A simple quantitative method analysing amikacin, gentamicin, and vancomycin levels in human newborn plasma using ion-pair liquid chromatography/tandem mass spectrometry and its applicability to a clinical study

Yuma Bijleveld*, Timo de Haan, Jan Toersche, Sona Jorjani, Johanna van der Lee, Floris Groenendaal, Peter Dijk, Arno van Heijst, Antonio W. D. Gavilanes, Rogier de Jonge, Koen P. Dijkman, Henrica van Straaten, Monique Rijken, Inge Zonnenberg, Filip Cools, Debbie Nuytemans, Ron Mathot

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

53 Citations (Web of Science)


Neuroprotective controlled therapeutic hypothermia is the standard of care for newborns suffering perinatal asphyxia. Antibiotic drugs, such as amikacin, gentamicin, and vancomycin are frequently administered during controlled hypothermia, which possibly alters their pharmacokinetic (PK) and pharmacodynamic (PD) profiles. In order to examine this effect an LC-MS/MS method for the simultaneous quantification of amikacin, the major gentamicin components (gentamicin C, Cl a and C2), and vancomycin in plasma was developed. In 25 mu L plasma proteins were precipitated with trichloroacetic acid (TCA) and detection of the components was achieved using ion-pair reversed phase chromatography coupled with electrospray ionization tandem mass spectrometry. The chromatographic runtime was 7.5 min per sample. Calibration standards were prepared over a range of 0.3-50 mg L-1 for amikacin and gentamicin and 1.0-100 mg L-1 for vancomycin. At LLOQ accuracy was between 103 and 120% and imprecision was less than 19%. For concentrations above LLOQ accuracy ranged from 98% to 102% and imprecision was less than 6%. Process efficiency, ionization efficiency, and recovery were acceptable. Samples and stock solutions were stable during the time periods and at the different temperatures examined. The applicability of the method was shown by analysing plasma samples from 3 neonatal patients. The developed method allows accurate and precise simultaneous quantification of amikacin, gentamicin, and vancomycin in a small volume (25 mu L) of plasma.
Original languageEnglish
Pages (from-to)110-118
JournalJournal of Chromatography B
Publication statusPublished - 1 Mar 2014


  • LC-MS/MS
  • Amikacin
  • Gentamicin
  • Vancomycin
  • Hypothermia
  • Neonates

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