3.1 Silica column chromatography
FAS inhibition activity guided purification of the extract prepared as described above was done by silica column chromatography (5 x 30 cm column) and 240 mesh size silica as stationary phase. Column was subjected to gradient elution using ethyl acetate to methanol, and finally by dichloromethane. The compound got eluted at dichloromethane:methanol, at 1:1 and precipitated in white waxy form, when excess methanol was added to the eluant. Further purification steps were carried out as described in the methods section to yield the purified compound. Final purification was carried out with HPLC provided with Extend-C18 column of 1.8 µm, 2.1 x 50 mm, Diode Array Detector in combination with Chem32, Chemstation software.