Acetone test and Flavokawain B content

[HeadHodge]

edit by kavadude: I moved this discussion into its own thread because I think it is worth talking about

@HeadHodge - Actually, the chemical analysis (HPLC/NIRS) is not the final proof. Vanuatu Dept of Ag currently includes acetone test results on their COA, since chemotype alone is not sufficient to exclude the presence of tudei.

Well if you don't mind... Could you remind me again for the reason why I care if it's tudei or not? I was under the impression that it contained higher concentrations of some substances more so than Noble kava.

 

[HeadHodge]

Why you ask? Flavokavain-B. It has demonstrated cytotoxicity to human liver tissue, and while it does not occur in Noble kava strains, it is detected in substantial amounts in tudei and wichmanii varieties. So, yes, tudei, Isa, etc. have a different balance of kavalactones, some of which have a cult following in the US and other markets due to perceived "strength" but they also contain flavokavains that simply do not occur in varieties that have seen kastom use for generations.

@Deleted User.... So that's my original point. Are you saying there are no chemical tests that actually can be used to test for ?? If so isn't that what we should be testing for, if possible??

 

[kavadude]

Do you mean the "smoking gun" in terms of the culprit behind liver damage? I don't think we'll ever get one of those, it is clear to me there needs to be a confluence of factors and bad practice at multiple steps in the kava business in order for kava to be implicated in liver damage. To me though the potential hepatotoxicity of FKB is the primary reason to develop and perform these tests. Sure it would be nice to know with 100% certainty what variety of kava I am consuming but frankly if the test does not correlate with FKB levels then it is more of a nice thing to have than something that vendors and exporters need to be held accountable for.

 

[Deleted User01]

Good Post Global Kava Exports and I might add that both you and GHK have managed to find Kava in Vanuatu that tested well. However there seems to be a lot of evidence (including admissions by them) that some farmers are adding Tudei to their Kava but not all Farmers. If all the Vendors made the same effort as you and GHK to find good 100% Noble Kava, then all the farmers would have to toe the line or lose business. But I agree that we can't paint every farmer with the same brush stroke.

@Deleted User, I have complete confidence in your tests. Also I would remind everyone that we don't have rock solid evidence concerning liver toxicity and Tudei but that is not what we are debating. We are debating "Truth in Advertising". Tell us if your kava is 'blended" and let everyone make an educated buying decision. If you don't tell us, then Deleted User will tell us.

 

[ApéroNoble]

Do you mean the "smoking gun" in terms of the culprit behind liver damage? I don't think we'll ever get one of those, it is clear to me there needs to be a confluence of factors and bad practice at multiple steps in the kava business in order for kava to be implicated in liver damage. To me though the potential hepatotoxicity of FKB is the primary reason to develop and perform these tests. Sure it would be nice to know with 100% certainty what variety of kava I am consuming but frankly if the test does not correlate with FKB levels then it is more of a nice thing to have than something that vendors and exporters need to be held accountable for.

It seems a bit elementary regarding choice though, Dr. Lebots already stated Noble had none, the high concentrations were in tudei, thus accountability for variety is of utmost importance here.

 

[kavadude]

You're preaching to the choir. I am not recommending that people drink tudei kava. I am concerned about the validity of your tests.

As far as I know, tudeis are now defined by their FKB content, both by the general consensus around here and by the Vanuatu Department of Agriculture. The simple acetone test was developed by comparing the results of the test to an HPTLC analysis of FKB content. In other words, I would hope that your test correlates with levels of FKB. Is it not possible that other molecules can cause a lesser color change, undetectable in the simple test, and that you are measuring something other than the nobility of the kava?

 

[kavadude]

I hope everyone understands that Deleted User is not testing for FKB content. He is measuring the color differences between kava samples when dissolved in acetone, in a way that is much more objective and sensitive than the simple test, the end goal being to detect "spiked" kava. However, unlike the simple test, to my knowledge the results are not backed up by an analysis of FKB content.

 

[kavadude]

@Deleted User You're gonna have to give me a bit to dig this up because some of it is from private correspondence or ancient posts, in the meantime here is the video where Lebot shows an HPTLC analysis of noble/tudei/wichmanii.


The test was never claimed to measure FKB; I *thought* it was developed by comparing the results to HPTLC analysis of samples. I'm not actually sure where I got this from so I'm going to have to look through my old mail.

Kava (Piper methysticum) is used to prepare the traditional beverage of the Pacific islands. In Europe, kava has been suspected to cause hepatoxicity with flavokavin B (FKB) considered as a possible factor. The present study describes an HPTLC protocol for rapid screening of samples. The objectives are: to detect the presence of flavokavins in extracts and to compare the FKB levels in different cultivars. Overall, 172 samples originating from four cultivars groups (noble, medicinal, two-days and wichmannii), were analysed. Results indicate that the ratio FKB/kavalactones is much higher in two-days (0.39) and wichmannii (0.32) compared to nobles (0.09) and medicinal cultivars (0.10). For each group, the ratios flavokavins/kavalactones do not change significantly between roots, stumps or basal stems and among clones, indicating that they are genetically controlled. This protocol has good accuracy and is cost efficient for routine analysis. We discuss how it could be used for quality control.

http://www.ncbi.nlm.nih.gov/pubmed/24423570

In other words, the gold standard (lol) for nobility is the FKB/kavalactone ratio as determined by HPTLC analysis. I would expect this ratio to be somewhere inbetween in your spiked samples, and the same should hold true for the spiked samples you've obtained from vendors if your test is indeed a measure of nobility.

 

[kavadude]

What makes you positive that the "orange molecule" is what you're measuring with the spectrophotometer? Could other differences in chemical composition between noble & tudei (and different varieties of noble kava) not cause less obvious changes in the color of the solution? Differences that, unlike FKB/kavalactone ratio might vary significantly between variety/harvest? Again, my understanding is that Lebot developed the simple test while double checking everything with HPTLC. I hope that @infraredz or @Kapmcrunk will jump in here as they are more in touch with what is going on at the Vanuatu Dept. of Agriculture.

 

[ObiWan]

I noticed that some of my samples have changed their colour after a few weeks. The Fu'u for example was in my test definitely noble with a bright yellow. Now some of the acteon has evaporated and the ratio is nearly 1:1. It looks now very orange. That leads me to the question: How important is it to use exactly the right ratio (1/3 Kava to 2/3 Aceton)? Should I measure the volume or the weight? 1 Teaspoon Kava can differ a lot in volume, depending if it is micronized or coarse grind.

 

[ObiWan]

The ratio should be kava/acetone 1:3. Acetone is by volume, kava by weight. Weighing isn't always practical, so volume is often substituted. Results should still be usable (when a reference noble/tudei is available), but it's not going to be as reliable as actually weighing the kava. A "level tablespoon", for instance, could vary quite a bit due to how much it is "packed" when leveled. The key is using the same technique for both the reference samples and the kava in question.
I always weigh the kava, and when comparing weight to volume across 200+ samples, I've found a pretty wide variation. I use 1,334mg kava to 4ml acetone, and that amount of kava varies from about 3ml to 5ml in volume, so it is considerable.

I have no measuring cup which is suitable for the Aceton, so my idea is to use the weight. When I understand it right, 1 ml of Aceton should be 0,79g. So if I use 5g of Kava, I would have to add about 19g of Aceton. Does this make sense?

 

[ObiWan]

Ah, I failed with a simple rule of three

I am frightend that the Aceton will be damaging the plastic. Or did you use a syringe made of glass?