Abstract: A reversed phase HPLC method for determination of hydroxylsafflower yellow A in safflower W/O cream was established. The column was Zorbax Eclipse C₁₈ column(4. 6 mm×250 mm, 5 μm), and the mobile phase was composed of methanol, acetonitrile and 0. 02% phosphoric acid solution(26 ∶2〓 ∶72). The flow rate of mobile phase was set at 1. 0 mL/min, and the column temperature was kept at 55 °C. The detection wavelength was 403 nm. Safflower W/O cream was successively demulsified with methanol at high temperature and followed by the addition of purified water for the extraction. The results showed that the excipients did not interfere with the chromatographic peak of hydroxylsafflower yellow A. Hydroxylsafflower yellow A presented a good linear relationship in the range of 1. 236- 12. 36 μg/mL(y=156. 17x+1. 198 3, r=0. 999 5), and the detection limit was 23. 6 ng/mL with the quantitative limit of 118 ng/mL. The percentage of extracting recovery was in the range of 99. 7% to 103. 3%. The precision RSD was 0. 12%(n=6), and the sample stability was acceptable when being stored at room temperature for 24 h. The developed method in this study was simple, rapid, accurate and reproducible, and can be used for the determination of hydroxylsafflower yellow A in safflower W/O cream.