Scrutinizing Strains with Science : An Objective Discussion

[spurr]

FWIW,

Here is a good YouTube'd video of TLC in action with explanation of steps, etc. Good intro to TLC I think:

Lec 3 | MIT 5.301 Chemistry Laboratory Techniques, IAP 2004
http://www.youtube.com/watch?v=EUn2skAAjHk

 

[MyGreenToe]

Nice stuff Spurr. I've been really busy with work lately and just havent had it in me to properly edit this thread. If you would like to write a few paragraphs about what TLC is and how you think it's useful for MJ, that would be awesome. Then I could just quote it in the first post, instead of waiting until I have the time to do it. You clearly understand it better than I do anyways....
Keep it Green
MGT

 

[funkfingers]

raw data....someone break it down in dumb person terms

raw data....someone break it down in dumb person terms

here is some raw test results the one I am specifically curious about is the sherpa...I know she is no slouch but the test results would almost make it seem as ditch weed.. can someone please break down what the hell these tests mean..lol I mean I have a grasp but would love to here some other thoughts..

 

[Forest20]

Bump...Great collective information going on.

 

[CARE giver]

The other day I was just thinking about scientifically creating the perfect strain for me. I mean Monsanto can genetically alter plants, why not cannabis?

 

[onegreenday]

notice the THC-V in the Mexican strain is albout 2.5 times the closest strain
in the amount. This may be part of Mexican uniqueness........

 

[onegreenday]

one thing FF is I don't see a test for THC-v which the other test
earlier in the thread shows.

the Sherpa is higher in CBD (may be what you feel)

 

[Hrpuffnkush]

My highest CBD's so far are form Bubbleberry X ECSD aka Berry Garcia
i will post others i have soon

 

[Thomkal Vwalaa]

The other day I was just thinking about scientifically creating the perfect strain for me. I mean Monsanto can genetically alter plants, why not cannabis?

That's coming bredren. It's only a matter of time... They may have GMO ganja already. InI don't think Monsanto, or any of those big corporations, they cyan't touch it while di ganja is illegal. American babylonian federal war on ganja keeps the plant in InI hands. Until it is safe, or the money is really worth it. Monsanto is basically a part of the government system. If they have it already, they won't go public until rescheduling.

 

[onegreenday]

hey kush puffer that's great info from Rm3 Labs LLC.
Sounds like a solid company

Can you post the rest of their info on CBD. The page
ends where they are explaining CBD. Thanks

 

[Hrpuffnkush]

1gDay its like 3 pages of descriptions, not of results , Do you guys want that Info??
ill try to scan the rest out for you guys in couple days

 

[onegreenday]

1gDay its like 3 pages of descriptions, not of results , Do you guys want that Info??
ill try to scan the rest out for you guys in couple days

is there more info on CBD?
It seemed like there was more.

results not important.
thanks.

 

[Scrogerman]

tagged!

 

[spurr]

dang it, I forgot about this thread, and I complied a bunch of papers and best practices for everyone that describes how to due testing with GC, TLC, HPLC, OPLC, etc., et al (ex. from the UN, etc). I will try to post tomorrow. I have a butt load of info to post for people who are interested.

 

[Thomkal Vwalaa]

is there more info on CBD?
It seemed like there was more.

results not important.
thanks.

Doctors to Study Effectiveness of CBD

http://www.medicalmarijuana411.com/2010/11/11/doctors-to-study-effectiveness-of-cbd/

This was too long to post here, but is worth following the link to read if you are into CBD.

Tod Mikuriya, MD, did not live to see it, but his dream of investigating the medical potential of compounds in the cannabis plant other than THC is now within the grasp of his successors.

The Society of Cannabis Clinicians, the group Mikuriya founded in 1999, has drafted a “Strain Evaluation Survey” to collect data from patients who medicate with cannabis in which cannabidiol (CBD) is predominant.

CBD-rich cannabis will be available at California and Colorado dispensaries by late summer —and soon thereafter, inevitably, in other states where patients can legally use cannabis as medicine.

Twelve strains rich in cannabidiol (CBD) have been identified in the year and a half since an analytic chemistry lab began testing cannabis samples provided by California dispensaries, growers, and edible makers. Buds from five of these strains have been available intermittently at Harborside Health Center in Oakland. Herbal Solutions in Long Beach also has provided CBD-rich cannabis to patients.

Eight of the CBD-rich strains are currently being grown out. The others cannot be reproduced because the growers hadn’t saved or couldn’t regain access to the genetic material that yielded their buds of interest.

More than 9,000 samples have been tested to date by the Steep Hill lab in Oakland. Other start-up labs in California, Colorado, and Montana have begun testing for the burgeoning industry. The Full Spectrum lab in Denver has tested some 4,500 strains and identified seven CBD-rich strains.

A strain that is roughly 6% CBD and 6% THC, “Cannatonic,” has been developed by Resin Seeds in Barcelona and is being grown from seed by several collectives. Its name may be misleading, since CBD supposedly cancels the sedating effects of THC.

For purposes of the data collection being planned by the Society of Cannabis Clinicians, “CBD-rich” cannabis is being defined as more than 4% cannabidiol by weight (without respect to THC content) or more than 2.5% CBD if CBD exceeds THC.

Until testing for cannabinoid content began, it was widely assumed that CBD, which is non-psychoactive, had been bred out of all the cannabis in California by generations of growers seeking maximum THC content.

Doctors in the SCC have watched with great interest in recent years as a British company, G.W. Pharmaceuticals conducted clinical trials of cannabis-plant extracts. G.W. has a license from the British government and backing from Otsuka, a Tokyo-based multinational.

G.W.’s flagship product, Sativex, is a plant extract that contains approximately equal amounts of CBD and THC. What benefits did G.W. scientists expect a CBD-rich extract to confer?

Various studies published in the medical and scientific literature suggest that CBD could be effective in easing the symptoms of rheumatoid arthritis, diabetes, nausea, and inflammatory bowel disorders, among other difficult-to-control conditions. CBD also has demonstrated neuroprotective effects, and its anti-cancer potential is being explored at several academic research centers.

An even wider market would emerge if the reduced psychoactivity of CBD-rich cannabis makes it an appealing treatment option for patients seeking anti-inflammatory, anti-pain, anti-anxiety, and/or anti-spasm effects delivered without disconcerting euphoria or lethargy.

The plant richest in CBD is a “True Blueberry/OG Kush” cross grown in the mountains south of Yreka by Wendell Lee of Full Spectrum Genetics (not to be confused with the lab in Colorado). Dried buds of TB/OGK have been sent for testing on four occasions by Harborside, the dispensary with which Lee is associated. Samples were consistently found to contain about 10% CBD (with THC levels around 6 to 7%). On the only occasion that a crop grown outdoors by Lee was tested by Steep Hill lab, it was found to contain 13.9% CBD.

Two other labs have confirmed the CBD content of Lee’s TB/OGK.

Lee is now working to “stabilize the genetics” and produce TB/OGK seeds. Several plants he provided to Project CBD (a nonprofit organized by writer/activist Martin Lee to promote research) are being grown out by experienced hands. Processed medicine and clones will be available at dispensaries in the months ahead. Details will be available on ProjectCBD.com, a website that will be launched by mid-August, according to Martin Lee (no relation to Wendell)

The California strain richest in CBD proportionally, “Women’s Collective Stinky Purple,” tested at 9.7% CBD and 1.2% THC. It was brought to Harborside by Grower #1 who also grows a strain called “Cotton Candy/Diesel” that was found to contain about 6% CBD and 6% THC. Grower #1 gets her starter plants from friends in Northeastern Mendocino County. Is there something in the genome of plants that have been swapped over the years by growers in those hills that encourages expression of CBD?

Another strain containing more than 8% CBD, grown indoors in the East Bay, was brought to Harborside in late April. “Omrita Rx3” is the name the grower has given it after learning that it was of special interest to SCC doctors.

A few weeks later a strain called “Harlequin” was found to contain about 8% CBD. And soon thereafter a pound of “Jamaican Lion” tested at 8.9% CBD. Clones of these strains are being grown out and will be available through Harborside and Project CBD in the months ahead, along with the Soma A+ that was first to be identified.

Pineapple Thai (5% CBD, 2.4% THC) is being grown out by Herbal Solutions in Long Beach.

 

[Sam_Skunkman]

spurr,
Have you actually done terpenoid analysis using just TLC? I never have. Terpenoid standards are easy to get and not controlled, and they are cheap to buy.
You mentioned Cannabinoids that get you high, like THC, CBN almost gets you high, but you can forget CBD, CBC, CBG, THCV and the rest of the propyls (3 carbon] They do not get you high, although they do have medical applications. I have tried them all as pure single Cannabinoids.
CBD, CBN, THCV are THC antagonists and they do modify THC's effects, like in the case of CBD which delays THC onset, reduces peak effects and lengthens the reduced effects time of activity.
CBD is also funny in that if you smoke 100mg of pure CBD before any Cannabis smoking, you can't get high from THC for several hours, even if you smoke the strongest hash.
While if you smoke a THC/CBD mixture first thing, you do get high, like almost all imported hash, that has considerable CBD with the THC.
But the high is different then THC only varieties. I prefer high THC only, but to each their own....
It is the terpenes with high THC that makes the subjective effects.

PM me I am curious where you live. I am in Amsterdam.

I prefer GC over TLC because I want quantitative accuracy. I did develop my own TLC methods, but quit years ago because I could only really use them for comparative analysis. Preparative HPLC I used to separate small amounts {1 gram} of pure Cannabinoids.
A GC with 2 injection ports, ( one column for THC and the rest of the Cannabinoids and a longer column for CBD separation) auto sampling is the way for me. Run 96 samples a night while I sleep.

-SamS

along with testing for normal secondary metabolites like cannabinoids, testing for some terpenoids and flavonoids should be the norm IMO. Terpenoids such as myrcene, etc., can be tested with GC, HPLC, TLC; as well as some flavonoids. Finding standards for those substances (for GC), or finding Rf values or spot color (for TLC) is the tricky part. I think I have info about extracting and testing myrcene with TLC but I am not sure.

The biggest obstacle I see in terms of testing for terpenoids and flavonoids is knowing what ones to test and the effect they have when smoked (or ingested). There are a few terpenoids known to have worthwhile medical efficacy, and a few that affect our high, etc., but I do not know of a large list.

I also think expanding the cannabinoids tested is a good idea, sometimes a freak of nature can happen worth trying to work with. Testing for the normal THC, CBD, CBN is fine, but I think labs and individuals should also test for CBC, CBG, etc., and THC-V, CBD-V, CBC-V, along with other varin (-V) cannabinoids.

 

[Thomkal Vwalaa]

There is so much good stuff but I just wanted to say:

I have tried them all as pure single Cannabinoids.

Recreational chemistry is fun. I once hit a joint with more than a few drops of pure THC added. Very interesting affects.

CBD is also funny in that if you smoke 100mg of pure CBD before any Cannabis smoking, you can't get high from THC for several hours, even if you smoke the strongest hash.

Sounds like the CB receptors are saturated, but I'm sure you know more than I.

It is the terpenes with high THC that makes the subjective effects.

I've heard rumblings about terpene infusion chambers for commercially produced flowers to alter the affects (read: attempt to improve taste/smell/affect and get more money). Many thanks for the knowledge.

 

[Dkgrower]

Very cool tread keep the information going

Good to see you here Sam together with spurr this could be one informative tread

 

[ghostly]

my lab results...

my lab results...

hope this is readable.
i havent seen many people post the results... whats up ICMAG?

 

[spurr]

spurr,
Have you actually done terpenoid analysis using just TLC? I never have. Terpenoid standards are easy to get and not controlled, and they are cheap to buy.

Not yet, but I agree it's a no-brainier. The trick is figuring out which ones to test for, I listed a couple from the two studies I previously posted.

You mentioned Cannabinoids that get you high, like THC, CBN almost gets you high, but you can forget CBD, CBC, CBG, THCV and the rest of the propyls (3 carbon] They do not get you high, although they do have medical applications. I have tried them all as pure single Cannabinoids.

I didn't mean they get you high alone, I meant they affect the high from THC in a synergistic or antagonist manner. Most cultivars grown today are lacking in decent quantities of CBC, THCV, etc., esp. CBG. I'm sure this info is nothing new to you. However, being there are currently 70 identified cannabioinds (4 new ones), with too little research thus far, it's hard to say none other than THC have strong psychotropic properties (ElSohly and Slade, 2005).

CBD, CBN, THCV are THC antagonists and they do modify THC's effects, like in the case of CBD which delays THC onset, reduces peak effects and lengthens the reduced effects time of activity. CBD is also funny in that if you smoke 100mg of pure CBD before any Cannabis smoking, you can't get high from THC for several hours, even if you smoke the strongest hash.

Yea, I've read a few papers on that, interesting stuff for sure. I wonder about the myriad of other cannabinoids found in small quantities in cannabis, too little research has been conducted on all 70 of them. I assume at least a couple are psychotropically active, besides THC.

While if you smoke a THC/CBD mixture first thing, you do get high, like almost all imported hash, that has considerable CBD with the THC. But the high is different then THC only varieties. I prefer high THC only, but to each their own....It is the terpenes with high THC that makes the subjective effects.

What do you mean by "subjective effects"? Anything non-quantitative, or none comparative (such as TLC spot density measurement without standards) is subjective, ex., smoking a bud. AFAIK, other secondary metabolites than cannabinoids and some terpenoids affect the high, ex., some flavonoids.

PM me I am curious where you live. I am in Amsterdam.

I'm in the US...for now.

I prefer GC over TLC because I want quantitative accuracy. I did develop my own TLC methods, but quit years ago because I could only really use them for comparative analysis. Preparative HPLC I used to separate small amounts {1 gram} of pure Cannabinoids.

I agree, but for most people (at least in the US) using a GC is not possible, only a few states have labs that will tests cannabis (usually using GC or HPLC). Thus using comparative TLC via. spot density measurement (i.e. densitometry) with "JustTLC", is a good runner up. Also, TLC can be used in a truly quantitative manner by using densitometry and standards, but it's more cumbersome than using other planar chromatography quantitative methods I list below (Raharjo and Verpoorte, 2004).

There are other, rather simple, and inexpensive planar chromatography methods that are quantitative and provide accuracy on par with HPLC and even GC, ex. using High-Performance Thin-Layer Chromatography (HPTLC), or Optimum-Performance Laminar Chromatography (previously known as OverPressured-Layer Chromatography; OPLC). Using either HPTLC or OPLC, are superior, and more trivial for quantitative assays vs. classical TLC.

I think using HPTLC, for the 'average Joe' grower, for quantitative measurement is the best bet, esp. because they can do so at home. And one can buy DEA-exempt cannabinoid standards now, thus there is little stopping a motivated and intelligent grower from carrying out truly quantitative (and accurate) assays in the US and abroad. Considering GC and HPLC are not available for most people, and that HPTLC is cheaper, and easier, then GC and HPLC, I think HPTLC is the way to go. Also using HPTLC one can quantify THC-COOH, unlike when using GC. That is why I think using HPTLC with DEA-exempt standards is the best option for most 'average Joes'; and TLC using spot density measurement comparison (via. "JustTLC") being a runner up for most people.

Using a standards and spot density measurement allows for quantitative assay of samples using classical TLC. Because one can buy DEA-exempt cannabinoid standards in the US, quantitative TLC is possible for nearly anyone to carry out for a few hundred dollars to setup the lab. That said, using classical TLC is not the best choice IMO, as I mentioned above, using HPTLC would be a better choice. I have a pretty good papers on quantitative planar chromatography (ex. HPTLC, OPLC and TLC). That said, GC is definitely the better method for people with lots of money, expertize and resources.

I am very interested in HPTLC and OPLC, vs. classical TLC, HPLC and GC. HPTLC offers benefits over GC and HPLC, such as "lower running costs [and setup costs], more rapid analysis time and the ability to analyse multiple samples simultaneously..." (Fischedick, Glas, Hazekamp, Verpoorte, 2009).

Did you use a staining/visualization reagent for TLC? Such as fast blue BB salt? Or did you rely upon UV? Did you use color (staining) reagent and scan spot density for measurement, or use spectrophotometry (UV) and/or Rf values?

A great read for newbies on TLC and GC, and cannabis assays in general, is the "Recommended methods for the identification and analysis of cannabis and cannabis products", United Nations Office on Drugs and Crime (UNODC) 2009 (link). That is probably the best source for a person to develop standardized TLC and GC assay methods.

Testing standardization is one worry I have about various tests from labs, not all labs follow the same methodology, or even use the same standards or methods (i.e. GC vs. HPLC, etc.). Thus I worry about the ability to normalize and compare results from various labs...I think we need much more standardization than we have today.

One reason I dislike GC is you can't quantify THC-A (THC-COOH), only total THC due do the decarb of THC-A from heat of GC (Raharjo and Verpoorte, 2004). This isn't a problem with some planar chromatography if following certain methods such as HPTLC or OPLC; HPTLC might be the best option.

I am interested not only in GC, classical TLC and HPLC, but other planar chromatography methods, esp. HTPLC and OPLC. I have a papers on OPLC, Automated Multiple Development (AMD) and HPTLC of cannabis, among other papers on GC, HPLC, et al.

A GC with 2 injection ports, ( one column for THC and the rest of the Cannabinoids and a longer column for CBD separation) auto sampling is the way for me. Run 96 samples a night while I sleep.

-SamS

Nice setup!



Various refs., and cites from above:

Mahmoud A. ElSohly, Desmond Slade. 2005. Chemical constituents of marijuana: The complex mixture of natural cannabinoid. Life Sciences, 78, (2005), 539 – 548

Justin T. Fischedick, Ronald Glas, Arno Hazekamp, and Rob Verpoorte. 2009. A Qualitative and Quantitative HPTLC
Densitometry Method for the Analysis of Cannabinoids in Cannabis sativa L. Phytochem. Anal. 2009, 20, 421–426

Tri J. Raharjo and Robert Verpoorte. 2004. Methods for the Analysis of Cannabinoids in Biological Materials: a Review. Phytochem. Anal. 15, 79–94 (2004)

B. Szabady, E. Hidv6gi, Sz. Nyiredy. 2002. Determination of Neutral Cannabinoids in Hemp Samples by Overpressured-Layer Chromatography. Chromatographia Supplement Vol. 56, 2002

Richard Johnsson, Gustav Träff, Martin Sundén and Ulf Ellervik. 2007. Evaluation of quantitative thin layer chromatography using staining reagents. Journal of Chromatography A, Volume 1164, Issues 1-2, 14 September 2007, Pages 298-305
​

^^^ that last paper is looks at efficacy of using JustTLC for (stained) spot density measurement with TLC for quantification of samples with a flat bed scanner (instead of doing so spectrophotometrically), using a reagent such as Fast Blue BB salt. That is the method I suggest most people use with classical TLC, in a comparative manner if one does not have standards, and is what I have been working on for the 'average Joe'.