Development of a UV spectrophotometric method for quantitative determination of a clopidogrel metabolite suitable for chemical and toxicological analysis

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Abstract

Introduction. In Russia, more than 50% of all diseases and a fifth of all cases of disability are due to coronary heart disease and stroke. According to the recommendations of the Russian Society of Cardiology (RKO) and the European Society of Cardiology, antiplatelet therapy with clopidogrel is included in the standards of treatment of patients with COVID-19. Clopidogrel is a prodrug, with the help of esterases, hydrolysis occurs with the formation of the main (inactive) circulating metabolite - a derivative of carboxylic acid, which accounts for 85% of all metabolites of this drug. According to the information given in the scientific literature over the past 10 years, clopidogrel is of interest from the point of view of chemical and toxicological studies, since this drug very often becomes the "drug of choice" for suicide purposes. Objective: to develop and validate a technique for the quantitative determination of the main metabolite of clopidogrel -clopidogrel carboxylic acid using an affordable and widely implemented chemical-toxicological analysis method of UV spectrophotometry. Material and methods. Objects of research - Clopidogrel carboxylic acid (SLA) substance-powder. The light absorption of solutions in the UV spectrum was measured with the SF-46 spectrophotometer (JSC LOMO, Russia), the spectral measurement range was from 200 to 350 nm. A standard solution of clopidogrel carboxylic acid in 0.1 M hydrochloric acid solution (100 mcg/ml) was used. Results. The optimal wavelength for the quantitative determination of the main metabolite of clopidogrel - clopidogrel carboxylic acid is 278 nm. The calibration graph for the UV spectrophotometric method was described by the equation: A=0.004128C - 0.008667; linearity was observed within the concentrations of clopidogrel carboxylic acid 20.0-200 mcg/ml; LOD and LOQ were, respectively, 1.655 mcg and 5.015 mcg in the sample. Conclusion. The developed method of quantitative determination of the main metabolite of clopidogrel - clopidogrel carboxylic acid using UV spectrophotometric method meets the requirements for methods recommended for use in forensic toxicology, which is confirmed by validation characteristics.

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About the authors

L. S Anosova

Donetsk National Medical University named after M. Gorky

Author for correspondence.
Email: apteka-nanya@yandex.ru

I. P Remezova

Pyatigorsk Medical and Pharmaceutical Institute - branch of the Volgograd State Medical University of the Ministry of Health of Russia

Email: i.p.remezova@pmedpharm.ru

A. M Agafonov

Donetsk National Medical University named after M. Gorky

Email: chuh2008@yandex.ru

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Supplementary files

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2. Fig. 1. Hydrolysis product, the main inactive metabolite of clopidogrel – clopidogrel carboxylic acid

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3. Fig. 2. UV spectrum of clopidogrel carboxylic acid in 0.1 M hydrochloric acid solution (concentration 100 mcg/ml; l=10 mm)

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4. Fig. 3. Calibration graph for UV spectrophotometric determination of clopidogrel carboxylic acid (λ=278 nm; l=10 mm)

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