Determination of phenylephrine in human plasma using ultra-performance liquid chromatography–tandem mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 915-916; pp. 28 - 32
• Main Authors: Feng, Sheng, Zhao, Qian, Jiang, Ji, Hu, Pei
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2013
• Subjects: acetonitrile; Acetonitriles - chemistry; Chromatography, High Pressure Liquid - methods; Determination; Drug Stability; Formates - chemistry; Human plasma; Humans; ionization; Least-Squares Analysis; monitoring; pharmacokinetics; Phenylephrine; Phenylephrine - blood; Phenylephrine - chemistry; Phenylephrine - pharmacokinetics; rapid methods; Reproducibility of Results; Sensitivity and Specificity; Solid Phase Extraction; tandem mass spectrometry; Tandem Mass Spectrometry - methods; ultra-performance liquid chromatography; UPLC–MS/MS; Human plasma; Determination; UPLC–MS/MS; Phenylephrine
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2012.12.019

► Analyte was quantifiable in amounts well below previously reported values. ► The total run time was much fast compared to the literature. ► The method was fully validated and applied successfully in a clinical study. This paper described a sensitive and rapid method based on ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC–MS/MS) for the determination of phenylephrine in human plasma. Plasma samples were pre-purified by solid-phase extraction (SPE). The chromatographic separation was achieved with BEH HILIC column using a mixture of 10mM pH 3.5 ammonium formate and acetonitrile (10:90, v/v) under isocratic conditions at a flow rate of 0.4mL/min. The mass spectrometry was carried out using positive electrospray ionization (ESI) and data acquisition was carried out in the multiple reaction monitoring (MRM) mode. The method was fully validated over the concentration range of 10.0–5000pg/mL. The lower limit of quantification (LLOQ) was 10.0pg/mL. Inter- and intra-batch precision was less than 15% and the accuracy was within 85–115%. Extraction recovery was 78.5%. Selectivity, matrix effects and stability were also validated. The method was applied to the pharmacokinetic study of phenylephrine hydrochloride in Chinese subjects.