The invention discloses melaleuca alternefolia total flavone extract and a preparation method thereof. The preparation method comprises the following steps of extracting essence oil from the melaleuca alternefolia branch and leave by water vapor, performing ethanol heating and refluxing extraction, reducing the pressure to recycle the ethanol, concentrating to remove the ethanol flavor, separating and purifying by macroporous absorption resin, and drying; measuring the content of total flavone in the melaleuca alternefolia by adopting an ultraviolet-visible spectrophotometry method. The preparation method has the advantages that the method is accurate and stable, the operation is simple, the requirement of industrial production is met, and the quality of the melaleuca alternefolia total flavone extract can be effectively controlled; the content of total flavone in the melaleuca alternefolia total flavone extract can reach more than 50 percent, the toxicity is little, and the pharmacological functions of resisting inflammation, inhibiting bacteria, alleviating pain, resisting allergy, resisting oxidizing and the like are realized; the preparation method is reasonable and stable, the production cycle is short, the preparation method is suitable for industrial production, the important meaning is realized on the deep development and utilization of melaleuca alternefolia resources, and the development prospect is good.