The invention belongs to the field of medicinal preparations and discloses fat emulsion pre-emulsifying concentrated solution for teniposide intravenous injection and a preparation method thereof. The fat emulsion pre-emulsifying concentrated solution for teniposide intravenous injection comprises the following components in percentage by weight: 0.01 to 10 percent of teniposide, 0 to 20 percent of oil phase, 10 to 80 percent of phospholipids and 10 to 99 percent of organic solvent for injection. The preparation method comprises the following steps of: dissolving the active ingredient, namely the teniposide, in the organic solvent or the oil phase or the mixture of the organic solvent and the oil phase, adding the other components in the formula into the mixture, and after stirring to uniformly mix the components, obtaining the transparent and clear pre-emulsifying concentrated solution.