Research Ethylene and other ways to increase biosynthesis in cacti experiment

[modern]

Researching the biosynthesis of mescaline in cacti I proposed the idea that ethylene would increase the production of mescaline my increasing the final step of conversion but took a year to get around to actually being able to test it due to lacking extraction experience my first attempts.

I first did this with Cielo and had precipitate vs almost none with the control. However due to not being able to get measurements it was only visual confirmation.

So I set up an experiment using the same exact clone from the same exact stump so completely uniform. I selected the largest as the control and the ethylene and another product as the two variables.
For two weeks I added fresh calcium carbide daily to a white plastic bag and the other two I kept laying in bright spot with one being the control and the other being the product.
Once complete I dried them all in the same manner in the microwave until cracker dry taking around 20-30 minutes each. Saving the liquid I decanted to reapply to each accordingly. I then added them all to glass jars and added equal amounts of lye water same concentrations in excess of 13ph after a day added salt in equal amounts and xyelne then shook. Every time I’d see them I’d shake them and allow them to seperate. Around 100ml lye solution and 100ml xylene. I replaced with fresh xyelne and repeated and after a week I pulled and salted each. I added excess hcl water to salt all the same concentration each reading ph of 1-2 after salted
Then I started to evaporate. The control had the normal red impurities with the mescaline the ethylene had very little red impurities only appearing for small portion and the product x had more red impurities then the control.

Control salts are 185mg with starting weight of 11.02 means yield of 1.68% red impurities

Ethylene salts are 184mg with starting weight of 9.26 means 1.99% with much less red impurities.

Product x salts are 216mg with starting weight of 9.31 means 2.32% with more red impurities than control



Perhaps combining product x with ethylene will increase even more? Need to test how much remains after isopropyl cleanup.

I have filtered each after adding fresh 30ml of water so they are evaporating once more. I’ll weight again and then do isopropyl 99 wash and weight each separately.

Ceilo would be a great test since so selective with mescaline.

 

[Transform]

Interesting stuff. Do you have any links to your original test. I remember this being discussed a while back.

Were you able to clarify whether the acetylene/ethyne from the calcium carbide gets metabolised by the plant into ethylene/ethene? Seems it's a ripening gas anyhow, but there are caveats (most notably, highly toxic phosphine):

Calcium carbide is sometimes used as source of acetylene, which like ethylene gas, is a ripening agent. However, this is illegal in some countries as, in the production of acetylene from calcium carbide, contamination often leads to trace production of phosphine and arsine.

These impurities can be removed by passing the acetylene gas through acidified copper sulfate solution, but, in developing countries, this precaution is often neglected.

 

[modern]

Interesting stuff. Do you have any links to your original test. I remember this being discussed a while back.

Were you able to clarify whether the acetylene/ethyne from the calcium carbide gets metabolised by the plant into ethylene/ethene? Seems it's a ripening gas anyhow, but there are caveats (most notably, highly toxic phosphine):

Calcium carbide here is different than some other countries as is our isopropanol 99%
Like with other chemicals there are different processes for the production and here because of industry scale and other regulations.
Isopropanol 99% has minimal benzene contamination due to route of production in the states with 80% production being made in that manner that may have some benzene. Here in Brazil the process for isopropanol 99% is different as with different compounds leading to not having this issue of benzene contamination with our iso.

Ethephon Is usually the source used for ethylene but there are also painted calcium carbide that are planted in the soil in a time release manner. Carbide is prohibited use but due to false advertising the fruits being lower nutritional values. Also there are inferior quality carbide so always a possibility so that extra step to clean the gas is worth doing since not difficult. Consuming constant amount of fruits that have been matured with carbide may be bad so everyone do your own research.

That being said I do suggest using bananas or apple bundle for the ethylene source being more readily available. There are also other sources like smoke from burning wood or even gas burning.

acetylene Is converted to ethylene by plants and even used to induce flowering out of season by some farmers. There are better and easier sources however that can be applied as a foliar but usually need to buy large volumes.

I made a post on proposed mescaline biosynthesis a while back on possible ways to increase each step in the process. With ethylene being the final step. (I may have been wrong)

@Loveall made a post as well on this.

The “product x” is a mixture of organic acids I plan on trying to sell as a foliar spray for greener flesh and higher nutrient uptake among other benefits.

I still plan on doing isopropanol wash and seeing yields of “washed mescaline” vs the other impurities. I also plan on doing TLC on the two to see if indeed my experience of “non mescaline alkaloids” is not just mescaline as some think.

 

[modern]

Here are the links
Ideas to increase biosynthesis of mescaline

Increasing Mescaline Biosynthesis to Genetic Potential

So I shared this idea on my YT but since discussion isn't very easy on that platform I'm posting this on a few forums so others can test this idea. This may all be bullshit but all seems viable and logical to me. I'd love if someone could test atleast the apple/banana theory next harvest of...

forum.dmt-nexus.me

@Loveall thread with some different info/references

Research - Ethylene gas experiment

Documenting a simple experiment here to test the following hypothesis: "Stressing cactus in the presence of ethylene gas increases mescaline content". This hypothesis has been brought up by user modern. See spoiler for more details as I understand the latest status. My attempt at...

forum.dmt-nexus.me

 

[modern]

Still finishing up but control has .39% yield
And ethylene has .67% washed with isopropanol until no more discoloration. The content saved is insoluble in isopropyl 99 and washed.

72% increase in mescaline content with control vs ethylene.

Only 23% of the total alkaloids is mescaline hcl that is completely insoluble in isopropyl 99. For the control

For 34% of the total alkaloids for the ethylene experiment.

There may be some mescaline that dissolved in the washes which I’ll test soon via TLC.


The first photo is the discoloration of the isopropyl washes done until clear in small addition. Second photo you can see some crystals that MAY be mescaline hcl with the washes but I’ll need to test. The 3rd photo is the clean mescaline hcl. Started out red. These are from the ethylene experiment. The control ended up a gray like color.

Should have the product results in another week. Good chance that the mescaline content didn’t increase and only the other impurities since it was the most red.

I have another pulls of the xylene to see if there is more content still in the cactus. Will test next year most likely.

 

[modern]

So I did a test run on TLC using a dilute lophophora extraction and had zero staining with my ninhydrin spray. I either need to heat the plate or need to concentrate the solution more. I also didn't use the mobile solvent since I still didn't get my methanol or ammonia. I just did a quick test with the rest of my tlc plates.

First I'll concentrate the extract I made from .5g hcl alcohol solution. Soaked for a week shaking then centrifuge with a fan to separate the liquid. Then I took a tiny capillary and made only single spots on my DIY plates. The ninhydrin spray I used .2g in 40mL ethanol 96 following a recipe I found online. They suggest adding hcl to acidify which I may just follow. The paper shared here did a lower concentration of ninhydrin which I may attempt first so I can spread my 10g further also skipping the acid since not added sometimes.

Lophophora vials under black light. I need to concentrate them since didn't show any staining with the ninhydrin.

EDIT I did a TLC but used 50/50 ethyl acetate/ isopropanol with hcl acidification. Didn't use the standard tlc solvent of methanol and ammonia

 

[modern]

Ok took longer than the paper that @An1cca took like 1-2H vs 20-30 minutes but maybe substitute using ethanol 96 vs acetone.

Despite my very DIY plates and methods seems that quantitative measurements is possible. These plates were spotted 1,2,3 times each with different solutions.

Just a first attempt and it was successful. I had some streaking but will try in the future using methanol and ammonia 100:1.5 ratio.

If plates are still stained tomorrow I’ll take a photo to try my hand at the photo calculations. In the future I’ll make a control spotting of 1mg/ml mescaline solution as the reference then calculate from there.

Another thing is I’ll be making only the thicker sílica plates as those seem to work better with staining. I got it to work with these but with mixed results.

Im the future I’ll make a plate and test if too concentrated then dilute as needed.

If I can get a somewhat reasonable result in quantitative measurements I hope to test lots of different ideas… pretty excited for the new year

 

[modern]

Control salts are 185mg with starting weight of 11.02 means yield of 1.68% red impurities

Ethylene salts are 184mg with starting weight of 9.26 means 1.99% with much less red impurities.

Product x salts are 216mg with starting weight of 9.31 means 2.32% with more red impurities than control


Control has .39% washed hcl
Ethylene has .67% washed hcl
Product X has .83% washed hcl
with isopropanol until no more discoloration. The content saved is insoluble in isopropyl 99 and washed.

72% increase in mescaline content with control vs ethylene.
113% increase in mescaline content with control vs product x
24% increase in mescaline content between ethylene vs product x.

Only 23% of the total alkaloids is mescaline hcl that is completely insoluble in isopropyl 99. For the control

Mescaline content is 34% of the total alkaloids for the ethylene experiment.

Only 35.6% mescaline content of the total alkaloid content of the product x.

The photos are of the product x mescaline and isopropanol “nonmescaline” alkaloids. All three experiments received identical treatments. I have combined all the non mescaline washes and will recrystalize and see if I can possible precipitate any mescaline hcl. I will preform a TLC in the future but will mês to make plates again.

 

[modern]

I'm not exactly sure what to make of this information. There is a 'true increase' regardless with a single treatment since starting Fresh cut weights are identical however weights after 1 month and dry are quite different and I'm unsure which yield is correct? 14.5% increase or 46% increase

So I just finished up the second experiment testing 'True Color' and while reviewing the data noticed a large 'discrepancy' in the final results depending on how you present the information.

I had an even larger skewed result when I only compared the insoluble washed crystals leading to 107% increase but found 40% total alkaloid increase more representative. This was how I presented the information the first experiment.

What's missing here and would be nice to have is 1 more extraction of the FRESH CUT to have results before the 1 Month experiment.

	Control	Single Treatment	Multiple Treatments
Fresh weight	395	396	381
Water Loss % +1 Month	-23%	-29%	-28%
+1 Month Fresh	304	281	275
Dry	23	18	16
Yield	393mg	451mg	389mg
Y% Fresh	.0995%	.114%	.102%
y% Fresh +1M	.129%	.161%	.141%
Y% Dry	1.71%	2.5%	2.43%
% Difference from Fresh	-	+14.5%	+2.51%
% Difference Fresh +1M	-	+24.8%	+9.3%
% Difference Dry Weights	-	+46.2%	42.1%

 

[modern]

So I went back to check fresh weight yields of my first ethylene and 'true color' experiment since I felt the dry weights were strange.The first experiment the dry and fresh % gains were very close to each other.

Experiment 1 Fresh W Yield F% yield F% Yield + Dry% Yield +
Control 300g 185mg .062% - -
'True Color' 252g 216mg .086% 38.71% ~40%
'Ethylene' 256g 184mg .072% 16.13% ~20%

Experiment 2
Control 395g 393mg .0995% - -
'True Color' 1x 396g 451mg .114% 14.5% 46.2%
'True Color' 7x 381g 389mg .102% 2.51% 42.1%

The second experiment the clone seems to already be stronger... there was a 1 month 'aging' period vs 2 weeks with the first experiment so maybe that has a larger influence. As always more experiments needed to draw more conclusions.

Many think the difference in yield is just the water loss that leads to increase content but these results o show a 'true' increase since comparing the starting fresh weights before any 'aging or treatment'