title: 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.
abstract: 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, C1a and C2), and vancomycin in plasma was developed. In 25μL 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 7.5min per sample. Calibration standards were prepared over a range of 0.3-50mgL(-1) for amikacin and gentamicin and 1.0-100mgL(-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 (25μL) of plasma.
authors.affiliation: Department of Pharmacy, Academic Medical Centre, Amsterdam, The Netherlands. Electronic address: y.a.bijleveld@amc.nl.
authors.forename: Yuma
authors.initials: Y
authors.lastname: Bijleveld
authors.affiliation: Department of Neonatology, Emma Children's Hospital, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Timo R
authors.initials: TR
authors.lastname: de Haan
authors.affiliation: Department of Pharmacy, Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: Toersche
authors.affiliation: Department of Pharmacy, Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: Sona
authors.initials: S
authors.lastname: Jorjani
authors.affiliation: Department of Paediatric Clinical Epidemiology, Emma Children's Hospital, Academic Medical Center, University of Amsterdam, The Netherlands.
authors.forename: Johanna
authors.initials: J
authors.lastname: van der Lee
authors.affiliation: Department of Neonatology, Wilhelmina Children's Hospital, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Floris
authors.initials: F
authors.lastname: Groenendaal
authors.affiliation: Department of Neonatology, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Dijk
authors.affiliation: Department of Neonatology, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands.
authors.forename: Arno
authors.initials: A
authors.lastname: van Heijst
authors.affiliation: Department of Neonatology, Maastricht University Medical Center, Maastricht, The Netherlands.
authors.forename: Antonio W D
authors.initials: AWD
authors.lastname: Gavilanes
authors.affiliation: Department of Neonatology, Erasmus MC-Sophia Children's Hospital, Rotterdam, The Netherlands.
authors.forename: Rogier
authors.initials: R
authors.lastname: de Jonge
authors.affiliation: Department of Neonatology, Maxima Medical Center Veldhoven, Veldhoven, The Netherlands.
authors.forename: Koen P
authors.initials: KP
authors.lastname: Dijkman
authors.affiliation: Department of Neonatology, Isala Clinics, Zwolle, The Netherlands.
authors.forename: Henrica
authors.initials: H
authors.lastname: van Straaten
authors.affiliation: Department of Neonatology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Monique
authors.initials: M
authors.lastname: Rijken
authors.affiliation: Department of Neonatology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Inge
authors.initials: I
authors.lastname: Zonnenberg
authors.affiliation: Department of Neonatology, Vrije Universiteit Brussel, Brussels, Belgium.
authors.forename: Filip
authors.initials: F
authors.lastname: Cools
authors.affiliation: PharmaCool Study, The Netherlands.
authors.forename: Debbie
authors.initials: D
authors.lastname: Nuytemans
authors.affiliation: Department of Pharmacy, Academic Medical Centre, Amsterdam, The Netherlands.
authors.forename: Ron
authors.initials: R
authors.lastname: Mathôt
mesh_terms: D000583:Amikacin / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods; D004355:Drug Stability; D005839:Gentamicins / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D006801:Humans; D007231:Infant, Newborn; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods; D014640:Vancomycin / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics
chemical_list: D005839:Gentamicins; D014640:Vancomycin; D000583:Amikacin
keywords: Amikacin; Gentamicin; Hypothermia; LC–MS/MS; Neonates; Vancomycin