Non-psychotropic plant cannabinoids: new therapeutic opportunities

Non-psychotropic plant cannabinoids: new therapeutic opportunities

elmanito said:

HPLC means High Pressure Liquid Chromatography.You can use it to get rid of some constituents which you dont wanna have.

Namaste

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I just gooled High Pressure Liquid Chromatography and what I got was High performance liquid chromatography...is that the beastie you're talking about?

muA

medi-useA said:

I just gooled High Pressure Liquid Chromatography and what I got was High performance liquid chromatography...is that the beastie you're talking about?

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Its the same!!!!

Namaste

elmanito said:

For identification if your strain does containCBD you can better use the TLC-method

Namaste

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TLC is pretty useless for determining the % of the Cannabinoids. It can be used to see if a plant has THC, CBD, CBG, CBN, etc. If you want the % use GC or MS/GC. A HPLC is only required if you want to know the ratio of acids, like THCA vs THC. A GC will give the total THCA + THC as the THCA is converted to THC by the heat of the GC analysis. HPLC's are not required for Cannabinoid analysis unless you need to know the acid % like THCA, CBDA, CBCA, etc. But that is not what you want as in a plant almost all Cannabinoids are acid during growth and for months after drying and storage at low temps.
A GC or GC/MS is what to use for Cannabinoid analysis.
I have both, and have used TLC in the past.
I use HPLC to isolate pure Cannabinoids, not for analysis, unless I need to know the acid %, which is never.
-SamS

Like i said before TLC is for identification not quantification, but for identification it is more accurate than GC and less expensive.(e.g. retention time of CBD & CBC)
To measure the quantity of the cannabinoids you can better use GC/MS than GC alone, unless you make the reference cannabinoid standards by your own every time you do a GC analyse.

Another method H-NMR can be used for quantitative analysis of Cannabinoids

http://www.jstage.jst.go.jp/article/cpb/52/6/718/_pdf

Namaste

I used a GC for thousands of analysis. I agree a MS/GC is better because once calibrated you don't need to use your analytical samples. But I did not have a MS/GC and I had zero problems. What is more important is two injection ports so you can have one column for the majority of the Cannabinoids and another just to separate CBD from CBC. As well an auto sample tray, mine holds 96 samples and can run un-supervised over night.
I don't believe a TLC is more accurate, specially in small amounts like .1-.001. And I have done hundreds of TLC tests.
As for H-NMR I know of no lab using this besides the folks that published the paper, maybe one or two others. What does a Nuclear Magnetic Resonance Spectroscopy set up cost? Sounds fast and easy if you happen to have a Nuclear Magnetic Resonance Spectroscopy set-up laying around, I don't.
-SamS

Sam_Skunkman said:

TLC is pretty useless for determining the % of the Cannabinoids. It can be used to see if a plant has THC, CBD, CBG, CBN, etc. If you want the % use GC or MS/GC. A HPLC is only required if you want to know the ratio of acids, like THCA vs THC. A GC will give the total THCA + THC as the THCA is converted to THC by the heat of the GC analysis. HPLC's are not required for Cannabinoid analysis unless you need to know the acid % like THCA, CBDA, CBCA, etc. But that is not what you want as in a plant almost all Cannabinoids are acid during growth and for months after drying and storage at low temps.
A GC or GC/MS is what to use for Cannabinoid analysis.
I have both, and have used TLC in the past.
I use HPLC to isolate pure Cannabinoids, not for analysis, unless I need to know the acid %, which is never.
-SamS

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Sam, I don't agree with you 100% on this one. GC/MS and HPLC are equivalent methods for identification of components of a sample when they are present in greater than ppm quantities. For the major cannabinoids either will give you very accurate % composition. Each has strengths and weaknesses which can better suit for a given application. I have some great papers comparing the two methods for cannabis if you are interested.

GC/MS

GC/MS has a better lower detection limit, and is much better for identifying unknown molecules. But it is also VERY costly to operate and running plant extracts through them requiers a very tight maintenance cycle. Cost per sample is usually higher than a HPLC system. As you said, it has trouble detection heat liable molecules, such as the acid components in cannabis. It is an excellent research tool.

HPLC

Has about a order of a magnitude higher detection limit, and it is more difficult to identify unknown molecules. However, for plant extracts and raw plant material we are not trying to find molecules in the ppb range. So sensitivity is not an issue. Also cost per sample is very low. Operational costs are a fraction of a GC/MS system. Cleaning is no problem, so the overall through put on a HPLC system will be higher. As you said, HPLC is good for detecting a heat liable components. Also, you can inject a larger sample through a HPLC system, which to an extent makes sample prep easier (IE, less tech time, less dilutions), and you can collect fractions and recover components through it. The real problem is developing good separations methods for the sample you are trying to analysis.


It should be noted that to my knowledge no analytical lab uses GC/MS for high volume sample analysis where a suitable HPLC method is available. All of your supplements and vitamins are certified by HPLC these days. I see the two as complimentary methods, no one really being better than the other, just better suited for a given task.

as to the TLC test, that is nice if no other option is available, but I have seen the kit you are talking about and trying to sell it as a "semi-quantative" method is really misleading. TLC can be semi qunatiative, but given the reagents and equipment provided in that kit, getting reproducible results through it is very hard. Also, the people providing that kit are charging a whole lot of money for a what amounts to a experiment from a freshman biology lab. Its great that it is out there, because there are alot of people who don't have access to anything thing better. Hopefully that will change soon.

Drop me a line Sam, I would love to pick your brain. I am still reading through your 200+ pages of posts here, but so far you have some really impressive posts in there. We are forming a scientific advisory board, and I need to fill some seats if you are interested.

veta

For Cannabis HPLC is only required if you need to know the acid components like the THCA, or extracting pure Cannabinoids, but it is not needed for determining the Cannabinoid %'s. I have been to all the big labs doing Cannabinoid analytical work, they all use MS/GC, why? Because a HPLC is not needed, it gives info that is not needed for determining Cannabinoid %'s. They do have HPLC's but they don't use them much for this kind of work. As I said I have done ten's of thousands of GC analysis for Cannabinoids, a few labs have done more, but only a few, and they all use GC not HPLC for the vast majority of their analysis.
You say "you have some great papers comparing the two methods for cannabis if you are interested". Let me know the titles and I can see if I have them already, I know most of the authors personally for years.
As for the TLC I used it was not the kit you refer to, I made an in-house kit we tried to develop for use in the field, but we were never happy with it, so we quit with it, it did work, but I prefer GC results.
Have you done Cannabinoid analysis on any sort of regular basis or is your preference of HPLC just an intellectual one? I have used both GC and HPLC for more then 15 years. I don't use my HPLC's much except for my production HPLC that can isolate small amounts of a single Cannabinoids for standards. The first few years we used the HPLC a lot until we understood that the plant produces the Cannabinoids in their acid form, un-decarboxylated, and if kept refrigerated or frozen they are very stable if kept from Oxygen.
I had already sent you a visitor note but not a PM as I thought you can't get one yet. Look at your public profile and check your visitor messages.
No I am not interested in your offer of the scientific advisory board position, I am way to busy with my work and life, thanks for the offer.
Send me a list of the labs doing lots of Cannabinoid analysis using HPLC for lots of Cannabinoid analysis, I can tell you if they use HPLC or GC for the majority of their work. I have been to all the major players labs, and worked with many of them.

Have you seen Arno Hazekamp's power point review of the acid and neutral Cannabinoids and their retention times etc with GC, MS, HPLC, TLC? A good place to start if you want to do Cannabinoid analysis seriously. Ask him and he will sell you a copy.
-SamS

I have read Hazekamp's work, its kind of hard to miss. I agree with you most of the research labs are using GC/MS for research purposes. It is the best fit for that application. I am interested in large scale open to the public commercial testing application.

MY post count is rising slowly, you can reach me at tortalk@gmail.com untill i can get pm's

veta

I did email you at gmail.
-SamS

veta said:

I have read Hazekamp's work, its kind of hard to miss. I agree with you most of the research labs are using GC/MS for research purposes. It is the best fit for that application. I am interested in large scale open to the public commercial testing application.

MY post count is rising slowly, you can reach me at tortalk@gmail.com untill i can get pm's

veta

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You have read the power point presentation?
Is it even legal to test Cannabis in the USA, for the public?
Have you ever done Cannabinoid analysis?

-SamS

check your mail sam

hmm what a coincidence..i am opening a THC/CBD testing facility also. just send those buds you want tested right to me, and my eager team will tell you just how they hold up. not only is the service free, it's two for the price of one day! you heard that right! our state of the art testing facility is located right here in the USA:

this one is good..

this one sucks.

cough, tastes like shit.

oh yeah! i can't feel my face and the air smells like when i was 5

<insert kallen two thumbs up photo here>

we now return you to your regularly scheduled thread..

xoox

elmanito said:

I thought that GW Pharmaceuticals had a strain in their possession of 98% CBD and a THC strain of 98% THC.
In researches i saw some hemp strains mainly from Turkey which had a CBD-content of 5% CBD.Even some varieties of Vavilov Institute were higher than 1% CBD than what you say above.The strain Finola can have a CBD content of 4-5%.
What can you tell about the strain USO-31?????

Namaste

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98% of all cannabinoids, not CBD by dry weight.. it gets confusing..

I would also think that it is 98% CBD of all cannabinoids and not dry weight.
Its indeed confusing, since i saw a video recording of a speech of Geoffrey Guy, where he talked about the strains which they were using.

Namaste

I have read "Medsins", is the name of G.W's strain,anyone worked with this strain?
Interesting thread!

sorry for the late post and i hope you guys are still around. i have a question about clarification regarding an earlier post, maybe Sam Skunkman can help?

elmanito quoted somebody (can't find the original) who said that in this publication there is an EU requirement for CBD:THC ratios to be 2:1. with a THC limit of 0.2% for industrial hemp, that would put the CBD limit at 0.4%. Thank you very much for any help. http://www.tandfonline.com/doi/abs/10.1300/J237v07n01_09#.U4t6Cijwr8g

Also, i can confirm that GW do use a strain called "medisins" and another called "grace".

There is no limit to CBD, as it is not scheduled.
For example Finola has under 0.2% THC and up to >2% CBD.

Halonium said:

There is no limit to CBD, as it is not scheduled.
For example Finola has under 0.2% THC and up to >2% CBD.

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Unfortunately, CBD and THC synthases aren't 'perfect' enzymes and hence, in a pure CBD plant, THC is still present at about 5 to 10% of the CBD content (the same is +/- true the other way round).
That means, if you somehow get your 'hemp' to produce 10 or 20% CBD you risk to get involuntarily over the legal THC limit. The law/cops won't care if this kind of weed can't get you high ;( .

GreenerGhost said:

...elmanito quoted somebody (can't find the original) who said that in this publication there is an EU requirement for CBD:THC ratios to be 2:1. with a THC limit of 0.2% for industrial hemp, that would put the CBD limit at 0.4%...

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BTW I have seen the notion of >2:1 CBD/THC or something alike as well... don't know where, though. This is important (in a legal sense) to guarantee that the plants used as industrial hemp don't express the THC synthase but only the CBD one (i.e. are BD homozygous). The notion 2:1 is not a fix ratio but a minimum requirement because all industrial hemp has a ration of approximately 10-20:1. Therefore, there's no direct limit to CBD but only indirectly because of the THC as stated in the beginning of my post .

finola when it's fresh and abit humid makes a fine smoke abit like tobacco, i had some industrial hemp in france and i was getting a clear headed mong from a spliff. not like a heady mong, more like a sedative one like a tiny dose of ketamine. i want to find the cbd ratio somewhere online but it's not clear.

I wasn't aware that there was a way to turn CBDA or CBD to THC. If so how is it done?

With decarb you can take any strain and boost the THC or convert the THC to CBN in order to make the CBD more easily available.

In my case I have a tumor in my brainstem so I'm always hunting for CBDs. Before (with high THC strains) I had to be borderline bad trip to feel the healing effects, after decarbing out the THC I was able to take the same dose without the associated head high. Same strain, different decarb profile.