| Summary: | <i>Artemisia cina</i> is a plant used in traditional Chinese medicine as a remedy for parasitic diseases. This study describes the isolation and chemical characterization of anthelmintic compounds of <i>A. cina</i> against <i>Haemonchus contortus</i> infective larvae (L<sub>3</sub>) through lethal testing. Previously, three extracts—<i>n</i>-hexane (HexAc), ethyl acetate (EtOAc) and methanol (MeOAc)—were evaluated at concentrations of 4 to 0.5 mg/mL, resulting in the HexAc extract with the greatest effect of 76.6% mortality of the larvae at 4 mg/mL. Then, this was chemically fractioned by polarity, obtaining seven fractions (C1F1–C1F7), and, when evaluated at concentrations from 2 to 0.25 mg/mL, the 2 mg/mL C1F5 fraction produced an effect against the nematode <i>H. contortus</i> of 100% mortality of the larvae. Thus, this fraction was fractionated again by column chromatography, obtaining twelve subfractions (C2F1–C2F12) which were evaluated from 1 to 0.125 mg/mL, with the C2F5 subfraction causing a nematicidal effect of 100% mortality. NMR analysis of one (<sup>1</sup>H, <sup>13</sup>C and DEPT) and two dimensions (COSY, HSQC and HMBC) and mass spectrometry of this fraction allowed us to identify the mixture of 3′-demethoxy-6-O-demethylisoguaiacin and norisoguaiacin. Therefore, it can be assumed that the mixture of these compounds is responsible for the anthelmintic effect. These results indicate that <i>A. cina</i> containing anthelmintic compounds and might be used as an antiparasitic drug against <i>H. contortus</i>.
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