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Abstract
A rapid, sensitive, and specific high-performance liquid chromatography tandem mass spectrometric method was developed for the simultaneous determination and confirmation of amoxicillin and clavulanic acid in plasma. Plasma sample was subjected to a simple deproteinization with acetonitrile, and then the supernatant was directly diluted by water. Analysis was performed on a Phenomenex Luna C8 reversed-phase column by detection with mass spectrometry in negative ions multiple reaction monitoring mode. A gradient elution program with 0.1% formic acid and acetonitrile was performed at a flow of 0.25 mL min−1. There is good linearity in the range of 0.5–500 ng mL−1 for both amoxicillin and clavulanic acid. The decision limits of amoxicillin and clavulanic acid were 0.06 ng mL−1 and 0.08 ng mL−1 in plasma, respectively, and the detection capabilities of two analytes were below 0.5 ng mL−1. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The extraction recoveries of amoxicillin and clavulanic acid were between 102% and 115% in plasma at three spiked levels of 0.5, 50, and 500 ng mL−1, with the relative standard deviations less than 15% for each analyte. The developed method was applied to pharmacokinetic studies of amoxicillin and clavulanic acid tablets in healthy beagles.
Acknowledgments
W. Xi and L. He contributed equally to this work.
Notes
*Represents quantitative ion; precursor ions are [M-H]−; DP represents declustering potential; CE represents collision energy; MRM ratio represents area ratio of the qualifier ion to the corresponding quantitative ion from 0.5 to 500 ng mL−1; value in parentheses is relative standard deviation (n = 18).
RSD, relative standard deviation.
SD, intra-day standard deviation; RSD, inter-day relative standard deviation (n = 15).