A method for preparing 13C labeled docosahexaenoic acid (DHA) represented by Formula A: The method comprises the conversion of 2-pentyn-1-ol to 13C labeled DHA by reaction with propargyl alcohol, 13C labeled propargyl alcohol and methyl pent-4-ynoate. The various steps involved include tosylation, coupling, bromination, selective hydrogenation and ester hydrolysis to obtain the final product.