PMID- 18041566 OWN - NLM STAT- MEDLINE DCOM- 20080115 LR - 20190917 IS - 0042-8450 (Print) IS - 0042-8450 (Linking) VI - 64 IP - 10 DP - 2007 Oct TI - [Determination of diazepam and its metabolites in serum by the use of liquid chromatography-mass spectrometry method]. PG - 659-62 AB - BACKGROUND/AIM: Diazepam is a benzodiazepine anxyolitic. Metabolism of diazepam takes place in liver which generates pharmacologically active metabolites N-desmethyldiazepam, temazepam and oxazepam. The aim of this study was to develop and validate the method of liquid chromatographymass spectrometry (LC-MS) for separation and determination of diazepam and its active metabolites in the serum of rats samples after i.p. application of diazepam in a dose of 10 mg/kg. METHODS: The serum samples taken from Wistar rats, were used in LC-MS analysis after the application of 10 mg/kg of diazepam i.p. RESULTS: After alkaline extraction from the serum samples with diethylether and separation on a C18 reversed-phase column by using mobile phase methanolglacial acetic acid-water (50:1:49 v/v), diazepam and its metabolites were quantified. Determination was performed in a selective ion monitoring (SIM) mode, thereby the other exogenous and endogenous compounds did not interfere with this assay. Diazepam, N-desmethyldiazepam, oxazepam and temazepam were eluted in 14 minutes. The standard curve was linear in the range from 10-2 000 ng/ml. The limits of detection for diazepam, N-desmethyldiazepam, oxazepam and temazepam were 4.37, 3.13, 4.38 and 7.31 ng/ml, respectively. The limits of quantitation for diazepam, N-desmethyldiazepam, oxazepam and temazepam were 14.58, 10.41, 14.59 and 24.36 ng/ml, respectively. CONCLUSION: The described LC-MS is a simple, sensitive, specific and accurate method and could be used for routine identification and quantification of small concentrations of diazepam and its metabolites in biological fluids. FAU - Dordevic, Snezana AU - Dordevic S AD - Vojnomedicinska akademija, Nacionalni centar za kontrolu trovanja, Odeljenje za toksikolosku hemiju, Beograd, Srbija. FAU - Kilibarda, Vesna AU - Kilibarda V LA - srp PT - English Abstract PT - Journal Article PL - Serbia TA - Vojnosanit Pregl JT - Vojnosanitetski pregled JID - 21530700R RN - Q3JTX2Q7TU (Diazepam) SB - IM MH - Animals MH - *Chromatography, Liquid MH - Diazepam/analogs & derivatives/*pharmacokinetics MH - Female MH - *Mass Spectrometry MH - Rats MH - Rats, Wistar EDAT- 2007/11/29 09:00 MHDA- 2008/01/16 09:00 CRDT- 2007/11/29 09:00 PHST- 2007/11/29 09:00 [pubmed] PHST- 2008/01/16 09:00 [medline] PHST- 2007/11/29 09:00 [entrez] AID - 10.2298/vsp0710659d [doi] PST - ppublish SO - Vojnosanit Pregl. 2007 Oct;64(10):659-62. doi: 10.2298/vsp0710659d.