OBJECTIVE: To establish a method for the plasma concentration determination of carboplatin, and study the pharmacokinetics of carboplatin in female rats after intravenous injection and intraperitoneal injection. METHODS: HPLC was performed on the column of Agilent TC-C18 with mobile phase of methanol-water (5 ∶ 95, V/V) at a flow rate of 1.0 mL/min, detection wavelength was 229 nm, and column temperature was 25 ℃. The inner standard was 5-bromouracil, and injection volume was 20 μL. 24 SD rats were randomly divided into 4 groups, 6 in each group. The rats were intravenously injected and intraperitoneally injected carboplatin 20, 40 mg/kg respectively. 0.5 mL blood sample was taken from eyes before administration and after administration of 0.25, 0.5, 1, 1.5, 2, 4, 6, 8, 10, 12 h. The plasma concentration of carboplatin was determined, and DAS 2.0 was used to calculate the pharmacokinetic parameters. RESULTS: The linear range of carboplatin in plasma was 0.30-60.00 μg/mL (r=0.999 1); RSDs of intra-day, inter-day precision were lower than 10% (n=5); RSD of peak area in stability test was lower than 10% (n=5); method recovery was 98.7%-102.4% (RSD≤6.08%, n=5), and extraction recovery was 83.38%-85.45% (RSD≤5.97%, n=5). AUC0-12 h of carboplatin 20, 40 mg/kg by intravenous injection and intraperitoneal injection in female rats were (15.503±4.172), (23.402±4.266), (6.716±2.306), (9.384±2.205) μg·h/mL; AUC0-∞ were (16.424±4.846), (23.404±4.266), (6.790±2.378), (9.765±2.095) μg·h/mL; t1/2z were (1.246±0.765), (0.394±0.058), (0.513±0.156), (0.884±0.460) h; and tmax were (0.700±0.274), (0.400±0.335), (0.542±0.368), (0.833±0.289) h, respectively. CONCLUSIONS: The method is simple, economic and accurate, with suitable internal standard, and can be used for the plasma concentration determination of carboplatin in female rats and the pharmacokinetic studies.