DIY Thin Layer Chromatography (TLC) of cannabinoids at home - tutorial

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ryanrayla

Member
I did try ordering from one supplier (Santa Cruz Biotechnology) and they emailed saying they couldn’t ship anything to a residential address. I’ve since checked with about 10 different companies and everyone advertises that they won’t ship to residential addresses or that they “inform law enforcement of any unusual purchases.” To my knowledge it’s not illegal to posses these chemicals, but I don’t want to work with any company that threatens its customers before even ordering.

I have been using the tiny sample of Fast Blue BB that came with a test kit I bought from cannalyticssupply.com. They are selling a replacement vial that holds 150mg of Fast Blue BB for $15, which I’ve since reluctantly purchased. That’s $0.10/mg from the test kit vendor versus the $0.005/mg from Santa Barbara Biotechnology. I’m still looking for a larger quantity to lower the per unit cost.
 
PhenoMenal

PhenoMenal

Well-Known Member
You're correct, it's not illegal to possess Fast Blue BB (it is not a drug, it is not a precursor, it is not an explosive, it is not a controlled substance, etc etc, it's essentially just a dye). When you email labs, mention that you want to use the Fast Blue BB "specifically for the visualisation of phenolic compounds in strawberries". [Google: "Fast Blue BB" strawberries]

ps. if you got a test kit, it's more than likely Fast Blue B (not BB) that you received, as B doesn't require being kept in the freezer like BB does (you should receive BB packaged with dry ice). I've never personally used B myself, but I do love the extra vibrancy of BB, and BB was created due to safety concerns about B, so definitely go for BB if you can ... B still makes a very good second option though!
 
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ryanrayla

Member
Thank you for the reply, @PhenoMenal. I ended up purchasing the 150mg of unrefrigerated "visualization dye" just to keep moving forward.

In almost every test I've ever done, I've had a hard time getting the spots to be round enough to accurately measure using some rulers I purchased that are calibrated for that purpose. I've attached two plates below as an example. Lane 1 on both plates was created from the same sample which was plated and developed under the same conditions 9 days apart. The THC spot on the first plate, resolved beautifully while the spot on the second plate was vertically compressed. I've seen this phenomenon on nearly every plate I've developed but haven't been able to understand why.

Eluent: 100% Chloroform
Solvent: 100% Hexane
Plate: Silica on Glass
Plated with: 2ul Micropipette.

Do you have any thoughts as to what might be causing the vertical compression in my spots?


TLC-1017.jpgTLC-1018.jpg
 
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cdsmith12

Member
@PhenoMenal So when Missouri passed medical marijuana and allowed patients to home grow, I built a grow room and started growing, mainly to make RSO/FECO because I have severe chronic back pain, and narcolepsy. Due to the narcolepsy I needed to grow low THC strains so I grew ACDC and Harlequin from regular seeds and had 4 females of each. When I started making FECO, I noticed that it really wasn't helping as much as the Hemp FECO I had been getting so I researched testing and decided to purchase a TLC kit from thctestkits. After being intrigued by the process, but scared to test too many due to the cost of the refill kits, I started doing some research on buying my own supplies and found your thread. Excellent information by the way.

After reading the entire thread, I realized that my kit, as well as Grow Buddy/Montana Bio Tech and Alpha Cat are all about the same. All of them have one bottle of liquid for both the eluent and solvent, and of course no label, but I am assuming its hexane. In one of your earlier post you mentioned that you were going to try hexane without the diethyl either for both, but unless I missed it, I didn't see the result. I was wondering if you ever tried this? Or do you know what they are using that would allow one chemical for both? The plate on the left was soaked in the extraction tubes for about 15 mins in accordance with their instructions for flower. The plate on the right is the same extraction tubes that I tested again the next morning. I screwed up and didn't let it dry before spraying the dye so the result is messy, but the THC and CBD are higher. On each plate, the two on the left are indica strains and the two on the right are Harlequin and ACDC.
 

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PhenoMenal

PhenoMenal

Well-Known Member
All of them have one bottle of liquid for both the eluent and solvent, and of course no label, but I am assuming its hexane.
Click to expand...
There are several variations of chemicals that people can use for TLC of cannabinoids at home, and as I've experienced and documented they all produce unique results.

When "reverse engineering" this TLC process it was helpful that they provided MSDS safety data sheets, which showed which chemicals/substances they were using.

I've already documented everything I've tried (mostly on the icmag TLC thread, as I was there while I was learning the process), so I don't want to waste time rehashing anything, so please check that, and the links in my signature

ps. You're getting some good results there, clearly showing your Harlequin and ACDC have high levels of CBD :) (and your indica obviously showing heavy levels of THC)

Your 1st and 2nd lanes are just typical beautiful heavy-THC indica bombs :)

Your 3rd lane suggests an approx 1:1 THC:CBD ratio, which I know my bro-in-law found to be a very helpful ratio during chemotherapy.

Your 4th lane suggests a lot more CBD than THC!, which might be preferable to people who want medicine with minimal 'stone effect'

Beautiful plates bro :)
 
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PhenoMenal

PhenoMenal

Well-Known Member
ryanrayla said:
Do you have any thoughts as to what might be causing the vertical compression in my spots?
Click to expand...
I'm not quite sure what you mean by 'compression', but if you mean it's in regards to the outer lanes having weirder effects than the middle lanes, yes that's the effect I got from using Chloroform rather than Hexane+Diethyl
 
PhenoMenal

PhenoMenal

Well-Known Member
I just wanted to say CONGRATULATIONS to all the people in this thread who've "taken the plunge" by taking the time and expense to learn Thin Layer Chromatography ... TLC is literally THE MOST POWERFUL ANALYSIS TOOL available to home growers, and it gives me warm fuzzies of people who've taken advantage of this tool, and to see people posting their TLC plates with clearly distinguishable CBD and THC etc, and seeing their reactions now that they have this new power or ability, it's beautiful :)

ps. Don't forget, if you use TLC you now have the ability to detect the cannabinoid profile of plants when they're only a few weeks old (see my signature - "Find a CBD mother in 3 weeks from seed")
 
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cdsmith12

Member
@PhenoMenal I am getting ready to order my own stuff instead of getting a refill from the company I got my kit from and I have read tons of post about Fast BB. I'm not sure if I'm allowed to post a link so I will describe what I'm seeing on Sigma. When I type Fast Blue BB it comes up with several, but under the CAS number you have given as reference it has two. One says "Dye Content >=80%" which is $66.40 for 5g, the other says "microscopy (Hist)", for $27.50 for 10g, but this one also says for Cannabis Analysis in the General Description. Is this the one to get, the price is what made me question it because I've seen many other members post they paid $65 and up for 5g. Thanks for all your doing, and I did read the other post all the way through that you referenced in my past post.
 
PhenoMenal

PhenoMenal

Well-Known Member
I can't really blindly recommend one or the other, but if the CAS number is correct then it should be good. 5gms goes a long way
 
PhenoMenal

PhenoMenal

Well-Known Member
DrTricoma said:
All this strains are from my garden, quite interesting to see how the cannabidols evolve with the decarboxilation time and temp
Click to expand...
Good luck with your decarboxylation/time tests!
Here are some of mine I prepared earlier lol ... I only tried at 125C/257F and 135C/275F though (which is about as low as my oven can go)
Two different strains (both with interesting cannabinoid profiles), both tested in two different eluents (you can see both eluents have their own unique pros and cons) ...







 
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Hlusaf

Hlusaf

Well-Known Member
PhenoMenal said:
Good luck with your decarboxylation/time tests!
Here are some of mine I prepared earlier lol ... I only tried at 125C/257F and 135C/275F though (which is about as low as my oven can go)
Two different strains (both with interesting cannabinoid profiles), both tested in two different eluents (you can see both eluents have their own unique pros and cons) ...







Click to expand...
Thank you for the outstanding walkthrough, I'm extremely grateful and eager to test some plants. This is one of the best threads I've seen! If you ever need or want anything, please let me know. Cheers.
 
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Broos

New Member
I I am very new to this and have read through the thread, and it is very helpful. I am having a very hard time getting a strong reading from coconut oil. An even harder time with alcohol tincture.
Does anyone have any specific directions, inclusing amounts of oil and reaction fluid?

The third and fourth lanes are the oil. I did a hot test on the first two lanes (flower) and no heat on the other two as they were already decarboxylated.

Oil was 0.1ml and solvent was 1ml
 

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Robin_hearts_Science

New Member
Really excellent tutorial! Thank you for taking the time to experiment and to share your results.

As a chemist, I'd like to add a couple small (hopefully helpful) suggestions.
1) You can purchase a box of glass pasteur pipettes (available on amazon or other sources) instead of using the micropipette (which is pricey to buy). Just dip the glass end of the pasteur pipette in your cannabis solution and dot it onto your TLC plate (just as you suggested using a metal pin head for).
2) Add your solvent to your TLC jar early and close the lid. Letting the solvent sit in the jar allows it to establish equilibrium with the evaporated vapors in the jar and will give you better/more precise capillary results.
3) You can draw a circle around the separated cannabinoids on your TLC plate and label them to review later if the dye is fading.

There is a lot more information about CBD and THC available now than a few years ago. CBD is present in small amounts in the plant but the majority of it is in an acid form called CBDa while it is in the plant. Heat is required to convert the CBDa to CBD (a process known as decarboxylation or decarb). It might give more accurate results of CBD content if you decarb your plant material prior to extraction for your TLC analysis. (Note that the same decarb process is necessary for THC as well.)
 
DancesWithWeeds

DancesWithWeeds

Well-Known Member
Hay, PhenoMenal,

I've been following you for about 6 or 8 months now and really have learned a lot from you.

I was just messing around in my lab and came up with something you might be interested in. I am looking for easier and less expensive ways to visulize the TLC plates. This didn't work on the TLC plates (f254) but I tried it on a filter paper and got unexpected results.

This was just a time passer while I wait on chemicals and no care was taken (on this plate). The paper was spotted with some old oil and put under a UV light after running. Not much happened. I soaked the paper in UV ink, the kind they use to stamp your hand. A little more showed up. THEN, about a half cup of water and about 15 drops of green food coloring and I got what you see in the picture. I think it was about half/half hexane and acetone.

Later experiments showed that the UV ink has to be put on after the plate has been run. After reading what you can with UV dip in in the green dye and the colors seem to come out fairly nice. Another thing, shin the UV light thru from the back and it makes it easier to read.

This may be nothing, but it's worth looking at.
20210528_155837.jpg20210528_163846.jpg20210528_160234.jpg20210528_155837.jpg

In about a week or so I'll try it again with the right solvents and a lot of TenderLovingCare.

DancesWithWeeds
 
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