The invention relates to the field of assisted reproductive technology, and discloses a composition and application thereof, and a sperm refrigerating fluid and a preparation method thereof. The spermrefrigerating fluid comprises a basic culture solution, the composition, a cryoprotectant, human serum albumin and a bactericide, wherein the composition is prepared from resveratrol with a concentration of 12 to 26mg/L, reduced glutathione with a concentration of 0.2 to 1.9 mg/L , lecithin with a concentration of 436 to 775 mg/L, cholesterol with a concentration of 0.5 to 1.6 mg/L, an ATP sodiumsalt with a concentration of 9 to 17 mg/L, heparin with a concentration of 11 to 28 mg/L, phosphatidylserine with a concentration of 93 to 299 mg/L and coenzyme Q10with a concentration of 5 to 25 mg/L; and the cryoprotectant comprises paclitaxel with a concentration of 0.3 to 1.1 mg/L of, trehalose with a concentration of 16 to 22 g/L, cane sugar with a concentration of 7 to 13 g/L and glycerol with a concentration of 80 to 250 mL/L. The sperm refrigerating fluid can reduce damage of sperms in low-temperature refrigeration and in freezing recovery, improve vitality of the sperms after freezing recovery, prolong the survival time of the sperms after freezing recovery, promote sperm capacitation and enhance penetrating fertilization capacity of the sperms to egg cells; and thus, the fertilization success rate and blastocyst formation rate of the ova are improved, and finally, the effect of improving the pregnancy success rate of an in-vitro fertilization assisted reproduction technologyis achieved.