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How to perform the acetonic test

Roaddog

Kava Who?
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Ok, ready for a newbie test? I was unable to find the right size containers, so I had to modify the test a bit. I basically cut the amount of Acetone, and kava, by 1/4. So instead of using 25 ml. of Acetone, I used 6.25 ml. Instead of using 2 level tea spoon, I used 1/2 level teaspoon. So the ratios are correct, just in smaller batches. As you can see (I hope you can see) I compared BKH Melo Melo, a known Nobel, with mood&mind Vanuats. As I hope you can see both samples came out, the exact same light yellow color, the look pretty identical. Hope the quality of the pictures makes the cut. I used regular lab test tubes.
 

Roaddog

Kava Who?
SANY0011.JPG



Here is a close up. I know its a bit rough. Who would have thought that the cheapest Amazon crap, would be a Nobel. I was kind of worried about mood&mind, but they seemed to pass the test. Still not as tasty as the BKH. But at least I know what I got. This test is awesome.Much love. Roaddog....
 

The Kap'n

The Groggy Kaptain (40g)
KavaForums Founder
Not for chemotype, but for testing on the question of being noble, the acetone test is the best, cheapest and easiest way to tell if the kava you're testing is noble or not.
 
D

Deactivated Account

I suspect this Fiji maybe isa, could you guys recognise it by itself if I do the test ?

I don't have a control to put it next to.
 

sɥɐʞɐs

Avg. Dosage: 8 Tbsp. (58g)
Review Maestro
I suck at this test...everything comes out the tudei color...and the proportions of root material to visible liquid appear way different than what I actually put in and different from each other. :banghead: I've been using 70% rubbing alcohol in place of acetone or any pure 99% solvent...but if anything, I would have assumed it would create a lighter hue since it's more watered down.
 

kavadude

❦ॐ tanuki tamer
Well, I'd use the amounts that infraredz said and make sure to use acetone. I think tests done with other solvents have produced similar results but most of them have been done with the acetone, so we don't know if there are any edge cases or anything.
 

sɥɐʞɐs

Avg. Dosage: 8 Tbsp. (58g)
Review Maestro
I've been using the suggested measurements but I guess I'm gonna get some Acetone. Lebot says it works with any solvent...but it's probably best if we all use the same one. I might need to filter my dry root through a strainer 1st too...to get the particle sizes more similar.
 
D

Deactivated Account

I've been using the suggested measurements but I guess I'm gonna get some Acetone. Lebot says it works with any solvent...but it's probably best if we all use the same one. I might need to filter my dry root through a strainer 1st too...to get the particle sizes more similar.
I did mine just 15 mins ago with acetone & its already settled to a nice bright pissy yellow. Ill leave a few more hours to draw a conclusion….but man alive for powdered kava its wickedly strong ! I can feel the gorn syndrome fast approaching. This is powdered kava i can drink.

I think we could all thank infraredz for his efforts here, seriously great work here mate !
 

infraredz

BULA!
The Results
Although it is not necessary to use more than one sample, but it can help to test a known "noble" kava as a reference, or "control" to compare your sample to.

My results: For my purposes, I figured we all know Boroguru is a common noble kava. I'm calling this the control (to show what the color should be).
"Koniak" is a kava that has been suspected to be tudei. Based on the technique Dr. Lebot outlines, 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.​

Kapm also used this technique to test "Hawaiian Kava Center Tudei" (on the Left) and Bula Kava House "Borogu" (on the right) by using one teaspoon of each kava and equal amounts of methanol, which can also be used instead of acetone though I think acetone is more common and probably cheaper though I'm not sure.​

Violet has also used this technique to test known "ISA" (tudei), Wakacon's "Vanuatu Kava" as well as "Papa Kea". The same method as mine was used and her results are below.

Chris from Gourmet Hawaiian Kava has also used this test to compare noble Hawaiian varieties and Hawaiian isa. He also has tested varying degrees of noble/isa ratios and the colors they give. More of his testing can be found here

Roaddog's has also used this to test Bula Kava House's "Melo Melo" and Mood and Mind's "Premium Kava Kava Powder".​

Below are my results (as well as other members' test results) for reference.
-My samples were backlit and photographed, though you obviously don't have to do this. This was only meant to help as a demonstrative measure.

-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).

View attachment 657

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.
View attachment 658

Kapm's Results:
View attachment 659
Left- Hawaiian Kava Center Tudei
Right - Bula Kava House Borogu

Violet's Results:
View attachment 660

Chris' Results:
View attachment 722 View attachment 723

Three varieties of Hawaiian Noble Kava and The results of an Hawaiian ISA

Roaddog's Results:
View attachment 777

"Melo Melo" on left and "Premium Kava Kava Powder" on right

Discussion on Results:
Something that might not be immediately apparent from the photograph is that Boroguru is the clearest of all the samples. I suspect that without a centrifuge, the smaller particles responsible for the cloudiness of the other two samples might not decant completely so cloudiness is not something to worry about.

The glass quality of my vials is poor as well as fairly thick, giving seemingly different tones from one side to the other.

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)

Original Thoughts for Color Difference:
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'm really 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.

My New Theory (decoded)
[UPDATED: Currently the theory being used by Dr. Lebot as of 03/21/14]
FKB, is a chalcone which is a natural biological compound and is an intermediary to flavonoids (which is another type of natural biological compound, common in fruit and vegetables and currently the subject of an abundance of positive health effects).
When chalcones undergo conversion (conjugate ring closure) in the plant they result in flavonoids.

I propose that it is not unreasonable to assume that the presence of FKB can cause an increase or difference in the formation of flavonoids (which then have color properties).

My supposition is that the yellow vs orange tincture could be due to the presence and/or abundance of flavones or flavonoids (in tudei kava). For instance, FKB might result in an orange tint or color whereas noble kava (that has far less FKB, and therefore less substrate [chalcones] being converted to a flavonoid with an orange color) results in a "lighter" color of yellow whereas a higher concentration of FKB corresponds to a higher concentration of a flavonoid that results in an orange color.

I was, at one time, a photography major and took 2 years of extensive coursework.
From that, I can say that these colors can be looked at this way:
  1. Red is the highest saturation "amount" of red (Red, Green and Blue being the "primary color")
  2. Orange is the same hue as red, but slightly lower saturation of red.
  3. Yellow is the same hue as red, but even slower saturation than orange.
Hue is the "color", and "saturation" is the amount of that "hue"

The essence of these theories are currently the operating hypotheses of Lebot as a possible mechanism behind the differences in supernatant color and method of distinguishing "suitable" supernatant color, though it will take some time before definitive information is available due to having to test hundreds of compounds.
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