Pyroligneous acid (PA), obtained from charcoal production, has been reported excellent capability in antioxidant by chemical examination. However, the biological effect of PA in living animal is still unknown. In this study, a simple model organism, the nematode Caenorhabditis elegans, was used as an in vivo system to assess the biological effects of PA treatment. The worms were exposed to concentrated pyroligneous acid (CPA) and extraction (CPAE) thereof in a 0.5-5.0 mg/mL concentration and their brood size and germline cell apoptotisis were examined. The results showed that CPA and CPAE rescued the germline cell apoptosis induced by paraquat, a reactive oxygen species (ROS) generator. Additionally, CPA and CPAE did not show negative effect on the brood size and germline cell apoptosis in wild type at normal culture condition, suggesting safety of PA in living worms. To further scrutinize the antioxidant ability of PA, the apoptotic cell of germline induced by parapuat was assayed after treatment of 0.5 mg/mL CPA, CPAE, L-ascorbic acid and alpha-tocopherol. Treatment with CPAE or L-ascorbic acid could almost completely rescue the germline apoptosis to control level, indicating a potential material in food and biomedicine of PA.