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Dinafem's Dinamed CBD thin layer chromatography

Discussion in 'Medicating' started by PhenoMenal, Nov 6, 2017.

  1.  
    PhenoMenal

    PhenoMenal Well-Known Member

    Hello, im helping a friend with terminal cancer with his grow. Even with terminal cancer he still won't let me grow more than 2 plants at a time (current limit here for "personal/non-dealer grow") which is frustrating, but it is what it is and he has a family and house etc.

    Anyway I got a dozen different CBD strains to try, Dinamed CBD by Dinafem is the first of the CBD ones we've tried. I also grew it alongside a Cinderella XX (Grimm Bros official feminised version of Cinderella 99).

    I did Beam's Test (Wiki) for the first time a couple days ago (5gm potassium hydroxide dissolved in 95mls of ethanol -- i used cheap methylated spirits which is 95% ethanol and it worked fine, and you can also use that "Everclear" stuff, and possibly others but I'm not a chemist and you don't want to mix the wrong stuff!) and that produced a very solid result for the Dinamed, and basically no reaction for the C99 - both results as to be expected.

    Beam's Test is very simple, quick, affordable, and no issues sourcing the two easy-to-buy materials, and after trying it myself I have to thoroughly recommend it to anyone who simply needs to know "does this bud have a significant amount of CBD?" it's basically a Yes/No test -- it turns violet/purple to CBD, but does not react at all to THC. Super handy!

    Unfortunately though, that's literally all Beam's Test tells you. A few months ago I spent literally two and a half months asking around trying to figure out how to do Thin Layer Chromatography (TLC) - you can buy kits like AlphaCat etc but they're quite expensive given how very few tests they give you, but by reading their MSDS and manuals etc I was able to figure out what they were using, and with the help of the one person I could find who'd done it before was able to figure out the puzzle.

    Beam's Test gave me confidence that I'd get a good result from the TLC, and sure enough the next day I was able to confirm that my Dinamed CBD is a 1:1 CBD:THC, and with very healthy levels of both, as well as strong levels of CBG and CBC (both those two in the C99 also!). I've tested about 7 strains using TLC before and only seen healthy CBG levels in 1 before, so it was a pleasant surprise to see it again - in both strains!

    It's important to note that Dinamed CBD is supposed to be a SATIVA-dominant thin-leaf plant which is advertised as stable at around 20% CBD to 1% THC. I got a very-much INDICA-dominant phenotype (made even more obvious when growing alongside the very-sativa-leafed C99!), so clearly I did not get "the main Dinamed CBD" ... disappointing in one respect that this pheno doesnt offer the flexibility of a 20:1, but at the same time it's probably also a relief to know that even with this other pheno it's still packing a healthy amount of CBD, so to me that's a good sign in regards to "the main sativa pheno". Unfortunately this indica pheno didn't yield very well though - about 2oz, compared to the 4oz from the C99. Smells and tastes really nice, seems like a complex terp profile, and reminded me of a "perfume flavor" that I hadn't experienced since trying a strain called Sherry a decade ago. Dinamed is a descendent from the Dancehall strain.

    Anyway here's the TLC result. The Dinamed was at 62 days and harvested, while the C99 was only at 48 days (snipped off a sample) and am not expecting to harvest that until ~58, so I'm guessing the C99 is still packing on a bit more THC:
    [​IMG]

    It's important to note that I'm an amateur and don't work in science/chemistry, and don't use precise measurements, so you cannot compare one lane to another, but you can compare all the dots in any ONE lane, as, being not a quantitative but qualitative test, the RATIOS are fairly reliable and don't discriminate. You can easily see for example this is very close to a 1:1 ratio, probably something like 12% THC 12% CBD give or take 5%, but I think it's pointless trying to use TLC to get accurate percentages. Really need a lab with proper HPLC or GC equipment etc for that (and then you can get terpene profile too!), but you can still see how useful TLC is for any home user needing to know the RATIO of CBD:THC in their sample. :) I think it's a fantastic tool, especially in combination with Beam's Test which you can use to help determine if you even want to do a TLC - for example if hunting for CBD you might only want to do a TLC if Beam's Test comes up positive first.
     
    Last edited: Nov 6, 2017
    Observe & Report likes this.
  2.  
    freemanjack

    freemanjack Well-Known Member

    Top post m8, I've been looking for a basic 'kitchen sink' test for ages. Made all the more annoying by having not troubled my lad coz his degree/phd lab work took all his time, he published his thesis this month and has decided 8 yrs in the lab is quite enough and he's looking to work in bloody marketing of all things!! bah! I'll email him a copy of your post, see if he can advise on ways of improving the reliability of the results or perhaps suggest a better type of test.
     
    vostok likes this.
  3.  
    PhenoMenal

    PhenoMenal Well-Known Member

    thanks mate :) Would love to hear if your lad has some more suggestions! there just aren't many options at the moment :(

    I've been looking for a basic 'kitchen sink' test for ages.​

    Beam's Test really is awesome as a "kitchen sink test" :) I've only used it once as mentioned above, but was very impressed by it... it made it very obvious to me that one sample had CBD and the other didn't, so I went into the TLC the next day with a feeling of confidence that the TLC would back that up. Sure enough it did. And it's cheap and quick and only need 2 easy-to-buy things, so if you're hunting CBD it's a fantastic tool! And when you've made the solution you can keep it on the shelf indefinitely, in other words you can do CBD tests immediately - just pour a tiny amount in a shotglass and drop your bud sample in. Wait a few minutes. Simple as that.

    But TLC also is a "kitchen sink test"!!! it's really quite simple - many primary/elementary schools introduce TLC to young students by using the ink of a black-coloured pen as the sample and water as the mobile-phase eluent ([example] "TLC for kids"), eg:
    [​IMG]

    TLC for cannabinoids is only slightly more complicated... you can't use water ... (out of curiosity I TRIED! Lol - this was with full spectrum CBD Paste as the sample):
    [​IMG]

    ... so for cannabinoid TLC you need:
    1) a good solvent to first extract the cannabinoids into a liquid suspension, i just use Hexane, but I'm tempted to try Chloroform next time alongside also to see what (if any) diff it'll make
    2) a good solvent-based mobile-phase (i used Chloroform in one, and Hexane: Diethyl Ether 4:1 in other ... please don't ask me about polarities haha)
    3) and finally to then visualise the result you need a special dye (the ink pen example above brings with it its own visualiser!), which for cannabinoids is either Fast Blue B or Fast Blue BB (which needs to be kept in the freezer). Try to get BB instead of B if you can (although it does cost more): 1) BB was created due to concerns about B being carcinogenic... there are concerns about BB too though, and 2) BB produces more vibrant results than B.
    4) also some more TLC-related lab equipment such as micro-pipette, eppendorf tubes, nothing major or expensive though

    And, you need to observe all safety precautions - the dye is carcinogenic, and my nostrils are no longer virgins after using Diethyl Ether, insane fumes even when well-ventilated! Chloroform is far gentler on the snout in that respect, but there are only a couple times when you need to hold your breath for 10 secs so it's really not a problem.

    But they're basically the only twists on basic TLC, so I encourage people to give TLC a go if they need more than Beam's "Yes/No" about the CBD in their buds and might not have access to, or don't want the expense of, or don't need a professional lab analysis.

    And of course feel free to buy the commercial TLC cannabinoid kits if you like - if you only want to do a few samples it might even work out better for your budget and you don't have to source the dozen or so various parts needed as theyre all included. But I think if you're planning to do more than half a years worth of testing you should just buy the raw materials yourself.

    btw you can read my 2.5-month struggle to learn how to do TLC with cannabinoids [here] lol, username sadpanda (hey i was desperate for help), in which you can see my various other experimental results with TLC including high-THVC/CBG strains etc - it's a beautiful tool.

    For me, as somebody hunting a natural source of CBD while somebody in my family fights terminal cancer, and they've had such AMAZING positive responses to THC and CBD, both Beam's Test and TLC are LIFESAVERS.♥
     
    Last edited: Nov 7, 2017
    freemanjack likes this.
  4.  
    vostok

    vostok Well-Known Member

    [​IMG]
    this ones popular back east but pricy (MINI Kit - 89 euros)

    • 1 alpha-CAT manual guide
    • 1 THC and 1 CBD calibration chart, standards that allow accurate % quantification
    • 2 Test plates
    • 2 Dye powder microtubes (0,06 gr), for storage in cool, dry, dark conditions
    • 1 Bottles of alpha-CAT test fluids (10 ml)
    • 8 Eppendorf tubes (1,5 ml)
    • 1 Developing jar
    • 1 Dipping tray
    • 1 Pipettes (3 ml)
    • 1 Syringe (1 ml)
    • 1 Vial with 10 capillary tubes (1 ul)
    • 4 Nitrile gloves
    • 1 Becher (25 ml)
    • 1 capillary pipette bulb
    (https://www.alpha-cat.org/shop/test-kits/mini-kit/)
     
  5.  
    PhenoMenal

    PhenoMenal Well-Known Member

    In other words they only give you 60mg of Fast Blue (B, not BB), and seemingly need 30mg per sample with that Fast Blue B ... BB is far more economic in that respect (only need a tiny, tiny amount - i use approximately 3 "pinheads" worth of BB per 25mL bath). Just buy it yourself and keep it in the freezer and protected from light and it'll last years.

    I'm not sure what their test fluid is, but you need to mix the Fast Blue with something like sodium hydroxide 1.0M to make a shallow solution you can lay the plate in for a few seconds, but 10ml would literally be only 1 bath worth, unless diluted. (i use 25ml in my bath, which is only just big enough to fit the plate with a good depth of 1-2mm liquid). Or perhaps the 'test fluid' is the mobile phase eluent? I'm not sure. And what do they provide to actually extract cannabinoids from the sample into a liquid suspension?

    And that list also shows they only provide "2 plates" in their kit ... I showed 2 plates worth in my first post!, but if you shop around you can buy 100 high-quality plates (eg Merck etc) for far less the cost of this kit. Likewise, "4 nitrile gloves" = only enough for 2 runs, but you can easily find boxes of nitrile gloves for cheap. MUST be nitrile btw, but they're still cheap so there's no excuse for not using them. "8 Eppendorf tubes", again ... you can buy hundreds very cheap, disposable plastic.

    In summary, their kit contains everything you need -- but only for 2 tests. Then they want you to buy 'refills' at hyperinflated prices.

    (These are the basic reasons I decided to try to figure out all the components for myself!) :)

    It might actually be a good starting point though in regards to buying all the lab equipment, but I don't know the quality of their equipment.

    But yes, the list shows that you do need quite a few items to properly do a TLC with cannabinoids, and why it took me 2 and a half fricken months to get everything figured out. Maybe i should post my procedure. Beam's Test on the other hand is very simple!

    I do take my hat off to the AlphaCat team though, if you check out the videos theyve released etc you can see they're a very professional French outfit, and without their groundwork I'm not sure I would've been able to do this myself. If you're leaning towards buying one of the commercial kits I would definitely go with AlphaCat, although I'm just saying that from what I've seen and my current understanding of the scene, as I haven't tried their kit personally. They're expensive yes, but seem the best quality out of what's available.
     
    Last edited: Nov 7, 2017
    freemanjack likes this.

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