Question

. In our column chromatography of spinach most students obtain three or four bands. However, a TLC of the same spinach extract can yield up to 7 spots on the TLC. Explain why one can not isolate the same number of bands from the column as there are on the TLC?

Answer 1

Solution:-

TLC showed that there are seven spots that means the difference between the R_f values of spots are very much low. The spots are very much close to each other and difficult to separate. Polarity of mobile phase is another reason for the mixture of products, may be the column run with high polarity, so run the column in a gradient way. Also solubility could be a reason for less separation. The height and diameter of the column and thickness of slurry bed also affect the separation.