Objective To establish an HPLC method for simultaneous determination of Quercetin , Luteolin, pigenin in Matricaria Chamomila L.. Methods HPLC analysis was performed on Agilent Zorbox SB-C18 column (4.6 mm×250 mm, 5 μm) with the methanol and phosphoric acid as mobile phase in equal degree model, and the column temperature was set at 25 ℃, and the flow rate was 1.0 ml.min-1 and the detecting wave length was at 350 nm. Results The linear response ranges were from 0.25-4.04 μg of Quercetin (r=1.000, n=6) and from 0.25-3.98 μg of Luteolin (r=1.000, n=6) and 0.25-4.02 μg of Apigenin (r=1.000, n=6);and the recoveries, the precision and the stability RSD of Quercetin, Luteolin and Apigenin meet the requirements. The average recovery rate of quercetin, luteolin and apigenin was 93.64%, 95.85% and 95.40%, respectively. Conclusions All 3 samples in Matricaria Chamomila.L were determined by HPLC. The method is simple, rapid, and reproducible. It can be used as reference for the quality control of Matricaria Chamomila L..