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

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.

1. Dorofeeva OA. Kliniko-farmakologicheskie zakonomernosti terapevticheskogo dejstviya raznyx lekarstvennyx form fenazepama u bol'nyx s trevozhnymi rasstrojstvami. Avtoreferat dissertacii na soiskanie uchenoj stepeni kandidata medicinskix nauk. Moskva, 2009; 24 s. Russian.

2. Kalekin RA. Sudebno-medicinskaya (ximicheskaya) ocenka nejroleptikov. Sudebno-ximicheskoe issledovanie i toksikologicheskaya xarakteristika nejroleptikov proizvodnyx benzamida. LAP LAMBERT. Academic Publishing GmbH & Co., 2012; 110 s. Russian.

3. Kalekin RA, Salomatin EM, Kalekina VA, Volkova AA. Laboratornaya diagnostika otravlenij nejroleptikami proizvodnymi benzamida v narkologii: vozmozhnosti i problemy. Narkologiya. 2008; 7 (4–76): 33–37. Russian.

4. Kalekin RA, Salomatin EM, Kalekina VA, Volkova AA. Problemy i oshibki v sudebno-ximicheskix issledovaniyax na narkoticheskie i psixotropnye veshhestva. V sbornike: V. V. Kolkutin, redaktor. O problemnyx voprosax v organizacii proizvodstva sudebno-medicinskix ehkspertiz. Materialy Vserossijskoj nauchno-prakticheskoj konferencii. 2009; s. 313–317. Russian.

5. Kalekin RA, Salomatin EM, Kalekina VA. Perspektivy razrabotki sudebno-ximicheskogo i ximiko-toksikologicheskogo analiza nejroleptikov zameshhennyx benzamidov. V sbornike: V. A. Klevno, redaktor. Sovremennye problemy mediko-kriminalisticheskix, sudebno-ximicheskix i ximiko-toksikologicheskix ehkspertnyx issledovanij. Sbornik materialov Vsesoyuznoj nauchno-prakticheskoj konferencii, posvyashhennoj pamyati professora Yu.M. Kubickogo. 2007; s. 212–218. Russian.

6. Instrukciya po medicinskomu primeneniyu lekarstvennogo preparata Fenazepam LP-005121-191018. Russian.

7. Orlova AM, Kalekin RA, Volkova AA, Nevmyatova SR, Polushkina NV. Obnaruzhenie klobazama v moche metodom tonkoslojnoj xromatografii. Vestnik VGU, Ximiya. Biologiya. Farmaciya. 2021; 4: 18–24. Russian.

8. Orlova AM, Kalekin RA, Savchuk SA. Osobennosti issledovaniya pregabalina dlya celej i zadach sudebno-ximicheskix i ximiko-toksikologicheskix issledovanij. Sudebnaya medicina. 2018; 4 (S1): 109. Russian.

9. Porseva NYu, Dvorskaya ON. Pravovye aspekty oborota snotvornyx preparatov, sposobnyx vyzyvat' lekarstvennuyu zavisimost'. Sovremennye problemy nauki i obrazovaniya. 2015; 3: 242. Russian.

10. Instrukciya po medicinskomu primeneniyu lekarstvennogo preparata Fenazepam R #003672/01-160609. Russian.

11. Instrukciya po medicinskomu primeneniyu lekarstvennogo preparata Fenazepam LSR-001772/09-100309. Russian.

12. Orlova AM, Kalekin RA, Pavlova AZ. Sudebno-medicinskaya ehkspertiza veshhestvennyx dokazatel'stv i vozmozhnosti issledovaniya pregabalina pri sudebno-ximicheskom i sudebno-toksikologicheskom issledovanii v kombinacii s drugimi sil'nodejstvuyushhimi veshhestvami. Sudebno-medicinskaya nauka i praktika. Materialy nauchno-prakticheskoj konferencii molodyx uchenyx i specialistov. (28 noyabrya, 2018). M.: ANO IC, YurInfoZdrav, 2019; 13: 97–100. Russian.

13. Kalekin RA, Pavlova AZ, Orlova AM, Bulygina IE. K voprosu ustanovleniya fakta otravleniya psixotropnymi i sil'nodejstvuyushhimi veshhestvami v biologicheskix ob"ektax ne invazivnogo otbora. Aktual'nye voprosy sudebnoj mediciny i obshhej patologii. Mater. nauch. prak. konf. Dekabr'skie chteniya v RUDN (21 dekabrya, 2018). M., 2019; s. 80–84. Russian.

14. Kalekin RA, Salomatin EM, Kalekina VA, Volkova AA. Differenciaciya nejroleptikov gruppy proizvodnyx benzamidov metodom vysokoehffektivnoj zhidkostnoj xromatografii (VEhZhX). V sbornike: Materialy Vserossijskogo soveshhaniya sudebno-medicinskix ehkspertov po primeneniyu pravil i medicinskix kriteriev opredeleniya stepeni tyazhesti vreda, prichinennogo zdorov'yu cheloveka. 2008; s. 226–8. Russian.

15. Helfer AG, Michely JA, Weber AA, Meyer MR, Maurer HH. Liquid chromatography-high resolution-tandem mass spectrometry using Orbitrap technology for comprehensive screening to detect drugs and their metabolites in blood plasma. Analytica Chimica Acta. 2017; 965: 83–95. Available from: https://doi.org/10.1016/j.aca.2017.03.002.

16. Helfer AG, Michely JA, Weber AA, Meyer MR, Maurer HH. Orbitrap technology for comprehensive metabolite-based liquid chromatographic-high resolution-tandem mass spectrometric urine drug screening — exemplified for cardiovascular drugs. Analytica Chimica Acta. 2015; 891: 221–33.

17. Barsegyan SS, Salomatin EM, Pleteneva TV, Maksimova TV, Dolinkin AO. V sbornike: R. A. Kalyokin, redaktor. Metodicheskie rekomendacii po validacii analiticheskix metodik, ispol'zuemyx v sudebno-ximicheskom i ximiko-toksikologicheskom analize biologicheskogo materiala. Metodicheskie rekomendacii. M.: RCSMEh, 2014. Russian.