Abstract: Content of α-linolenic acid in nutritional eggs has been determined using capillary gas chromatography. The total fat in nutritional eggs was extract by hydrochloric acid for acid hydrolysis. Then the extract of the total fat was treated with a 0.5 mol/L potassium hydroxide―methanol solution for methyl esterification. The esterified sample, including α-methyl linolenate, was finally determined by a capillary gas chromatograph―flame ionization detector (FID), using a FFAP capillary column with temperature programming. The intro-day and inter-day precision of the method were 1.56% and 2.72%, respectively. The average recovery rate of the standard amount added to the sample was 97.3% (RSD was 2.9%). The content of α-linolenic acid in the detected nutritional egg was 0.49%. The result showed that the developed method was accurate, repeatable and easy to operate. It is suitable for the determination of α-linolenic acid and other polyunsaturated fatty acid in nutritional eggs.