Background: Ticagrelor is a highly recommended new antiplatelet agent for the treatment
of patients with acute coronary syndrome at moderate or high ischemic risk. There is a real need for
rapid and accurate analytical methods for ticagrelor determination in biological fluids for pharmacokinetic
studies. In this study, a sensitive and specific LC-MS method was developed and validated
for quantification of ticagrelor and its Active Metabolite (AM) in human plasma over expected clinical
Methods: Samples were handled by Liquid-Liquid Extraction (LLE). A linear gradient was applied
with a mobile phase composed of formic acid 0.1% and acetonitrile with 0.1% of formic acid using a
C18 reversed-phase column. MS spectra were obtained by electrospray ionization in negative mode
and optimized at 521.4→360.9 m/z, 477.2→361.2 m/z and 528.1→367.9 m/z transitions for ticagrelor,
AM and ticagrelor-d7, respectively.
Results: This method allowed rapid elution, in less than 4 minutes, and quantification of concentrations
as low as 2 ng/mL for ticagrelor and 1 ng/mL for AM using only 100 μL of human plasma.
LLE using hexane/ethyl acetate (50/50) was an optimal compromise in terms of extraction recovery
and endogenous compounds interference. Trueness values of 87.8% and 89.5% and precisions of
84.1% and 93.8% were obtained for ticagrelor and AM, respectively. Finally, the usefulness of the
method was assessed in a clinical trial where a single 180 mg ticagrelor was orally administered to
healthy male volunteers. Pharmacokinetic parameters of ticagrelor and its active metabolite were
Conclusion: A sensitive and specific quantification LC-MS-MS method was developed and validated
for ticagrelor and its active metabolite determination in human plasma. The method was successfully
applied to a clinical trial where a single ticagrelor 180 mg dose was orally administered to
healthy male volunteers. The described method allows quantification of concentrations as low as 2
ng/mL of ticagrelor and 1 ng/mL of the metabolite using only 100 μL of plasma.