The invention discloses a donkey embryo flushing fluid and an application method thereof. The donkey embryo flushing fluid comprises the following main ingredients: sodium chloride, potassium chloride, dipotassium phosphate, magnesium sulfate, sodium bicarbonate, calcium chloride dehydrate, sodium lactate, EDTA (Ethylene Diamine Tetraacetic Acid), sodium pyruvate, glutamine, HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), penicillin-streptomycin and BSA (Bull Serum Albumin). The donkey embryo flushing fluid has the following beneficial effects that the problem that an existingembryo flushing fluid is poor in pertinence, expensive, poor in embryonic development and high in a hysteritis incidence rate after embryo transplantation can be solved. The donkey embryo flushing fluid is very suitable for the mass production of donkey non-operative method embryo transplantation, and is favorable for the promotion and the application of an embryo transplantation technology.