Quantitative determination of a selective alpha-1a receptor antagonist in human plasma by high-performance liquid chromatography with tandem mass spectrometric detection.
Solid-phase extraction, utilizing a 96-well plate format, was used to isolate an alpha-1a receptor antagonist and internal standard from human plasma. Following the isolation procedure, the analyte and internal standard were separated and detected using reversed-phase HPLC coupled with atmospheric pressure chemical ionization (APCI) mass spectrometry operated in the positive ion multiple reaction monitoring (MRM) mode. Based upon the peak area ratio (analyte: internal standard) the analyte was quantified over a concentration range of 0.02-2 ng/ml. Assay validation results including parameters such as precision and accuracy are presented. The validated method was subsequently used to support human pharmacokinetic studies.
Department of Drug Metabolism, WP26-372, Merck Research Laboratories, P.O. Box 4, West Point, PA 19486, USA. dick_simpson@merck.com
Richard C
RC
Simpson
Amanda
A
Skarbek
Bogdan K
BK
Matuszewski
D058668:Adrenergic alpha-1 Receptor Antagonists; D000317:Adrenergic alpha-Antagonists / Q000097:blood* / Q000493:pharmacokinetics; D001274:Atmospheric Pressure; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D018340:Receptors, Adrenergic, alpha-1; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
C547713:ADRA1A protein, human; D058668:Adrenergic alpha-1 Receptor Antagonists; D000317:Adrenergic alpha-Antagonists; D018340:Receptors, Adrenergic, alpha-1