Novel oral anticoagulants (NOACs) are, as an alternative therapy to vitamin K antagonists, used frequently to treat and prevent thromboembolism. Their precise quantitation is necessary to identify the presence/absence of an anticoagulant effect or to determine the concentration of drug that may be helpful for patient management. Furthermore, anticoagulants screening is required for interventional emergency, emergency bleeding and programmed surgery for the elderly. Such analysis is mainly done by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). To streamline the workflow, we developed a complete solution including stable isotope labeled standards for better precision and accuracy, and investigated the use of a fully automated sample preparation system coupled online with LC-MS/MS.