Abstract: In addition to high concentrations of unsaturated fatty acids in Camellia oil, Camellia oleifera also contains polysaccharides, saponins, polyphenols and other bioactive ingredients. On the premise of the extraction and preparation of bioactive substance from C. oleifera, with simple and effective extraction of oil as the goal in this study, oil content as index, the orthogonal design was carried out to investigate the effect of extraction time, extraction temperature, extraction solvent kinds and weight of single sample after single factor test with soxhlet extraction method.The results showed that the optimum conditions of soxhlet extraction were as follows: extraction time 6 h, temperature 80 ℃, organic solvent acetone, single sample weight 4 g.Under this condition, the content of C. oleifera seed oil was more than 43.24% ± 0.53%. The oil contents of both whole seeds and seed kernels from high to low were followed by C. semiserrata Chi, C. oleifera Abel, C. meiocarpa Hu and C. gauchowensis Chang.The highest oil contents of both whole seeds and seed kernels were from C. semiserrata Chi, while oil content of seed cakes was the lowest, which were 49.7%, 67.5% and 6.1%, respectively. The optimized soxhlet extraction conditions were applicable for the effective extraction and the quantitative analysis of C. oleifera seed oil.