The invention discloses a preparation method of pleurotus nebrodensis health vinegar, belonging to the field of food processing. The preparation method is characterized by comprising the following steps: carrying out liquid culture on pleurotus nebrodensis strain, carrying out treatment on fermented mash liquid, carrying out solid culture, carrying out digestion, carrying out acetic fermentation, thus obtaining a semi-finished product, adding with auxiliary materials, carrying out cold storage for removing impurities, carrying out sterilization, and carrying out packaging, and obtaining a finished product. The preparation method has the beneficial effects that the obtained product is transparent liquid which is clear and transparent, is aromatic in flavor, pure and natural in taste, and has the unique faint scent of pleurotus nebrodensis. The pleurotus nebrodensis health vinegar is rich in nutrition and delicious in taste, can regulate the human physiological equilibrium and enhance the human immune function, has the health efficacies of removing food retention and relieving cough, is convenient to take, is pleasantly acid in mouthfeel, is unique in flavor, is suitable for people of all ages, is pollution-free, and is beneficial for the health.