The invention discloses pumpkin, spirulina and vinegar fragrance beef jerky and a preparation method of the pumpkin, spirulina and vinegar fragrance beef jerky. The pumpkin, spirulina and vinegar fragrance beef jerky comprises the following raw materials, by weight ratio, 50-60 parts of beef jerky, 0.3-0.5 part of grape leaves, 0.2-0.3 part of fructus amomi, 0.3-0.4 part of herba schizonepetae, 0.1-0.3 part of saffron crocuses, 0.2-0.3 part of pepper, 0.1-0.2 part of sweet wormwood herbs, 0.2-0.3 part of clove, 5-8 parts of pumpkins, 4-5 parts of abrus herbs, 3-4 parts of paniculate microcosm leaves, 2-3 parts of sunflower discs, 2-3 parts of pomegranate rind, 5-10 parts of spirulina powder, 4-6 parts of apple vinegar, 1-1.5 parts of salt, 1.5-2 parts of soybean sauce, 1-1.5 parts of monosodium glutamate, 5-8 parts of white sugar and 1-2 parts of yellow rice wine. The pumpkin, spirulina and vinegar fragrance beef jerky has the advantages of being rich in nutrition, red and bright in color and luster, tasty and special in flavor, having high elasticity and chewiness, sweet pumpkin fragrance and apple vinegar fragrance, and being novel, and is leisure food which is green, healthy, non-additive, suitable for people of all ages and both sexes, and capable of satisfying requirements of different crowds.