Validation of assay method of cardiazole by highly efficient liquid chromatography

Keywords: validation parameters, Cardiazole, high-performance 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.

References

Analyst® Software Version 1.5.2. http://www.appliedbiosystems.com

Andrade EL, Bento AF, Cavalli J. Non-clinical studies in the process of new drug development - Part II: Good laboratory practice, metabolism, pharmacokinetics, safety and dose translation to clinical studies. Braz. J Med Biol Res. 2016; 49(12):e5646. https://doi.org/10.1590/1414-431x20165646

Bagirova VL, Grizodub AI, Chibilyaev TH etc. Guide to the validation of methods of analysis of drugs ed. N.V. Jurgel. M .: Pharm. prom., 2007. 58 p. Russian

Calibration and Validation of Analytical Methods. A Sampling of Current Approaches / еd. Mark Stauffer. 2018. doi:10.5772/interchopen.69918.

Diksha Rajput, Vaishnav Rajat, Anju Goyal. Validation of Analytical Methods for Pharmaceutical Analysis. International Journal of Pharmaceutical Erudition. 2013;3(1):31-40.

Drapak IV, Seredynska NM, Pinyazhko OR et al. Synthesis and cardiotropic action of [3-allyl-4- (41-methoxyphenyl) -3H-thiazol-2-ylidene] - (32-trifluoromethylphenyl) amine hydrobromide in white rats. Pharmacology and Drug Toxicology. 2019; 13 (4): 255-262. Ukrainian

Drapak IV, Zimenkovsky BS, Seredynska NM, at al. [3-Allyl-4- (41-methoxyphenyl) -3H-thiazol-2-ylidene] - (32-trifluoromethylphenyl) amine hydrobromide (Cardiazole), which exhibits cardioprotective and antioxidant activity. Patent UA №140537; declared 07/22/2019; publ. 10.03.2020, bul. 5. Ukrainian

Drapak I, Zimenkovsky B, Bezruk I, Ivanauskas L, at al. Development of metod of quantative determination of Cardiazol substance with using highly efficient liquid chromatography. Scientific Journal "ScienceRise: Pharmaceutical Science". 2019;2(18):33-38. https://doi.org/10.15587/2519-4852.2019.165763

Drapak I, Bezruk I, Ivanauskas L. Development of assay method for analysis of a novel Kardiazol substance by HPLC. Аbstracts of 22nd international scient.-pract. Conf. Baltpharmforum - 2019. Pharmacists as drug experts: their role in health care system; 2019 April 13-14; Kaunas, Lithuania; 2019. 44

European Pharmacopoeia, 9th ed., European Directorate for the Quality of Medicines & Health Care, Strasbourg 2017.

Georgiyants VA, Evifeeva OA. Validation of analytical methods in pharmacy: theory, normative aspects, problems of practice Pharmaceutical Journal 2007. 2: 13-18. Ukrainian

Grizodub AI Standardization of the procedure for validation of methods of quality control of drugs. Analytical chemistry in the creation, standardization and quality control of drugs: in 3 volumes. Ed. Corresponding member. NAS of Ukraine VP Georgievsky. Kharkiv: NTMT, 2011, 3. 934-1063. Russian.

Gupta PC. Method Validation of Analytical Procedures. Pharma Tutor. 2015; 3(1): 32-39.

Haleem RM, Salem MY, Fatahallah FA, Abdelfattah LE. Quality in the pharmaceutical industry - a literature review. Saudi Pharmaceutical Journal. 2013; 23(5): 463-469. https://doi.org/10.1016/j.jsps.2013.11.004.

Marsona BM, Concentinoa V, Junkerta AM et al. Validation of analytical mathods in a pharmaceutical quality system: an overview focused on HPLC methods. Quim. Nova. 2020; XY:1-14, https://doi.org/10.21577/0100-4042.20170589

Medicines. Good manufacturing practice. ST-instruction of the Ministry of Health 42-4.0: 2016. Ukrainian

Nikolin B, Imamovi? B, Medanhodzi?-Vuk S, Sober M. High perfomance liquid chromatography in pharmaceutical analyses. Bosnian Journal Basic Medical Sciences. 2004; 4(2):5-9. https://doi.org/10.17305/bjbms.2004.3405

State Pharmacopoeia of Ukraine: in 3 volumes - 2nd ed. Kharkiv: State Enterprise "Ukrainian Scientific Pharmacopoeial Center for Quality of Medicines", 2015. Vol. 1, 1128.; 2014, Vol. 2, 724p. Ukrainian

Sibel A. Ozkan. Analytical Method Validation: The Importance for Pharmaceutical Analysis. Pharmaceutical Sciences. 2018;24:1-2. doi: 10.15171/PS.2018.01. https://doi.org/10.15171/PS.2018.01

Sullivan JO, Blake K, Berntgen M at al. Overview of the European Medicines Agency's Development of Product Specific Bioequivalence Guidelines. Clin Pharmacol Ther. 2018; 104(3): 539-545. doi: 10.1002/cpt.957. https://doi.org/10.1002/cpt.957

Published
2020-12-20
How to Cite
Drapak, I. (2020). Validation of assay method of cardiazole by highly efficient liquid chromatography. Acta Medica Leopoliensia, 26(4), 80-90. https://doi.org/10.25040/aml2020.04.080