Simultaneous high-performance liquid chromatographic analysis of flunitrazepam and four metabolites in serum.
A high-performance liquid chromatographic method for the simultaneous determination of flunitrazepam and four metabolites, desmethylflunitrazepam (DMF), 7-aminodesmethylflunitrazepam (7-NH2DMF), 7-aminoflunitrazepam (7-NH2F) and 3-hydroxyflunitrazepam (3-OHF), in serum is described. The method involves a simple extraction from alkalinized plasma (pH 9.5) into diethyl ether-chloroform (80:20, v/v). Prazepam was used as an internal standard for the quantification of the five compounds. Separation was achieved with a 10 microns RSil CN column (300 x 3.9 mn I.D.). The detection wavelength was set at 242 nm. The limits of detection ranged from 2.5 to 5 micrograms/l with a limit of quantification of 10 micrograms/l for all analytes.
Berthault
F
F
Institute of Legal Medecine, Strasbourg, France.
Kintz
P
P
Mangin
P
P
D014151:Anti-Anxiety Agents / Q000097:blood* / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005445:Flunitrazepam / Q000097:blood* / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D011222:Prazepam / Q000097:blood; D012015:Reference Standards; D012044:Regression Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D014151:Anti-Anxiety Agents; D005445:Flunitrazepam; D011222:Prazepam