Medical Marijuana Analysis

[cindy99]

Hi I did a quick search and didn't find a whole lot of discussion on this subject. I'm hoping it will be interesting enough to some users here to help with my dilemma.

I am working on developing products for non-smoking medical marijuana patients. I need to regularly test samples for cannabinoid levels. My problem is this is relatively new to the area I am in and the closest lab where I can take these samples in for testing is a 2 hour drive away. Due to the nature of the product, shipping is not an option. Because of the cost and travel time, it's prohibitive to do anything other a very limited amount of testing. So, here I am, trying to figure out if it is a realistic option for me to do this kind of testing myself.

I have a VERY limited level of knowledge when it comes to discussing HPLC equipment and procedures. What I do know, is that it's very important for me to know the levels of THC-A, THC, CBD-A and CBD in the material I am working with. A huge bonus, would be the ability to test for pesticides and solvents like hexane, butane, etc. What I don't know, is what machine will work, what won't or how to operate any of them to get the data I need. At this point, I could probably get as far as plugging one in and finding the power button.

I have looked into the costs of new machines and pretty much eliminated that as a possibility. I have a very limited budget to work with.

So, I guess this is where my questions start. Is there older equipment that will give me the results I need and can be found for under, say a few grand? Is there a specific model I can look for? And is it even realistic to find training for older equipment in that price range?

[Vlad Orlovsky]

If you want we can look at this for your free of charge. contact me to discuss the ways to do this. Here is Free method Development Submission form:
http://www.sielc.com/Services_MethodDev ... tForm.html

[mattmullaney]

Hi Cindy99,

Looks like there are a number of possibilities on ScienceDirect (Elseveir publisher) web site for both HPLC and GC separations for the cannabinoids...my favorite one after a quick look is this reference...this will give you a start with a separation, at least. Don't know what you have in your lab...HPLC/DAD (or HPLC/PDA, depending on the vendor of the instrument), if you don't have one already, unfortunately will cost far more than a few thousand dollars...Good Luck in your work!

http://www.sciencedirect.com/science/ar ... 3209007545

Journal of Chromatography B, Volume 877, Issue 32, 15 December 2009, Pages 4115–4124. Innovative development and validation of an HPLC/DAD method for the qualitative and quantitative determination of major cannabinoids in cannabis plant material, Benjamin De Backera, Corresponding author contact information, E-mail the corresponding author, Benjamin Debrusb, Pierre Lebrunb, Laetitia Theunisa, Nathalie Duboisa, Lies Decockc, Alain Verstraetec, Philippe Hubertb, Corinne Charliera.

Abstract

GC is commonly used for the analysis of cannabis samples, e.g. in forensic chemistry. However, as this method is based on heating of the sample, acidic forms of cannabinoids are decarboxylated into their neutral counterparts. Conversely, HPLC permits the determination of the original composition of plant cannabinoids by direct analysis. Several HPLC methods have been described in the literature, but most of them failed to separate efficiently all the cannabinoids or were not validated according to general guidelines. By use of an innovative methodology for modelling chromatographic responses, a simple and accurate HPLC/DAD method was developed for the quantification of major neutral and acidic cannabinoids present in cannabis plant material: Δ9-tetrahydrocannabinol (THC), THC acid (THCA), cannabidiol (CBD), CBD acid (CBDA), cannabigerol (CBG), CBG acid (CBGA) and cannabinol (CBN). Δ8-Tetrahydrocannabinol (Δ8-THC) was determined qualitatively. Following the practice of design of experiments, predictive multilinear models were developed and used in order to find optimal chromatographic analytical conditions. The method was validated following an approach using accuracy profiles based on β-expectation tolerance intervals for the total error measurement, and assessing the measurements uncertainty. This analytical method can be used for diverse applications, e.g. plant phenotype determination, evaluation of psychoactive potency and control of material quality.

[sepscientologist]

I don't know where you are geographically but I would suggest you find your local college and find
an professor of organic chemistry. They could tell you about TLC or you can read about it online.
You might be able to use TLC (thin layer chromatography) to test for cannabinoids. Cost is under
$100 but validation (verifying that it works) might be a one time cost that is higher.

[mattmullaney]

I am a dunce...sepscientologist makes a good point, and I re-read the first post...here's a link to a TLC method that separates all of your analytes of choice, so long that they are cannabinoids. See what you think, and best of luck cindy99.

http://www.bing.com/search?q=tlc+of+can ... &FORM=AWRE

Thanks for waking me up, sepscientologist!

Oh, here is a link to a TLC-test kit, too:

http://www.alpha-cat.org/cannabinoids-analysis

[tom jupille]

I'll chime in here: TLC is great for identification, not so hot for quantitation unless you have appropriate equipment -- which makes just about as complicated/expensive as HPLC.

Is there older equipment that will give me the results I need and can be found for under, say a few grand?

Maybe, if you were experienced and willing/able to troubleshoot and upgrade. Given what you've told us about your background and skills, I'd say "no way".

Is there a specific model I can look for?

For the cannabinoids, almost any reasonably current HPLC system from a major vendor (e.g., Agilent, Waters, Thermo, Shimadzu -- in no particular order). For the pesticides, you're looking at LC-MS or LC-MS/MS, and if by "several" you mean "several tens", then you can find something.

And is it even realistic to find training for older equipment in that price range?

Probably not at an affordable price, unless you find a local university prof or grad student to moonlight. And, in that case, you might be better off simply paying them to do the analyses for you. Even if you have to hire someone to drive samples a couple of hours each way, that would still come out cheaper than doing it yourself.

[Vlad Orlovsky]

if you desperate enough you can assemble HPLC system under $500, buy solvents in Walgreens or Walmart (Alcohols, distilled water, acids), you jus need to be creative and have some knowledge of chromatography.
Something like this will work:
http://www.ebay.com/itm/Millipore-Water ... 565659a8e5 (pump)
http://www.ebay.com/itm/NEW-Waters-Atla ... 4d0c5f9b34 (HPLC column)
http://www.ebay.com/itm/ABI-Kratos-Spec ... 1e7ac12812 (detector)
http://www.ebay.com/itm/Pharmacia-LKB-R ... 3cd14b2ed5 (chart recorder)
TOTAL $370 (plus shipping of these items)

When we started SIELC 10 years ago our first system was under $500 and we used 3 of them for QC of the columns for a few months. After that we spent 2500-3000 to built current QC systems. Our QC is still using this inexpensive equipment and it works like a clock.

[cindy99]

Thanks so much for the tips! I am going to have to continue using the lab I have been for now, but will be investigating all of this much further and taking up chromatography as a new hobby. I am in Tucson, AZ and very close to the University of Arizona. I think I'll start looking for someone in the area to hire as a consultant. Something like what Vlad is talking about is what I would REALLY like to start with.

[Vlad Orlovsky]

I have one of our customers/friend in University of Arizona. Let me know if you need a name and email (send inquiry through our website:
http://www.sielc.com/AboutUs_ContactUs.html

[cindy99]

Awesome! I will do that, thank you!

[Vlad Orlovsky]

sent you email it may end up in spam folder

[HippyLabRat]

Hi Cindy, I may be able to help you. Where are you located?

[ROC]

Cindy 99, glad you posted this. I'm in a similar situation and don't want to pull the trigger on something foolishly. I have been pointed strongly toward HPLC over TLC and GC, this coming from a couple of chemist friends.

Wondering if some of the guru's in this community can point me in a good direction as far as detectors go. Currently I'm looking at VWD vs. DAD with an 1100. Any advice out there on which would be better for the consistent detection of cannabinoids quantitatively?

I have a chemist consultant (friend) who will be teaching my team how to use, maintain, calibrate, etc. Looking for accuracy above ease of use.

Thanks in advance for any input.

[Kuzdzal]

Hi Cindy,

There are many, many labs that perform cannabis testing. For a good testing directory in your state, check:
http://mjbizdaily.com/industry-director ... -services/

Also, turn-key LCs with methods are available from several manufacturers. See:
http://www.shimadzu.com/an/journal/cont ... 09-113.pdf

Best,

Scott

[Shralping]

If you are testing raw or recently room temp dried flowers rather than concentrates and edibles (that you don't know if they are aged or de-carboxylated) you could go with GC and get a test or two done in a lab on the same sample material via HPLC so you can see a trend of what to expect in way of acid forms of compounds under the same post harvest methods. For a new system in similar class, HPLC might cost 15 K more than a new GC. For HPLC keep in mind that you will need a suitable program to display the data and probably run the equipment/controller.
Its easy to buy the unit components off ebay or something and discover you cant get software with key and need to fork out thousands more. Lets say 2.5 -3K USA.
You can find an old GC with integrator or even analogue to digital converter for cheap. Keep in mind that there is not a guarantee it will work for those prices. You will also want reference standards to run into you machine to create a calibration curve and then to quantify the peaks you are seeing from your samples and see what time they are eluting from your column so you know what compound you are looking at.
This all comes down to how many samples you want to run as to what pathway you should take. If you want to go with GC the HP 5890 are a good choice.