Determination of the anti-platelet-activating factor BN-50727 and metabolites in human urine by high-performance liquid chromatography using solid-phase extraction.
A sensitive and selective HPLC solid-phase extraction procedure was developed for the determination of platelet-activating factor antagonist BN-50727 and its metabolites in human urine. The procedure consisted in a double solid-phase extraction of the urine samples on cyanopropyl and silica cartridges, followed by an automated solid-phase extraction of the drug and metabolites on CBA cartridges and posterior elution on-line to the chromatographic system for its separation. The method allowed quantitation in the concentration range 10-2400 ng/ml urine for both BN-50727 and the main metabolite, the O-demethylated BN-50727 product. The limit of quantitation for both compounds was 10 ng/ml. The inter-assay precision of the method, expressed as relative standard deviation, ranged from 1.9 to 4.5% for BN-50727 and from 2.5 to 9.0% for the metabolite. The accuracy, expressed as relative error, ranged from -2.4 to 4.2% and from 0.2 to 6.2%, respectively. This paper describes the validation of the analytical methodology for the determination of BN-50727 in human urine and also for its metabolites. The method has been used to follow the time course of BN-50727 and its metabolites in human urine after single-dose administration.
Pruñonosa
J
J
S.A. Lasa Laboratorios, Pharmacokinetic Department, Barcelona, Spain.
Solà
J
J
Peraire
C
C
Pla
F
F
Lavergne
O
O
Obach
R
R
D001381:Azepines / Q000008:administration & dosage / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D010975:Platelet Aggregation Inhibitors / Q000008:administration & dosage / Q000652:urine*; D012015:Reference Standards; D012016:Reference Values; D015203:Reproducibility of Results; D058924:Thienopyridines; D014230:Triazoles / Q000008:administration & dosage / Q000652:urine*
D001381:Azepines; D010975:Platelet Aggregation Inhibitors; D058924:Thienopyridines; D014230:Triazoles; C067429:Setipafant