Lec 10 | MIT 5.301 Chemistry Laboratory Techniques, IAP 2004



Column Chromatography

It takes considerable practice to master the art of “running a column”. This video will get you started, with tips on picking appropriate conditions, packing and running a column, monitoring separation, and even making a micro-column from a pipet.

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23 replies
  1. ChemistDrummer
    ChemistDrummer says:

    @gshivashimpi77 you are wrong my friend.

    hexane,EtOAc or a mixture of them and likewise chloroform and methanol are the most common solvent systems used but it all depends on the mixture you want to separate…

    Reply
  2. Silverweed
    Silverweed says:

    @HelloBrainMind An automated system like biotage.

    I wish I had this video when I was an undergraduate. Its every bit of knowledge I eventually sucked up while running tons of horrible columns separating cis trans isomers that would barely separate and took forever. One thing that I didnt notice, she may have mentioned, is that dissolving silica in polar solvent is exothermic and can heat the silica enough to make fissures and ruin the column. Watch out!

    Great video!

    Reply
  3. Silverweed
    Silverweed says:

    Having to collect 100+ fractions and spot all of them on TLC only to realize you didn't get enough separation. Having the column fissure right after adding your precious sample and having to recover every bit you can rotovapping down tons and tons of solvent praying you can get everything back. ughhhh never again please!! 😛

    Reply
  4. ChemistDrummer
    ChemistDrummer says:

    You forgot to add another "Caution!". Never clamp the pressure tube in place when having the stopcock close like shown in the vid. Danger of explosioni if you dont release the pressure quickly enough.

    Reply
  5. Bryce B
    Bryce B says:

    so if you determine the right amount of solvent to use for good separation, let's say 45% petroleum ether, 50% ethyl acetate, and 5% methanol from doing TLC, is that the solvent ratio I would put into the column?

    Reply
  6. daveyboy2112
    daveyboy2112 says:

    Yes, TLC is pretty much directly applicable to the column as you have the same stationary phase….you would generally run a gradient of solvent though through to the optimal solvent ratio….can't remember exactly but you can go from something below the optimal up to 5/4 of the strong solvent (where 4/4 is the optimal ratio giving you the best separation/Rf value).

    Reply
  7. Frédéric Diotte
    Frédéric Diotte says:

    I had a "science integration activity" course in college where I had to run an experimental project on my own, and I'm now really disappointed that I didn't use this technique. These videos have an awesome pedagogy, thanks!

    Reply
  8. Phillip Allen
    Phillip Allen says:

    she didn't explain how the sample got between the silica and the sand, or the reasons for doing this.  The sample was loaded on top of the second sand layer (which was loaded directly on top of the silica); but once the sample was completely loaded, it was shown that the sand was on top of the sample.

    Reply

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