A high-performance liquid chromatographic method (HPLC) with evaporative light scattering detection (ELSD) has been developed for fingerprint analysis of Flos Lonicerae. The samples were separated with an Agilent C18 column using acetonitrile (A) and 0.1% formic acid solution under gradient conditions (0→10 min：11.5%→15% A, 10→12 min：15% →22% A, 12→18min：22%→33% A; 18→30min: 33%→45% A) as the mobile phase at a flow rate of 0.8ml min−1 within 30 min. The ELSD conditions were optimized at nebulizer-gas flow rate 2.7 L min−1 and drift tube temperature 105°C. Precision experiments showed relative standard deviation (R.S.D.) of peak area and retention time were better than 2.5%, inter-day and intra-day variabilities showed that R.S.D. was ranged from 0.66% to 4.17%. Accuracy validation showed that average recovery was between 95.33% and 104.10%. The method was validated to achieve the satisfactory precision and recovery. Relative retention time and relative peak area were used to identify the common peaks for fingerprint analysis. There are nine common peaks in the fingerprint. The quality of sixteen batches of Flos Lonicerae samples was evaluated by hierarchical clustering analysis, which classfied all bacthes into two clusters. Furthermore, the contents of important medicinal compounds (chlorogenic acid, macranthoidin B and dipsacoside B) in different batches of Flos Lonicerae samples were determined simultaneously using the developed HPLC-ELSD method. The developed analytical procedure was proved to be a reliable and rapid method for the quality control of Flos Lonicerae.