To separate spots resolved with DCM in TLC, use a column chromatography solvent gradient starting with hexane/heptane and increasing polarity with DCM or ethyl acetate. Alternatively, mix DCM with ethyl acetate (e.g., 90:10 or 80:20) to enhance separation based on compound polarity.

To separate your three spots using column chromatography, use a DCM-based gradient:

Monitor with TLC and adjust the solvent ratio as needed.

To separate spots lifted with DCM in TLC using column chromatography, start with a less polar solvent like hexane and gradually add a more polar solvent such as ethyl acetate. Test different ratios using TLC to find the optimal mixture that provides distinct separation before applying it to column chromatography.

You can use a mixed solvent of cyclohexane and ethyl acetate for column chromatography. And constantly adjust the proportion of solvents.