In this study, water-soluble crude polysaccharides from Tussilago farfara L. and the polysaccharides, deproteinized and decolorized from the crude polysaccharides were obtained. We evaluated the antioxidant activities of crude polysaccharides and deproteinized polysaccharides in vitro by flow injection analysis chemiluminescence (FIA-CL), based on the inhibition effect of their natural antioxidants on the free radical-induced luminol chemiluminescence. Ascorbic acid was used as positive control. At the optimum operational conditions, the antioxidant activity was measured by plotting the inhibition vs. the content of samples to calculate the IC50. The results showed that deproteinized polysaccharides showed higher scavenging activities on the reactive oxygen species such as superoxide anion radical, hydroxyl radical and hydrogen peroxide than that of crude polysaccharides, but less than that of ascorbic acid. This suggests that Tussilago farfara is a good natural antioxidant and has the potential to be used in the manufacturing of foods and medicines.