My organic compounds have a molecular weight in the range of 2500g/mol and other is 1066.22 g/mol. aThe TLC results show three spots. But the main problem with my compounds are acid sensitivity of each of them and also I'm getting the final products in a very tiny amount (approx. 20mg). Can someone please provide me with the exact protocol size exclusion chromatography? Can I use Organic solvents such as Methanol instead of PBS? From the literature, I have selected the particular bead size of Sephadex and selected Sephadex G-25. Please provide me with .the Protocol if any has used it in the past. Kindly do let me know how to separate. Thanks in advance
SEC works best for molecules having large molecular weight differences. Surely you can try it, but you will be forced to inject small volumes of sample and large column volumes. Adsorption chromatography is not an option for your system?
If you can resolve them by TLC, then you should scale up the separation by column chromatography, using the exact same chemistry. I don't think SEC is the right way to do this.
Agree with others. The mass difference is not so large for SEC. Depending on the structure, you may try prep-LC (RP or HILIC).
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