Eluants Amount of solvent (in ml) No. of collected beakers
100% n-hexane 300 1-3
25% Petroleum ether in hexane 100 4
50% Petroleum ether in hexane 100 5
100% Petroleum ether 200 6-7
2.5% Ethyl acetate in petroleum ether 100 8
5% Ethyl acetate in petroleum ether 300 9-11
7.5% Ethyl acetate in petroleum ether 100 12
10% Ethyl acetate in petroleum ether 300 13-15
12.5% Ethyl acetate in petroleum ether 100 16
15% Ethyl acetate in petroleum ether 100 17
20% Ethyl acetate in petroleum ether 100 18
25% Ethyl acetate in petroleum ether 100 19
30% Ethyl acetate in petroleum ether 200 20-21
40% Ethyl acetate in petroleum ether 100 22
50% Ethyl acetate in petroleum ether 100 23
60% Ethyl acetate in petroleum ether 100 24
70% Ethyl acetate in petroleum ether 100 25
80% Ethyl acetate in petroleum ether 100 26
90% Ethyl acetate in petroleum ether 100 27
100% Ethyl acetate 100 28
5% Methanol in ethyl acetate 200 29-30
15% Methanol in ethyl acetate 100 31
50% Methanol in ethyl acetate 200 32-33
100% Methanol wash 100 34
The fractions were combined on the basis of their preliminary TLC examination to give combined fractions A, B, C, D, E, F (Table.2). Each combined fraction was evaporated to dryness under reduced pressure.
Table 1: Solvent used for column chromatography.