Azerbaijan has abundant raw material resources for biodiesel production, one of which is cotton oil. Taking into account the high cost and toxicity of methanol, a mixture of methanol, ethanol, propanol-1, and propanol-2 has been used for the first time in the production of biodiesel. In the presented work, various operational indicators (viscosity, density, ignition temperature, etc.) for biodiesel based on mixtures of cottonseed oil and low-molecular-weight alcohol tested by ASTM standards and also found optimal conditions, comparison of fuels, and biodiesel mixers have been carried out. The results of the research indicate that the obtained fuel mixtures are more efficient and have high performance characteristics. Additionally, by using NMR spectroscopy, the activity problems of alcohol mixtures in the transesterification reaction have been investigated.