The　metabolic　differences　between　patients　with　the　anticancer　drug　daunorubicin　(DRN)　pose　difficulties　in　providing　clinical　delivery　strategies　for　different　patients.　Therefore,　it　is　urgent　to　develop　a　pharmacokinetic　platform　that　can　real-time　monitor　drug　levels　in　vivo　to　achieve　precise　drug　delivery　and　reduce　drug　side　effects.　To　this　end,　an　electrochemical　aptamer-based　(E-AB)　sensor　was　designed　to　realize　real-time　and　continuous　monitoring　of　DRN　levels　in　vivo　and　obtain　DRN　pharmacokinetics　of　different　individuals　for　the　first.　Specifically,　the　binding　free　energy　of　the　aptamer　to　the　target　was　calculated　by　molecular　docking　simulations.　Using　nanoporous　gold　working　electrodes　with　immobilized　aptamers　as　E-AB　sensors,　which　monitored　DRN　in　the　blood　of　different　rat　models　continuously　for　many　hours　with　nanomolar　precision　and　second　resolution,　and　hundreds　of　DRN　concentration　values　were　obtained,　based　on　which　important　pharmacokinetic　parameters　were　simulated　and　computed,　including　half-life　of　drug　DRN　distribution　and　elimination,　and　peak　concentration　in　blood.　These　parameters　help　to　determine　the　metabolism　of　drug　DRN　in　different　patients　and　adjust　the　administered　dose　in　time　to　achieve　precise　treatment.　Therefore,　this　E-AB　platform　may　provide　a　valuable　new　tool　for　clinicians　to　monitor　pharmacokinetics　in　different　patients.　©　2024　Elsevier　B.V.