Provided are a composition comprising 2-palmitic acid glyceride and a preparation method thereof, and further provided are a structural fat comprising 1, 3-diunsaturated fatty acid-2-palmitic acid and a preparation method. The preparation method of the composition comprises: (1) providing a glyceride comprising a palmitic acid residue; (2) mixing the glyceride comprising the palmitic acid residue with palmitic acid, a salt thereof or a palmitic acid nonglyceride to conduct ester exchange so as to obtain an oil A; (3) conducting Sn-1, 3 site enzymatic directed interesterification on the oil A and the unsaturated fatty acid or the nonglyceride thereof so as to obtain a mixed oil B; and (4) refining the mixed oil B to obtain the composition comprising 2-palmitic acid glyceride.
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