OK guys,

I just sent a sample of the freebase before Manske to EC.

Also, I put both powders under the microscope.

first picture: Freebase to acetate to freebase in situ (=on the microscopic slide with vinegar and NaOH)
second picture: Har.HCl (after Manske of freebase) to acetate to freebase in situ

I don't see any difference.
When comparing with the pictures of sample 1 in post #342 (=the same batch), it becomes clear that it isn't easy. Some regions are full of 'oily debris', while others are clear and only contain harmine-needles. I searched for DHH-type crystals in the current slides, but couldn't find any.

So if the FB-analysis of EC confirms the HCl-results, one might conclude that indeed DHH was converted to HAR. If it says 78/4% (like 3 years ago), it looks like Manske does selectively crystallise HAR out of a mixture with DHH. So in both ways, results are going to be informative
Thanks Endlessness!

I also found no difference between before and after manske, as expected.
The dhh survived the manske glamorous and actually looked a tad cleaned up by the manske.
In attachment a little vid that voyages though the drop so we have more to look at than a single snapshot. When a fern is on it's side it does look like a spike, but is not like a spiking harmine crystal. So this is 3 year old DHH FB --> manske --> FB --> in situ inspection.

I've some dhh in the freezer to see if that does something, that's the only immediate pointable difference between my batch and yours An1cca. So far my dhh (and thh) seemed to have survive quite well considered in FB form and at room temp.

Most interesting, Jees! Your DHH was clearly well preserved. Congrats on your director-skills as well ...

Obviously DHH can easily be transformed by Manske in the case of a mostly DHH-sample. However, it might turn out that when DHH is only a few percent of a mostly HAR-sample, the Manske does selectively pull out the main 95% of 'Manskeable' alkaloids, leaving DHH in solution? If this is confirmed by the EC analysis, then this might be a way of cleaning up mostly pure harmine. Following this principle, in the case of a HAR+DHH to HAR+THH Zn-reduction, a Manske could be used to clean up the HAR fraction?

Let's see what comes out of the analysis. It might turn out that I got the topic of this thread totally wrong ...

My THH doesn't seem to survive well at all. The batch I reduced a couple of months ago seems to be mostly DHH now (based on bioassays), and I've stored it in the fridge.

I wonder if storing THH as acetate salts would be a better long-term solution? (no pun intended)

It's not years but 3 days of DHH FB in the freezer did not change the visual signature of the DHH crystals when recrystallized in situ.

I feel trepidation to ask but An1cca are you positive that there was no mix up of labels or lids with ID on? Such a hard roll back is quite outspoken.

Yes Jagube a salt form is expected to store better, maybe better to go that way.
Is your THH starting to taste bitter? Did you taste it at the beginning when fresh?

It started to taste bitter a long time ago, but now it feels like pure harmaline, no THH effects.

Ok Guys, thanks tot Endlessness & EC, we have the results of the HAR-freebase analysis. It turns out to contain 88% HAR and 1,5% DHH (harmaline).

This is a rather confusing result, as the EXACT SAME freebase sample was sent 3 years ago and tested as being 78% HAR and 4% DHH. I have not dried it any more than it was then. It was stored compacted in a glass vial with metal lid.

So, it might still be that a part of the DHH converted to HAR. Even then, the remaining 7,5% extra HAR remains unexplained.

Also, this result would suggest that a further fraction of the remaining DHH was lost in the Manske process as well. Therefore, Manske could be a way of getting rid of small impurities in a relatively pure single harmala-sample...

Could the measuring error of a HP-LC + MS technique (7,5%-10%) be that large in the experience of others? Or do we see any other explanation?

I will ask them to test it again to see if we can at least eliminate the possibility of error.

Possibilities are loss of volatiles (water or organic solvent) from your sample or some error or inconsistency with the standards used by ec. Not criticising here, just thinking out loud.
You say your sample was stored air tight, therefore loss of water is not possible.
What if the sample you sent to ec this time around was allowed to sit out in the open for a day, and water evaporation occurred then. But three years ago, this was not allowed to occur.

If I understand correctly (I hope so, please correct if mistaken), that was a recent manske right? So the sequence is:

* Har FB 3 years ago = 78%har + 4%dhh = 82% alks total
* wait/storage for 3 years
* perform a manske
* move back to FB
* new analyse: 88%har + 1.5%dhh = 89.5% alks total

If that's correct then an elevation of total alk purity due a manske could be normal I think.

The fall back of dhh from 4 to 1.5 could come also partly due the 3 years waiting time, so not necessarily due the manske alone. Maybe they both work in same direction, the storage time and the manske, hard to know their respective dues.

Or maybe the ambient humidity three years ago was higher and more ambient moisture got absorbed during handling back then?

Comparing the DHH to harmine ratios - 4:78 (=1:20.5) vs. 1.5:88 (=1:58.67) - they tantalisingly suggest that DHH was indeed converting into harmine over time - if this really was the exact same alkaloid sample.

Certainly possible. Both theories could be tested. If anicca takes another sample out of storage and weighs it on a decent set of scales, then leaves the sample out in the open for a couple of weeks, then re-weighs the sample. The drying or absorbing theory will be tested.

@ Endlessness: I have been doing some quantitative TLC lately and my respect for chromatographers has become as big as my first standard deviations ... If they would like to repeat the analysis, at least one explanation can be ruled out (measuring error this time). A measuring error 3 years ago still remains a possibility.

@ Jees: the thing is, only a part of the original freebase-batch was converted to HCl-salts (I thought this form would be better for room-temperature storage and easy access). The FB sample I sent a few weeks ago was from the exact same FB-batch, unmodified. So there was no Manske step to raise purity (if that is even possible beyond 88%??).

@ leratiomyces: as it happens, I have a very decent scale (0.1mg). So as you suggest, I will do the experiment and let a batch of FB dry over CaO for 2 weeks, measure 5,000g and then place it over water under cover for 2 weeks and weigh again. I don't want to insert a heating step that might bring other uncertainties with it...

Thanks for thinking along, guys! Nexians rule