PROCEDURE FOR THE STUDY OF VALIDATION PARAMETER “DILUTION INTEGRITY” OF ANALYTICAL METHODOLOGY OF QUANTITATIVE DETERMINATION OF SOME ANTIHYPERTENSIVE ANALYTES IN BIOLOGICAL LIQUIDS FOR PHARMACOKINETIC STUDIES
Introduction. Bioanalytical methodology is the founder of research on bioequivalence and therapeutic drug monitoring, so the focus of the study of pharmacokinetics and bioequivalence should be given to the development and validation of bioanalytical methods of analysis.
The aim of the study – to learn experimentally the validation parameter “dilution integrity” of analytical methods for quantitative determination of amlodipine, bisoprolol and enalapril in blood plasma for pharmacokinetic studies.
Research Methods. The bioanalytical method for the determination of amlodipine, bisoprolol and enalapril is based on HPLC/MS/MS analysis of analytes in investigated solutions obtained from plasma samples after pre-precipitation of proteins.
Results and Discussion. This paper describes the validation parameter “ dilution integrity”. Developed electronic protocols using Microsoft Excel, which provide fields for data entry. To develop a procedure for determining and evaluating dilution integrity a model experiment was performed. In this work, the validation parameter “dilution integrity” is described. We developed electronic protocols using Microsoft Exсel, which provides fields for data entry. To develop a procedure for determining and evaluating dilution integrity we performed a model experiment. Blank samples of plasma diluted 2- and 4-fold dilutions with six determinations. The dilution of the samples did not affect the accuracy and precision of the developed methodology. Validation parameter "dilution integrity" is undeservedly deprived of attention in the guides on the validation of bioanalytical techniques – the possibility of dilution of samples allows narrowing the range of application of the method and apply for calibration a simpler model and receive better indicators of linearity, correctness and precision; to develop a technique for the field of low concentrations and use it as a universal one.
Conclusions. The study “dilution integrity” of the quantitative determination of amlodipine, bisoprolol and enalapril in blood plasma for pharmacokinetic studies was researched. Conclusion regarding the developed method according to the validation parameter “dilution integrity” is correct.
Braggio, S., Barnaby, R.J., Grosi, P., & Cugola, M. (1996). A strategy for validation of bioanalytical methods. Journal of Pharmaceutical and Biomedical Analysis, 14 (4), 375-388.
Singh, U.K., Pandey, P., & Keshri, P.K. (2000). Bioanlytical method development and validation. Biorg. Chem., 2, 34-45.
Causon, R. (1997). Validation of chromatographic methods in biomedical analysis viewpoint and discussion. J. Chromatogr. B., 689 (1), 175-180.
Sharma, A., & Rathore, S. (2012). Bioanalytical method development and validation of drugs in biological fluid. Int. J. of Pharm & Research Sci., 1 (4), 216-226.
James, C.A., Breda, M., & Frigerio, E. (2004). Bioanalytical method validation: a risk-based approach. Journal of Pharmaceutical and Biomedical Analysis, 35 (4), 887-889.
Murugan, S., Pravallika, N., Sirisha, P., & Chandrakala, K. (2013). Bioanalytical method development and validation by using LC-MS/MS. Journal of Chemical and Pharmaceutical Sciences, 6 (1), 41-45.
(2013). Validatsiya bioanaliticheskogo metoda: metod. rekomendatsii [Validation of the bioanalytical method: method. recommendations]. Kyiv: GP “Gosudarstvennyy ekspertnyy tsentr” [in Ukrainian].
Logoyda, L., Korobko, D., & Kovalenko, S. (2018). The methods for determination of combination antihypertensive drugs in human plasma by HPLC MS/MS. 23nd Dubai International Pharmaceuticals & Technologies Conference & Exhibition, (February 27 – March 01), Dubai, p. 40.