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Development and Validation of an LC-MS-MS Method for the Detection of 40 Benzodiazepines and Three Z-Drugs in Blood and Urine by Solid-Phase Extraction.
- Source :
-
Journal of analytical toxicology [J Anal Toxicol] 2020 Oct 12; Vol. 44 (7), pp. 708-717. - Publication Year :
- 2020
-
Abstract
- An analytical method for the detection of 40 benzodiazepines, (±)-zopiclone, zaleplon and zolpidem in blood and urine by solid-phase extraction liquid chromatography-tandem mass spectrometry was developed and validated. Twenty-nine of 43 analytes were quantified in 0.5 mL whole blood for investigating postmortem, drug-facilitated sexual assault (DFSA) and driving under the influence of drugs cases (DUID). The four different dynamic ranges of the seven-point, linear, 1/x weighted calibration curves with lower limits of quantification of 2, 5, 10 and 20 μg/L across the analytes encompassed the majority of our casework encountered in postmortem, DFSA and DUID samples. Reference materials were available for all analytes except α-hydroxyflualprazolam, a hydroxylated metabolite of flualprazolam. The fragmentation of α-hydroxyflualprazolam was predicted from the fragmentation pattern of α-hydroxyalprazolam, and the appropriate transitions were added to the method to enable monitoring for this analyte. Urine samples were hydrolyzed at 55°C for 30 min with a genetically modified β-glucuronidase enzyme, which resulted in >95% efficiency measured by oxazepam glucuronide. Extensive sample preparation included combining osmotic lysing and protein precipitation with methanol/acetonitrile mixture followed by freezing and centrifugation resulted in exceptionally high signal-to-noise ratios. Bias and between-and within-day imprecision for quality controls (QCs) were all within ±15%, except for clonazolam and etizolam that were within ±20%. All 29 of the 43 analytes tested for QC performance met quantitative reporting criteria within the dynamic ranges of the calibration curves, and 14 analytes, present only in the calibrator solution, were qualitatively reported. Twenty-five analytes met all quantitative reporting criteria including dilution integrity. The ability to analyze quantitative blood and qualitative urine samples in the same batch is one of the most useful elements of this procedure. This sensitive, specific and robust analytical method was routinely employed in the analysis of >300 samples in our laboratory over the last 6 months.<br /> (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Subjects :
- Alprazolam analogs & derivatives
Azabicyclo Compounds blood
Azabicyclo Compounds metabolism
Azabicyclo Compounds urine
Benzodiazepines blood
Benzodiazepines urine
Chromatography, Liquid methods
Diazepam analogs & derivatives
Forensic Toxicology
Humans
Hypnotics and Sedatives analysis
Hypnotics and Sedatives blood
Hypnotics and Sedatives urine
Limit of Detection
Piperazines blood
Piperazines metabolism
Piperazines urine
Sleep Aids, Pharmaceutical blood
Sleep Aids, Pharmaceutical metabolism
Sleep Aids, Pharmaceutical urine
Solid Phase Extraction methods
Tandem Mass Spectrometry methods
Zolpidem blood
Zolpidem metabolism
Zolpidem urine
Benzodiazepines metabolism
Hypnotics and Sedatives metabolism
Substance Abuse Detection methods
Subjects
Details
- Language :
- English
- ISSN :
- 1945-2403
- Volume :
- 44
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Journal of analytical toxicology
- Publication Type :
- Academic Journal
- Accession number :
- 32808043
- Full Text :
- https://doi.org/10.1093/jat/bkaa072