Simultaneous Determination of Amoxicillin and Clavulanic Acid in Dog Plasma by High-Performance Liquid Chromatography Tandem Mass Spectrometry

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 C 8 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 À1 . There is good linearity in the range of 0.5-500 ng mL À1 for both amoxicillin and clavulanic acid. The decision limits of amoxicillin and clavulanic acid were 0.06 ng mL À1 and 0.08 ng mL À1 in plasma, respectively, and the detection capabilities of two analytes were below 0.5 ng mL À1 . 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 À1 , 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.

Electrospray ionization collision‐induced dissociation tandem mass spectrometry of amoxicillin and ampicillin and their degradation products

“…In both mass spectra there two abundant fragment ions formed as a result of the loss of a molecule of CO 2 ( m/z 320 and 304) and as a result of the breaking of three bonds of the bicyclic system ( m/z 223 and 207). Such interpretation was already presented in the literature and we agree with it. In the ESI‐CID‐MS/MS spectrum of ion [ Amox –H] – there are also less abundant ions at m/z 303, 206 and 129.…”

“…Also in the positive ion mode, a number of detailed studies of mass spectrometric fragmentation pathways of these antibiotics and their degradation products (by using ESI‐MS/MS experiments) have been published . In the negative ion mode, LC/ESI‐MS was also used for determination of amoxicillin and ampicillin, as well as their degradation products (although more rarely than in the positive ion mode), and mass spectrometric fragmentation pathways in the negative ion mode were also discussed . Mass spectrometric decomposition is used for the determinations of the organic compounds (including the title compounds), when the so‐called multiple‐reaction monitoring (MRM) is applied during LC/ESI‐MS analysis.…”

Pharmacokinetics of the amoxicillin–clavulanic acid combination after intravenous and oral administration in cats

“…The plasma levels of AMX and CLV were measured simultaneously by HPLC‐tandem mass spectrometry (LC‐MS‐MS) using a method previously validated for canine plasma (Xi, He, Guo, Cai, & Zeng, ). The system was equipped with an Agilent 1200 Series HPLC (Agilent Technologies, Wilmington, DE, USA) and a triple quadrupole mass spectrometer (Applied Biosystems API 4000; AB SCIEX LLC., Milford, MA, USA) with Analyst 1.5 software.…”

“…The mass spectrometer was operated under the multiple reaction monitoring (MRM) mode. More details about the other optimized parameter values and the MRM transitions used for quantification of AMX and CLV can be found in our previous study (Xi et al, ).…”

Validated LC-MS/MS Method for the Concomitant Determination of Amoxicillin and Clavulanic Acid from Human Plasma

“…There are several methods known in the literature for the individual and/or simultaneous determination of amoxicillin and clavulanic acid in human plasma and other biological matrix using UV [2,6,11,13] and MS/MS methods [3][4][5][7][8][9]12]. All MS/MS methods involve polarity switching, for monitoring the clavulanic acid in negative electrospray ionization mode.…”