To establishthe HPLC fingerprint of Nymphaeae Flos and Quantitative analysis of multi-components by single-marker (QAMS) for seven contents.The HPLC fingerprint was established by Phenomenex Gemini NX-C₁₈ (250 mm×4.6 mm,5 μm).The mobile phase was acetonitrile-0.2% phosphoric acid aqueous solution.The flow rate was 1.0 mL/min.The column temperature was 30 ℃.The detection wavelength were 266 nm. The data were studied by Similarity analysis,cluster analysis and principal component analysis.Gallic acid was used as an internal standard to calculate the relative correction factors of the other six constituents.A total of eleven common peaks were identified and seven components were identified for the analysis of 17 batches of Nymphaeae Flos.The similarity of 17 batches of samples was greater than 0.9.The HPLC similarity of two commercial samples and S16 was 0.568 and 0.730,which the results show that commercial Nymphaeae Flos were different from cultivated and wild Nymphaeae Flos.Principal component analysis,cluster analysis and orthogonal partial least squares discriminant analysis (OPLS-DA) were confirmed this conclusion.With gallic acid was the reference,the relative correction factors of methyl gallate,isostrictiniin,geraniin,ellagic acid,nicotiflorin,pentagalloylglucose were 0.965 0,0.974 0,1.201 3,0.175 4,1.160 3 and 2.117 3,RSD were 0.37%,0.38%,0.32%,1.76%,0.37% and 0.08% (n = 8).At the same time,the result obtained by QAMS approximated those obtained by external standard method (ESM).The established fingerprint and QAMS content determination method of Nymphaeae Flos were feasible and provide a reference for the improvement of the quality control method of Nymphaeae Flos.