Determination of 7-amino-flunitrazepam (Ro 20-1815) and 7-amino-desmethylflunitrazepam (Ro 5-4650) in plasma by high-performance liquid chromatography and fluorescence detection.
A high-performance liquid chromatographic method for the simultaneous determination of 7-amino-flunitrazepam (Ro 20-1815) and 7-amino-desmethylflunitrazepam (Ro 5-4650) in plasma is described. After extraction with an organic solvent, the compounds and their internal standard (7-amino-methylclonazepam or Ro 5-3384) are derivatized with fluorescamine and chromatographed on a reversed-phase muBondapak C18 column using pH 8 buffer solution-acetonitrile (3:1) as mobile phase. The detection is performed by a fluorometer at excitation and emission wavelengths of 390 and 470 nm, respectively. The sensitivity limit is about 1 ng/ml of plasma for both 7-amino-flunitrazepam and 7-amino-desmethylflunitrazepam. The method has been applied to the determination of plasma levels of these substances during pharmacokinetic studies of flunitrazepam, desmethylflunitrazepam and 7-amino-flunitrazepam.
D000818:Animals; D014151:Anti-Anxiety Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005445:Flunitrazepam / Q000031:analogs & derivatives / Q000097:blood*; D005450:Fluorescamine; D006801:Humans; D008253:Macaca mulatta; D013050:Spectrometry, Fluorescence
D014151:Anti-Anxiety Agents; D005450:Fluorescamine; D005445:Flunitrazepam; C036343:7-aminoflunitrazepam; C036342:7-aminodesmethylflunitrazepam