title: High-performance liquid chromatography coupled with electrospray tandem mass spectrometry (LC/MS/MS) method for the simultaneous determination of diazepam, atropine and pralidoxime in human plasma.
abstract: A high-performance liquid chromatography coupled with electrospray tandem mass spectrometry (LC/MS/MS) procedure for the simultaneous determination of diazepam from avizafone, atropine and pralidoxime in human plasma is described. Sample pretreatment consisted of protein precipitation from 100microl of plasma using acetonitrile containing the internal standard (diazepam D5). Chromatographic separation was performed on a X-Terra MS C8 column (100mmx2.1mm, i.d. 3.5microm), with a quick stepwise gradient using a formate buffer (pH 3, 2mM) and acetonitrile at a flow rate of 0.2ml/min. The triple quadrupole mass spectrometer was operated in positive ion mode and multiple reaction monitoring was used for drug quantification. The method was validated over the concentration ranges of 1-500ng/ml for diazepam, 0.25-50ng/ml for atropine and 5-1000ng/ml for pralidoxime. The coefficients of variation were always <15% for both intra-day and inter-day precision for each analyte. Mean accuracies were also within +/-15%. This method has been successfully applied to a pharmacokinetic study of the three compounds after intramuscular injection of an avizafone-atropine-pralidoxime combination, in healthy subjects.
authors.affiliation: Université d'Angers, UFR médecine, Angers cedex F-49045, France. chadi.abbara@gmail.com
authors.forename: Chadi
authors.initials: C
authors.lastname: Abbara
authors.forename: Isabelle
authors.initials: I
authors.lastname: Bardot
authors.forename: Annie
authors.initials: A
authors.lastname: Cailleux
authors.forename: Guy
authors.initials: G
authors.lastname: Lallement
authors.forename: Anne
authors.initials: A
authors.lastname: Le Bouil
authors.forename: Alain
authors.initials: A
authors.lastname: Turcant
authors.forename: Pascal
authors.initials: P
authors.lastname: Clair
authors.forename: Bertrand
authors.initials: B
authors.lastname: Diquet
mesh_terms: D001285:Atropine / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003975:Diazepam / Q000097:blood*; D006801:Humans; D011220:Pralidoxime Compounds / Q000097:blood*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011220:Pralidoxime Compounds; D001285:Atropine; C028797:pralidoxime; D003975:Diazepam