In this study, Xanthium sibiricum Patr oil, a non-edible oil, was investigated for the first time to produce biodiesel. X. sibiricum Patr has very good environmental adaptability and thus has plenty of wild resources. Its seed has a high oil content (42.34%) which gives potential annual output of 100,000 tons just in China. And the oil acid value is pretty low (1.38 mg KOH/g). Under the optimal conditions, the FAME content and yield of X. sibiricum biodiesel were 98.7 wt.% and 92.0%, respectively. The properties of the biodiesel product were tested and most properties were in accordance with EN 14214-08, ASTM D6751-10 and GB/T 20828-07 standards, except cetane number and oxidative stability. The results indicated that X. sibiricum Patr is a promising species as a biodiesel feedstock in China.