Friscia M, Jolliff HA, Logan BK
Society of Forensic Toxicology- 2016 Conference MeetingThe recreational drug market has seen a proliferation of synthetic drugs, many of which are analogs or minor chemical modifications of existing compounds. This has led to a large number of synthetic cannabinoids, cathinones and beta-keto amphetamines, and most recently opioid receptor agonists entering the illicit drug market. The latest pharmacological class to be impacted in this way are the benzodiazepines. Various novel benzodiazepines, which have never been available legitimately in the United States, are now appearing in the illicit drug supply. Among the first benzodiazepines identified in controlled substance casework were phenazepam and etizolam, followed more recently by flubromazepam, flubromazelam, bromazepam, and clonazolam. Clonazolam is a potent triazolo-analog of clonazepam, and is also known as clonitrazolam, with a 0.5 mg oral dose leading to strong sedation and amnesia.
Objective
The purpose of this presentation is to describe the identification of clonazolam in “Pinzor” tablets, and report a method for screening and confirmation of clonazolam in urine from two individuals who appeared in the emergency room following the alleged ingestion of the tablets.
Methods
Screening analysis was performed using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF) (Xevo® G2-S with an Acquity I-class UPLC, Waters®; Milford, MA) to establish the presence of drugs and their accurate mass. The method has been previously described and validated against an in house database for approximately 1200 drugs and their common metabolites, including medicinal, therapeutic and novel psychoactive substances.
Confirmatory analysis for clonazolam was performed by liquid chromatography tandem mass spectrometry (LC-MS/MS) (Agilent 6430 with a 1200 Series HPLC, Santa Clara, CA). Confirmatory methods for urine were validated according to SWGTOX guidelines, including determination of linear range, limits of detection and quantitation, precision, accuracy, stability, and interferences. Additional confirmatory analysis for clonazolam in the tablets was performed by gas chromatography mass spectrometry (GC/MS) (Agilent 5975; Santa Clara, CA).
Results
The analysis of the “Pinzor” tablets revealed the presence of a single major peak with an accurate mass of 354.0779, consistent with C17H12ClN5O2, the molecular formula for clonazolam. This identification was confirmed by verification of retention time, and GCMS analysis relative to a clonazolam reference standard.
The methods were used to screen urine samples from two subjects reporting to an emergency room with symptoms of acute intoxication. Subject 1 was a 20-year-old male, who has a reported history of anxiety and depression. He was found unconscious in his car. He admitted to taking three 0.7 mg tablets of a “Pinzor” that had been purchased on-line. Upon arrival to the hospital he had stable vital signs, but quickly became bradycardic and hypotensive. The urine drug screen (UDS) performed at the hospital had produced a positive result for benzodiazepines. We detected and confirmed the presence of clonazolam in the urine at a concentration of 15.3 ng/mL.
Subject 2 was an 18-year-old male from the same vehicle who admitted to taking two 0.7 mg “Pinzor” tablets. He described becoming drowsy after taking the pills and was awakened by emergency medical services. His vitals were significant for tachycardia and hypertension. The UDS performed in the hospital produced positive results for benzodiazepines, THC and opiates.
We detected and confirmed the presence of clonazolam in the urine at a concentration of 30.7 ng/mL.
Conclusion/Discussion
This is the first known clinical report of a clonazolam intoxication in the emergency room in the United States and confirms concerns that novel benzodiazepines may now be available in the United States. The potency of this drug creates concerns about forensic implications for its use and should be considered as a possibility in cases with positive immunoassay benzodiazepine results that do not confirm for more common benzodiazepines.
Keywords: Designer Benzodiazepines, Clonazolam, Analysis
abstract
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