We aimed to build population pharmacokinetics of cefuroxime lysine in normal, liver injury, and kidney injury rats. After intravenous injection of 67.5 mg/kg cefuroxime lysine, the plasma samples were collected at specific time points and determined by a validated ultra fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method. Non-compartmental analysis (NCA) method was applied to analyze the data using DAS software, and the AUC and MRT of renal-impaired group were significant higher than these of normal group. The first-order conditional estimation with interaction (FOCE-I) approach was used to build the population model by NONMEM software. The basic pharmacokinetic model was best described by a two-compartment model. Renal function has significant impact on the drug clearance (Cl) and uea was incorporated in the final model. The typical population pharmacokinetic parameters of cefuroxime lysine in all rats were as follows: Cl: 0.87 L/h/kg, central volume of distribution (V-c): 039 L/kg, peripheral volume of distribution (V-p): 0.27 L/kg, clearance between central and peripheral compartments (Q): 0.13 L/h/kg. relational coefficient of urea on Cl (f(Urea)): 0.89. This study demonstrated that renal impairment attended to reduce renal clearance significantly, which should be paid more attention for the proper dosage regimen for patients in kidney injury.