Scrutinizing Strains with Science : An Objective Discussion

[shaggyballs]

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[Sam_Skunkman]

Thanks to everyone who has contributed to this thread, it's an incredibly interesting and informative read.

I have a few questions based on some topics brought up throughout this thread, and note I'm a layman and have no scientific background so please be nice





I've always understood (or possibly misunderstood) that curing cannabis after drying will:
* allow the slow escape of moisture.
* allow chlorophyll to break down for a smoother smoke.
* allow for decarboxylation of THCA to THC where COOH is removed. (THIS IS NOT CORRECT)
* allow chemical changes in terpenoids/flavonoids to occur to refine the effect and flavor.

All in the name of creating a better product. I've also known that heating the THCA during consumption (either vaporization or burning) will covert it to THC via immediate decarboxylation so do we really care about that aspect of curing? (FOR TASTE YES)
I always assumed that maybe by burning/vaporizing we don't decarboxylate all of it so curing allows more of it to be readily available thus increasing potency. Am I completely wrong? (MOSTLY YES)
Also has anyone done GC tests before and after curing to see changes in levels of THCA and THC? (YES I HAVE, LITTLE DIFFERENCE) How about terpenoid profile before and after?
(SOME OF THE MORE VOLATILE TERPENES ARE LESS, ALL OF THE TERPENES LEVELS ARE A LITTLE BIT LOWER)




SamS, I'm curious since you mention that only THC got you "high", I'm curious since you tested pure forms of various cannabinoids and terpenoids, can you comment on the effects felt if any from the other cannabinoids/terpenoids in their pure forms?

(THEY DID NOT GET ME HIGH, THAT WAS MY FOCUS) (THEY ALSO AFFECTED THE HIGH IN A NEGATIVE WAY TO ME) (BUT MAYBE SOMEONE LIKES IT)

Also there's been some discussion on about CBD and counteractive effects on THC. I've long used high cbd or mellow cultivars (see that spurr?), which I assumed have higher CBD:THC ratios, to counteract ill effects of high anxiety and stressful cultivars.

(DID YOU LAB TEST THE VARIETIES WITH MAYBE HIGH CBD? OR ARE YOU JUST GUESSING) (I ASK BECAUSE IN THE PAST MOST COUCHLOCK EFFECTS WERE MEDIATED BY TERPENES NOT CBD)

I've always wondered about doing the same to counteract effects of edibles when they're too potent. I believe it should work the same but whenever I have edibles that are tripping me out bad I'm too scared to try it even though I feel like it will help.
(EAT LESS OR MADE FROM A DIFFERENT VARIETY)
I heard that with edibles the liver converts the delta-9 THC to delta-11 THC which affects you differently than delta-9, if this is true does the different delta bond somehow change the effect?
(YES IT IS TRUE 11HYDROXY THC HAS DIFFERENT EFFECTS THEN SMOKED THC)

Also, I don't quite understand the mechanism in which THC and CBD affect us differently, and this is probably due to my lack of knowledge in neurochemistry. Lets say you have a bunch of CB1 and CB2 recepters and introduce THC or CBD. Someone here said CBD doesn't bind well to CB1 recepters and better to CB2, so is it the different recepters that they bind to that changes their effects in the brain? Or if THC binded to the same recepter as CBD would they have the same effects? (NO THEY HAVE DIFFERENT AFFINITY FOR THE DIFFERENT RECEPTORS AND THE EFFECTS ARE VERY DIFFERENT)

I've read lots of misinformation out there and i'm hoping get some information.

(DIGEST THIS AND MAYBE ASK SOME MORE, I WILL ANSWER IF I THINK I KNOW)
-SAMS

 

[WelderDan]

Here's the science method I go by. I sample it. If I like it, I grow it again. If I don't like it, I don't grow it again. My criteria are effect and taste. All the GC/MS data be damned. We are all wired different and have different likes and expectations for the herb.

What one may find as sub-par, another may think is perfect.

There is no quantifying the quality of herb in a lab. It's all about the way it makes each individual feel and how it tastes.

 

[Sam_Skunkman]

So just to be clear, you have no interest in knowing which Cannabinoids and which terpenes at what %'s you prefer or dislike?
It seems the info would help you find new varieties you like if they were also tested for Cannabinoids and terpenes before you even bought them?
And while every smoker may prefer different effects or tastes, the "quantifying the quality of herb in a lab" can be done, and will be the future. Why should GC/MS data be damned? Isn't it better to use every tool, be it a lab or mans taste, smell, and experience. Together they are both stronger.
I still remember when everyone believed that THCV was the Cannabinoid that was better then THC, until I made pure THCV, tried it and said it does not even get you high. Later it was proven by Dr. Roger Pertwee to be antagonistic to THC. With out science and lab results we are just ignorant, of course to some ignorance is bliss....

-SamS

Here's the science method I go by. I sample it. If I like it, I grow it again. If I don't like it, I don't grow it again. My criteria are effect and taste. All the GC/MS data be damned. We are all wired different and have different likes and expectations for the herb.

What one may find as sub-par, another may think is perfect.

There is no quantifying the quality of herb in a lab. It's all about the way it makes each individual feel and how it tastes.

 

[The Hatter]

A very nice thread.

I have read from several sources that letting a plant flower longer will increase CBN. From what I can remember off the top of my head CBN is the result of one of the other Cannabanoids oxidizing. I believe it is THC which oxidizes into CBN.

 

[The Hatter]

Hello all. I'm not sure if anyone is still adding to this thread since the last post was in March. Either way though, I really love the general idea of this thread and the topics and content that has been specifically discussed. Being a neuroscience and pharmacology researcher, I could provide a few thoughts/references regarding a couple things. Naturally, and in the spirit of the thread, if I have made any errors please feel free to point out. If I have added anything redundant or already posted, my apologies. Here we go...

1.)Some non-CB1/non-CB2 receptor effects of certain cannabinoids can be non-receptor mediated (i.e THC and CBD being antioxidants) but as far as other important receptors for Endo/Phyto/Synthetic cannabinoids, one that has only recently been investigated is GPR55. Important for our discussion, THC binds to GPR55 and I believe CBD may be a GPR55 antagonist. GPR55 is an orphan GPCR(G protein coupled receptor) that was discovered by Big Pharma when fishing for novel ligands with medical applications. I have kept a vigilent eye on pubmed for any new papers on this receptor's role in physiology and there have thus far only been a few which I will post below.

Then of course there is TRPV1, which is mainly a CBD thing.

Also as far as a correlation between neurochemistry and subjective effects of cannabis/cannabinoids etc, there is a lot of crosstalk between the cannabinoid system and the opioid system. In certain systems, THC can augment opiate function of the u-opiate receptor (the euphoric one) and in other instances can augment the K-opiate receptor (the dysphoric one). This effect is dose dependent and biphasic (as are many of the effects of cannabis). Low doses may be more anxiolytic/euphoric which would relate to the u-opiate system, and higher doses with potential anxiety/paranoia may be related to K-opiate system. As far as terpenes and opiates, I believe that Myrcene has shown some mild opiate activity in vivo and in vitro.

Another biphasic effect neurochemically (and perhaps also cognitive/subjective wise) is the cholinergic system. Low doses of THC augment the release of acetylcholine in the hippocampus, while high doses inhibit release. Related to terpenes, many of them (such as a-pinene) are acetylcholinesterase inhibitors.

Ok I have a few refs posted below, the rest I'm still looking for(but I know I have them all).

The existence of opiate cross talk effects from Cannabanoids is further supported by how closely the physical withdrawal from Cannabanoid agonists mimic opiate withdrawal. The reason natrual Cannabanoids do not produce this sort of physical withdrawal is likely linked to their extremely lengthy elimination times from the body and the fact that they are partial rather than full agonists at the cannabanoid receptor sites.

I read an interesting medical report about a man hospitalized due to sever cannabanoid withdrawal after he quit smoking one of the synthetic spice type products after chronic heavy use. His symptoms essentially exactly mimicked moderate opiate withdrawal which was noted by the clinician treating the patient. Unlike natural cannabis the synthetics that are being made are full agonists that tend to be rather rapidly eliminated from the body both of which are likely contributing factors to the severity of the withdrawal symptoms.

 

[RegisPhilburn1]

Seems fitting to post this here

SPR World Class lecture series presents Dr. Justin Fischedick, PhD
Cannabis – Cannabinoid and Terpenoid Profiling & Medicinal Properties

See ya there

 

[Only Ornamental]

Hi there,

Just wanted to add my own two cents to the discussion (sorry for eventual repetitions of earlier posts).

To clarify some things:
- Many/some of you talk about quantitative and qualitative methods with regard to GC, MS (generic term), HPLC and TLC. That in principle is wrong; these are separation methods and don't govern whether the analysis is quantitative or qualitative (comparative). It's the detection afterwards which makes them one and/or the other. There are different detection techniques which can often be combined with the different separation strategies . Quantitative detection is for example possible with a FID or ELSD whereas semi-quantitative and qualitative detection is possible with GC (detector), UV and fluorescence detectors or TLC visualisation reagents. (GC may be regarded as something special as it often combines separation and detection.)
- TLC spot density is not reliable and not linear (at least, one has to include a calibration curve on each plate). Properly done and applied in lines (and not spots as proposed by commercial THC kit vendors) a TLC is at best semi-quantitative. It's stupid to give concentrations of for example 12.54%: That would rather be something like 10-15%. But for that, you don't need a scanner and a software, just a standard curve and estimate . Better results come from HPTLC...
- HPTLC is nothing one can easily do at home: You may use a HPTLC plate but lacking the technique to apply the sample in a fine line (and not a spot), the small gain in resolution is not worth the money. You will only safe time and space due to a shorter running distance (and pay a lot more for the plates). To do it properly, you need an automated approach with maybe a CAMAG HPTLC machine (actually 3 to 4 separate modules): Only like this, you may profit from the full advantages of HPTLC (intra- and inter-lab and day to day reproducibility, Rf-reliability, semi-quantitative analysis etc.). That gets pretty costly and time consuming!
- Cannabinoid (apart from CB1/2 ligands) and terpene analysis for finding a new 'strain' to grow or for strain-to-strain comparison doesn't make much sense to me. Why? Well, knowing which percentage of a certain terpene is in a sample doesn't mean you get the same absolute or relative amount in your harvest. And both are responsible for eventual modulations of the experience (but no one knows which terpenes may do that). Additionally, flavour and taste (of essential oil in general) depend on the whole mixture and seldom on one or two selected terpenes. Testing for THC gives a good impression on the overall strength of the strain but that's it. I'm also critical with regard to CBD if it's only present in a few percent and not at ~1:1 with regard to THC. Anyone an idea on how much you need to modulate THC effects? I think constituent analysis (again, apart from THC/CBD chemotype analysis) is good for industries (maybe commercial growers too) to test batch to batch consistency and homogeneity. But there, the analysed constituents are only markers (the actual pharmaceutical effects of them may be of no importance at all). In a growers life (and likely for commercial growers too?), test consuming seems still the way to go.

Uppsss... have to hurry, work's calling .

 

[Sam_Skunkman]

Hi there,

Just wanted to add my own two cents to the discussion (sorry for eventual repetitions of earlier posts).

To clarify some things:
- Many/some of you talk about quantitative and qualitative methods with regard to GC, MS (generic term), HPLC and TLC. That in principle is wrong; these are separation methods and don't govern whether the analysis is quantitative or qualitative (comparative). It's the detection afterwards which makes them one and/or the other. There are different detection techniques which can often be combined with the different separation strategies . Quantitative detection is for example possible with a FID or ELSD whereas semi-quantitative and qualitative detection is possible with GC (detector), UV and fluorescence detectors or TLC visualisation reagents. (GC may be regarded as something special as it often combines separation and detection.)
- TLC spot density is not reliable and not linear (at least, one has to include a calibration curve on each plate). Properly done and applied in lines (and not spots as proposed by commercial THC kit vendors) a TLC is at best semi-quantitative. It's stupid to give concentrations of for example 12.54%: That would rather be something like 10-15%. But for that, you don't need a scanner and a software, just a standard curve and estimate . Better results come from HPTLC...
- HPTLC is nothing one can easily do at home: You may use a HPTLC plate but lacking the technique to apply the sample in a fine line (and not a spot), the small gain in resolution is not worth the money. You will only safe time and space due to a shorter running distance (and pay a lot more for the plates). To do it properly, you need an automated approach with maybe a CAMAG HPTLC machine (actually 3 to 4 separate modules): Only like this, you may profit from the full advantages of HPTLC (intra- and inter-lab and day to day reproducibility, Rf-reliability, semi-quantitative analysis etc.). That gets pretty costly and time consuming!
- Cannabinoid (apart from CB1/2 ligands) and terpene analysis for finding a new 'strain' to grow or for strain-to-strain comparison doesn't make much sense to me.

Why? Well, knowing which percentage of a certain terpene is in a sample doesn't mean you get the same absolute or relative amount in your harvest. And both are responsible for eventual modulations of the experience (but no one knows which terpenes may do that).

THAT IS NOT QUITE TRUE, SURE THE %'s DO GET ALTERED WITH MATURATION, SO DO SEVERAL TERPENE ANALYSIS, ONE REAL EARLY, ONE MID FLOWERING, AND ONE VERY LATE HARVEST. YOU WILL SEE THE CHANGES CLEARLY AND CAN HARVEST WHEN YOU PREFER. AS FOR KNOWING WHICH TERPENES ARE RESPONSIBLE FOR MODULATIONS OF THE EXPERIENCE, YOU DON'T KNOW, BUT OTHERS DO.

Additionally, flavour and taste (of essential oil in general) depend on the whole mixture and seldom on one or two selected terpenes. Testing for THC gives a good impression on the overall strength of the strain but that's it. I'm also critical with regard to CBD if it's only present in a few percent and not at ~1:1 with regard to THC. Anyone an idea on how much you need to modulate THC effects?

TO MODULATE 25 MG THC YOU ONLY NEED 10-25 MG CBD.
TO MAKE IT SO YOU GET ALMOST ZERO EFFECTS FROM THC TRY SMOKING 100MG CBD BEFORE YOU SMOKE THE THC.
-SAMS

I think constituent analysis (again, apart from THC/CBD chemotype analysis) is good for industries (maybe commercial growers too) to test batch to batch consistency and homogeneity. But there, the analysed constituents are only markers (the actual pharmaceutical effects of them may be of no importance at all). In a growers life (and likely for commercial growers too?), test consuming seems still the way to go.

Uppsss... have to hurry, work's calling .

-SAMS

 

[shaggyballs]

SURE THE %'s DO GET ALTERED WITH MATURATION, SO DO SEVERAL TERPENE ANALYSIS, ONE REAL EARLY, ONE MID FLOWERING, AND ONE VERY LATE HARVEST. YOU WILL SEE THE CHANGES CLEARLY AND CAN HARVEST WHEN YOU PREFER.
-SAMS

I am just learning of this first hand.
I have a blueberry cross.
Early in the harvest window it has a distinct blueberry smell and taste......no cough(good kind)
Mid harvest blueberry is fading to a earthy tone .....a little cough(what is being called god weed I think is the base for earthy terpenes)
By late late harvest the blueberry is gone and replaced by the earthy flavor...extra cough.

My smoke lounge report

 

[Only Ornamental]

THAT IS NOT QUITE TRUE, SURE THE %'s DO GET ALTERED WITH MATURATION, SO DO SEVERAL TERPENE ANALYSIS, ONE REAL EARLY, ONE MID FLOWERING, AND ONE VERY LATE HARVEST.

TO MODULATE 25 MG THC YOU ONLY NEED 10-25 MG CBD. -SAMS

Hi Sam,

Thanks for the answer, haven't seen it first cause it was in the quote.

What I meant was that usually a plant is not something fixed and especially when grown outdoors the secondary metabolite composition (in all medical plants) fluctuates. One knows which constituents should be present and in which concentration range they usually fall but it always varies more or less. Phytocompanies therefore standardise their extracts etc. The best 'hydro' terpene profile at Greenhouse seeds won't do any good for an outdoor guy like myself .

Concerning CBD: That's what I supposed. Thanks for the confirmation.
This really means, cannabinoid testing of a drug type cannabis strain (genotype BT/BT) for more than THC concentration (and maybe CBN for quality/age) is, with our current knowledge, pointless. Claims like 'good medical use due to high CBD' for a variety containing 20% THC and 2% CBD is just marketing as the CBD won't do much.
Sure enough, testing hash is something completely different as one could have all possible proportions.

 

[pinkus]

Bumping because I'm reading the thread....and taking the plunge on learning DIY TLC. Great thread.

 

[Only Ornamental]

....and taking the plunge on learning DIY TLC....

Do you have some literature on TLC or do you 'expect' to learn it here?
I could explain it but believe me, doing nice TLCs is way easier and more efficient by showing it in live .
Just one little trick for now: You can apply the solution as spot (proposed by those THC quantifying kit sellers) or as band (line).
The former sometimes works better for semi-quantitative things like the THC test with a selective detection reagent (spray), the latter gives better resolution (if you want to test for example terpenes with a non-selective reagent like vanilline-sulfuric acid).
Spotting bands may be difficult -> apply as spot and let run first with methanol but only slightly higher than the upper part of the applied spot (~1 cm high). Dry and repeat 2-3 times. Methanol has a high elution capacity and nearly all constituents will migrate with it thereby forming a nice fine line at its front. That's now your new starting point in the form of a perfect line resulting in a high and good separation when you run it with the real eluent (= solvent mixture).

Got to go...

 

[pinkus]

you're going to teach me right? kidding
I'm buying a kit and literature. Right now I'm trying to figure out what classes I should take locally that would get me in the proper lab while I'm learning technique and concurrently doing my own DE TLC @home. I have worked in many undergrad labs but they have all been animal-centric bio labs.

I know I've got my work cut out for me. Even simple staining slides takes practice.

 

[Only Ornamental]

Take practical courses in chemistry or pharmacy; in Europe (well, depends a little on the country), students start making TLC in the second year and good ones in the third.
'Hire' a final year or doctoral student for a crash cours. Shouldn't take longer than 3 times an hour or a half of private lessons. Especially when you're about cannabinoids, you already have the eluent and the spray reagent. That makes everything way easier. Basically, you need to learn how to prepare a TLC; understanding the why and predicting or optimising eluent systems is helpful but can be learned from even easy literature and you can ameliorate that by yourself at home.

Maybe a bit more which may not be found in books:
- Don't touch the TLC plate with your fingers, only with proper gloves but better just don't (on the sides and the back its obviously okay ).
- Go with aluminium sheet silica and not glass supports. Cutting is done with a cutter, a scale and on the aluminium side (silica side towards the worktop) and not with scissors; gently carve (without cutting all the way through the sheet) 2 to 3 times so that you get a nice 'nick' all along the cutting line but with the two parts still holding together and then bend them like a butterfly. The alu will smoothly break without detaching the silica layer from it.
- When applying your solution keep 1 cm from the borders (left/right and bottom).
- Migrate over at least 7 cm.
- Apply ~10 micrograms of pure compounds or ~100 micrograms extract per spot. This works for most detection reagents and gives a good resolution. That means: prepare a stock solution of 1 mg per ml and spot 10 microliter for pure compounds and go with 1 to 10 mg of an extract per ml depending on solubility. Obviously, you got to adjust the microliters.
- Use the most apolar solvent possible to apply your samples (limitation is solubility): for THC and terpenes hexane should work at 1 mg/ml, diethyl ether or ethyl acetate may be better for 10 mg/ml. Try to avoid water because it doesn't dry well (you need to dry the spot/line prior to migrating). Methanol on the other hand works for most applications but gives big spots (rather circles) and doesn't work well for application as lines (go with the tip in my previous post).
- Avoid acetone as eluent, usually doesn't work.

For terpenes and a lot of other plant metabolites, go with a good book like THIS one here (maybe the best in my biz and available as free PDF ) and THAT one (lacks the cover but it's one of the classics).

 

[pinkus]

Thank you so much! It's on the tablet now.

I also have a bunch of cacti of Andean lineage I'd like to assay, any thoughts on those slimy guys?

 

[Only Ornamental]

I had around a hundred species as a teen .

There's often not much inside 'em ;( .
Lophophora sp.(peyote) and Trichocereus/Echinopsis sp. (San Pedro) need natural climate to produce enough mescaline (they won't do it in Europe or indoors). Besides that, there aren't many other 'interesting' alkaloids present...

Most cacti are edible from tip to toe (but taste like immature cucumbers); many fruits are delicious, though!

 

[pinkus]

My cacti all went back outside a few days ago and it seems likely I won't have to pull them in again until next December. I've often wondered about alkaloid production and climate. I'm lucky on that front but have a friend who very successfully grows and enjoys them (the Andean family) in Michigan... plenty of alkaloids by all accounts. That's yet another reason I want to learn this; do I need to keep verified specimens, or do I just need to fulfill their requirements? or both?

 

[Only Ornamental]

You only need a known botanical species like L. williamsii (there's not much known genetic variation between indivduals) and the appropriate environment. Especially peyote produces the more the slower (more natural) it grows; pump it up with ferts will often result in no mescaline.
I think San Pedro is way better in that regard as it is one of the naturally fast growing species accepting a broader range of climates.

 

[pinkus]

The cacti I was thinking of assaying are all of the peruvian type, though I live very close to peyote's natural range. I go to a ranch about once a year that should have them by geography, but I haven't spotted any. I wouldn't touch them anyway... well, i'd touch them in a loving way .