LC-MS/MS analysis of 13 benzodiazepines and metabolites in urine, serum, plasma, and meconium.
We describe a single method for the detection and quantitation of 13 commonly prescribed benzodiazepines and metabolites: alpha-hydroxyalprazolam, alpha-hydroxyethylflurazepam, alpha-hydroxytriazolam, alprazolam, desalkylflurazepam, diazepam, lorazepam, midazolam, nordiazepam, oxazepam, temazepam, clonazepam and 7-aminoclonazepam in urine, serum, plasma, and meconium. The urine and meconium specimens undergo enzyme hydrolysis to convert the compounds of interest to their free form. All specimens are prepared for analysis using solid-phase extraction (SPE), analyzed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), and quantified using a three-point calibration curve. Deuterated analogs of all 13 analytes are included as internal standards. The instrument is operated in multiple reaction-monitoring (MRM) mode with an electrospray ionization (ESI) source in positive ionization mode. Urine and meconium specimens have matrix-matched calibrators and controls. The serum and plasma specimens are quantified using the urine calibrators but employing plasma-based controls. Oxazepam glucuronide is used as a hydrolysis control.
Marin
Stephanie J
SJ
ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT, USA.
McMillin
Gwendolyn A
GA
D000525:Alprazolam / Q000031:analogs & derivatives / Q000097:blood / Q000652:urine; D001569:Benzodiazepines / Q000097:blood* / Q000652:urine*; D002853:Chromatography, Liquid / Q000379:methods*; D002998:Clonazepam / Q000031:analogs & derivatives / Q000097:blood / Q000652:urine; D005479:Flurazepam / Q000031:analogs & derivatives / Q000097:blood / Q000652:urine; D006801:Humans; D007231:Infant, Newborn; D008470:Meconium / Q000737:chemistry*; D003708:Nordazepam / Q000097:blood / Q000652:urine; D010076:Oxazepam / Q000097:blood / Q000652:urine; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014229:Triazolam / Q000031:analogs & derivatives / Q000097:blood / Q000652:urine
D001569:Benzodiazepines; C020860:7-aminoclonazepam; D014229:Triazolam; C032318:alpha-hydroxyalprazolam; C028638:alpha-hydroxytriazolam; D002998:Clonazepam; D003708:Nordazepam; C041538:hydroxyethylflurazepam; D010076:Oxazepam; D005479:Flurazepam; D000525:Alprazolam