Buffalo milk fat was first separated by thin layer chromatography (t.l.c.) into high (HMT), medium (MMT) and low (LMT) molecular weight triacylglycerols. The triacylglycerol (TG) fractions thus obtained were further resolved by argentation t.l.c. into saturated, trans-monoene, cis-monoene, diene and polyene TG species. Whereas 18:1 was concentrated in HMT, 6:0, 8:0 and 10:0 tended to be located in MMT. LMT was unique in containing almost all 4:0. HMT, MMT and LMT showed great variations in their levels of different TG species. The distribution of fatty acids among the TG species was found to be characteristic of each TG species.