To establish HPLC fingerprint of Gentiana macrophylla, which was expected to be standards of quality control and identification of the Chinese crude drug.The HPLC method was used, chromatography conditions were Shimadzu C18 (150 mm x 4.6 mm, 5 microm) column with gradient mobile phase of methanol-0.4% Phosphoric acid, UV detection wavelength was at 230 nm and the column temperature was at 25 degrees C with the flow rate of 1.0 ml/min.5 peaks were identified as the characteristic fingerprints of Gentiana macrophylla and the fingerprints of the 14 samples had high similarities. The similarities of disqualified and fake medicinal materials were low.This HPLC fingerprint of Gentiana macrophylla can be used as a standard of quality control and identification of the Chinese crude drug.