The simultaneous determination of diazepam and its three metabolites in dog plasma by high-performance liquid chromatography with mass spectroscopy detection.
A fast, sensitive and specific LC/MS/MS method for the simultaneous determination of diazepam and its three metabolites, oxazepam, temazepam and desmethyldiazepam, in dog plasma is described. The method consists of an automated 96-well solid phase extraction procedure and electrospray LC/MS/MS analysis. D(5)-Diazepam is used as the internal standard for all the compounds. Intra-day and inter-day assay coefficients of variations are less than 12.7%. The lower limit of quantitation (LLOQ) is 1 nM for each analyte, based on 0.1 ml aliquots of dog plasma. The analytical run time was 5 min. Linearity is observed over the range of 1--500 nM. This method has been used to support the discovery of pharmacokinetic studies.
Liu
Z
Z
Department of Drug Metabolism and Pharmacokinetics, The DuPont Pharmaceuticals Company, Stine-Haskell Research Center, PO Box 30, Newark, DE 19714, USA. peter.z.liu@dupontpharma.com
Short
J
J
Rose
A
A
Ren
S
S
Contel
N
N
Grossman
S
S
Unger
S
S
D000818:Animals; D014151:Anti-Anxiety Agents / Q000097:blood* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003975:Diazepam / Q000097:blood* / Q000378:metabolism; D004285:Dogs; D013058:Mass Spectrometry / Q000379:methods; D003708:Nordazepam / Q000097:blood* / Q000378:metabolism; D010076:Oxazepam / Q000097:blood* / Q000378:metabolism; D015203:Reproducibility of Results; D013693:Temazepam / Q000097:blood* / Q000378:metabolism
D014151:Anti-Anxiety Agents; D003708:Nordazepam; D010076:Oxazepam; D013693:Temazepam; D003975:Diazepam