Scrutinizing Strains with Science : An Objective Discussion

[Donn]

I have tried THCV both pure and with pure THC, THCV does not get you high, and is more like VBD in that it delays onset, reduces peak experiences of THC, you do not want it for recreational use it is a THC antagonist. I introduced S African Durban Poison in the 70's, in California, I know it well.
-SamS

Yes, maybe that is in a way how it worked out with me, that 1% or so THCV was enough to knock down some psychoactive aspects a little - but not all of them. So it wasn't for me, but some people love the stuff. That's cool, with data to look at, we can each choose our favorite high. Or of course, with various medical purposes in mind you might even be looking for even more elevated levels.

 

[Siskiyou Sam]

I'm thinking that all these terpene profiles can be used to see if there are dominant and recessive terpenes. For example I just collected all the Girl Scout Cookie terp tests I could find, and knowing that GSC is a cross between OG and Durban Poison, I collected all of those profiles...currently analyzing.

 

[Bubbleblower]

I did try Ed's stupid idea of eating a mango for Myrcene with Cannabis or THC, zero difference that I could see.

Same here.
Try a drop of peppermint oil in a can of tea (3 drops a liter).

I'm thinking that all these terpene profiles can be used to see if there are dominant and recessive terpenes. For example I just collected all the Girl Scout Cookie terp tests I could find, and knowing that GSC is a cross between OG and Durban Poison, I collected all of those profiles...currently analyzing.

Rich in humulene like GG#4.

 

[Donn]

So it does matter how it gets into the system.

Though if you eat your cannabis and lemonade, looks like the route should be exactly the same. Takes a while, too - weirdly long time, caffeine for example takes just a couple minutes, but THC in eats takes at least 10 times as long to settle in. No idea how long for limonene, going to try a little lime juice this minute.

 

[Donn]

Rich in humulene like GG#4.

See 3mg/g alpha-bisabolol in Gorilla Glue #4? That's what jumps out at me, in an SC analysis online. One of the principle active ingredients in chamomile, various physiological properties.

 

[MrBelvedere]

Here are some sample attributes that can be logged, please add more if you can think of them...

<xml>

<SampleUUID> <! always generate a fresh UUID for every sample >

<PlantUUID> <! the UUID of the plant, if clone of another plant put that same plantUUID here, if submitting samples off multiple branches leave plantUUID same. !!If unknown, generate fresh UUID!! >

<CloneCuttingOfPlantUUID> <! the PlantUUID of plant the clone was taken from. Blank if not a clone. Possibly useful for establishing drift.>

<FatherPlantUUID> <! if known, put the PlantUUID of the father >

<MotherPlantUUID> <! if known, put the PlantUUID of the mother >

<SiblingOfPlantUUID> <! there can be multiple records here, put the PlantUUID of all siblings >

<PhotoPeriodChangeDateTime> <! put the datetime the photoperiod was changed for the Sample/PlantUUID >

<CutDateTime> <! datetime the SampleUUID was cut >

<SampleTakenFrom> <!top of topmost cola, top of side branch cola, bottom of topmost cola, bottom of side branch cola, middle of topmost cola, there could be hundreds of new and different data fields here- request for comments on them. ie North/South/East/West direction the side branch cola grew in, LatitudeLongitude, etc. >

<DaysDrying> <! number of days sample dried before submission>

<DaysCuring> <! number of days curing in storage before AnalysisDateTime >

<THC>

<Limonene>

<CBD>

<UsedForTesting> <! describes the mechanism used for analysis. For example: HomegrownTLC, LabBasedTLC, WatersHPLC, AgilentHPLC, etc. be verbose. >

</xml>


What else besides the obvious noids and terps should be logged?

Software for translating old propietary MS file formats, has screenshots. Alternatives are available. Used for illustration.
http://www.sisweb.com/software/csw/gcmsfile.htm

UUID generator for illustration
https://www.uuidgenerator.net/

Attempt at open MS format- to facilitate dataset sharing and collabo. "Replaced" by mzML open standard with input from manufacturers.
http://tools.proteomecenter.org/wiki/index.php?title=Formats:mzXML
mzXML is an open data format for storage and exchange of mass spectroscopy data, developed at the SPC/Institute for Systems Biology. mzXML provides a standard container for ms and ms/ms proteomics data and is the foundation of our proteomic pipelines. Raw, proprietary file formats from most vendors can be converted to the open mzXML format.
Patrick Pedrioli was the primary original author;

http://en.m.wikipedia.org/wiki/Mass_spectrometry_data_format

Sample simplified mzXML file
<mzXML xmlns="http://sashimi.sourceforge.net/schema_revision/mzXML_3.0" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://sashimi.sourceforge.net/schema_revision/mzXML_3.0 http://sashimi.sourceforge.net/schema_revision/mzXML_3.0/mzXML_idx_3.0.xsd">
<msRun scanCount="3113" startTime="PT5.0005S" endTime="PT119.982S">
<parentFile fileName="file://PROTEOMICSDEV01/Inetpub/wwwroot/ISB/data/class/day1/runsearch/ICAT_test4.RAW" fileType="RAWData" fileSha1="71be39fb2700ab2f3c8b2234b91274968b6899b1"/>
<msInstrument>
<msManufacturer category="msManufacturer" value="ThermoFinnigan"/>
<msModel category="msModel" value="LCQ Deca"/>
<msIonisation category="msIonisation" value="ESI"/>
<msMassAnalyzer category="msMassAnalyzer" value="Ion Trap"/>
<msDetector category="msDetector" value="EMT"/>
<software type="acquisition" name="Xcalibur" version="1.3 alpha 8"/>
</msInstrument>
<dataProcessing centroided="0">
<software type="conversion" name="ReAdW" version="1.2"/>
</dataProcessing>
<scan num="19" msLevel="1" peaksCount="1313" polarity="+" scanType="Full" retentionTime="PT353.43S" lowMz="400.39" highMz="1795.56" basePeakMz="445.347" basePeakIntensity="120053" totIonCurrent="1.66755e+007">
<peaks precision="32" byteOrder="network" contentType="m/z-int" compressionType="zlib" compressedLen="782">
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
</peaks>
<scan num="20" msLevel="2" peaksCount="43" polarity="+" scanType="Full" retentionTime="PT356.68S" collisionEnergy="35" lowMz="223.089" highMz="531.078" basePeakMz="428.905" basePeakIntensity="301045" totIonCurrent="764637">
<precursorMz precursorIntensity="120053">445.3466797</precursorMz>
<peaks precision="32" byteOrder="network" contentType="m/z-int" compressionType="zlib" compressedLen="55">
eJw7wAABDQzYAbr4ARzyB9DoBhw0OnDAYQ66uAOaPDqNbh6x7kC3DxeA6UO3D10cxgcA3f0RQQ==
</peaks>
</scan>
</scan>
</msRun>
<index name="scan">
<offset id="19">1209</offset>
<offset id="20">2577</offset>
</index>
<indexOffset>31713417</indexOffset>
<sha1>e83e234ed25a2e675ad8a3fbcd56f16585a237c2</sha1>
</mzXML>

a limonene rich variety is likely always limonene rich but one time it's more, another time less, and the whole profile of volatiles will differ (mostly in relative amounts) rendering standard statistical analyses beyond feasible.

If I understand you-the terpene/noid attribute levels will change between samples? The SQL db algorithm post #141 takes into account this variance, maybe?



Thx

 

[Only Ornamental]

...
If I understand you-the terpene/noid attribute levels will change between samples? The SQL db algorithm post #141 takes into account this variance, maybe?

I don't know what your program really does, so I really help you with that. You would have to try with a dozen real life samples preferably having 20 or so terpenes.
Depends on a lot of things: maths and statistical methods (e.g. outlier test for single constituents), if you use absolute and/or relative amounts, which variance you tolerate... It might actually work in some instances to attribute a sample to a certain population or even a specific variety but I still highly doubt it. Hillig and others tried that just to see if they could distinguish C. sativa, C. indica, and C. ruderalis. It barely worked for C. sativa v.s. C. indica thanks to a very few constituents only present in one or the other (most of all sesquiterpenes in C. indica) but didn't for C. sativa v.s. C. ruderalis. Whether or not this is due to both being the same species, a too small sample population, or chemotaxonomy being useless in this regard, we don't know.
Genetic markers or the like would be very useful as an additional tool (even if it's only to prove the authenticity of the samples).

I propose to treat terpene and cannabinoid profiles separately.
Mostly because THC/CBD are determined by one single locus and influence also other cannabinoids; the BT allele gives THCA which results in THC and finally CBN, the three are linked and for a chemotype profile only the direct biosynthetic products should be used i.e. one single value for THCA+THC+CBN.

Finally, you would have to attribute the single samples (not varieties!) to certain illnesses or effects. How this is going to work...???

 

[MrBelvedere]

What I am trying to say (not very well) is that *before anything* a tiny baby step needs to be taken, not a leap... The first baby step is to simply see how the samples grow, harvest, and handling environments will change quantities of noids and terps.

For example test a thousand samples of the same plantUUID and analyze the large dataset to see exactly which levels (noids, terps, etc) have the highest level of variance. Ie X levels vary little and Y levels change a lot- even with identical environment. This baby step of knowledge would help crudely "calibrate" future tests and analysis. In all these "calibration tests" the environment (grow, harvest, handling) would be highly controlled to be uniform. Same feeding, same harvest date, same handling etc.

The program is doing just that, it is capable of finding the variance of levels of the same plant... As the amount of records in the dataset increases by the thousands so does its accuracy.

 

[Only Ornamental]

Ah, that way! Sorry, I misunderstood you before.
You're absolutely right. Although, I'm not so sure if your approach really is such a baby step.
Your dataset would certainly be very helpful for breeding and growing too.

 

[CannaBrix]

I have tried THCV both pure and with pure THC, THCV does not get you high, and is more like VBD in that it delays onset, reduces peak experiences of THC, you do not want it for recreational use it is a THC antagonist. I introduced S African Durban Poison in the 70's, in California, I know it well.
-SamS

And how about the effects of THCV along with other cannabinoids, and terpenes? Would myrcene counter THCV?

Could cannabis with certain levels of THCV be good for a person who wants less of a high? Or even a different high?

I'll quote a friend of mine "...and maybe one day you can go to the store and pick up a pack of joints, but they won't get you high. It'll taste and smoke like bud, but just doesn't get you high...or at least as high. You know, so if you don't want to smoke tobacco, there is an alternative."

 

[Bubbleblower]

They should test mouse motility to see if it is coughlock or energizing.

 

[Donn]

Could cannabis with certain levels of THCV be good for a person who wants less of a high? Or even a different high?

Sure! Ask someone who likes Durban Poison, Chernobyl, Jack the Ripper, no doubt a few others. We're talking levels like a couple percent, order of magnitude less than THC levels in the same sample, but considerably higher than normal. I don't think "less high" is the attraction with those strains, or even if there's any consistent effect across the whole bunch. I've tried Durban Poison from one source. Jack the Ripper apparently gets the trait from Jack's Cleaner, whose parentage is a longer story, and Chernobyl gets it from Jack the Ripper. What do you think, do the high THCV strains in this Jack's Cleaner family work out the same as Durban Poison?

Now, if the THCV levels were comparable to THC levels, or greater, I guess that might be more in the medical applications category. Someone probably has that, but I don't see any evidence of anything that's actively marketed. Another plus for "other cannabinoid" test data numbers: if there's something like that on the shelf, you don't want to be talking "energetic" or even "high THCV." If it's 12% THCV, you want to know that. Whether we know exactly what it does or not!

 

[Donn]

Mostly because THC/CBD are determined by one single locus and influence also other cannabinoids; the BT allele gives THCA which results in THC and finally CBN, the three are linked and for a chemotype profile only the direct biosynthetic products should be used i.e. one single value for THCA+THC+CBN.

That makes sense to me, but where does CBG fit in that picture? Usually present at noticeable levels in the Streep Hill analyses. It's a precursor to various cannabinoids, do I have that right? but in cured material it would be stable?

 

[Sam_Skunkman]

Really what I am saying is that people have tested various terpenes with pure THC, there is no doubt that THC's effects are modulated and potentiated by the right terpenes being smoked or vaporized with the THC. I have not heard of the same tests with oral pure THC with terpenes. So any presumed modulation is pure conjecture, I would not be surprised but it is not proven.
An easy test would be to first take .25mg pure THC orally then the next day try .25mg THC with a spike of one terpene orally, then the next day repeat with a different terpene, etc.
One of the reasons I did not try them orally was concerns about the safety of the terpenes orally, and the length of time for the effects to become known vs smoked or vaporized which is fast!
I thought by now someone would have tried pure THC and various terpenes one by one orally, but if anyone has I have not heard, until then, I will just not know if terpenes are active modulating THC when taken orally.
-SamS

@Sam: Mono- and most sesquiterpenes are readily resorbed when eaten and even applied to the skin, roughly 50% are bioavailable (the rest 'evaporates') and will cross the blood-brain-barrier. Though the AUC is similar (cumulative dose over time), the kinetic is different: it takes more time to achieve maximal plasma concentration which is lower but longer lasting. Combining smoked THC with eaten essential oil will not be the same as the two curves (concentration over time) aren't identical (though, they're pretty similar when applied the same way).

For me, orange and lemon juice brings me down and clears my head (when taken after the 'peak'). Back in my wilder days it was our first aid for 'overdoses' and worked with most of the clique astonishingly well.

I tried the combination of cannabis 'chai' with eaten mango, oranges, and candied ginger, respectively: No difference noticed.

 

[Sam_Skunkman]

You are presuming that the THCV is the reason, not just other terpenes found in the Durban Poison S African modulating the THC?
What I can say is you do not want a high THCV variety for recreational use, I have them that are 90% THCV, of the Cannabinoids, you do not get high, and 25% THCV, it just ruins the high.
-SamS

Sure! Ask someone who likes Durban Poison, Chernobyl, Jack the Ripper, no doubt a few others. We're talking levels like a couple percent, order of magnitude less than THC levels in the same sample, but considerably higher than normal. I don't think "less high" is the attraction with those strains, or even if there's any consistent effect across the whole bunch. I've tried Durban Poison from one source. Jack the Ripper apparently gets the trait from Jack's Cleaner, whose parentage is a longer story, and Chernobyl gets it from Jack the Ripper. What do you think, do the high THCV strains in this Jack's Cleaner family work out the same as Durban Poison?

Now, if the THCV levels were comparable to THC levels, or greater, I guess that might be more in the medical applications category. Someone probably has that, but I don't see any evidence of anything that's actively marketed. Another plus for "other cannabinoid" test data numbers: if there's something like that on the shelf, you don't want to be talking "energetic" or even "high THCV." If it's 12% THCV, you want to know that. Whether we know exactly what it does or not!

 

[Donn]

You are presuming that the THCV is the reason, not just other terpenes found in the Durban Poison S African modulating the THC?

It's a theory - I'd need a lot more data to say with any confidence! But it's the distinctive thing in the "fingerprint", though that has only a few terpenes, and I think it's consistent with your observation about effects of higher THCV - here we're talking about an order of magnitude less, and it more selectively suppressed certain psychoactive aspects - for me. So I'm not presenting this very weak evidence really to say anything about THCV, so much as to just plead for its inclusion in test reports. I was just looking at a Werc Shop presentation of a Cannabaceutical label that, I'm pleased to say, has THCVA. But no CBC. And terpenes - they just list the top 7 terpenes for the sample, so it's more compact if less digestible.

In this presentation - you guys have probably all seen this talk at Humboldt by Jeffrey Raber - he presents some scatter plots from principal component analysis of a lot of strain cannabinoids & terpenes. And talks about publishing all these strain "chemotypes", though I sure can't find any of that. This stuff is all 45-50 minutes in. He talks about it mostly from a taxonomic viewpoint, looking for effective phenotype relationships. Conventional sativa vs. indica labels don't seem to have much value with respect to the chemistry (I guess this is not news.)

 

[MrBelvedere]

Really what I am saying is that people have tested various terpenes with pure THC, there is no doubt that THC's effects are modulated and potentiated by the right terpenes being smoked or vaporized with the THC. I have not heard of the same tests with oral pure THC with terpenes. So any presumed modulation is pure conjecture, I would not be surprised but it is not proven.
An easy test would be to first take .25mg pure THC orally then the next day try .25mg THC with a spike of one terpene orally, then the next day repeat with a different terpene, etc.
One of the reasons I did not try them orally was concerns about the safety of the terpenes orally, and the length of time for the effects to become known vs smoked or vaporized which is fast!
I thought by now someone would have tried pure THC and various terpenes one by one orally, but if anyone has I have not heard, until then, I will just not know if terpenes are active modulating THC when taken orally.
-SamS

Thanks, What is a good "pure" source of these compounds ?

Linalool, Myrcene, Limonene, Terpinolene, α-Pinene, β-Pinene, Humulene, Caryophyllene, and Caryophyllene oxide

I googled for a while and could find some pure sources, some are in essential oil extract I'm not sure if that would be the same? I would rather smoke it with cannabis than eat it, for research. If you used an essential oil in your test, did you dry it out before smoking? Just put a drop on a big bud, let it dry, and smoke a big bong hit?

https://www.icmag.com/ic/showthread.php?t=260175

Limonene looks easy to fine, but all the others are not so easy to locate online...
http://www.perfumersworld.com/datasheet/3cq00276-limonene-d-.html?lang=en

http://www.sigmaaldrich.com/catalog/product/aldrich/53676?lang=en&region=US humulene is 300USD for 100g food grade! And not sure if it is pure?

http://www.perfumersworld.com/datasheet/3ws00089-caryophyllene-beta-.html?lang=en humulene

Perfumersworld seems to have a few? And sigma Aldrich?

 

[Bubbleblower]

I have not heard of the same tests with oral pure THC with terpenes.

You listen to the wrong people.

Just put a drop on a big bud, let it dry, and smoke a big bong hit?

Or put a drop in your tea or a drop of lemon in a fresh banana milkshake.
Ingesting has more effect than inhaling it.

OO should be able to tell us a lot about the effects of different terpenes; Germany is a decade ahead in this field and has the best doping doctors.

 

[Sam_Skunkman]

x

You listen to the wrong people.

No problem, who are the right people to listen to that have tried pure THC and single terpenes orally?
I am curious of their results.
All the best,
-SamS



Or put a drop in your tea or a drop of lemon in a fresh banana milkshake.
Ingesting has more effect than inhaling it.

OO should be able to tell us a lot about the effects of different terpenes; Germany is a decade ahead in this field and has the best doping doctors.

 

[Bubbleblower]

x

THC plus terpenes has been tested (typically 300% more effect) and single terpenes have been tested, but I am not sure about single terpenes with THC.
Obviously they should test that too.
IMO they should test anything oïds and make as many multivariate analyses as possible.
It seems anything that passes the blood-brain barrier is psycho-active one way or another and most of these substances have synergy.

Who we should probably really listen to are the Chinese.
They are proving rapidly what TCM has claimed all along to get all the valuable patents; they already have hundreds of patents on cannabis. Ayurveda similar story.
They have been breeding for the best cannabis for thousands of years and probably knew what they were doing.

I am an expert on your DP too, just check my avatar It was from a freebee I got from GN when he still had his shop.
What makes DP so special could be related to liquorice. Somehow that is addicting especially to children.
If you ask Dutch people abroad what they miss most many will answer liquorice -which is a weird answer- and they order it specially back home.

“In traditional Chinese medicine, liquorice (मुलेठी, 甘草) is believed to "harmonize" the ingredients in a formula and to carry the formula to the 12 "regular meridians".”
It does make you feel warmer, they are probably getting the patent as we speak.