In this study, quantitative and pattern recognition analysis for the quality evaluation of Magnoliae Flos using HPLC/ UV was developed. For quantitative analysis, eleven major bioactive lignan compounds were determined. The separation conditions employed for HPLC/UV were optimized using ODS C18 column (250 × 4.6 mm, 5 µm) with isocratic elution of acetonitrile and water with 1% acetic acid as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 278 nm. These methods were fully validated with respect to the linearity, accuracy, precision, recovery, and robustness. The HPLC/UV method was applied successfully to the quantification of eleven major compounds in the extract of Magnoliae Flos. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of twenty one reference samples corresponding to seven different species of Magnoliae Flos and nine samples purchased from market. The results indicate that the established HPLC/UV method is suitable for the quantitative analysis and quality control of multi-components in Magnoliae Flos.