In this study, buffalo spermatozoa were in-vitro capacitated and acrosome reacted using modified Tyrode's bicarbonatebuffered medium (sp-TALP media). Lipids were extracted from freshly ejaculated, in-vitro capacitated and acrosome reacted buffalo spermatozoa. 8 cells in fresh, capacitated and acrosome reacted spermatozoa respectively. The results generated by this study will be one step further to understand the lipid changes in buffalo spermatozoa during capacitation, acrosome reaction and fertilization process.