Development and validation of a liquid chromatography–tandem mass spectrometry method for the determination of phenylephrine in human plasma and its application to a pharmacokinetic study

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2007-10, Vol.858 (1), p.263-268
• Main Authors: Ptáček, P., Klíma, J., Macek, J.
• Format: Article
• Language: English
• Subjects: Adrenergic alpha-Agonists - blood; Adrenergic alpha-Agonists - pharmacokinetics; Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, Liquid - methods; Etilefrin; Fundamental and applied biological sciences. Psychology; General pharmacology; HILIC; Humans; Liquid chromatography; Mass spectrometry; Medical sciences; Molecular Structure; Pharmacology. Drug treatments; Phenylephrine; Phenylephrine - blood; Phenylephrine - pharmacokinetics; Phenylephrine - standards; Reference Standards; Reproducibility of Results; Solvents - chemistry; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry - methods
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2007.09.011

A sensitive and reliable method was developed to quantitate phenylephrine in human plasma using liquid chromatography–electrospray tandem mass spectrometry. The assay was based on solid-phase extraction with C 18 cartridges and hydrophilic interaction chromatography performed on a pentafluorophenylpropylsilica column (50 mm × 4 mm, 3 μm particles), the mobile phase consisted of methanol–10 mM ammonium acetate (90:10, v/v). Quantification was through positive-ion mode and selected reaction monitoring at m/ z 168.1 → 135.0 for phenylephrine and m/ z 182.1 → 135.0 for internal standard etilefrin, respectively. The lower limit of quantitation was 51 pg/ml using 0.25 ml of plasma and linearity was observed from 51 to 5500 pg/ml. Within-day and between-day precision expressed by relative standard deviation was less than 12% and inaccuracy did not exceed 8% at all levels. The assay was applied to the analysis of samples from a pharmacokinetic study.