Novel and Nonroutine Benzodiazepines and Suvorexant by LC-MS-MS

Area of Science:

• Forensic Toxicology
• Analytical Chemistry
• Pharmacology

Background:

• Benzodiazepines are frequently prescribed but carry abuse potential.
• Increasing prevalence of novel/nonroutine benzodiazepines in drug seizures from 2017-2019 prompted method development.
• Existing methods may not adequately detect emerging benzodiazepine analogs.

Purpose of the Study:

• To develop and validate a sensitive analytical method for novel and nonroutine benzodiazepines and suvorexant in biological specimens.
• To address the growing challenge of identifying and quantifying emerging psychoactive substances.
• To provide a robust tool for forensic laboratories analyzing drug seizures.

Main Methods:

• Liquid chromatography-tandem mass spectrometry (LC-MS-MS) was employed for analysis.
• The method targets 16 novel/nonroutine benzodiazepines and suvorexant in 200 µL of whole blood.
• Analysis involved protein precipitation with an 8-minute run-time per sample.

Main Results:

• The method achieved limits of detection (LOD) of 1 or 5 ng/mL and limits of quantitation (LOQ) of 5 or 25 ng/mL.
• Quantitative analysis was validated for 15 out of 17 target analytes, with flubromazepam and prazepam validated for qualitative identification only.
• A quadratic calibration model (r² > 0.990) with 1/x weighting was utilized, with coefficients of variation within 14% and bias within ±14%.

Conclusions:

• The developed LC-MS-MS method is sensitive, efficient, and robust for detecting novel and nonroutine benzodiazepines and suvorexant.
• This method provides crucial support for forensic investigations by enabling accurate identification and quantification of emerging drug threats.
• The method's ability to analyze a wide range of analytes with minimal sample volume makes it highly practical for forensic toxicology labs.