Tlc silica gels 60 f254

Ten microliters of methanol dissolved extract (200 mg mL -1 ) were loaded to TLC plates (TLC silica gels 60 F254, 2×10 cm, layer thickness 0.25 mm, Merck, Darmstadt, Germany) and separated using eluent system: acetone: chloroform (v: v = 1: 1) in the TLC chamber. The TLCdirect bioautography test was conducted using agar diffusion method. Mycelia of P. oryzae race 173 was harvested from 7 days old of fungal culture (10 6 cfu mL -1 ) which previously grew on PDB medium and mixed with 15 mL of PDA medium (48°C). Furthermore, the developed TLC plates were placed aseptically in the Petri dishes and covered with PDA broth medium that contained fungal mycelia. After 10 days of incubation, antifungal activity of crude extract was observed. The active fraction with antifungal activity was characterized by clear zone around the developed TLC plate.

The crude extract from isolate STGG 14 was separated using TLC plates (TLC silica gels 60 F254, 20×20 cm, layer thickness 0.25 mm, Merck, Darmstadt, Germany). The crude extract was dissolved using methanol to final concentration of 200 mg mL -1 . Afterward, the dissolved extract was injected into TLC plates using CAMAG Linomat 5 and developed using eluent system: acetone: chloroform (v/v = 1/1) as optimum eluent. Eluent residue was removed by incubating the TLC plates at room temperature for 15 minutes. The separated compounds were visualized under UV light with 254 nm and 366 wavelength using CAMAG TLC Visualizer 2.