Simple validated LC-MS/MS method for the determination of atropine and scopolamine in plasma for clinical and forensic toxicological purposes.
A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of atropine and scopolamine in 100μL human plasma was developed and validated. Sample pretreatment consisted of protein precipitation with acetonitrile followed by a concentration step. Analytes and levobupivacaine (internal standard) were separated on a Zorbax XDB-CN column (75mm×4.6mm i.d., 3.5μm) with gradient elution (purified water, acetonitrile, formic acid). The triple quadrupole MS was operated in ESI positive mode. Matrix effect was estimated for deproteinised plasma samples. Selected reaction monitoring (SRM) was used for quantification in the range of 0.10-50.00ng/mL. Interday precision for both tropanes and intraday precision for atropine was <10%, intraday precision for scopolamine was <14% and <18% at lower limit of quantification (LLOQ). Mean interday and intraday accuracies for atropine were within ±7% and for scopolamine within ±11%. The method can be used for determination of therapeutic and toxic levels of both compounds and has been successfully applied to a study of pharmacodynamic and pharmacokinetic properties of tropanes, where plasma samples of volunteers were collected at fixed time intervals after ingestion of a buckwheat meal, spiked with five low doses of tropanes.
Institute of Forensic Medicine, Faculty of Medicine, Korytkova 2, SI-1000 Ljubljana, Slovenia. Electronic address: gordana.kozelj@mf.uni-lj.si.
Gordana
G
Koželj
Department of Infectious Diseases and Environmental Risk, National Institute of Public Health, Trubarjeva 2, SI-1000 Ljubljana, Slovenia. Electronic address: Lucija.Perharic@ivz-rs.si.
Lucija
L
Perharič
Institute of Pharmacology and Experimental Toxicology, Faculty of Medicine, Korytkova 2, SI-1000 Ljubljana, Slovenia. Electronic address: lovro.stanovnik@mf.uni-lj.si.
Lovro
L
Stanovnik
Faculty of Chemistry and Chemical Technology, Aškerčeva 5, SI-1000 Ljubljana, Slovenia. Electronic address: Helena.Prosen@fkkt.uni-lj.si.
Helena
H
Prosen
D000328:Adult; D001285:Atropine / Q000097:blood* / Q000633:toxicity; D002853:Chromatography, Liquid / Q000379:methods*; D004311:Double-Blind Method; D053593:Forensic Toxicology / Q000379:methods; D006801:Humans; D057230:Limit of Detection; D015203:Reproducibility of Results; D012601:Scopolamine / Q000097:blood* / Q000633:toxicity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D001285:Atropine; D012601:Scopolamine
Atropine; LC–MS/MS; Plasma; Scopolamine; Validation