Determination of dapoxetine, an investigational agent with the potential for treating depression, and its mono- and di-desmethyl metabolites in human plasma using column-switching high-performance liquid chromatography.
A column-switching high-performance liquid chromatographic (HPLC) method is described for the determination of dapoxetine and its mono- and di-desmethyl metabolites in human plasma. The analytes, including an internal standard, were extracted from plasma at basic pH with hexane-ethyl acetate. The organic extract was evaporated to dryness and the residue reconstituted with acetonitrile. The analytes were separated from late-eluting endogenous substances on a Zorbax RX-C8 pre-column. The front-cut fraction containing the analytes was further separated on a second RX-C8 column. The analytes were detected by their native fluorescence, using excitation and emission wavelengths of 230 and 330 nm, respectively. The limit of quantitation was determined to be 20 ng/ml, and the response was linear from 20 to 200 ng/ml. The method has been successfully applied to human plasma samples in a Phase I study.
Hamilton
C L
CL
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285.
Cornpropst
J D
JD
D000928:Antidepressive Agents / Q000097:blood* / Q000493:pharmacokinetics; D001596:Benzylamines / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D015507:Drugs, Investigational / Q000493:pharmacokinetics; D006801:Humans; D007202:Indicators and Reagents; D009281:Naphthalenes / Q000097:blood* / Q000493:pharmacokinetics; D017367:Selective Serotonin Reuptake Inhibitors / Q000097:blood* / Q000493:pharmacokinetics; D013050:Spectrometry, Fluorescence
D000928:Antidepressive Agents; D001596:Benzylamines; D015507:Drugs, Investigational; D007202:Indicators and Reagents; D009281:Naphthalenes; D017367:Selective Serotonin Reuptake Inhibitors; C080600:di-desmethyldapoxetine; C080599:mono-desmethyldapoxetine; C080598:dapoxetine