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Determination of acyclovir in horse plasma and body fluids by high‐performance liquid chromatography combined with fluorescence detection and heated electrospray ionization tandem mass spectrometry
Author(s) -
Maes A.,
Garré B.,
Desmet N.,
van der Meulen K.,
Nauwynck H.,
De Backer P.,
Croubels S.
Publication year - 2009
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1093
Subject(s) - chemistry , chromatography , electrospray ionization , mass spectrometry , tandem mass spectrometry , electrospray , fluorescence , direct electron ionization liquid chromatography–mass spectrometry interface , liquid chromatography–mass spectrometry , extractive electrospray ionization , sample preparation in mass spectrometry , ionization , chemical ionization , ion , organic chemistry , physics , quantum mechanics
Two methods are presented for the determination of ‘respectively’ the plasma protein unbound and total concentration of acyclovir in horse plasma and body fluids: first, a liquid–liquid extraction was performed on plasma, combined with HPLC‐fluorescence detection for the total plasma concentration; second a more sensitive method using high‐performance liquid chromatography combined with heated electrospray ionization tandem mass spectrometry (LC‐HESI‐MS/MS) was described for plasma and for body fluids analysis. To obtain the unbound concentration of acyclovir in plasma, a simple deproteinization step using a Microcon® filter was performed. Ganciclovir was used as an internal standard. Analysis was carried out on an Inertsil 5 ODS‐3 column for the HPLC‐fluorescence method. For the LC‐HESI‐MS/MS method a PLRP‐S column was used. The limit of quantification (LOQ) for the total concentration was set at 50 and 2 ng mL −1 for the HPLC‐fluorescence method and the LC‐HESI‐MS/MS method, respectively. The limit of quantification for the unbound concentration was set at 5 ng mL −1 and at 2 ng mL −1 for body fluids. The methods were successfully used to perform pharmacokinetic and clinical studies in horses after intravenous and oral dosage of acyclovir and its prodrug valacyclovir. Copyright © 2008 John Wiley & Sons, Ltd.