Abstract: 0.100 0 g of the crushed sample was added into 10 mL of water, and the mixture was ultrasonicated for 20 min and then diluted to 25 mL with water. After the solution was filtrated with 0.22 μm organic filter membrane, Thermo Hypersil Gold C₁₈ column (100 mm×2.1 mm, 1.9 μm) was used for seperation, with acetonitrile-0.1% (volume fraction) formic acid solution as the mobile phase for gradient elution. A diode array detector was used for detection at the wavelength of 403 nm. A linear relationship was found between the peak area and the mass concentration of hydroxysafflor yellow A in the range of 42-336 mg·L^-1, with the detection limit (3S/N) of 19.3 μg·L^-1. Values of recovery obtained by standard addition method ranged from 92.7% to 95.2%. The relative standard deviation (n=6) of the measured values was less than 1.0%. The method was used to determine the content of hydroxysafflor yellow A in Carthamus tinctorius L. from different producing areas in Xinjiang, giving results between 24.00 and 46.43 mg·g^-1. The results of cluster analysis showed that Carthamus tinctorius L. samples could be clustered into 4 major groups.