Development and validation of a methodology for HPLC analysis of clopidogrel

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Abstract

Aim – to develop a methodology for the chemical and toxicological analysis of clopidogrel using HPLC.

Material and methods. The study was performed on Clopidogrel bisulfate substance-powder, manufacturer: PJSC "Farmak", Ukraine, corresponding to the European Pharmacopoeia with an active substance content of 99.31% (LM2504208 series). For the chromatographic analysis we used a micro-column liquid chromatograph "Milichrome A-02" (CJSC "Econova", Russia). The analysis was done according to the unified HPLC technique developed by G.I. Baram.

Results. The highest spectral ratio by HPLC was observed at 280 nm / 210 nm (maximum clopidogrel peak area R = 1.0787). The calibration curve for the HPLC method was described by the equation: у = 0.0005912⋅х – 0.001385 (r = 0.9998; S02 = 25.29⋅10-7); linearity within 1−400 mcg/ml, which corresponded to the content of clopidogrel in the sample from 2 ng to 800 ng, respectively. Based on the values of the calibration curve, the LOD and LOQ were calculated: 2,334 mcg/ml and 7,074 mcg/ml, respectively.

Conclusion. A technique for the quantitative determination of clopidogrel by HPLC has been developed that allows for determining the substance in a mixture. The methodology was validated according to the following criteria: specificity, limit of detection, limit of quantitative determination, linearity, correctness and precision.

About the authors

Ludmila S. Anosova

Donetsk National Medical University named after M. Gorky

Author for correspondence.
Email: Apteka-NaNya@yandex.ru
ORCID iD: 0000-0002-9380-4619

Assistant of the Department of Pharmaceutical and Medical Chemistry

Russian Federation, Donetsk

Irina P. Remezova

Pyatigorsk Medical and Pharmaceutical Institute – branch of the Volgograd State Medical University

Email: i.p.remezova@pmedpharm.ru
ORCID iD: 0000-0003-3456-8553

PhD, Professor, Department of Toxicological and Analytical Chemistry

Russian Federation, Pyatigorsk

Alexey M. Agafonov

Donetsk National Medical University named after M. Gorky

Email: Chuh2008@yandex.ru
ORCID iD: 0000-0002-0682-1366

Assistant of the Department of Pharmaceutical and Medical Chemistry

Russian Federation, Donetsk

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5. Figure 1. Chromatogram of clopidogrel (concentration 10 μg/ml).

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6. Figure 2. A calibration curve of the quantitative determination of clopidogrel by HPLC (λ=280 nm, sample volume 2 µl).

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Copyright (c) 2022 Anosova L.S., Remezova I.P., Agafonov A.M.

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