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.