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I did a test on the beads that are turning greenish blue after refusing to elute anything into room temp alkali solutions:


Boiled them in alkali water. They released a bunch of stuff. The beads stayed discolored, but much less intensity. I filtered the elute, acidified it and put it on fresh beads. Surprisingly, it smelled funky, with a shroomy odor. Then, I filtered this out, rinse the beads and extracted in alkali water at room temp. They released a bunch of stuff into solution.


So by boiling the beads that had stopped eluting into a base at room temp, some stuff was released. When this stuff was acidified and put through a set of fresh cation beads, at least two fractions could be recovered: one that did not bind to the beads in acid conditions and was stinly, and another one that did bind and could be released into alkali solution at room temp (before boiling this was not happening).


I think this is consistent with some kind of change during the boil that makes the extract more friendly to ion exchange elution? Seems consistent with protein denaturalization through heat?


I don't know, maybe I'm wrong. One next test as mentioned before is to boil the next tea after the enzymatic conversion and see how that behaves.


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