Sample pretreatment and HPLC determination of anti-platelet drug ticagrelor in blood plasma from patients with acute coronary syndromes.

A simple HPLC method for the determination of anti-platelet drug ticagrelor in blood has been developed. Sample preparation and extraction conditions were investigated and optimized. The preparation of blood plasma was investigated by protein precipitation using perchloric acid, methanol, acetonitrile and trifluoroacetic acid. Protein precipitation with acetonitrile was found to be the most suitable. Chromatographic separation of ticagrelor was performed on a C18 column with a mobile phase consisted of acetonitrile and 15 mM ammonium acetate buffered at pH 8.0. The method was applied to determination of ticagrelor in blood plasma of patients who had a heart attack. Blood samples were collected 1.5 hours after the administration of the initial loading dose of anti-platelet drug. The average concentration of ticagrelor was found to be 0.97±0.53 μg/mL. The developed method proved to be very selective, without interferences from other endogenous substances and the influences of possible co-administered drugs. The limits of detection and quantification estimated by the signal-to-noise ratio in real samples were 0.24 μg/mL and 0.4 μg/mL respectively. The developed method is simple and can be easily applied in clinical conditions and emergency cardiac situations after initial loading dose of ticagrelor in the first few hours of a heart attack.