Detection of ultra-low concentrations of bromodihydrochlorophenylbenzodiazepine (phenazepam) and its metabolites in biological objects

Volkova AA1,2, Kalekin RA1,2, Moskaleva NE1,3, Astashkina OG1,4, Orlova AM1, Markin PA1,3
About authors

1 Russian Center of Forensic Medical Expertise, Moscow, Russia

2 Peoples’ Friendship University of Russia, Moscow, Russia

3 Sechenov First Moscow State Medical University, Moscow, Russia

4 Bureau of Forensic Medical Examination, Moscow, Russia

Correspondence should be addressed: Roman A. Kalekin
Polikarpova, 12/13, Moscow, 125284, Russia; ur.ems-cr@ajimih

About paper

Author contribution: Volkova AA, Kalekin RA, Orlova AM, Astashkina OG — data acquisition, manuscript writing; Volkova AA, Kalekin RA, Moskaleva NE, Markin PA — experimental procedure.

Compliance with ethical standards: the study was planned and conducted in accordance with the requirements of the Declaration of Helsinki of the World Medical Association (2000) and subsequent revisions thereto; the informed consent was submitted by all study participants.

Received: 2022-02-23 Accepted: 2022-03-11 Published online: 2022-03-25

In extreme situations, reliable detection of the minimum therapeutic concentrations of psychotropic substances is important, since this allows one to provide adequate resuscitation. The group of benzodiazepine derivatives, which includes bromodihydrochlorophenylbenzodiazepine (phenazepam), is widely used in clinical practice. Along with the positive clinical effect, phenazepam has numerous side effects, capable of causing poisoning, even death. The study was aimed to develop the method for detection of the phenazepam metabolites by high-resolution HPLC–TMS suitable for achieving the aims and objectives of forensic medical expertise in case of the ultra-low urine substance concentrations. Urine of six patients (males and females aged 28–40), who were prescribed phenazepam and took the drug at minimum therapeutic concentrations on an ad hoc basis, was used during the study. Optimum conditions for the analyte chromatography after the urine sample preparation were defined with the phenazepam retention time of 7.05 ± 0.06 min; specific ions (m/z) 179, 183, 206, 242, 271, 285, 320, 348 (main) were defined for identification of phenazepam.

Keywords: high-performance liquid chromatography, phenazepam, bromodihydrochlorophenylbenzodiazepine, 3-hydroxyphenazepam, forensic chemistry research, chemical toxicological analysis, urine