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Simultaneous assay of sildenafil and desmethylsildenafil in neonatal plasma by ultra‐performance liquid chromatography–tandem mass spectrometry
Author(s) -
Witjes Bregje C.,
Ahsman Maurice J.,
van der Nagel Bart C.,
Tibboel Dick,
Mathot Ron A.
Publication year - 2010
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.1268
Subject(s) - sildenafil , chemistry , chromatography , pharmacokinetics , tandem mass spectrometry , liquid chromatography–mass spectrometry , analyte , mass spectrometry , extraction (chemistry) , calibration curve , high performance liquid chromatography , detection limit , pharmacology , medicine
Sildenafil is used to treat pulmonary hypertension in neonatal and pediatric patients. Pharmacokinetic studies in these patients are complicated by the limited sample volume. We present the validation results of an assay method to quantitate sildenafil and desmethylsildenafil simultaneously in 50 µL of plasma. Deuterated sildenafil was used as an internal standard. After liquid–liquid extraction, analytes were separated on an ultra‐performance liquid chromatography (UPLC)‐column and quantified via tandem mass spectrometry. The calibration range was linear, with acceptable accuracy and a precision of <15% for both compounds. The lower limits of quantification were 1 ng/mL. Matrix effects were present, but inter‐plasma batch variability was under 12%. The method was successfully applied to samples from a pharmacokinetic study into sildenafil pharmacokinetics in neonates, making maximum use of the limited number and amount of plasma samples available. Copyright © 2009 John Wiley & Sons, Ltd.