Chemical and Toxicological Diagnosis of Acute Poisonings with Phenazepam
https://doi.org/10.23934/2223-9022-2018-7-4-319-324
Abstract
Background. The relative availability of Phenazepam makes it a frequent cause of overdose, suicide and non-medical use. At the same time, it remains insufficiently studied in chemical and toxicological terms.
The aim of study. to create an accessible, rapid method for detecting Phenazepam in biological matrices of patients with acute poisoning.
Materials and methods. We used thin-layer chromatography (TLC), gas chromatography with a mass selective detector (GC-MS), high performance liquid chromatography with a tandem mass-selective detector (LC-MS/MS) and immunochromatographic analysis (ICA). The preparation of samples of intact urine with the addition of standard solutions of Phenazepam and real urine samples of patients with acute poisoning with Phenazepam was carried out using liquid-liquid extraction or precipitation of related components of the sample with acetonitrile. Hydrolysis and derivatization were also added in GC-MS analysis.
Results. The analysis of statistics of the Department of Acute Poisonings of the N.V. Sklifosovsky Research Institute for Emergency Medicine in 2014-2016 showed that Phenazepam poisonings averaged 9.2% of the total number of admissions and mainly occurred as suicidal attempts. A technique has been developed for the detection of Phenazepam by TLC, which gives more objective results than ICA. For confirmatory analysis, it is advisable to use LC-MS/MS method for the native substance and GC-MS for the products of hydrolysis after derivatization. Compared to confirmatory methods, the developed TLC-screening technique is expressive, does not require the use of expensive high-tech equipment, difficult sample preparation, and makes it possible to reliably detect toxic and lethal concentrations of Phenazepam.
About the Authors
M. V. BelovaRussian Federation
Belova Maria Vladimirovna - Docent, Doctor of Biological Sciences Leading Researcher N.V. Sklifosovsky RIEM, Associate professor I.M. Sechenov FM SMU.
3 Bolshaya Sukharevskaya Square, Moscow 129090; 2 Bolshaya Pirogovskaya St., b. 4, Moscow 119991
E. A. Klyuyev
Russian Federation
Klyuyev Evgeny Aleksandrovich - Clinical Laboratory Physician.
3 Bolshaya Sukharevskaya Square, Moscow 129090
E. S. Melnikov
Russian Federation
Melnikov Evgeny Sergeyevich - Candidate of Pharm. Sci., Assistant of the Department of Pharmaceutical and Toxicological Chemistry.
2 Bolshaya Pirogovskaya St., b. 4, Moscow 119991
D. M. Yeliseyeva
Russian Federation
Yeliseyeva Daria Mikhaylovna - student of Pharmacy Faculty, Department of Pharmaceutic and Toxicological Chemistry
2 Bolshaya Pirogovskaya St., b. 4, Moscow 119991
References
1. Avedisova A.S. On the issue of dependence on benzodiazepines. Psikhiatriya i psikhofarmakoterapiya. 1999; (1): 12-15. (In Russian).
2. Phenazepam. Available at: http://www.netnarkotik.ru/fenazepam (Accessed 13 Feb 2018) (In Russian).
3. In Russia preoccupied statistics application fenazepama “without appointment”. Available at: https://regnum.ru/news/2356075.html (Accessed 13 Feb 2018) (In Russian).
4. Kartashov V.A., Chernova L.V., Velichko G.P. TLC-screening and toxic substances retention indices. Novyye tekhnologii. 2012; (2): 240-247. (In Russian).
5. Pospelova A.A., Karpova L.N., Malkova T.L. The estimation of a possible use of reversed-phase thin-layer chromatography in chemo-toxicological studies of certain groups of pharmaceutical and chemical compounds. Toksikologicheskiy vestnik. 2012; (2): 20-23. (In Russian).
6. Saitman A., Park H. D., Fitzgerald R. L. False-positive interferences of common urine drug screen immunoassays: a review. JAnalytic Toxicol. 2014; 38(7): 387-396. PMID: 24986836. DOI: 10.1093/jat/bku075.
7. Mikel C., Pesce A.J, Rosenthal M., West C. Therapeutic monitoring of benzodiazepines in the management of pain: current limitations of point of care immunoassays suggest testing by mass spectrometry to assure accuracy and improve patient safety. Clin Chim Acta. 2012; 413(15-16): 11991202. PMID: 22484396. DOI: 10.1016/j.cca.2012.03.017.
8. Bertol E., Vaiano F., Borsotti M., et al. Comparison of immunoassay screening tests and LC-MS-MS for urine detection of benzodiazepines and their metabolites: results of a national proficiency test. J Analytic Toxicol. 2013; 37(9): 659-664. PMID: 23943436. DOI: 10.1093/jat/bkt063.
9. Yegles M., Mersch F., Wennig R. Detection of benzodiazepines and other psychotropic drugs in human hair by GC/MS. Forens Sci Int. 1997; 84(1-3): 211-218.
10. Maurer H., Pfleger K. Identification and differentiation of benzodiazepines and their metabolites in urine by computerized gas chromatography— mass spectrometry. J Chromatogr. 1987; 422: 85-101. PMID: 3437028.
11. Valentine J.L., Middleton R., Sparks C. Identification of urinary benzodiazepines and their metabolites: comparison of automated HPLC and GC-MS after immunoassay screening of clinical specimens. J Analytic Toxicol. 1996; 20(6): 416-424. PMID: 8889678.
12. Kratzsch C., Tenberken O., Peters F.T., et al. Screening, library-assisted identification and validated quantification of 23 benzodiazepines, flumazenil, zaleplone, zolpidem and zopiclone in plasma by liquid chromatography/mass spectrometry with atmospheric pressure chemical ionization. J Mass Spectrom. 2004; 39(8): 856-872. PMID: 15329838.
13. Quintela O., Sauvage F.L., Charvier F., et al. Liquid chromatography-tandem mass spectrometry for detection of low concentrations of 21 benzodiazepines, metabolites, and analogs in urine: method with forensic applications. Clin Chem. 2006; 52(7): 1346-1355. PMID: 16709625. DOI: 10.1373/clinchem.2005.065631.
14. Belova M.V., Lisovik Zh.A., Klyuyev A.E., et al. Chemical-toxicological analysis of acute chemical poisoning. Moscow: Grafikon Print, 2007: 66-67, 75-79. (In Russian).
15. Arzamastsev A.P., Ramenskaya G.V., Rodionova G.M., Kuznetsova N.I., Petukhov A.E. TLC screening of toxicologically significant compounds, isolated by extraction and sorption. Moscow: GEOTAR-Media Publ., 2010: 128-132. (In Russian).
16. Rust K. Y., Baumgartner M.R., Meggiolaro N.Kraemer T. Detection and validated quantification of 21 benzodiazepines and 3 “z-drugs” in human hair by LC-MS/MS //Forensic science international. 2012; 215(1-3): 64-72.
Review
For citations:
Belova M.V., Klyuyev E.A., Melnikov E.S., Yeliseyeva D.M. Chemical and Toxicological Diagnosis of Acute Poisonings with Phenazepam. Russian Sklifosovsky Journal "Emergency Medical Care". 2018;7(4):319-324. https://doi.org/10.23934/2223-9022-2018-7-4-319-324