Im sorry but d8 isnt made through a enzymatic pathway both papers say it is made through oxidation
And i quote
" Compounds of this class might be isolation artifacts resulting from Δ9-THC by acid- or oxidatively promoted shift of the endocyclic double bond, or from CBD by electrophilic cyclization. The Δ8 location is thermodynamically more stable than the Δ9 location, and this drives the isomerization."
CBDA-synthase and THCA-synthase have been cloned from the storage cavity of the glandular trichomes of cannabis,20,21 and they exclusively produced their corresponding phytocannabinoids
Which is wrong. Both make both. At least when tested in vitro.
Proposing that ALL D8 produced in planta is produced via UV based degradation doesn’t fit the data.
At least not without proposing some other difference (intra cellular pH).
If it doesn’t fit the data, the hypothesis is wrong….
If this were true youd always have cbd in d9 disty which isnt true
THCA synthase has also been crystallized, and the FAD and substrate-binding sites identified.22 Apparently, the enzyme selectively produce one of the two isomeric THC acids present in nature, THCA-A.22 THCA-
Either way d8 isnt produced by the plant its a result of environmental factors whether its from a hydride shift or oxidation
This proves nothing and can be contradicted by the fact that analyzing d9 strains the same way results in no d8. Plus the fact that extracting samples for extended times, or using some heat, does not results in more d8 (and less d9, or more d8+d9 and less cbd…)
And saying youve tested hundreds of samples and that because of that theres more d8 in hemp then in d9 strains doesnt mean anything either especially when d8 is made from d9 in ALL cannabis strains
would it not then create a similar ratio of D8 in his high D9 samples?
a BETTER fit to the data would be the proposal that @Dr_Jebril’s method turns some of his CBD to D8
that accounts for ratio with D9 not being the same…which your hypothesis does not.
IMO that doesn’t fit all the data…
we actually have the molecular modelling software and appropriate structures required to look at the problem…it should be relatively easy to show I’m wrong computationally.
You need to turn cbd into d9 first though you cant go straight from cbd to d8 everyone knows this
Which is why if you have a high d9 cultivar it is more likely to have higher d8 concentration then a cbd cultivar because its double the work to make cbd into d8
So youre basically saying every paper thats ever been written on the subject of this is wrong?
Im sorry what you’re saying doesnt fit at all especially when you look at the ring structure and where the double bond lies
Theres literally a bunch of papers on this yet you refute them by saying theyre all wrong
In my HPLC experience from 2 facilities, one marijuana and one hemp, there is a greater likelihood of having D8 in hemp varieties. I can’t say why, but the samples I’ve tested and seen tested have shown this correlation, and I can say I also agree with @Dr_Jebril’s statement:
I’m not in D8 so I don’t have a pony in this race, just wanted to comment on a trend I’ve also noticed.
This is the opposite we have found with our in house testing along with 3rd party testing. We have found thc biomass to have more d8 than our cbd biomass that we extract at our facility. I feel like storage conditions could have something to do with this.
show me a single paper where folks have specifically asked if CBDA systhase is making D8…
@seth gave us one where they are looking at what the enzymes make, and one might assume the authors could tell D8 from D9…but we have plenty of precedent for folks unable to make that distinction, and the authors do not identify all the products of their reaction.
no, the plant doesn’t have an enzyme whos job is to make D8, but both “hemp” and “marijuana” have enzymes that are making off target cannabinoids. THCAS can “accidentally” making CBD. CBDAS can “accidentally” make D9.
You’re stating that you’re CERTAIN CBDAS isn’t responsible for making D8.
so why is the D8/D9 ratio higher in a CBDAS plant than a THCAS plant? were it environmental, those ratios would be the same.
we decided the world was a sphere because the data simply didn’t work with “flat”. sure looks flat. but it’s not. the hypothesis needs to fit the data. if it doesn’t you need to re-evaluate.
Could be. The hemp I’ve tested is from many places all over the country. The marijuana was all grown at the same facility it was tested at. I can’t say how the hemp was treated but the marijuana was relatively fresh (<6 mos from harvest usually).
Do you have any proof of what youre saying because other people have found the opposite of what youre saying (post above)
The levels wont be the same when theres different levels of d9 content in both cultivars
This is something i also believe to be true
MJ (especially when in metrc) gets treated alot better when it comes to storage conditions. Also hemp is more likely to go full term which would definitely contribute to enviromental factors and conversion
@seth can you chime in here on if you guys tried to test the cbd or thc enzyme pathways for d8? D8 is pretty common to test for now adays so I’d be surprised if you didnt
I don’t disagree that treating your high D9 poorly will likely make D8 from D9, at a level dependent on D9 level and level of abuse.
However, @Dr_Jebril’s data suggest; treat your CBDAS only plant any old way, you get fairly tight D8/D9 ratio. suggesting a different mechanism might be in play.
how does “UV induced hydride shift” explain that disconnect? ratio should be constant across all cultivars if it’s not genetically encoded.
I have no attachment to my hypothesis other than it fits the data better than yours. show me where mine doesn’t, and explain how yours fits better
@Gail is reporting absolute D8 level. @Dr_Jebril is reporting d8/d9 ratio.
I dont see d8 being made by the enzyme. Its not going to shift the double bond on cbg like that and youre not going to synthesize cbg with the double bond in the wrong place so it makes d8 instead of d9
it’s goal wasn’t to close the ring. that is an off target reaction. having the bond in the D8 position is energetically favorable and easily achievable with an acid catalyst. Are you certain there isn’t anything in/near the active site of CBDAS that could act in the same manner?
oh wait, there HAS to be…because it’s accidentally making D9…and D8 is “easier”
certainly not an enzyme that does it as it’s primary reaction.
given that one can shift the bond without need of an enzyme, the contention that the same could not be achieved by an enzyme is pretty hard to support.
" The electrophilic cyclization step is highly specific in terms of termination. In one version of the process, the C-8 cation (menthane numbering) behaves as a Broensted acid, and is quenched by loss of a proton from C-9 to generate the exocyclic double bond of CBDA (Scheme 2). In the alternative version of the termination, the C-8 menthyl cation behaves as an electrophilic sink for one of the two ortho-hydroxyls, generating Δ9-THCA-A from the hydroxyl para- to the carboxylate, and Δ9-THCA-B from the other phenolic hydroxyl."
You do realize to do what youre talking about youd need to make d9 first… and also the same paper i just quoted says that the plant doesnt have a way to turn cbd into thc
" In the alternative version of the termination, the C-8 menthyl cation behaves as an electrophilic sink for one of the two ortho-hydroxyls, generating Δ9-THCA-A from the hydroxyl para- to the carboxylate, and Δ9-THCA-B from the other phenolic hydroxyl. The oxidative- and the electrophilic cyclase activities are closely associated, and the menthyl cation is not released or leaking from the enzymatic cleft where it is generated, making the two termination process biogenetically orthogonal. This is consistent with the paradoxical observation that, while CBD is easily converted into Δ8- and Δ9-THC by acidic treatment under laboratory conditions, CBDA is not converted into THCA in cannabis tissues"
CBD plants do produce d9-THCA, likely from CBGA. What would preclude them to further convert part of this d9 to d8 (as it does for CBNA or HHCA, and as any guy with some acid can do in his garage, as many lurkers here…) ?
@EHO_AZ@Gail, the hemp samples Im dealing with are of any sort (flowers, extract, indoor/outdoor/greenhouse, aged, fresh, even right freeze dried from the field…). And there is a systematic. The THC samples are generally indoor, also fresh and treated with care, showing no d8. Low quality flowers will show some, but still lower than in CBD strains. Moroccan hashs, which are have been through rougher conditions (heat, travel, time etc…) shows a little more, but still much lower than the 1-2% CBN (and the ysual 1-4% CBD).
