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Rapid quantification of the aminoglycoside arbekacin in serum using high performance liquid chromatography-tandem
Autumn R Breaud1, Claudia L Henemyre-Harris, Sabitha Schools
1The Johns Hopkins School of Medicine, Department of Pathology, Clinical Chemistry, Baltimore, MD 21287, USA.
Background:
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.
Methods:
Following a protein precipitation with 0.3 mol/l perchloric acid containing internal standard dibekacin at a concentration of 0.6 μg/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 3.25 minute 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.
Results:
The limit of quantification for arbekacin was 0.1 μg/ml. All simple and total precision CV's were less than 6.2%. The method was linear from 0.1 μg/ml to 45.9 μg/ml (slope of 0.973). The mean recovery ranged from 94.7 to 103.8%. No matrix effects were detected.
Conclusions:
This developed and validated LC-MS/MS method allows for the quantification of arbekacin in serum following protein precipitation.
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