Purpose To establish a HPLC method for determination of the content of trifluoroacetic acid in bivalirudin.Methods Kromasil 100-5 C_(18) column(4.6 mm×200 mm,5 μm)and 0.07% phosphoric acid solution(pH 3.0)-methanol(98:2)as the mobile phase at a flow rate of 1.0 mL/min were used with a column temperature of 30 ℃,a detection wavelength of 210 nm and a injection volume of 20 μL.Results The standard curve of trifluomacetic acid was linear in the range of 198.4-992.0 μg/mL,r=0.999 9.The detection limit Was 40 ng.The average recovery rate of method was 99.9%,and RSD was 0.41%.Conclusion The method of content determination was practicable and accurate.It can be used for content determination of trifluoroacetic acid in bivalirudin.