The invention relates to a method for separating and purifying mogroside V by subcritical water desorption technology. The macroporous adsorption resin enriched with mogroside V is subjected to desorption under a subcritical condition of water using water as a solvent, to give an aqueous solution rich in mogroside V. The method not only improves the content of mogroside V in product, but also effectively removes bitter impurities and residual pesticides, greatly improves the taste adaptability of the product, and improves the safety and quality of the product. The method reduces the processing steps and reduces the use of organic solvents in the prior art, and reduces total production costs.