There is growing interest in the antioxidants and antibacterial activity from natural substances. The purpose of the research was to gain and distinguish phenolic substances and organic acids in the Camellia oleifera cake, and to study their antioxidant and antibacterial activities. The extraction and purification of them were achieved by solvent extraction and column separation, respectively. The conclusions displayed that purity of the phenolic substances was 94.1 ± 0.5% w/w and that of organic acid was 96.0 ± 0.3% w/w; Fifteen phenolic substances were certificated using HPLC–ESI–MS technology; oxalic, citric, acetic, malic, and succinic acids are discovered to be main organic acids. In addition, the phenolic substances and organic acids both have good antioxidant activity and obvious inhibition against six species of bacteria. These conclusions can be useful in the reuse of the waste of Camellia oleifera oil industry in the future.