This study was aimed to establish an UFLC fingerprint of Tibetan medicine Pterocephalus hookeir samples from different habitats. UFLC-PDA was adopted to analyse 21 batches of P. hookeir samples from different habitats. The chromatographic condition was as follow: Agilent proshell 120 SB-C18 column (4.6 mm x 100 mm, 2.7 microm) eluted with the mobile phases of acetonitrile and 0.2% phosphoric acid water in gradient mode. The flow rate was 1.0 mL x min(-1), and the detection wavelength was set at 238 nm. The fingerprints of 21 batches P. hookeir were carried out by similarity comparation, and 15 chromatographic peaks were extracted as the common peaks of fingerprint, of which 5 peaks were identified as chlorogenic acid, loganin, sweroside, sylvestroside III, triplostoside A. The similarity degrees of 18 batchs of samples were above 0.9, and the other 3 batchs of samples were below 0.9. This is the first established fingerprint of P. hookeir by using UFLC-PDA. This method has good precision, stability and repeatability that it could provide basis for quality control and evaluation of P. hookeir.