Development and validation of technique of new antithrombotic drug quantitative determination in rats blood plasma by HPLC
Abstract
botic events, along with hypertension and ischemic heart and brain tissue are the main causes of such
life-threatening conditions such as: acute coronary syndrome, myocardial infarction, stroke.
Currently, a search of new medicines for the treatment of thrombosis and prevention is relevant. One
of the main aspects of the study of innovative drugs is to study its pharmacokinetic properties, conducted in
order to identify characteristics of absorption, distribution, conversion in the body, and excretion of a drug.
These studies are obligatory for all new drugs. The most important step in the study of pharmacokinetics is
the development and validation of bioanalytical methods of quantitative determination of the drug in the
blood plasma of laboratory animals.
The aim of this work was the development and validation of bioanalytical methods of quantitative
determination of a new innovative drug - platelet aggregation inhibitor (GRS) in the blood plasma of rats by
HPLC.
Quantitative determination of the GRS in rat plasma was carried out on the chromatograph Milichrom-A02
(CJSC "EcoNova", Novosibirsk) column measuring 75.0×2.0 mm filled with sorbent ProntoSIL
C18, 5 micron, and spectrophotometric detection at 360 nm. The mobile phase A used 0.01 M ammonium
acetate solution of pH equal to 4.0 and mobile phase B - methanol. Eluent flow rate was 0.15 ml/min, column
temperature - 35 ° C, sample injection volume - 20 microliters. Liquid-liquid acetonitrile extraction method
was used for GRS extraction from plasma in accordance with QuEChERS principle.The validation was carried out to confirm the suitability of the developed method. Selectivity of
techniques and its linearity were proven in the concentration range from 10 to 750 ng / ml. Correctness of the
method is from 93.0 to 99.9%, and its accuracy - from 95.2 to 97.1%.
The developed method can be used to quantify the GRS content in blood plasma of rats at different
pharmacokinetic studies.
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