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The development of Dai medicine is relatively slow, and <i>Zanthoxylum</i> has great economic and medicinal value. It is still difficult to obtain medicinal components from the low-polarity parts of <i>Zanthoxylum</i> belonging to Dai medicine. In this study, we introduced one simple and quick strategy of separating target compounds from the barks of <i>Z. acanthopodium</i> var. <i>timbor</i> by high-performance countercurrent chromatography (HPCCC) with an off-line anti-inflammatory activity screening mode. The development of this strategy was based on the TLC-based generally useful estimation of solvent systems (GUESS) method and HPCCC in combination. This paper presented a rapid method for obtaining target anti-inflammatory compounds. Three lignins were enriched by HPCCC with an off-line inhibition mode of nitric oxide production in lipopolysaccharide-stimulated RAW264.7 macrophage cells, using petroleum ether–ethyl acetate–methanol–water (3:2:3:2) as the solvent system. The results showed that this method was simple and practical and could be applied to trace the anti-inflammatory components of the low-polarity part in Dai medicine.