Standards Used in Testing

[Chimera]

Gobble

northstate- we aren't going to debunk any thing here... we're going to blow this mofo right outta the water. It hasn't even gotten good yet. Patience.

And this, my friends, is what separates Chimera Genetic Resource Management from the rest of the herd. Any and all seedsters are more than welcome to come and offer their contributions to this thread. Takers?

 

[Chimera]

One more for good measure

 

[mofeta]

Hey this is a good thread!

I don't have much time right now but I thought I would add a couple of things.

On the question of wide-spectrum testing for things like VOCs and pesticide residues, it is the type of detectors that are hooked to your column(s) that does the trick. The pesticide guys use NPD (nitrogen phosphorus detector) a lot. There are a number of selective detectors that can all be ganged on to your column besides the normal FID and MS.

Hi Chimera

Excellent contribution as usual.

Just a note, it is trivial really, but I know you are a stickler for strict terminology, as am I.

chromatograph= piece of equipment used to produce a chromatogram

 

[legalizeDK]

For this reason, it's really important (moreover, necessary) to have a pure sample of the known chemical to run through your GC and column with the internal standard (usually squalene), to identify the unique signature of your compound, and create a literal 'standard' against which future runs through your GC/column setup can be compared. Also the NIST-like databases are a good approximation, and may in many cases lead to proper identification of the compounds of interest, but they are not entirely %100 correct and can mis-identify molecules that have similar retention times in most columns. I suspect that this is really common in cannabis analytics, because CBD and CBC have incredibly similar retention times with most columns, the peaks are almost indistinguishable and one is often mis-identified as the other. Put that in your pipe and smoke it.
-Chimera

this sums up the problem quite well.

i work in lab and we used to do drug tests on a lcms. The lab that took over drugtests didn´t have the same standard as we did, because its also about how it is calibrated/tuned

 

[mofeta]

As to standardization:

The need for reference standard has already been well addressed in this thread.

The hardest part, though, are the protocols used to prepare the sample:

*Selection of the correct part of the submitted material- results could vary wildly from the same submission depending on what part of the bud the tech used on each run, and how much attention was paid to the relative amounts of the various structures selected for the sample (ie how much stem/bract/calyx etc). I think most techs would just mill the whole bud, but any smoker would realize that although this would homogenize the individual bud, the relative amounts of the individual structures vary from bud to bud. Special effort would have to be made not to leave resins behind if a mechanical milling method was used.

*The type of extraction done to prepare the analyte. There are myriad ways to seperate your analytes from the matrix. Soxhlet? SFE? Subcritical Water? SPME?

*The skill of the tech. This is huge. Can he/she properly calibrate the machine? Did they run a blank since the last sample? Are they fastidious in their technique?

The point would be to develop a standardized protocol specifically suitable for the analytes we are interested in, and take steps to assure this standardization (centralized training/accreditation of techs/labs).

 

[Gray Wolf]

Thank ya'll for the enlightenment!

Sure changes one's perspective, doesn't it?

 

[Green Supreme]

Wow I went to sleep for a bit, come back and its poppin off in here. Thanks all for your input. Peace GS

 

[Nondual]

Chimera - since you're in Canuckerville have you ever worked with Philippe Lucas from VICS? I talked to him a bit about 5 years ago when a buddy and I were looking at setting up some testing equipment. He said it was possible to acquire standards and he had set up in-house testing back then...pretty sure.

I'm familiar with things like HPLC and GC Mass Spec as that's what the labs I use test non cannabis samples with for certain parameters. I worked with an organic chemist in the late 90's and he taught me how to use the HPLC but we never got into the GC Mass Spec equipment.

I'm guessing you're using lab analysis in your breeding work now...eh?

 

[DTFuqua]

The most obvious question for non scientific minds is if a sample of weed has 30%THC, that's 30%m of what. If an ounce of weed is tested at 30%, do I have 3/10 of an ounce of THC? I think not. That is something that needs to be straighteened out and set to standard where some of the ignorant drug warriors can't make it look like something is worse than it is by talking about how potent things are.

 

[Gray Wolf]

The most obvious question for non scientific minds is if a sample of weed has 30%THC, that's 30%m of what. If an ounce of weed is tested at 30%, do I have 3/10 of an ounce of THC? I think not. That is something that needs to be straighteened out and set to standard where some of the ignorant drug warriors can't make it look like something is worse than it is by talking about how potent things are.

For scientific minds as well bro, regardless of how twisted and deranged some of us'n may be otherwise!

I have heard tell of strains producing 30% oil by weight, as compared to the weight of the starting material, but personally have never achieved more than around 25% before winterizing to remove the waxes, et al, with a record 23.7% BHO Absolute yield.

I believe that 25% would cover most strains, even if some produce 30%. The high CBD Catalyst strain that we raised, only produced 5.7% oil by weight.

If we extract that oil from one ounce (28.3 grams) of bud at 25% yield, we would have ~7 grams of unrefined BHO oleoresin extract.

We could further refine it by winterizing to remove inert waxes and end up closer to maybe 6 grams Absolute.

With most current strains, if you tested the Absolute, it would most likely be primarily THC and THCA, which combined can be in the high seventies to mid nineties as a percent of total composition.

If 90% of the 6 gram Absolute oil yield was THC/A, that would be 5.4 grams, which is about 19% of the starting 28.3 gram weight.

Plug in different numbers and get different answers, but that is how I would do the math.

 

[Mia]

Got my popcorn, er skillet out, chillin.
Muy interesante!
Seems comprehensive testing at this point in time is problematic to say the least....

 

[G.O. Joe]

CBN and CBD can be used as THC standard for GC; both are crystalline, so were extracted in their pure form long ago. THCA and - most impractically of them all for most - crystalline THC derivatives can also be prepared. Those with the organic chemistry background to operate GC's, and enough desire, should be able to handle any of this after doing some reading.

Solutions of analytical standards can be purchased from any number of chemical suppliers that trade in controlled substance standards. They'll probably want verification that they can legally do so, any that don't are probably in China or something. Real labs for drug analysis needing an analytical reference have real drug analysis licenses, though these licenses may not be easy to obtain. While having genuine standards from Fluka or whoever and all necessary operation licenses is no guarantee at all of the most important factor - competent operation - they should have all these things.

These are external standards, and are not the only calibration method. Internal standards are most important, and need not have anything to do with cannabinoids. Internal standards are a known amount of something added to the analyte. I've never read anything where cannabinoids were tested and an internal standard was not used. Internal standards are inherently more reliable than external, because experimental deviations affect a correctly-chosen internal standard equally.

 

[Green Supreme]

Do do you believe that the places{In Cali and Co. for instance} making all the cash for testing, have authentic standards. I have mentioned before, that the community settling on a common method and technique would also give testing validity.

DNA testing will also add validity. Once a database is created to compare against, we can watch much of the name game fall apart.

Thanks for everyone's input. Peace GS

 

[mofeta]

While having genuine standards from Fluka or whoever and all necessary operation licenses is no guarantee at all of the most important factor - competent operation - they should have all these things.

This is the nut of the issue. Obtaining standards, licenses, equipment etc. is the easy part. Developing the protocol is harder. Finding competent personnel to do the work is yet harder. Getting even the better techs to do EXACTLY the same thing EVERY time, without cutting corners on your carefully designed protocol- the most difficult part.

These are external standards, and are not the only calibration method. Internal standards are most important,...

So true. The proper use of internal standards would have to be a part of the protocol. Even the best technician is still human, it is basically impossible to inject exactly the same amount into the port every time, etc.

 

[G.O. Joe]

Like most college, chemistry classes do not prepare students in any way for actual work in the real world, at least this is how it is in the US. I think this even includes welding and truck driving at the community college.

Those interested in GC could find pdfs on the subject, also not preparing students for the real world, using terms such as gas chromatography at sites such as www.4shared.com or http://library.nu
http://ifile.it/s8lzma/__Basic_Gas_...l_Chemistry___2009_Ed__.l_61x3339o9x79x5o.pdf
http://www.4shared.com/office/mjzFo4J-/Modern_Practice_Of_Gas_Chromat.htm

 

[mofeta]

Yes you are right G.O. Joe, the state of higher education in the US is deplorable. They trick kids into going into massive debt, and then don't deliver the goods. People wonder why all the physicians and engineers etc. are from India or wherever.

That last link you gave is a good one, Bob Grob was the man when it came to GC.

Edit: I forgot to add this: Now that I have slammed US higher education, I'll point out that some of the best resources on the web for practical use of GC apparatus are the guides universities make for the non-majors and undergrads taking lab classes involving GC. There are lots of these on the web, most stress not doing stuff to mess up the equipment.

Here is a OK one from Wake Forest University with good illustrations.

 

[Gray Wolf]

Yes you are right G.O. Joe, the state of higher education in the US is deplorable. They trick kids into going into massive debt, and then don't deliver the goods. People wonder why all the physicians and engineers etc. are from India or wherever.

Edit: Good points!

 

[KiefSweat]

I'm going to see if I can add a little bit of my experience with getting some of my stuff tested.

Have been working on my own extracts for a while. The whole process has been trying to figure out how to make an almost standardize extract to get a 1mg/1ml solution.

I've used a few labs and now work with one I am very comfortable with. Some points have already been brought up, but I think the biggest issue is who's operating the equipment.

As far as standards go, I know the lab I use buys them from 4 different pharm/lab companies in both the USA and Europe.
I do believe they have access to both CBG and THC-V standards but the cost is extra so they are not run on most samples.

In regards to THC/THC-A. I think this is where the different machinery comes into play. The GC/MS is just used for the major 3 thc/cbd/cbn. Its pretty quick and as reliable as long as the machine is calibrated correctly. Usually this is used for buds and hash.
The HPLC machine can detect both acid and decarboxilated forms of the cannabinoids. So the GC machine for example would read 1% thc, the HPLC machine could tell you that 40% is thc-a and the other 60% already activated thc. HPLC is used more for smaller qtys and edibles. I also believe they would use this to test for the other cannaboinds.

As far as reliability, I don't worry about the actually numbers as much. As long as the ratios stay the same between cannabinoids, or even to compare potency between different tests and personal observations its quite useful.

I don't think I've seen inflated numbers from the lab. I've tested a wide variety of products and fell pretty comfortable with those numbers and other research I've done that shows similar results.

 

[GreenintheThumb]

Real labs for drug analysis needing an analytical reference have real drug analysis licenses, though these licenses may not be easy to obtain.

Out here Full Spectrum asked for a Schedule 1 license...they were swiftly raided by DEA.

 

[Nondual]

Out here Full Spectrum asked for a Schedule 1 license...they were swiftly raided by DEA.

What about Industrial Labs in CO? I thought I read they were setting up testing. They've been around forever and used them for some conventional product testing over 10 years ago.