I am working on Boraginaceae family. Now I have n-hexane fraction, I want to run column for this fraction. From which solvent system should i Start?? And then at what rate I change the polarity?
Hi Muhammad
What kind of molecules do you want to analyze? n-Hexan is not a strong solvent, so you will probably just get lipidic molecules in your extract. The choose of your solvents will depend on the molecules you want to separate. Alkanes usually have no chromatographical specifity and not much intermolecular effects that you can use for a chromatographic separation, therefore, you can also use Pentane, Cyclohexane and Heptan for extraction, they have far better toxic datas then n-Hexane which is quite critical in risk, without visible difference in analytical usage.
A simple system can be: extraction in Cyclohexan and TLC in Cyclohexane- Ethylacetate Mixtures
You can pre-check via tlc if you have any kind of movement or what amount of ethylacetate you might have to add. Be aware that your desired molecules dont run to far on tlc, if the polarity is to high, your separation on coumn can be poorer.
Usually a location in the first 30% of your total retention is safe.
Try to start from an isocratic mix, if possible...
Good luck
There are a variety of solvents with different selectivities for such a non-polar extract.
Try hexane and the following solvents: toluene, ethyl acetate, dichloromethane, methanol, and diethyl ether. For TLC plates, try silica, alumina, and diol (use a water spray to visualize the spots)
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