THE STUDY OF VALIDATION PARAMETER “LINEARITY/CALIBRATION MODEL” OF ANALYTICAL METHODOLOGY OF QUANTITATIVE DETERMINATION OF UROCARB IN BLOOD PLASMA FOR PHARMACOKINETIC STUDIES
Introduction. The analysis of publications in the leading world chemistry-analytical and pharmaceutical journals allows us to conclude that the researchers are of priority interest in the validation of bioanalytical techniques, as evidenced by the constant study of validation parameters, including “linearity/calibration model”. Thus, with the definition of the validation parameter “linearity/calibration model” used in preclinical pharmacological research of medicinal products and development of standardization their approach to such validation work for the original substance.
The aim of the study – to experimentally learn the validation parameter “linearity/calibration model” for quantitative determination of urocarb in human plasma for pharmacokinetic studies.
Research Methods. The bioanalytical method for the determination of urocarb is based on HPLC/MS/MS analysis of analytes in investigated solutions obtained from plasma samples after pre-precipitation of proteins. Samples were chromatographed using Discovery C18 chromatography column, 50×2.1 mm, with a particle size of 5 μm and gradient elution.
Results and Discussion. The suitability of the bioanalytical technique was confirmed by the validation characteristics that are advanced to the bioanalytical methods. In this paper, the validation parameter “linearity/calibration model” is described. We developed electronic protocols using Microsoft Exсel, which provides fields for data entry. When constructing a calibration curve, the following conditions must be fulfilled: for lower limit of quantification (LLOQ), the deviation from the nominal concentration should be no more than ± 20 %; for calibration solutions with concentrations more than LLOQ, the deviation from the nominal concentration should be no more than ± 15 %. A linear relationship was found between the concentration and the area of the chromatographic peaks of urocarb in the concentration range of 1 ng/ml – 100 ng/ml. The regression equation is y=0.00365x+0.000177, the correlation coefficient is r2 0.9993.
Conclusions. The study of the validation parameter “linearity/calibration model” of analytical method of quantitative determination of urocarb in plasma is conducted for pharmacokinetic studies. The conclusion on the developed methodology according to the validation parameter “linearity/calibration model” is correct.
(1995). ICH Harmonised Tripartite Guideline. Validation of Analytical Procedures: Text and Methodology Q2 (R1). Geneva: ICH, 1995. 13 p.
Grizodub A. I. (2011). Standartizovanye protsedury validatsii metodik kontrolya kachestva lekarstvennykh sredstv. Analiticheskaya khimiya v sozdanii, standartizatsii i kontrole kachestva lekarstvennykh sredstv: v 3 t. [Standardized procedures for the validation of quality control methods for medical produkts. Analytical chemistry in the creation, standardization and quality control of medicines]. Georgiyevskiy, V.P. (Ed.). Kharkov: NTMT [in Russian].
(2001). Guidance for Industry: Bioanalytical Method Validation. U.S. Department of Health and Human Services, Food and Drug Administration (FDA), Center for Drug Evolution and Research (CDER), Center for Veterinary Medicines (CVM). Washington, DC: U.S. Government Printing Office.
Guideline on bioanalytical method validation. European Medicines Agency (EMEA/CHMP/EWP/ 192217/2009). 2011. Retrieved from: http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2011/08/WC500109686.pdf.
(2013). Validatsiya bioanaliticheskogo metoda: metod. rekomendatsii [Validation of the bioanalytical method: method. recommendations]. Kiev: State Expert Center [in Russian].
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.
Drapak, I. (2019). Syntez, doslidzennia diuretychnoi aktyvnosti ta QSAR-analiz N-(1,3,4-tiadiazol-2-il) zamishchenykh amidiv kyslot alkankarbonovoho riadu [Synthesis, study of diuretic activity and QSAR analysis of N-(1,3,4-thiadiazol-2-yl) substituted amides of acids of the alkanecarboxylic acid series]. Farmatsevtychnyi zhurnal – Pharmaceutical Journal, 2, 55-65 [in Ukrainian].