METHODS OF ENZYMATIC HYDROLYSIS FOR BLOOD SAMPLE PREPARATION FOR LABORATORY DIAGNOSTICS OF POISONING WITH GAMMA-AMINO BUTYRIC ACID DERIVATIVES
DOI:
https://doi.org/10.11603/mcch.2410-681X.2022.i4.13577Keywords:
gamma-aminobutyric acid derivatives, blood, enzymatic hydrolysis, proteases, hyaluronidase, validation parameteAbstract
Introduction. New potentially dangerous psychoactive substances, which are not included in the list of narcotic drugs and psychotropic substances, the circulation of which is prohibited or restricted in Ukraine, include ten groups of substances of different structures, among which a group of GABA-A and GABA-B receptor agonists and their derivatives can be distinguished. Abuse of new dangerous psychoactive substances, including GABA derivatives, has been recorded in 94 countries of the world, including Ukraine. In connection with the recent increase in the number of people addicted to these drugs, as well as the number of acute and even fatal poisonings, the issue of chemical and toxicological research, which includes joint detection and quantitative determination of analyzed substances in biological bodies, is becoming more and more relevant projects.
The aim of the study – to develop methods for the isolation and determination of GABA derivatives (phenibut and baclofen) in biological fluids (blood).
Research Methods. The research was conducted using the substance phenibut - (±)-4-amino-3-phenylbutanoic acid hydrochloride and baclofen tablets. Enzymes were used for hydrolysis: papain, chymotrypsin, chymopsin and hyaluronidase, trilon B substance, cysteine substance
Results and Discussion. A significant difficulty in conducting research on biological objects for GABA derivatives (phenibut, pregabalin, gabapentin, baclofen, etc.) is that these substances are able to produce internal salt in the environment of biological fluids, which significantly complicates conducting laboratory studies on them. In our work, we used the method of extraction freezing, determining the selective conditions, pH 2 with acetonitrile at a temperature of -18 ... -20 C.
Conclusion. In the course of the conducted research, it was shown that the method of enzymatic hydrolysis by proteases of low specificity can be recommended for blood sample preparation in the diagnosis of non-medicinal use of drugs from the group of GABA derivatives.
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