Quantification of Ceftriaxone in Rat Plasma Using Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry

Quantification of Ceftriaxone in Rat Plasma Using Hydrophilic Interaction Chromatography-Tandem Mass Spectrometry

Ceftriaxone is a parenteral cephalosporin with a broad spectrum of activity against gram-positive and gramnegative pathogens. For the quantification of ceftriaxone in rat plasma, a selective and sensitive hydrophilic interaction chromatography-tandem mass spectrometric method was developed and validated using protein precipitation with acetonitrile as a sample clean-up procedure. The analytes were separated in a Luna HILIC column with a gradient elution of 5% acetonitrile in 100 mM ammonium formate and 100% acetonitrile in 0.1% formic acid, and mass-to-charge ratios (m/z) were determined in selective reaction monitoring mode using tandem mass spectrometry with m/z 554.9>395.7 for ceftriaxone and m/z 172.2>154.2 for gabapentin (internal standard). The standard curve was linear over the concentration ranges of 5− 5000 ng/mL using 30 µL rat plasma sample. The coefficient of variation and relative error for intra- and inter-assay at four QC levels were 5.2 to 11.6% and −7.3 to 14.0%, respectively. The overall recovery of ceftriaxone from rat plasma using protein precipitation was 90.9±3.8%. This method was successfully applied to the pharmacokinetics of ceftriaxone after its intravenous administration at a dose of 10 mg/kg in male Sprague-Dawley rats.