OBJECTIVE: To establish a method for content determination of gallic acid,eugenol,alantolatone and isoalantolactone in Tibetan medicine Jiebai pills simultaneously. METHODS: HPLC method was adopted. The separation was performed on CAPCELL PAK MGⅡ C18 column with of mobile phase composed of methanol-acetonitrile-0.2% phosphoric acid solution (gradient elution) at the flow rate of 0.8 mL/min. The detection wavelengths were 220 nm for alantolatone and isoalantolactone, 280 nm for gallic acid and eugenol. The column temperature was at 30 ℃, and sample size was 10 μL. RESULTS: The linear ranges of gallic acid,eugenol,alantolatone and isoalantolactone were 46.50-2 325.00, 21.28-1 064.00, 4.38-219.20 and 4.17-208.40 μg/mL, respectively (all r≥0.999 1). The limits of detection were 0.26, 0.09, 0.28,0.35 μg/mL, and the limits of quantitation were 0.74, 0.25, 0.95, 1.06 μg/mL, respectively. RSDs of precision, stability (48 h) and reproducibility tests were all lower than 2.0% (n=6 or n=7). The recoveries were 98.5%, 97.6%, 97.1% and 101.6%, and RSDs were 1.4%, 1.9%, 1.6%, 1.7% (n=6), respectively. The content of gallic acid,eugenol,alantolatone and isoalantolactone in 6 batches of Baijie pills were 4.28-7.02, 1.09-4.90, 0.25-0.60, 0.30-0.72 mg/g, respectively. CONCLUSIONS: The established method can be used for content determination of gallic acid,eugenol,alantolatone and isoalantolactone in Tibetan medicine Jiebai pills simultaneously.