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

well, hopefully analysis will prove that it is what it says it is. When I was hunting for CBD only 2-3 years ago i bought five seemingly "reputable high-CBD" strains, yet most of them showed ZERO CBD, and only one of those strains resulted in consistent and good levels of CBD in every plant (i was growing approx 5 plants each from the 5 strains).

I asked the AI model ChatGPT "how do I do Thin Layer Chromatography on cannabis" ... yep, it knows how, but its first result was the alternative version that requires UV lights to detect the spots:

[ChatGPT response]
Thin Layer Chromatography (TLC) can be used to analyze the cannabinoid content in cannabis samples. Here are the steps to perform TLC on cannabis:

1. Obtain a TLC plate: You will need a silica gel TLC plate with a fluorescent indicator. Cut the plate into the desired size for your analysis.
2. Prepare the sample: Grind the cannabis sample to a fine powder and extract the cannabinoids using a suitable solvent, such as ethanol or methanol. Filter the extract to remove any plant material or impurities.
3. Prepare the TLC solvent: Prepare a solvent mixture that is appropriate for separating the cannabinoids. For example, a 9:1 mixture of n-hexane and ethyl acetate is commonly used. Add a few drops of this solvent to a clean TLC chamber to saturate the chamber with solvent vapor.
4. Apply the sample: Apply a small amount of the cannabis extract to the bottom of the TLC plate using a micro-pipette. It is important not to overload the plate, as this can lead to poor separation.
5. Develop the plate: Place the plate in the TLC chamber and allow it to develop until the solvent front reaches the top of the plate. This can take anywhere from a few minutes to an hour depending on the solvent system used.
6. Visualize the spots: Remove the plate from the chamber and dry it with a stream of air or a heat gun. To visualize the cannabinoid spots, expose the plate to UV light or spray it with a suitable reagent, such as a solution of vanillin in sulfuric acid.
7. Interpret the results: Compare the cannabinoid spots on the TLC plate with standards to determine the identity and quantity of the cannabinoids present in the sample.

It is important to note that TLC is a qualitative technique, meaning that it can be used to identify the presence of cannabinoids, but it cannot provide precise quantitative data. For accurate quantitative analysis, a more sophisticated analytical method such as high-performance liquid chromatography (HPLC) or gas chromatography (GC) is required.

So... i changed my prompt to "how do I do Thin Layer Chromatography on cannabis using "Fast Blue BB" " ... but sadly it still thinks we need a "silica gel TLC plate with a fluorescent indicator". No we do not.

[ChatGPT response]
Fast Blue BB is a dye that can be used as a visualization reagent in Thin Layer Chromatography (TLC) to identify the presence of cannabinoids in cannabis. Here are the steps to perform TLC on cannabis using Fast Blue BB:

1. Obtain a TLC plate: You will need a silica gel TLC plate with a fluorescent indicator. Cut the plate into the desired size for your analysis.
2. Prepare the sample: Grind the cannabis sample to a fine powder and extract the cannabinoids using a suitable solvent, such as ethanol or methanol. Filter the extract to remove any plant material or impurities.
3. Prepare the TLC solvent: Prepare a solvent mixture that is appropriate for separating the cannabinoids. For example, a 9:1 mixture of n-hexane and ethyl acetate is commonly used.
4. Apply the sample: Apply a small amount of the cannabis extract to the bottom of the TLC plate using a micro-pipette. It is important not to overload the plate, as this can lead to poor separation.
5. Develop the plate: Place the plate in the TLC chamber and allow it to develop until the solvent front reaches the top of the plate. This can take anywhere from a few minutes to an hour depending on the solvent system used.
6. Visualize the spots using Fast Blue BB: Once the TLC plate has been developed, remove it from the chamber and allow it to dry. Then, prepare a solution of Fast Blue BB by dissolving 0.1 g of the dye in 10 ml of ethanol. Spray the TLC plate with the Fast Blue BB solution, making sure to cover the entire plate. Wait a few seconds for the plate to absorb the solution, then rinse it briefly with water to remove excess dye. The cannabinoids on the plate will appear as blue spots.
7. Interpret the results: Compare the cannabinoid spots on the TLC plate with standards to determine the identity of the cannabinoids present in the sample.

It is important to note that TLC is a qualitative technique, meaning that it can be used to identify the presence of cannabinoids, but it cannot provide precise quantitative data. For accurate quantitative analysis, a more sophisticated analytical method such as high-performance liquid chromatography (HPLC) or gas chromatography (GC) is required.

So i changed the prompt to "how do I do Thin Layer Chromatography on cannabis using "Fast Blue BB" without any UV or ultraviolet " ... strangely enough it STILL recommends a TLC plate with a "fluorescent indicator". No, just get any normal silica TLC plate.

[ChatGPT response]
Fast Blue BB is a commonly used visualization reagent in Thin Layer Chromatography (TLC) for identifying the presence of cannabinoids in cannabis samples. However, if you do not have access to a UV or ultraviolet light source, you can still use Fast Blue BB to visualize the cannabinoids on the TLC plate. Here are the steps:

1. Obtain a TLC plate: You will need a silica gel TLC plate with a fluorescent indicator. Cut the plate into the desired size for your analysis.
2. Prepare the sample: Grind the cannabis sample to a fine powder and extract the cannabinoids using a suitable solvent, such as ethanol or methanol. Filter the extract to remove any plant material or impurities.
3. Prepare the TLC solvent: Prepare a solvent mixture that is appropriate for separating the cannabinoids. For example, a 9:1 mixture of n-hexane and ethyl acetate is commonly used.
4. Apply the sample: Apply a small amount of the cannabis extract to the bottom of the TLC plate using a micro-pipette. It is important not to overload the plate, as this can lead to poor separation.
5. Develop the plate: Place the plate in the TLC chamber and allow it to develop until the solvent front reaches the top of the plate. This can take anywhere from a few minutes to an hour depending on the solvent system used.
6. Visualize the spots using Fast Blue BB: Once the TLC plate has been developed, remove it from the chamber and allow it to dry. Then, prepare a solution of Fast Blue BB by dissolving 0.1 g of the dye in 10 ml of ethanol. Dip the TLC plate in the Fast Blue BB solution, making sure to cover the entire plate. Wait a few seconds for the plate to absorb the solution, then remove it from the solution and allow it to dry. The cannabinoids on the plate will appear as blue spots.
7. Interpret the results: Compare the cannabinoid spots on the TLC plate with standards to determine the identity of the cannabinoids present in the sample.

It is important to note that without UV or ultraviolet light, the visualization of the cannabinoid spots may not be as clear or intense as with UV light. Therefore, it is recommended to use a UV light if available for the most accurate and reliable results.

PLEASE NOTE that I was only probing ChatGPT here - I don't know how well your results will turn out if you strictly follow its instructions (they may be great, they may be bad, I don't know). However you WILL get great results if you simply follow the (more comprehensive) instructions in my tutorial that I posted at the start of this thread.

However, it's best to view my tutorial in the internet archive as it has all the images that have since been lost here: https://web.archive.org/web/20180406085710/https://rollitup.org/t/diy-thin-layer-chromatography-tlc-of-cannabinoids-at-home-tutorial.953423/

Thin Layer Chromatography is the best way for home growers to get an accurate picture of what cannabinoids your cannabis plant is composed of, and you can do it from home, using a very affordable setup, and it's quite simple to do.

In summary ChatGPT sucks nuts!

Anyone tried the kit from TLC lab supply?

ChemDogLover said:

Anyone tried the kit from TLC lab supply?

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Seems to be pretty much the same as any of the other kits, but it looks like their kits come with a Bic lighter lol, not quite sure what that's for.

They also come with those 'size' charts, but again TLC is a qualitative test - not quantitative. That is, it can tell you "there's no CBD" or "there's a significant amount of CBD" etc, but it can't tell you "there's 5.5% CBD".

Unfortunately they don't seem to provide any MSDS safety sheets so they don't specify which dye they use, but from their images it seems pretty clear that they use Fast Blue B. It seems all the kits use Fast Blue B simply because it transports easier (no dry ice required).

I much prefer Fast Blue BB - it's much more vibrant, results are clearer and easier to interpret, and a slightly better safety profile - BB was developed due to carcinogenic concerns about B.

You CAN simply buy any kit, and then buy some Fast Blue BB to use instead.

For example, here is TLC Lab Supply's sample image, clearly using Fast Blue B:



Here's a few of my images, using Fast Blue BB:




This strain has no CBD, but high in THC (the red spots), but also very significant levels of CBG (the orange spots):


This is a rare strain that shows high levels of THCV which you don't see too often (if I recall correctly it was South African Durban Poison):


The Fast Blue BB i got is made by Wako (FUJIFILM Wako Pure Chemical Corporation), its 5 gm (which is HEAPS in this context! but they also offered a 1 or 2 gram option, as well as 25 gram etc), it was very affordable - especially considering you only need a tiny amount with each sample, and simply labelled "Fast Blue BB Salt", and CAS number is "5486-84-0" (that's the important bit). I can't recall which country I ordered it from, but I had no problems getting it sent to Australia (it's not a controlled substance, it's perfectly legal - it's just a dye) and it was packed in dry ice (keep it stored in your freezer and it'll last for years).
Here's a google if you want to find some to buy: https://www.google.com/search?q=buy+"5486-84-0"

PhenoMenal said:

Seems to be pretty much the same as any of the other kits, but it looks like their kits come with a Bic lighter lol, not quite sure what that's for.

They also come with those 'size' charts, but again TLC is a qualitative test - not quantitative. That is, it can tell you "there's no CBD" or "there's a significant amount of CBD" etc, but it can't tell you "there's 5.5% CBD".

Unfortunately they don't seem to provide any MSDS safety sheets so they don't specify which dye they use, but from their images it seems pretty clear that they use Fast Blue B. It seems all the kits use Fast Blue B simply because it transports easier (no dry ice required).

I much prefer Fast Blue BB - it's much more vibrant, results are clearer and easier to interpret, and a slightly better safety profile - BB was developed due to carcinogenic concerns about B.

You CAN simply buy any kit, and then buy some Fast Blue BB to use instead.

For example, here is TLC Lab Supply's sample image, clearly using Fast Blue B:

Here's a few of my images, using Fast Blue BB:

View attachment 5346489

This strain has no CBD, but high in THC (the red spots), but also very significant levels of CBG (the orange spots):
View attachment 5346490


The Fast Blue BB i got is made by Wako (FUJIFILM Wako Pure Chemical Corporation), its 5 gm (which is HEAPS in this context! but they also offered a 1 or 2 gram option, as well as 25 gram etc), it was very affordable - especially considering you only need a tiny amount with each sample, its number thingy is "062-05463", and simply labelled "Fast Blue BB Salt", and CAS number is "5486-84-0" (that's the important bit). I can't recall which country I ordered it from, but I had no problems getting it sent to Australia (it's not a controlled substance, it's perfectly legal - it's just a dye) and it was packed in dry ice (keep it stored in your freezer and it'll last for years). Here's a google if you want to find some to buy: https://www.google.com/search?q=buy+"5486-84-0"

Click to expand...

Very interesting, thank you for the reply

PhenoMenal said:

Im confused! how have you run 100 TLC's without Fast Blue B/BB?

btw you don't need to necessarily find a LOCAL lab supplier, just one able to order it in for you in your country. There are plenty of such labs in America, so you just need to keep emailing.

Click to expand...

PhenoMenal said:

Im confused! how have you run 100 TLC's without Fast Blue B/BB?

btw you don't need to necessarily find a LOCAL lab supplier, just one able to order it in for you in your country. There are plenty of such labs in America, so you just need to keep emailing.

Click to expand...

Retired from MilliPore Sigma and still can't find a company that will sell to a someone not in academics or a company. Should have taken some fast blue before reitirement. Give a few names of suppliers willing to sell to us.

PhenoMenal said:

well, hopefully analysis will prove that it is what it says it is. When I was hunting for CBD only 2-3 years ago i bought five seemingly "reputable high-CBD" strains, yet most of them showed ZERO CBD, and only one of those strains resulted in consistent and good levels of CBD in every plant (i was growing approx 5 plants each from the 5 strains).

Click to expand...

Thank you for sharing-so much to learn. I have saved your posts from 2017. I did my first TLC test several weeks ago, as a test, now I am ready to try again…thanks again