Date of Award

Spring 6-2019

Document Type


Degree Name

Master of Science (MS)


Forensic Science



First Advisor or Mentor

Marta Concheiro-Guisan

Second Reader

Gail Cooper

Third Advisor

Madeleine Swortwood


The misuse of designer benzodiazepines, as an alternative to prescription benzodiazepines and for drug-facilitated sexual assaults, has emerged as a growing threat, due in part to the ease of purchasing these drugs on the internet at low prices. Causing concern for safety, is the lack of dosage information resulting in users self-medicating, often leading to unintended overdoses, coma, or death at higher doses. With limited published data regarding quantification of designer benzodiazepines in forensic cases, a method was validated for the determination of thirteen and screening for two designer benzodiazepines in blood, in addition to a limited number of common designer benzodiazepines in the inhouse method. The developed method analyzed 3-hydroxyphenazepam, 4-chlorodiazepam, clobazam, clonazolam, delorazepam, deschloroetizolam, diclazepam, flualprazolam, flubromazepam, flubromazolam, flunitrazolam, meclonazepam, nifoxipam, pyrazolam, and zolazepam in 0.5 mL blood using liquid chromatography-tandem mass spectrometry. The analytes were treated with solid phase extraction before undergoing separation on a column and analyzed on the mass spectrometer in electrospray positive mode using multiple-reaction monitoring. The linear range of the calibration curve was 1-200 ng/mL, and up to 500 ng/mL for 3-hydroxyphenazepam, clobazam, flubromazepam, and pyrazolam. 4-chlorodiazepam and zolazepam were tested for qualitative analysis only. The limits of detection and quantitation were 0.5 ng/mL (S/N>3) and 1 ng/mL, respectively. Other parameters tested included bias, precision, matrix effect, recovery, carryover, stability, interference, and dilution integrity which all yielded acceptable results. With the application of this method to blood specimens from the NYC Office of Chief Medical Examiner, this validated method proved to be simple, reproducible, sensitive, and robust.



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