This study compared the lipid content, total and sn-2 fatty acids, and the levels of 1, 3-dioleoy-2-palmitoyl glycerol (OPO), 1-oleoyl-2-palmitoyl-3-linoleoyl glycerol (OPL), and 1, 3-linoleoyl-2-palmitoyl glycerol (LPL) in colostrum and mature milk (n = 122) from seven ethnic groups in China. Compared with the Han breast milk, the content of saturated fatty acid (SFA) and trans fatty acid (TFA) in the breast milk of ethnic groups (especially in Uygur) was higher, while monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) were lower. This disparity may be attributed to the higher consumption of meat and diary products. In terms of the distribution of fatty acids and the content of triacylglycerol (TAG), ethnic groups and Han showed consistency. The OPO, OPL, and LPL in colostrum (0.16 ± 0.06, 0.17 ± 0.08, 0.09 ± 0.04 g 100 mL−1) were significantly lower than those in mature milk (0.28 ± 0.11, 0.26 ± 0.09, 0.16 ± 0.07 g 100 mL−1). The results have shown that the composition and content of total and sn-2 FAs in breast milk were affected by lactation stage, ethnic minority background, and maternal diets. This study contributed to the enhancement of the Chinese breast milk lipid database and offered a scientific foundation for formulating lipid compositions in infant formulas tailored to specific ethnic groups.