Manually running a column can be a fun science experiment to show people. For example, separating the colors of a mixed color solution is usually a fun project for the kids to watch. Pour in the black liquid, and then watch as the red, yellow, and blue colors separate in the column. You see everyone’s eyes light up as the colors start to separate. But separating colorless 5-MeO-DMT from colorless DMT is a lot more difficult. It’s not something you can simply eyeball. There are ways of forcing human detectable colors by the addition of other chemicals. For example, a specific salt form of both alkaloids might be red for one, but colorless for the other. This way you can visually see them separating in the column. But you need color reaction data on both DMT and 5-MeO-DMT before this is possible. Where can this be found? Another option is that the collected batches are simply analyzed later by some other technique. There are many different ways to tackle this problem.
The thing SWIM hates about running a manual column is all the preparation. You have to first clean the column very well, which is hard because of its shape. Then you have to put a plug at the bottom, which is also hard sometimes. Then you need to add the stationary phase, eluent, etc. And there can’t be any air bubbles present. Then when it’s all ready you pour in your pre-dissolved alkaloids. Then you need to sit and watch it, and collect all these different batches. Then after you’re finally done, you need to remove the stationary phase, plug, etc., and rinse the column again. Oh what a pain. Sometimes it’s really hard to get the plug back out especially if anything you poured in became sticky.
With plant extractions sometimes there are all sorts of unknown things present that mess up the process. Sometimes the column becomes clogged or runs really slowly. Sometimes the alkaloid you want comes out along with other plant compounds unless the right solvent or solvent mix is used. It can take forever to figure out the right solvent mix if there are no references to follow for a particular alkaloid. Usually the fraction is just run again in a longer column.
I’m sure you know all of this, but I think most people hear “just run a column” and think it’s a pretty simple lab procedure. It’s not, it’s a lot of work and you do run into problems with the procedure sometimes.
There are cases when column chromatography is the only procedure that works well.
Column chromatography is definitely very useful when you need to separate dozens of alkaloids from a plant alkaloid extract. For example, an Anadenanthera colubrina alkaloid extract contains about a dozen different alkaloids. Column chromatography is the only good method for separating all the alkaloids present. But if you’re just targeting one or two alkaloids, then alkaloid precipitation using a solvent + acid or solvent + base mix is usually much easier if possible.
The disadvantage of using acetone citrate precipitation is that there are alkaloids other than 5-MeO-DMT citrate that will precipitate out. This is primarily good for separating 5-MeO-DMT from DMT. The DMT citrate remains in the acetone, but the 5-MeO-DMT precipitates out. It will not separate 5-MeO-DMT from bufotenine. Bufotenine will precipitate out along with the 5-MeO-DMT. There are other methods for separating 5-MeO-DMT from bufotenine. The one SWIM uses is to dissolve freebase MeO-DMT and freebase bufotenine in heptane (or naphtha). Freebase bufotenine is insoluble in heptane (or naphtha) but freebase 5-MeO-DMT is soluble in it. The bufotenine sinks to the bottom of the heptane and is easily filtered out.
