STUDY OF THE VALIDATION PARAMETER “LINEARITY /CALIBRATION MODEL” OF ANALYTICAL METHODOLOGY OF QUANTITATIVE DETERMINATION OF KARDIAZOL IN BIOLOGICAL LIQUIDS FOR PHARMACOKINETIC STUDIES
Introduction. Preclinical and clinical trials are one of the most important stages in the development of medicinal products, the proper conduct of which ensures in the future the safety and high therapeutic effectiveness of the developed medicines. A key element of preclinical research is a variety of pharmacological methods, in which a number of analytical measurements are carried out on certain biological objects. Thus, issues related to the identification of the peculiarities of the validation process of bioanalytical methods used in preclinical pharmacological research of medicinal products and the development of standardized approaches to such validation work for original substances become relevant.
The aim of the study – to make an experimental study of the validation parameter “linearity/calibration model” for quantitative determination of Кardiazol in human plasma for pharmacokinetic studies.
Research Methods. The bioanalytical method for the determination of Кardiazol 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. When constructing a calibration curve, the following conditions must be fulfilled: for lower limit of quantitative determination, the deviation from the nominal concentration should be no more than ± 20 %; for calibration solutions with concentrations more than lower limit of quantitative determination, 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 cardiogenol in the concentration range of 1 ng/ml – 100 ng/ml. The regression equation is y = 0.0141x + -0.00146, the correlation coefficient is r2 0.9985.
Conclusion. The conclusion regarding the developed methodology according to the validation parameter “linearity/calibration model” is correct.
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