrix effects, carryover, linearity, recovery, precision, and limit of quantification were carefully evaluated. The limit of quantification for arbekacin was 0.1μg/ml. All simple and total precision CV's were less than 6.2%. The method was linear from 0.1μg/ml to 45.9μg/ml (slope of 0.973). The mean recovery ranged from 94.7 to 103.8%. No matrix effects were detected. This developed and validated LC–MS/MS method allows for the quantification of arbekacin in serum following protein precipitation. ► We developed and validated an assay to quantify arbekacin in human serum. ► Serum samples must undergo a brief protein precipitation prior to analysis. ► The assay uses liquid chromatography coupled with tandem mass spectrometry. ► Arbekacin LOQ was 0.1μg/ml and the method was linear from 0.1μg/ml to 45.9μg/ml. ► The arbekacin assay will be used for TDM support of a future clinical trial.
ISSN:0009-8981
1873-3492
DOI:10.1016/j.cca.2013.01.004