Simultaneous assay of sildenafil and desmethylsildenafil in neonatal plasma by ultra-performance liquid chromatography-tandem mass spectrometry

BC Witjes, Maurice Ahsman, BC van der Nagel, Dick Tibboel, RAA Mathot

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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 mu L 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 (C) 2009 John Wiley & Sons, Ltd.
Original languageUndefined/Unknown
Pages (from-to)180-185
Number of pages6
JournalBiomedical Chromatography
Issue number2
Publication statusPublished - 2010

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