Antimicrobial activity analysis were regarding to disc diffusion. 10,11 The antimicrobial activity was measured by diameter of clear zone minus diameter of disc diffusion.
Thin Layer Chromatography (TLC)TLC analytical, TLC-bioautography and semipreparative separation were performed on aluminium silica TLC F254 plates. The TLC plates were developed with mobile phase chloroform: methanol (9:1) (F1, F3, F4, and F5) and hexane: ethyl acetate (7:3) (F2). Screening for antimicrobial compound conducted by TLC-bioautography. 12 Positive result reflected on clear zone developed by Rf. Further TLC-separation on clear zone-Rf were developed with chloroform: ethyl acetate (5:5) (F1, F4, and F5).