AbstractA method has been devised that gives the distribution of various fatty acids of pure and adulterated buffalo milk with cow milk. Gas chromatography (GC) was used for the qualitative and quantitative determination of fatty acids of authentic buffalo milk, cow milk and buffalo milk adulterated with cow milk. The milk fat was separated by fractional crystallization at ‐20 C into 2 fractions, i.e., semisolid and mother liquor. The concentration of fatty acids in the mother liquor changed significantly for 14:0, 16:0 and 18:1 as adulteration levels were increased. The fatty acids of the semisolid fractions change in the proportion of 16:0, 18:0 and 18:1 when cow milk is mixed with buffalo milk. By applying simple regression equations for these acids, adulteration of buffalo milk with 5% cow milk could be distinguished.