To reveal the characteristics of pyrolytic oil production from the thermal cracking of tobacco stem waste at different temperatures and oxygen concentrations, experimental studies of tobacco stem thermal cracking at 300-800°C and oxygen concentrations of 0-8% were carried out using a fixed-bed reactor, and the pyrolytic oil composition was analyzed by the gas chromatography-mass spectrometry (GC-MS) and the ultraviolet fluorescence spectroscopy (UV-F). The results showed that: 1.Thermal cracking of tobacco stem could produce about 23.25-39.50 wt% of pyrolytic oil, and the oil yield reached the highest at 500°C; 2.Increasing oxygen concentration could promote the oxidative consumption of volatiles, leading to the decrease of pyrolytic oil yield; 3.Nicotine yield reached the highest value at 500°C when oxygen concentration was 0% and 4%, 7.0 mg and 9.5 mg (per g of tobacco), respectively. When the oxygen concentration was 8%, the nicotine yield reached the highest value of 7.8 mg (per g of tobacco) at 400°C. 4.Under an inert atmosphere, high temperature inhibited the production of acids, ketones, aldehydes, alcohols, and heterocyclic substances and promoted the production of phenols, while oxidizing atmosphere helped to produce phenols, heterocyclic substances, and aromatic substances. 5.High temperature and oxidizing atmosphere significantly promoted the production of macrocyclic aromatic compounds. The results of this paper clarify the effects of reaction temperature and oxygen concentration on the generation characteristics of pyrolytic oil in the thermal cracking process of tobacco stem and provide theoretical guidance for the pyrolysis utilization of tobacco stem waste.