Objective To develop a reversed phase high performance liquid chromatographic (RP-HPLC) analysis system for hydroxysafflor yellow A (HSYA) in rat model of cold coagulation and blood stasis (CCBS), and to investigate the influence of Sappan lignum on the pharmacokinetics of HSYA in CCBS rats. Methods Rat CCBS models were randomly divided into two groups with each containing 6 animals. Rats were orally given Carthami flos extract or Carthami flos extract combined with Sappan lignum (The dosage: 20.0 g/kg crud drug of Carthami flos). Plasma samples were collected in heparinized tubes from the oculi chorioideae vein at 5, 10, 20, 30, 45, 60, 90, 120, 150, 210, and 270 min after drug administration; and the plasma proteins were precipitated with 20% trichloroacetic acid aqueous solution. Plasma concentrations of HSYA were detected by RP-HPLC at different time points after drug administration. The data were processed by DAS 2.0 software to calculate the pharmacokinetic parameters. Results Compared with the Carthami flos group, the pharmacokinetic parameters V1/F and CL/F of HSYA in the Carthami flos combined with Sappan lignum group were significantly decreased (P<0.01), and the AUC_0-t, C_max, and t_1/2α of HSYA were significantly increased (P<0.01). Conclusion Sappan lignum can promote the absorption of HSYA in rat model of CCBS and reduce the distribution of HSYA, thus exercise an efficacy-enhancing effect.