How to make a cannabinoid analysis - Alchimia blog https://www.alchimiaweb.

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Probably, many of you wonder what kind of test is performed to determine if a marijuana strain has significant rates of the main cannabi
noids, as well as the method used to know the precentatge (more or less accurate, as we will see) of each cannabinoid contained in the
tested sample. In this post we will explain step by step how to do a thin-layer chromatography (TLC), an analisis that will show us
the approximate percentages of the main cannabinoids detected in each sample. To do this, we used eight different samples of the
marijuana strain Fruity Jack / Jack el Frutero from Philosopher Seeds, with the simple purpose, in this case, to know which of the ana-
lyzed phenotypes have high amounts of the cannabinoid CBD.

The medical use of marijuana is constantly gaining ground in the past few years, as well as the interest from breeders and consumers
to know the exact amounts of cannabinoids present in their favourite cannabis strains. Thats why expressions such as thin-layer chro-
matography or gas chromatography are becoming more and more familiar for the cannabis community. To learn more about these
analysis systems, we met with a good friend and collaborator who explained us a system based on the thin-layer chromatography with
which we get reliable results quickly and safely for our health.

TLC Plates

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We will need the following materials for the analysis process of the samples:

Plates for thin layer chromatography, which can be made of aluminum, glass or plastic, always coated with a solid and adsorbent
material (silica gel or alumina)
Solvent, we used Hexane for this post
Laboratory Micro-Pipettes
Distilled water
Reagent, in this case Fast Blue B
Eluent. We used Chloroform in this analysis
Blotting Paper, Eppendorf tubes, glass jar, Pyrex tray, pasteur pipette , graphite pencil

Before starting, we should know be aware of the basic security measures in laboratories when using chemicals, and of course follow
them. Also, this goes for any kind of experiment, you should try to do it in place as clean and sterile as possible, thus avoiding contami
nation that could alter or distort the results of the test.

Chloroform being slowly absorbed by the plate

For the preparation, you should mix 1ml of solvent (Hexane) for each 0.1 gr of cannabis sample in an Eppendorf tube. Shake for 20
seconds and let it rest a minimum of 3 minutes before continuing. During the whole process, our partner warns us of the importance of a
correct and cautious labeling of each sample, because otherwise its easy to get confused and mix up results in case you want to
make several analyses. Once the sample is prepared, collect 0.2 micro-liter with the micro-pipette and deposit them in the point that we
have previously marked with the pencil on the underside of the plate. For our analysis, we will use a plate for each four phenotypes of
Fruity Jack / Jack el Frutero to be tested, using a total of two plates per 8 samples. We put the eight micro-drops on the two plates
(4+4), so we already have our samples prepared for the next step.

Now, we must proceed to decarboxylate the samples. To do this, we place the two plates with the eight droplets in a Pyrex tray, which
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are is in contact with the eluent. Its important to observe how the eluent is abdorbed by the plate; in case it doesnt do it in a uniform
way, we probably should discard the results of this sample; this is often the case when old or damaged plates are used. When the chlo-
roform has rose up, it leaves a trace in the form of spots (still not visible) for each of the main cannabinoids. Later on, these spots will
indicate us the approximate percentages of each cannabinoid in each sample.

Place the plates on the blotting paper

Allow the eluent to completely cover the plate until it reaches the upper edge (about 30 minutes). Remove the plates carefully and
leave them 5 minutes on the blotting paper in vertical position, with the pencil marks on the lower part. Meanwhile, we msut prepare the
mixture with the reagent to make the spots visible. For this process there are different methods, from UV light to specific reagents that
allow us to colour the spots. We have chosen the most common option: pour 20 ml of distilled water in a plastic tray and add 0.1 g of
a reagent salts called Fast Blue B (C14H12N4O2Cl2 ZnCl2) . Shake the mixture until its even and all salts are dissolved in the
distilled water.

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Distilled water and reactive salts

Now all you have to do is soaking each plate in the tray for just two seconds (you also can spray the mixture over it, but its much better
for your health to submerge them, even if you use more liquid). Take it out and leave it again on the blotting paper, always in vertical
position and with the spots at the bottom. After a few seconds, a number of spots will appear along the plates, showing the presence
or absence of certain cannabinoids (THC, CBD, CBN, CBG, THCV, CBC), as well as its approximate content. Each cannabinoid has
its own colour and position on the plate, so its easy to know if a specific cannabinoid is present in the sample. Its content is determined
by measuring the diameter of the spot and comparing it with a conversion table that will give us an approximate content (%). We use
the table included in the cannabinoid test kit from Alpha-Cat.

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TLC Plate soaked in reactive salts

Despite this system is not as accurate as the gas chromatography, it seems a good method for an approximate determination of the
cannabinoid content present in a sample. In our case, for example, it has been very useful because the only thing we wanted was to
know which of the eight analyzed samples contained high CBD rates. For the happiness of many medicinal marijuana users, six of the
eight analyzed phenotypes have shown remarkable CBD contents. The system is especially useful in processes of selection and
breeding, since it allows us to know which phenotypes we want to keep with no need for costly gas chromatography analysis.

With this method, anyone can do its own analysis at home and know, for example, what phenotypes contain CBD or which of them
has higher THC content. The more you think about it, the more applications seem to have this type of analysis, especially for the selec-
tion of individuals in relation to their specific cannabinoid content. It is also useful to test different flowering times, to know the perfect
time to harvest marijuana plants, to test fertilisers or substrates, etc. You can perform test crops with clones and see which contents are
obtained in each sample, find a phenotype that has a certain balance between its different cannabinoids, etc.

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Fruity Jack / Jack el Frutero, 6 of 8 in CBD

The possibilities, as we say, seem abundant; although it should always be kept in mind that the results dont have the accuracy of gas
chromatography analysis. Either way, they can be useful to know which specific phenotypes we want to study more in-depth.

We wish you happy analysis and good vibes.

Rosin Hash Dab-a-Doo 2015 Synthetic cannabis

Barcelona and cannabinoids

August 3, 2015 | Medical marijuana 6 Comments

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Papa Indica Well, for myself, it sounds like a lot of extra work, not to mention finding all of that stuff. Its difficult for me
August 3, 2015 to keep up with all I do now, it seems like Im constantly busy with something to do with my grow. For me, I
either like what Im growing, or I dont. If I dont, I move on. But, for someone like my cousin, this may be a big help to him. His son has
Autism and the state approved his petition to use medical marijuana to treat his son, so he is constantly working at fine-tuning things for
those treatments. I know at least once hes brought samples of the stuff hes growing for his son to someone else to be analyzed, I imag-
ine hed like to have a way to do it himself. Ill be telling him about this post.
Thanks for the info!

Dani Alchimia Hi Papa Indica,

August 5, 2015

Thanks for telling your cousin about this method, it is much easier and cheaper than gas chromatography and, while not being as accu-
rate as the GC, it tells you what cannabinoids are found in your sample, with an approximate % content. Very useful for both breeders
and medical patients.

All the best!

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Zac Hey Im sorry but what would the chemical be thats the same color as CBD is but its popping up below
January 3, 2016 THC instead of above it ??? Theres acids then purple then light orange then dark red then random
marks I know the dark red is thc .. but all the pics and examples I see say cbd is above but CBG is yellow ? I dont get this I used can-
nalytics ?

Dani Alchimia Hi Zac,

January 10, 2016

Sorry for the late reply.

With this test, you should see cannabinoids in this order:

CBD
THC
THCV
CBG
CBC

THCV is pink, and CBG is orange.

Hope it helped!

Max Wendroff Where can I purchase the supplies?

April 9, 2016

Dani Alchimia Hi Max,

April 11, 2016

Here you can purchase the Alpha-Cat analysis kit, with which you have everything you need to analize your samples.

Thanks for your confidence.

Alchimiaweb S.L.

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