Top

Published in:

01-07-2015 | Short Communication

Characterization of the four designer benzodiazepines clonazolam, deschloroetizolam, flubromazolam, and meclonazepam, and identification of their in vitro metabolites

Authors: Laura M. Huppertz, Philippe Bisel, Folker Westphal, Florian Franz, Volker Auwärter, Bjoern Moosmann

Published in: Forensic Toxicology | Issue 2/2015

Abstract

In 2012, the first designer benzodiazepines were offered in Internet shops as an alternative to prescription-only benzodiazepines. Soon after these compounds were scheduled in different countries, new substances such as clonazolam, deschloroetizolam, flubromazolam, and meclonazepam started to emerge. This article presents the characterization of these four designer benzodiazepines using nuclear magnetic resonance spectroscopy, gas chromatography–electron ionization-mass spectrometry, liquid chromatography–tandem mass spectrometry, liquid chromatography–quadrupole time-of-flight-mass spectrometry, and infrared spectroscopy. The major in vitro phase I metabolites of the substances were investigated using human liver microsomes. At least one monohydroxylated metabolite was identified for each compound. Dihydroxylated metabolites were found for deschloroetizolam and flubromazolam. For clonazolam and meclonazepam, signals at mass-to-charge ratios corresponding to the reduction of the nitro group to an amine were observed. Desalkylations, dehalogenations, or carboxylations were not observed for any of the compounds investigated. Furthermore, for clonazolam and meclonazepam, no metabolites formed by a combination of reduction and mono-/dihydroxylation were detected. This knowledge will help to analyze these drugs in biological samples.

Available only for authorised users

Kudo K, Usumoto Y, Usui K, Hayashida M, Kurisaki E, Saka K, Tsuji A, Ikeda N (2014) Rapid and simultaneous extraction of acidic and basic drugs from human whole blood for reliable semi-quantitative NAGINATA drug screening by GC–MS. Forensic Toxicol 32:97–104CrossRef

Wood AJ, Shader RI, Greenblatt DJ (1993) Use of benzodiazepines in anxiety disorders. N Engl J Med 328:1398–1405CrossRef

Browne TR, Penry JK (1973) Benzodiazepines in the treatment of epilepsy. A review. Epilepsia 14:277–310PubMedCrossRef

O'Brien CP (2005) Benzodiazepine use, abuse, and dependence. J Clin Psychiatry 66(Suppl 2):28–33PubMed

Huppertz LM, Kneisel S, Auwärter V, Kempf J (2014) A comprehensive library-based, automated screening procedure for 46 synthetic cannabinoids in serum employing liquid chromatography-quadrupole ion trap mass spectrometry with high-temperature electrospray ionization. J Mass Spectrom 49:117–127PubMedCrossRef

Uchiyama N, Shimokawa Y, Kawamura M, Kikura-Hanajiri R, Hakamatsuka T (2014) Chemical analysis of a benzofuran derivative, 2-(2-ethylaminopropyl)benzofuran (2-EAPB), eight synthetic cannabinoids, five cathinone derivatives, and five other designer drugs newly detected in illegal products. Forensic Toxicol 32:266–281CrossRef

Moosmann B, Hutter M, Huppertz LM, Ferlaino S, Redlingshöfer L, Auwärter V (2013) Characterization of the designer benzodiazepine pyrazolam and its detectability in human serum and urine. Forensic Toxicol 31:263–271CrossRef

Moosmann B, Huppertz LM, Hutter M, Buchwald A, Ferlaino S, Auwärter V (2013) Detection and identification of the designer benzodiazepine flubromazepam and preliminary data on its metabolism and pharmacokinetics. J Mass Spectrom 48:1150–1159PubMedCrossRef

Moosmann B, Bisel P, Auwärter V (2014) Characterization of the designer benzodiazepine diclazepam and preliminary data on its metabolism and pharmacokinetics. Drug Test Anal 6:757–763PubMedCrossRef

10.

Hester JB, Rudzik AD, Kamdar BV (1971) 6-Phenyl-4H-s-triazolo[4,3-a][1,4]benzodiazepines which have central nervous system depressant activity. J Med Chem 14:1078–1081PubMedCrossRef

11.

Abdul-Ghani R, Loutfy N, Hassan A (2009) Experimentally promising antischistosomal drugs: a review of some drug candidates not reaching the clinical use. Parasitol Res 105:899–906PubMedCrossRef

12.

Ansseau M, Doumont A, Thiry D, von Frenckell R, Collard J (1985) Initial study of methylclonazepam in generalized anxiety disorder. Psychopharmacology 87:130–135PubMedCrossRef

13.

Darragh A, Scully M, Lambe R, Brick I, O'Boyle C, Downie WW (1981) Investigation in man of the efficacy of a benzodiazepine antagonist, Ro 15–1788. Lancet 318:8–10CrossRef

14.

Abe M, Mikashima H, Moriwaki M, Tahara T (1988) Thieno(triazolo)diazepine compound and medicinal application of the same. Patent WO1988009333A1

15.

Sternbach LH, Randall LO, Banziger R, Lehr H (1968) Structure–activity relationships in the 1,4-benzodiazepine series. In: Burger A (ed) Drugs affecting the central nervous system, vol 2. Edward Arnold, London

16.

Bradley CM, Nicholson AN (1984) Activity of the chloro- and triazolo-benzodiazepines: behavioural studies in the monkey (Macaca mulatta). Neuropharmacology 23:327–331PubMedCrossRef

17.

Hester JB, Von Voigtlander P (1979) 6-Aryl-4H-s-triazolo[4,3-a][1,4]benzodiazepines. Influence of 1-substitution on pharmacological activity. J Med Chem 22:1390–1398PubMedCrossRef

18.

Cayman Chemical (2014) Cayman Spectral library. https://www.caymanchem.com/app/template/SpectralLibrary. Accessed December 2014

19.

Maurer HH, Pfleger K, Weber AA (2011) Mass spectral and GC data of drugs, poisons, pesticides, pollutants and their metabolites, 4th edn. Wiley-VCH, Weinheim

20.

Nakamae T, Shinozuka T, Sasaki C, Ogamo A, Murakami-Hashimoto C, Irie W, Terada M, Nakamura S, Furukawa M, Kurihara K (2008) Case report: etizolam and its major metabolites in two unnatural death cases. Forensic Sci Int 182:e1–e6PubMedCrossRef

Title: Characterization of the four designer benzodiazepines clonazolam, deschloroetizolam, flubromazolam, and meclonazepam, and identification of their in vitro metabolites
Authors: Laura M. Huppertz
Philippe Bisel
Folker Westphal
Florian Franz
Volker Auwärter
Bjoern Moosmann
Publication date: 01-07-2015
Publisher: Springer Japan
Published in: Forensic Toxicology / Issue 2/2015
Print ISSN: 1860-8965
Electronic ISSN: 1860-8973
DOI: https://doi.org/10.1007/s11419-015-0277-6

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Characterization of the four designer benzodiazepines clonazolam, deschloroetizolam, flubromazolam, and meclonazepam, and identification of their in vitro metabolites

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 2/2015

Identification and quantitation of 5-fluoro-ADB-PINACA and MAB-CHMINACA in dubious herbal products

Evaluation of the neurochemical effects of methoxetamine using brain microdialysis in mice

Postmortem distribution of MAB-CHMINACA in body fluids and solid tissues of a human cadaver

A case of fatal intoxication due to the new designer drug 25B-NBOMe

Structure–activity relationships of synthetic cannabinoid designer drug RCS-4 and its regioisomers and C4 homologues

Simultaneous plasma and oral fluid morphine and codeine concentrations after controlled administration of poppy seeds with known opiate content