The invention discloses a preparation method and application of reduced glutathione lipidosome, relates to a preparation method and application of lipidosome, and aims at solving the technical problem of low inclusion rate of the lipidosome prepared by the existing method. The method comprises the following steps: 1, weighing soya bean lecithin and fatty acid glycerides, dissolving into absolute ethyl alcohol, and heating in a water bath to prepare an oil phase; 2, weighing reduced glutathione and a surfactant to be dissolved into distilled water, and heating in the water bath to prepare a water phase; 3, mixing the prepared oil phase with the water phase, emulsifying, carrying out vacuum rotary evaporation to remove absolute ethyl alcohol, adding distilled water, pre-cooling after ice-bath, and then carrying out freeze drying to obtain lipidosome powder. The reduced glutathione lipidosome disclosed by the invention is applied to intravenous injection, oral medication, transdermal drug delivery, and a drug delivery system for the lung and eyes. The encapsulation efficiency of the reduced glutathione lipidosome disclosed by the invention is 68.15%+/-1.53%. The preparation method is applied to the field of preparation of the lipidosome.