The invention relates to the technical field of juniperus sabina, and in particular relates to a juniperus sabina total flavonoid extract as well as a preparation method and an application thereof. The juniperus sabina total flavonoid extract is obtained according to the following method: crushing juniperus sabina branches and leaves, performing degreasing after drying, performing reflux extraction after degreasing by using an ethanol solution, performing concentration after extraction, removing impurities by virtue of a silica gel column, and performing enrichment and purification by virtue of macroporous resin to obtain the juniperus sabina total flavonoid extract provided by the invention. The method, in comparison with a traditional method, has the following advantages that (1) the use of acid and lead salts is avoided, the components are stable and cannot be hydrolyzed easily, and no heavy metal is remained; (2) after removing oil-soluble impurities by using silica gel, the pollution of the macroporous resin can be effectively reduced, and the macroporous resin is easy to regenerate; and (3) the enrichment and purification of flavonoid components in juniperus sabina can be realized in a closed environment, the total flavonoid content is high, the process method is simple to operate, is low in cost and is pollution-free, and meanwhile, the juniperus sabina total flavonoid extract provided by the invention has a relatively good effect of resisting inflammation and easing pain, and can be used for treating arthritis diseases.