Acta Medica Leopoliensia (Dec 2020)
Validation of assay method of cardiazole by highly efficient liquid chromatography
Abstract
Aim. Validation of the method of quantitative determination of Cardiazole substance by high-performing liquid chromatography technique. Materials and Methods. The object of the study was the [3-allyl-4-(41-methoxyphenyl)-3H-thiazol-2-ylidene]-(32-trifluoromethylphenyl)-amine hydrobromide (Cardiazole) which possesses cardioprotective, anti-inflammatory, analgesic, hypolipidemic, antihypoxic, and antioxidant properties. The compound is patented and involved in the development plan of "Farmak" pharmaceutical company for further preclinical studies workflow. High-performance liquid chromatography was used for the quantitative determination of the Cardiazole substance. Validation of the proposed methodology was performed in accordance with the requirements of the State and European Pharmacopoeias requirements. The obtained data were analyzed using Analyst 1.5.2., as well as Statistica 10.0 and Microsoft Excel software. Results and Discussions. The high-performance liquid chromatography analytical method for the quantitative determination of Cardiazole substance was validated based on the main parameters according to the pharmacopeia requirements. The specificity of the technique was confirmed by comparing the chromatograms of the comparison solution, the test solution, and the blank solution. The linearity parameters are set over the entire range of the analyzed Cardiazole concentrations. The parameters of the correctness of the 9 prepared test solutions within the range of the analytical method application have met the following criteria: requirements for statistical and practical insignificance. The study of the parameters of intra-laboratory precision of 3 tests of the same sample was carried out by two analysts on different days during one week using different measuring ware. Thus, compliance with the criteria was confirmed. The obtained results of the experimental determining of the validation characteristics confirmed the correctness of the technique while reproduced in other laboratories. The results of the stability study showed: for optimal chromatography conditions it is necessary to use a freshly prepared solution of Cardiazole (within 24 hours). Conclusions. The evaluation of the validation parameters of the method of quantitative determination of Cardiazole by high-performance liquid chromatography was carried out. It was shown that the presented method of analysis of Cardiazole meets the requirements for specificity, linearity, accuracy, precision, and stability. The validated analytical method for the quantitative determination of Cardiazole by high-performance liquid chromatography can be used for standardization of the Cardiazole substance, as well as for studying the pharmacokinetics and pharmacodynamics, bioequivalence, and pharmaceutical development of dosage forms.
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