A solid phase extraction-high performance liquid chromatog. method for the detection of vitamin D content in food was established, which integrated the detection process of the national standard method, and the sample was purified by saponification-n-hexane extraction-silica gel matrix solid phase extraction, reversed-phase high-performance liquid chromatog., and quantified by external standard method.The detection limit of the test milk samples was less than 0.07 μg/100 g, the precision was less than 2%, the recovery rate was between 91.3%∼100.1%, the detection limit of the tested nutritional rice flour was less than 0.7 μg/100 g, the precision was less than 4%, and the recovery rate was between 82%∼100.9%, which provided a more economical, simple, stable and accurate method for the determination of vitamin D in food.