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Kava Science Lebot explains simple qualitative type test in lab

Discussion in 'Kava Lounge' started by Andrew Procyk, Sep 25, 2013.

  1. Andrew Procyk

    Andrew Procyk Noble Kava Kava Vendor

    Ideally, soon we will know what causes the different colors, and be able to easily determine noble-non vs. noble strains at home! Here's the scoop...

    Crunked, violet, LocalBliss and 3 others like this.
  2. Kapmcrunk

    Kapmcrunk The Kaptain of Crunk KavaForums Founder

    This is great!! I eagerly look forward to the rest of your Lebot videos. Talk about coming from the guy who wrote the book on kava...he did.

    Edit: Also moved this over to the research and studies forum. I believe if anything qualifies for this, these kinds of post do.
    Last edited: Sep 25, 2013
  3. Both Lebot videos were outstanding. I'm a little leary that these videos were submitted by a vendor of noble Vanatua Kava but I think his heart is in the right place. So what about Fiji, Tongan, Hawaiian Kava ... are they noble or not? Anybody want to stick some acetone in their Koniak and give us the result. Somebody who is familiar with the Scientific Method (Infraredz?). Hey Judd, you wanna put some Acetone in your Fiji and give us the result before I order? So many questions ....
  4. kavadude

    kavadude ❦ॐ tanuki tamer

    Nice...so who's going to sacrifice some kava and nail polish remover
  5. I read the studies though and through
    and I composed this song for you.
    So don't worry, drink Kava.

    You drink your Koniak night and day,
    Because you likes some nice Tudie,
    So don't worry, be Krunky.

    The Koniak is no good for you,
    You better switch to Borogu
    So don't worry, be happy now.

    The mean Lebot has burst your bubble
    I'd switch to Fuu on the double,
    So Don't worry, drink Kava.

    The Noble Kava is good for you,
    I have some her to sell to you.
    So Don't worry, be happy.

    You made Kava with Acetone
    Your silly neighbor drank it down.
    Don't worry, be grateful.

    The Scaly Zombies are attacking
    With yellow eyes and teeth a clacking,
    Don't worry, start running.
    violet, KavaGurl, infraredz and 2 others like this.
  6. infraredz

    infraredz BULA!

    Alright, so I went for it (except for the nail polish remover, I used straight acetone).

    I figured we all know Boroguru is a common noble kava. I'm calling this the control. "Koniak" as shown by the following report was reported to be tudei based on qualitative analysis. Based on the above technique, I decided to determine if BKH's Koniak is a tudei (based on the standard in the video [dark color]) and if Solomon's Island by Nakamal at Home is Piper wichmanii (based on the standard in the video [very dark color]) as some have theorized.

    Not knowing the process Lebot used, I unknowingly seemed to actually (almost) replicate the procedure. I was later informed that this test is generally performed with 10g of powder, and 30ml of acetone.

    My procedure:
    25mL of acetone was added to 10mL of each kava and agitated for 4 minutes then left to settle (as I don't have a centrifuge like Lebot seemed to use).
    (Side note: mL is a unit for liquid, not solid matter but it gives an easy "guide" as to how much kava you should use)

    Attached Files:

    Last edited: May 26, 2014
    Crunked, Deleted User and kavadude like this.
  7. infraredz

    infraredz BULA!

    It's important to note that the mechanism behind the color change has not be discovered.

    Also, Lebot stated that this sort of test could be conducted with ethanol, chloroform and acetone which all have different polarities. It would seem that the solvent used could have an impact on the result depending on what compounds are causing the color change (and consequently, would a non-polar solvent [eg. chloroform] differ from an a polar protic [eg. ethanol] or polar aprotic [eg. acetone] solvent).
    Last edited: Sep 25, 2013
  8. If you did the acetone test, then what color was the mixture on both (per the example on the video)?
  9. infraredz

    infraredz BULA!

    The suspension hasn't completely separated yet and I don't have a centrifuge like Lebot.
  10. Ok, thanks. Just let us know when the egg finally hatches.
  11. infraredz

    infraredz BULA!

    @Andrew Procyk Do you know which solvent Lebot used in the video?
  12. Andrew Procyk

    Andrew Procyk Noble Kava Kava Vendor

    So, you don't want to trust a fellow who was a university professor before he got into the kava business, and who is still (obviously) working with other researchers, because he also sells kava? How then, can you trust anything at all from anyone at all who sells anything? (Nice song by the way!) ;-)

    In all honesty: I can indeed see how your concern has some validity, but my intent in posting the qualitative type test for the Koniak was not to take issue with another vendor. I have spoken to Judd many times, and respect the model that he has applied to kava bars. Rather, my intent was to make people aware of the "controversy" surrounding the consumption of high-FKB kavas, and prevent any averse issues that could hurt all of us. I may have mentioned, there was just recently an injury report related to kava filed with the FDA. This is not good for ANYONE in the business, no matter what they sell. I know that Judd tries to sell the best kavas he can get, and everything he gets from Vanuatu is noble, even though testing for it is not technically required.

    As per your concerns because of my selling Vanuatu noble strains - Couldn't this test, when perfected, be used on any kava from anywhere? Or is this test capable of being performed only on kava from Vanuatu? Anyone can benefit from this, and it is a great and easy way to verify strains, and might actually function to protect the industry as a whole. That is where I was going with it. Not to mention, with the amount of online sales I have done in the past several months (when my web page has been down for the past several months) is completely negligible. Frankly, in the interest of disclosure, I have sent people to Judd's website before - even customers at my kava bar for items I did not carry or did not have in stock!

    Frankly, I work with Vanuatu because I know people there very well, both residents and Peace Corps volunteers that are/were serving there, have extremely close connections to villages, and more than anything - their economy needs capital infusions much more than Fiji, who gets tons of surfer-tourists, backpackers and honeymooners to throw their dollars around. (Or Hawaii who is part of a developed nation you may have heard of.) I am sure some people are in things for a quick buck, but I doubt anyone who bothers to spend time on here and cites their sources is trying to hoodwink anyone to sell an extra bag or two. I also recognize that not all vendors have the availability of resources I have as a former academic.

    Regarding other nations' kava, I would venture to guess that most kava exported from Fiji, Tonga, etc. are predominantly noble. They have a long history of drinking it, it is an important part of their economies, and I assume they would not want to risk the entire market. Vanuatu just stipulates it in the Kava Act. I know Fu'u is noble, and chances are anything 'graded' in Fiji (waka, lewena) comes from noble stock. (I just wish they'd mention the cultivar too.) PNG is another story, because some Borogu was brought in by missionaries, but I am pretty sure what they have growing there endogenously is mostly junk.

    I have an interest in protecting the industry more so than protecting my own profits. I have many more friends and family in faraway places who would see a much more serious decline in their standard of living if something were to happen with kava than I would. They are dependent, whereas I could simply get a different job if I needed to.

    As per the type test, in my lact email from Vincent, I was informed that this test is generally performed with 10g of powder, and 30ml of acetone. The purity of the acetone is also important. Using a different ration could get a darker/lighter color than what is expected, generally speaking.

    Lastly, my GREATEST concern is the plethora of tinctures available at health food stores. I have purchased every brand I could get, and am sending them to labs for testing right now, and look forward to publishing the results. I have recently found out one brand touts their kava as "wild harvested." There are no drinkable OR medicinal wild kavas, so that is very disconcerting. Additionally, even knowing that different ratios could cause different colors, I have yet to come across a tincture in the US that was yellow in color. Given the total ml in a bottle, and the total amount of kavalactones per serving, there should be some yellow ones based on the assumptions of ratios, extraction effectiveness of solvents, and general % of kavalactones in kava by weight. So, you have MUCH more reason to be wary of tinctures than vendors selling kava for aqueous extracts.

    Again, hope that was helpful and informative. More videos to come soon
    Cheers!

    Andrew/Iahi
    Ouroboros, infraredz and KavaGurl like this.
  13. kavadude

    kavadude ❦ॐ tanuki tamer

    I very much appreciate the work that went into these videos. I trust both you and Judd to sell quality kava. I just thought it would be interesting to see how easy it is for us to replicate the results.
  14. infraredz

    infraredz BULA!

    I echo kavadude's sentiments. The efforts both of you are doing for everyone is truly commendable and noble (pun intended).

    We all just want the benefits of kava to continue to grow throughout the world and making sure that it is safe is paramount.

    I too trust that you, Judd, Adil and the folks at N@H sell good kava and want to take this opportunity to thank all of you for being moral and ethical people. We truly have something unique in the kava world.
  15. Bula Kava House

    Bula Kava House Portland, OR Kava Vendor

    Just thought I'd throw my two cents in regarding my products because I figure these videos would (rightly) make people question the kava they're drinking, which is also how I make my living. I'm dedicated to selling the best kava out there at reasonable prices so here's a little summation of my thoughts about the info in this video, and some info about my products.

    Noble kava is the name given to a handful of varieties from Vanuatu. They are high quality daily drinking kavas. They were called noble long before any testing was possible. They all tend to be high in certain kavalactones, low in others. There are Vanuatu kavas that are neither noble or tudei.

    To say that only "noble" kava is safe is completely incorrect because that would mean that only kava grown in Vanuatu is safe. Any kava without high levels of flavokavain B and/or mold infection, devoid of stems and leaves is just as safe. It just won't be called "noble" if it's not from Vanuatu. For example, there are a few strains in Fiji that people simply call "white" kava. These strains would likely be deemed "noble" if they were Vanuatu grown.

    I'd be interested in knowing if Lebot tested all non–noble kavas in Vanuatu to check the flavokavain B levels. And could we assume that trace levels of that compound are more or less safe? I also wonder why he said that he didn't know for sure if flavokavain B caused hepatoxicity. It sounds like hepatoxicity is caused by either a, b, or c. Or is it a combination? I suppose I could just shoot an email over.

    Now, about our root:

    As Andrew stated, all four of the Vanuatu varieties we sell are noble tested varieties. Exportation of anything else is not allowed.

    Waka is from lateral root only, and is definitely NOT a tudei.

    Koniak is not described as a tudei by my supplier or the farmers, and is made with a combination of about 40% lateral roots and 60% chips. In PNG "Koniak" is simply the pidgin word used for kava. It actually doesn't define a specific cultivar. We just like the sound of it so that's how it's branded.

    Koniak and Waka will likely be tested someday, but they've both been safely enjoyed for decades, which for me is the best test.

    Fu'u has been tested by my supplier and I've seen the results. It too would likely fit into the "noble" category if it were grown in Vanuatu.
    All of our kava comes from FDA certified processing centers, and receives a phytosanitary certificate before importation. There is definitely no mold or other contaminants.

    I would never sell a kava I thought could be dangerous. I have a high moral standard for doing business. Besides that, hurting customers simply isn't in my best interest.
  16. infraredz

    infraredz BULA!

    I figured we all know Boroguru is a common noble kava. I'm calling this the control. "Koniak" as shown by the following report was reported to be tudei based on qualitative analysis. Based on the above technique, I decided to determine if BKH's Koniak is a tudei (based on the standard in the video [dark color]) and if Solomon's Island by Nakamal at Home is Piper wichmanii (based on the standard in the video [very dark color]) as some have theorized.

    Materials and Methods:
    25mL of acetone was added to 2 teaspoons (approximately 10mL of mass, but mL is measurement of a liquid but this gives a "clearer" ratio) of each kava and agitated for 4 minutes then left to settle for 24 hours. Visual examination of the acetonic portion of the suspension for hue, saturation and darkness is described by Lebot as the method of determination of tudei, wichmanii and noble kavas. Samples were backlit and photographed.

    The ratios of solvent to sample were kept constant. The reason for seemingly different amounts of kava at the bottom is due to the differing grinds (upon close inspection, the larger [most dense] particles settle lower).
    Results:
    acetonic_extraction.jpg

    Comparison between Boroguru on the left and Koniak on the right. Both samples were isolated and photographed with black flags on either side. Photos were then merged into Photoshop and 'split' then combined. Any white balance or curve adjustment was done on the combined image as a whole. The reason why the foreground of the Boroguru sample shows yellow is due to the sample being less opaque (therefore allowing more light from backlight through) than the Koniak sample which, as stated above is more opaque.
    acetonic_extraction_comparison.jpg

    Discussion:
    Something that might not be immediately apparent from the photograph is that Boroguru is the clearest of all the samples, and I suspect that without a centrifuge, the smaller particles responsible for the cloudiness of the other two samples might not decant completely but I will wait another 8-12 hours and see.

    The glass quality is poor as well as fairly thick, giving seemingly different tones from one side to the other. I'm waiting on some lab quality glassware. Just speaking from photo-geekness, it would be best to evaluate the central part of each sample for the most accurate representation of the actual color (due to refractivity of different wavelengths through the glass)

    It's interesting to note that flavokavain A, B, and C are all chalconoids and conjugate ring closures result in flavonoids which are responsible for a wide variety of plant pigmentation (eg. red vs yellow, for example). These chalconoids (FKA, FKB, FKC) reportedly possess antibacterial, antifungal, antitumor and anti-inflammatory properties (sound similar to kava?). Some chalconoids have demonstrated the ability to block voltage-dependent potassium channels (which is a proposed hypothesis regarding the MOA of kava, and in my opinion, sounds very likely to play a significant role in kava's effects). [http://www.ncbi.nlm.nih.gov/pubmed/18032041]

    I wonder if these secondary metabolites are in fact more important in the overall biosynthesis of different compounds than I thought. Flavonoids are responsible for a significant amount of color in a plant, and so I'mreally interested if the reason we are seeing tudeis (which have FKB, unlike nobles) show a red color vs. the "control" of yellow is because of the conjugation of this phenyl ring which would cause it to to become a flavonoid, which in turn possesses specific color properties. Chalconoids by themselves don't necessarily have color properties, but they serve as the intermediate in the biosynthesis to the flavonoids that do.

    This is something that is being investigated as a possible mechanism behind the differences in supernatant color.
    Last edited: May 26, 2014
  17. sɥɐʞɐs

    sɥɐʞɐs ‾‾‾‾‾‾‾‾‾‾‾ Review Maestro

    glad to see that solomon's might actually be 'noble'.
  18. Kapmcrunk

    Kapmcrunk The Kaptain of Crunk KavaForums Founder

    He said in the video that nearly any solvent would work. I've got about a gallon of 99.9% pure methanol that's left over from my instant days when I needed something that was guaranteed anti-freeze. I'll try it with that and the types I have in my pantry.
  19. kavadude

    kavadude ❦ॐ tanuki tamer

    Good stuff, guys, it's been a while since we did any mad science on the board.
  20. infraredz

    infraredz BULA!

    Any opportunity for mad science is a good one in my book!

    Sharpie circles on my dry flaking skin, hell yeah. Strong solvent extractions of kavas, of course.
    :D

    I'll be interested to see what Kapm's results are given the different type of solvent. Lebot said it didn't make a difference, but because of the differing polarities, I would imagine it should have some effect, but probably negligible. Just for information's sake, I got those containers at a local Target, but they're just spice bottles with a screw cap. Acetone can be purchased in high purities at hardware stores.
    Deleted User01 and kavadude like this.