The invention discloses a biebersteinia heterostemon total flavone extract as well as an extraction method and an application thereof. The extraction method comprises the following steps: mixing biebersteinia heterostemon raw material powder with a solvent and carrying out solid-liquid separation after Soxhlet extraction to obtain a filtrate A; concentrating to dry the filtrate A to obtain the biebersteinia heterostemon total flavone extract, wherein the time of the Soxhlet extraction is 2-24h; the solvent is an organic solvent or a mixed solvent of the organic solvent and water; in the mixed solvent of the organic solvent and water, the percent by volume of the organic solvent is above 40%; the organic solvent is one or more of alcohols solvent, chlorinated hydrocarbon solvent, esters solvent and alkanes solvent; the volume mass ratio of the solvent to the biebersteinia heterostemon raw material powder is 1-60mL/g. The extraction method provided by the invention is high in extraction rate, good in antibacterial ability of the total flavone extract, simple to operate and good in equipment universality, recyclable in extraction solvent, low in production cost and more suitable for industrial production.