Hello there
We started with 10mg and 5mg dry leaves. They were boiled in 1% acetic acid. The resulting tea was based with sodium carbonate and pulled with DCM. The DCM fraction evaporated and washed with methanol with ammonia. I forgot the exact measurements but I think I wrote them in previous posts in this thread

10mg and 5mg dry... thanks for that info. And you will be quantifying using TLC I presume? And selecting plants and breeding from there.

10mg isn't much, I could take samples from patches of wild phalaris for quantification. Do you know (roughly) the concentration of dmt/5meo you used as your reference spot?

I'm not the guy who is performing the TLC but a German friend of mine who is not a member on the Nexus.

He uses miniscule amount of DMT and 5-MEO-DMT for standards just enough to have a visible spot on the TLC.

TLC as a method is only semi-quantitative as it cannot tell you exactly how much product you have like LC-MS does. however the size and darkness of the spots can help give you a vague idea about how much of an alkaloid there is on the plate.

Luckily I have an active and good yielding strain that we will use was standard to compare with the other strains to pick out the most interesting ones for breeding.


I have sent you a pm.

Page won't let me reply to your PM because I am a new member. That breeding program sounds fun and interesting. I'm keen to get involved.

Your active and good yielding strain – how many plants did you have to go through to find it?

The active aquatica i have is a local cultivar reported to be 98% true to seed.. genetically pretty stable in my experience both quality and yield wise. Most of my bioassay reports are on the thread (phalaris project) I have tried both clones and seedlings of this cultivar to both have delivered the same yield and alakloid profile.


I have over 50 clones covering 3m Square area. Keep posting and sharing and I'm sure your membership will be upgraded soon. Looking forward to collaborate with you.

Here's a summary of all seedlings TLC analysis so far. Please feel free to share your interpretations.


Personally I think the sardinian seedling (last spot on the first tlc plate) is the most interesting as it has the cleanest alkaloids profile (no tyranines whatsoever) seems like it only contains either DMT or 5-meo-dmt plus gramine at lower proportion.


Uruguay seedling (last spot on third TLC plate) is also very promising as it's also substantially clean profile and has a deep big sspot for DMT.

Blue fluorescence was observed on tlc plates with nndmt but not with 5meodmt when using 275nm led but this fluorescence vanishes as the eluent (meoh + nh3) evaporates.

Italian strains 1, 2 and 3 look interesting as their alkaloid profiles are similar (dominant/stable chemotype) and clean. Are they the same due to high genetic stability or just selection based on lots of trial and error? Do we have any idea how common/rare a high yielding (say 0.5%) wild type Phalaris Aquatica is?
If wild high yielding strains were common, people would only need to sample a few local Phalaris plants, run quantitative TLC, then transplant and clone it at home. This way, people could have a consistent DMT source in about as little as 2 months.

Judging from the Phalaris project thread, it seems to be a good ayahuasca analogue so long as you find a clean strain, though the effects are most likely due to the activity of 5-meo. From what I have seen, extractions of random wild grasses seem to be hit and miss. Selection of a high yielding strain first seems necessary.

Regarding selection for a high yielding strain, take a look at the graph attached for an idea on how DMT concentration may change under selective breeding. Of course only a guide.

All seedlings sampled by tlc above are wild accessions brought from the internet. No breeding attempts have been made yet but is expected to start around February.

There are as many active high yielding wild strains as there are inactive/law yielding ones.. all kinds phenotypes exist and seem widely and randomly distributed throughout the native range of this species.

Selective breeding for higher tryptamine is so much easier than selecting for low tryptamine since it's the innate specie behaviour to produce these alakloids for defence. We found some variability within wild accessions not just amongst accessions. Cloning the strains of favourable traits and breeding them amongst each other will narrow down the generic pool in the direction we want..further selective breeding cycles will narrow it down even further so that less and less proportion of low alkaloid seedlings will emerge from a seed batch.

The larger number of wild accessions used in the breeding program the larger the starting genetic pool which will prove useful for future long term selective breeding. For other traits like vigorous growth.

Once a satisfactory yield And alkaloid ratio has been reached the next step would be to stabilize this hybrid to become a cultivar which can be achieved through back crossing this hybrid with an already established commercial cultivar like CV australis.

Even a single favourable mutation in a single seedling from a large field of a phenotype such as seed retention can be incorporated through breeding to become a dominant trait such is the case for the UNETA cultivar a line of CV australis.

I have already reached the 0.5% yield from my local commercial cultivar.. ans I didn't perform any breeding on it which I will do soon enough.

The literature on aquatica breeding is very plenty coursing through decades of documentation provided by CSIRO.

The most notable paper to read after AQ1 would be (the first century of phalaris aquatica breeding) by oram.

We're currently building an improvised HPLC and a UV spectrometer system to replace TLC. So far we've built and operated a crude prototype to get more acquainted with the principles of a chromatography column and UV detection. It kind of works but the signal quality still leaves much to be desired.

Quartz tubing has been ordered for the spectrometer flow cell and a high pressure pump is likely to be purchased as well to build higher resolution cappilary column.

Any suggestions where I can open such thread? Perhaps in advanced chemistry? Below I added some resources.

https://waveandsignal.blogspot.com/2012/?m=1

https://labsmith.com/lab...d-chromotography-system/

http://www.ionsource.com/tutorial/capillary/uv.htm

Tlc on basic A/B extraction of 32g of 2nd regrowth of light and water stressed (2 weeks) wild type phalaris aquatica (notice the leaves beginning to turn yellow). 3x30min boils, combined, based, naptha added and heated, then left to sit overnight. Yielded approx 1-4mg from this first pull (.003-.01%). Even assuming I pulled as little as 50% from what is in there, it still would be as little as .025%.

The phalaris in my area are just coming out of anthesis, will just have to continue doing this on wild strains until I find a good one I guess.

KEY:
C=caffeine
P=phalaris grass
A=longifolia extraction
B=Acetic Acid

You can see a faint spot on P column for where DMT should be which is a good sign.