The invention discloses a preparation method of high content conjugated linoleic acid by using safflower oil as a raw material. The method is as below: using safflower seeds as the raw material, first performing high temperature sterilization while introducing nitrogen into the safflower seeds, and conducting pressing and conducting fine filtering; then carrying out enzymatic hydrolysis on the obtained fine filtrate; adjusting the pH of the solution after enzymatic hydrolysis, and carrying out an acidification reaction; and finally, carrying out extraction to separate conjugated linoleic acid. The preparation method provided by the invention is simple, low in preparation cost, and short in preparation time, and the prepared conjugated linoleic acid is high in content, and high in purity and can be directly eaten.