This study systematically investigated a method for extraction of safflower (Carthamus tinctorius Linnaeus) colorants by ultrasonic treatment. Compared to pigments productivity and cell wall structures of safflower after general and ultrasonic method, the ultrasonic method showed high extraction efficiency of safflower pigments due to destruction of safflower cell wall caused by high vibration energies. Microscopic analysis confirmed the hypothesis that the ultrasonic treatment of safflower caused its cell wall structure loosened and made efficient extraction of safflower pigments. And also, LC-MS/MS analysis revealed that productivities of the yellow and red safflower pigments by ultrasonic method were 21.9% and 14.6% higher, respectively, than those of pigments extracted by general method. The ultrasonic extracted yellow and red colorants could be used to dye not only natural fibers like cotton, silk and wool, but also synthetic fiber like nylon, and generally gave a better color tone than the general extracted colorants from safflower due to the affinities of red and yellow colorant on different fibers. As the yellow and red colorant were extracted by ultrasonic treatment in water, the K/S value on of 550/440nm of cotton and rayon was increased but in the case of silk and wool the change of this value was almost not detected. Finally, this technique might provide a solution to establish reproducibility and standardization for the extraction and dyeing methods on fabrics.