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pinkoyd
#21 Posted : 3/14/2017 2:10:20 AM

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Sorry for not mentioning it but yes you have to be a member there.

It's well worth it though. Not the most active community anymore but it's a treasure trove of good cactus info.
I already asked Alice.

 

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urtica
#22 Posted : 5/26/2017 5:52:43 PM

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#12 Trichocereus sp. 'SS01 x SS02'

Ok, folks, moving on to hybrids.

I got these cuttings from Cactus Arizona. The plants are seed grown from Sacred Succulents seed. Both SS01 and SS02 are known to be nice alkaloid producers.

The cuttings are skinny, look more like a bridgesii.

A one foot cutting and a ten inch cutting together weighed 1000g.

Yield was 475 mg of a yellowish crystalline powder.

The marquis reaction was very nice and orange, looks to me like only one alkaloid in there, but I will have to wait for further testing.

TLC ran nicely although it barely moved up the plate.

So, 0.0475% fresh, 0.095% presumed dry of the crude extract.

Pretty good!

ENERGY CONTROL RESULTS: 86% of this extract was mescaline, with one unknown compound, MW = 163. So fresh mescaline 0.0409%, presumed dry mescaine 0.817. This is higher than Lumberjack folks.
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urtica
#23 Posted : 5/26/2017 6:01:34 PM

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#13 Trichocereus peruvianus "John"

This is a big, fat, woody cut that I got from Misplant. It has the blue color and the fat, fuzzy aereoles that I associate with an active peruvianus. The spines do look a little long maybe? Slight swelling at the base of some of them...

I am sick of only testing inactive peruvianus so I am trying to look at some of the nice, fat, blue 'Matucana' types, or at least named clones in the hopes that whoever named them did so because they are active not because they are trying to sell them for more money.

A ten inch cutting weighed 1060g.

It took some muscle to cut it up. Also please note that this cutting sat on a shelf in low light conditions for over a month before I got around to processing it.

Yield was 187 mg of a tannish powder.

So, 0.018% fresh. 0.35% presumed dry.

I am unimpressed altho way better than the cuzcos!

The marquis reaction was nice looking, looks like 1 alkaloid only.

The TLC also ran well with no plant goo visible.

ENERGY CONTROL RESULTS: 38% of this extract was mescaline, so 0.0068% in fresh plant material, 0.133% presumed dry. 68 mgs mescaline per kg fresh plant.
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urtica
#24 Posted : 5/26/2017 6:11:10 PM

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#14 Trichocereus peruvianus JS209 "Poots #2"

Sticking with peruvianus. This is a HUGE fat cutting from Cactus Affinity. It is very blue, has the fuzzy areoles, and has the honey colored at the bottom, black at the tips spines that look like a winner to me.

Six inches weighed 1230g! It was the bottom six inches of a tip cut, so it was not woody, cut easily. This cut also sat for over a month.

Yield was 380 mg of a tannish powder.

So, 0.031% fresh. 0.62% presumed dry.

Nice! That is a workable cactus!

Marquis and TLC were both nice & clean.

ENERGY CONTROL RESULTS: 38% of this extract was mescaline, so 0.0118% fresh, 0.236% presumed dry. 117.8 mgs mescaline per kg fresh plant.
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urtica
#25 Posted : 5/26/2017 6:28:12 PM

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#15 Trichocereus pachanoi P.C Hutchinson 1597 Peru 57.0884

I got this pachanoi from Cactus Affinity. I did not really pay attention to where it came from originally until I finished the extraction (spoiler, I was impressed!). This clone was collected as live material in the late 1950's in the Huancabamba region.

See more about it here:
http://troutsnotes.com/p-c-hutchison-1597/

Since I cant get my hands on an Ogunbodede cut or a Landfill, I am searching for a nicely active pachanoi.

This 7.5 inch cutting weighed 990g.

It is fat, and has the longer spines of a classic 'wild type' pachanoi. Honey colored spines with dark tips. This was the bottom of a tip cut that came from a fallen column. It had only been cut off the mother plant a week or so before the extraction. It cut up easy and was not woody at all.

Yield was 433 mg of really clean looking crystally powder.

That makes for a fresh % of 0.0437%. Presumed dry is 0.87%

I like this clone! Move over Landfill! Luckily I kept the tip to plant out. I think that especially considering that this cutting was not stressed at all that we have a nice alkaloid producer here.

P.S I have noticed that when the cactus tea forms a thick emulsion that is hard to break with the xylene, that is often a sign that it is alkaloid rich.

I forgot to take pictures of the cut that I extracted, so shown is the tip that was left over to plant.

Marquis and TLC were both nice, no goo.

ENERGY CONTROL RESULTS: 88% of this extract was mescaline, so 0.0385% fresh, 0.76% presumed dry. 385 mgs mescaline per kg fresh plant. This is the highest percentage mescaline of any extract that has been produced this far.
urtica attached the following image(s):
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DansMaTete
#26 Posted : 5/26/2017 10:59:23 PM

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Thanks for all your work Thumbs up




pinkoyd wrote:
Highly interesting stuff from our friends over at The Nook:
...
This is an increase of more than 100%, and this just because of storing the column for 3 month in the dark before drying!
...



I was runing my own experiment when this post popped out. In this thread you can find my results which, in most of case, give the same tendancy (increase of mescaline content) but not each time.


« I love the smell of boiling MHRB in the morning »
 
urtica
#27 Posted : 5/28/2017 2:44:17 AM

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DanMasTete, your research is great! I am curious about the couple that seemed to lose some potency.

Also I saw the post where you extracted your homegrown Mimosa tenuiflora & that was a wonderful thing to see. Way to go from seed to alkaloid with your research!
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urtica
#28 Posted : 6/1/2017 8:59:00 PM

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#16 Trichocereus spp. "Ohlone"

This is a big fat sorta peruvianus looking one from Cactus Affinity. He got it from someone who planted cacti around Ohlone College. This cut was made about a month ago.

This 1 foot cutting weighed 1820 g! Big fat guy, pretty blue. It does have big fuzzy areoles but it does have some kinda cuzco-y spination, some swollen bases.

I extracted as usual and ended up with 299 mg of a tannish crystalline powder.

Marquis reaction was present but was on the murkier side. TLC ran nice and did have a positive mescaline spot tho it was light.

So fresh is 0.017%, presumed dry is 0.33%. This puts it on par with T peruvianus "John". So based on the chemisry, I want to call this one a peruvianus? So far all the cuzcos have given me goo only with very little alkaloid in the goos... Seems like a workable clone.

My camera was away for the end of this extraction so I only have pics of the cutting, but you can imagine them by going back over the past entries.

ENERGY CONTROL RESULTS: 55% of this extract was mescaline, so 0.00935% fresh, 0.182% presumed dry, 93.5 mgs per kg fresh.
urtica attached the following image(s):
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An1cca
#29 Posted : 6/3/2017 10:19:08 PM

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Nice work Urtica!

I'm doing some research on running TLC on homogenised raw cactus material, 'sap'. This method seems good enough to give a relative idea of strenghts of different cacti and allows for semi-quantification if a standard is put on the plate as well. Possibly, the use of ninhydrin can help as well together with image manipulation software. I tried 'TLC analyzer' with promising results using UV-light.

BTW, do you know how big your extraction-error is? If you divide your cutting in 3 equal parts and run your extraction-procedure on it, how big is the range of extract weights?

Keep going, very valuable work!
 
urtica
#30 Posted : 6/4/2017 2:20:05 AM

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Hey An1cca!

That sounds really cool! You just smoosh up the cactus/blenderize it, then cut it with MeOH/NH4 & run that on the plate?

Then put a reference standard in one lane, and compare the strength of the color reaction or reaction to UV light?

I have not checked my extraction error and was not familiar with the idea. Do you think that extracting the same weight of the same batch of dried cactus skins three times would perform the same test? Also do you think that the variance in alkaloid content between different parts of the cactus would cause some of that error, not necessarily all the result of the extraction?

When you get the three weights is there some math that you do which gives you your extraction error % or something like that?

I would be so curious to see any results from the work you are doing!
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An1cca
#31 Posted : 6/4/2017 8:32:34 AM

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urtica wrote:
Hey An1cca!

That sounds really cool! You just smoosh up the cactus/blenderize it, then cut it with MeOH/NH4 & run that on the plate?


Actually, what I do is take a 'punch biopt' with a dermatological punch of 3mm. The sample is 7mm long. This way, the sample procedure is standardized. Then I freeze the sample in an 'Eppendorf' vial of 1,5ml for a few hours and then, upon thawing, I mash it up with a steel rod that fits snugly in the vial for a few minutes. I then fill about 1/8th of a spotting capillary from bunkpolice and aim to make as small a spot as possible. After thorough drying I run the plate with MeOH/NH4OH. Next to the cactus juice, I spot some different volumes of a M-SO4 solution. I'll post a picture of such a run. All ideas are welcome, I'm still just starting with this...
An1cca attached the following image(s):
TLC analyzer cactus juice.jpg (208kb) downloaded 833 time(s).
TLC UV cactus juice and reference.jpg (251kb) downloaded 824 time(s).
 
endlessness
#32 Posted : 6/4/2017 1:53:23 PM

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An1cca, that sounds like a great way to go about this.. The biopsy punch is a very good idea!

So the TLC Analyzer is free, I suppose? Is that the one you've been using?

Also, how do you scan the plates, do you just use a UV light and take a pic of the plate? How do you set up so the light is more uniform, at what angle is the light and the camera?

I've done this 'semi quantifying' for DMT in mimosa, except I was just using my eyesight instead of a software, and it seemed to work decently well.. Instructions and a test example in pictures below:



 
urtica
#33 Posted : 6/6/2017 6:25:59 PM

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That is all great, you guys!

I do think that making tests using tiny amounts of plant material sounds really nice & like a lot less work than always chopping up big cactus columns...

How long does it take the straight cactus juice spots to dry?
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urtica
#34 Posted : 6/6/2017 6:33:50 PM

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#17

Trichocereus bridgesii 'SS02'

This is a classic clone, originally from Sacred Succulents who gave it the 'SS02' name. This clone is known to be active, it was highlighted in The Entheogen Review years ago by K Trout.

These cutting that I analyzed came from Cactus Affinity, however. They were a little messed up, some healed over rot spots, and had been cut for at least a few weeks when I got them, then sat in my house in ambient light for about a month.

Three cutting totaling 31 inches weighed 1130g. I forgot to take a picture of the actual cuttings so I am including a picture of the clone growing in my greenhouse.

The emulsions during extraction were sticky, which implies to me alkaloids, and indeed I ended up with 904 mg of crude extract, which is the best so far!

This makes for a fresh yield of 0.08%, presumed dry yield of 1.6%.

The extract does seem a little oily/dirty, and the Marquis reaction was a little muddy looking as if there are multiple alkaloids in there.

TLC run was nice, looked pretty clean actually.

ENERGY CONTROL RESULTS: 75% of this extract was mescaline, so fresh 0.06%, presumed dry 1.2%. 600 mgs mescaline per kg fresh plant. The dry % is as good as some Lophophora percentages I have heard.
urtica attached the following image(s):
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An1cca
#35 Posted : 6/7/2017 2:06:02 PM

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endlessness wrote:
So the TLC Analyzer is free, I suppose? Is that the one you've been using?

Indeed

endlessness wrote:
Also, how do you scan the plates, do you just use a UV light and take a pic of the plate? How do you set up so the light is more uniform, at what angle is the light and the camera?


For the moment, I just hold Bunk Police's UV-C light 50 cm above the plate in my toilet and take a perpendicular picture of the plate on 'auto'. So not standardised at all (for the moment). Then I cut out the portion of the picture that contains the spots, set maximum saturation and contrast, outline the spots in GIMP and put it into TLC-analyzer.

Below I attached a picture from a plate that was spotted on both sides with a reference solution of M. The 3 center spots are taken from 3 different punches taken right beside eachother. I did this to evaluate the reproducibility of sample-taking.

Another way of approaching the picture is to use GIMP's histogram set on green channel to 'scan' 10x10 pixel spots of the picture. This seems to provide a promising semi-quantification as well and allows to determine a difference between the spot and its 'tail', allowing for relative quantification vs background. For example, the cactus spots in the uploaded pictures below have a 'relative intensity' of only 24, while the M-reference spots have a relative intensity of 44. So, although the cactus spots may seem to be dark, in fact they are only half as 'intense' as the M-reference spots. What is very interesting, is that these relative intensities as drawn from the histogram are comparable between pictures. The pictures I uploaded above give for the same M-reference a difference of 10,5 while the cactus-extract reads 5,5. The ratio stays the same. This is good news for comparing large batches of TLC-plates from many many cactuses taken in different lighting conditions. In fact, when we could put this next to a caffeïne-reference, this would allow all Nexians to compare the relative strengths of their cactuses (if further experimentation is favorable of course).


endlessness wrote:
I've done this 'semi quantifying' for DMT in mimosa, except I was just using my eyesight instead of a software, and it seemed to work decently well..

Nice work, Endlessness!

urtica wrote:
How long does it take the straight cactus juice spots to dry?

I use a heat gun set to 50°C to dry the spots in between spotting. Takes about 10 seconds.


This method works well for the presumed bridgesii I used. It's mashed-up sap is liquid enough to be sucked up into a spotting capillary. However, I tried to do this with a peruvianus and it's slimy sap makes this method impossible to use. I'm trying a micro-extraction with methanol for the moment...

An1cca attached the following image(s):
Triple sample + 2 ref.jpg (206kb) downloaded 753 time(s).
Triple sample + 2 ref 2.jpg (202kb) downloaded 747 time(s).
GIMP histogram.jpg (368kb) downloaded 749 time(s).
 
DGB
#36 Posted : 6/16/2017 8:40:04 AM
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Just wanted to say you're doing great work. Exactly the type of data I've been looking for and trying to collect. I hope you don’t mind but I linked to your thread and posted an abbreviated result summery on my thread on the shroomery, I also took the opportunity to retype the graph to include the new data that wasn’t in the first one. I also added two new categories and plugged in the numbers from the known clones from the 14 taxa papers.

 
An1cca
#37 Posted : 6/22/2017 7:08:21 PM

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Ok guys,
exit UV-C and TLC analyzer
enter ninhydrin and Fiji (=ImageJ)
More research coming in the next weeks...
An1cca attached the following image(s):
ninhydrin + Fiji.jpg (126kb) downloaded 606 time(s).
 
urtica
#38 Posted : 6/22/2017 8:22:53 PM

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DGB- thanks! And yea get the info out there! Maybe you could post a link here or PM one to me so I can check it out? I would like to be credited/mentioned if someone is presenting the data, which I am sure you did.

This is a work in progress & I will have more data here soon.

An1cca--

YEAH bring it! I am hoping to work with your techniques to analyze some species of Lophophora that I have been growing. I can't bring myself to extract the whole plants for the sake of data but your methods seem promising!
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urtica
#39 Posted : 6/22/2017 8:38:00 PM

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DGB-

I found your thread & yes I love what you did. Thank you for being sure to give me credit for the work that I have done. I love to see it getting spread around!
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DGB
#40 Posted : 6/23/2017 1:40:15 PM
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updated my thread and the graph.
 
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