A ganoderma lucidum spore oil authentication method is established by using near-infrared spectroscopy. The sunflower oil, barley oil, peanut oil, corn oil and blend oil are mixed into the ganoderma lucidum spore oil respectively. These mixed oil has a volume percentage from 0 to 100. The near-infrared spectroscopy is used for determination. After the optimal spectral bands are selected, a quantitative analysis model is established by using Partial Least Squares (PLS). The model has a relative prediction error in the range from -0.357 to 0.435 for the mixed oil with different adulterated percentages. The research result shows that this analysis method based on chemometrics and near-infrared spectroscopy is a simple, rapid, low-cost and accurate method for the authentication of oil kinds and the detection of mixer content.