Wisdom

Mushroom Dual-Extractions: How and Why

Mushroom Extracting

One thing I like to do here is try my best to make all of the important information I come across in my learning process available to everybody else for free. In a world where knowledge is power, and all the power is in the wrong hands, we need to re-distribute the power back to the people! One definite power grab is to become sovereign in your health. That is one reason I began experimenting with alchemical plant medicines to support myself. In this learning process it became quite clear that modern scientific research into traditional medicinal plants is actually quite few and far in between believe it or not. Now, we are no stranger to extractions here at the pharmacopeia however (and well assume you aren't either), there is very little in the way of comparative extraction methodologies for herbal practitioners with scientific backing. Some of the few studies discussed here are the forerunners in this field. The cited literature is some of the only published scientific research on mushroom comparative extraction methodologies, and the scope was rewarding albeit somewhat limiting nonetheless. The author of this article is currently attending the first ever undergraduate program designed to study medicinal plant chemistry. Seeing the lack of (although rapidly expanding) scientific research in a very necessary field of life, applied plant medicines, I am compelled to discuss the current research and scientific standing and to circulate new ideas for natural medicine scientific enquiry with the aims of creating more effective and readily available mushroom medicines.

Why a Dual-Extraction?

With that being said, when attempting to produce medicinal quality extractions of mushrooms, a few questions arise. First is, why do we need a dual-extraction? Well the answer is simple and kind of irrelevant to the facts of science but ill explain anyways. Essentially, mushrooms are host of hundreds of known chemical compounds and thousands of unknown compounds. Certain fractions of compounds can only be extracted with certain solvents (a.k.a. a liquid that acts as a solvent). Despite a vast majority of herbal medicines being effectively extracted with high-proof alcohols, mushrooms do not fall with in the vast majority of herbal medicines. They are novel creatures of their own and each one requires individual attention. For example (1) Chaga extracts have exceedingly high antioxidant activity in aqueous and ethanolic extracts yet Lions Mane show statistically insignificant anti-oxidant activities for water/aqueous extraction methods! But that doesn't negate the fact it carries water soluble medicinal compounds partly responsible for its nerve growth factor (2). So there is no one answer. Hence the dual-extraction. Multiple solvents allow us to capture multiple active constituents for the full therapeutic effect.

Fig. 2
Data showing aqueous extractions greater total phenolic content from 5 of 6 medicinal mushrooms
Fig. 5
Radical Oxygen Scavenging capacity in aqueous v ethanolic extracts of 6 common medicinal mushrooms

How to

The next question that arises in my mind is, how to? We need both water and alcohol to perform a dual extraction. The higher percentage the alcohol the better. Your water should be distilled. Now our methodology here comes from research in to the question of:

"Which solvent do we extract with first?"

This is an important question because some solvents may "de-activate" or de-nature medicinal complexes and desired compounds from our mushroom. Ethanol for example will cause the cell wall to undergo lysis, rendering the membrane bound compounds, some of the most important medicinal compounds, unusable and bio-unavailable through denaturing. Ethanol, high heat, unbalanced percent Hydrogen conditions, can all have a negative affect on these membrane bound compounds. Water on the other hand, acting as an acid and a base at 7.0 pH, readily cleaves off membrane bound compounds without lysing the cell wall, thus extracting the highly valuable beta-glucans, alphag-glucans, triterpenes, anticancer, immunoregulatory, anti-inflammatory compounds abundant in the polysaccharides, proteins, and lipids of the circumfusing cell walls.

Cold Water

The preliminary research of the Journal of Agriculture and Food Research suggests that aqueous extracts do not need significantly long periods of extraction times. Hot water extractions over 72 hours performed equally as well in anti-oxidant assays as the ones extracted for multiple weeks.

Hot water extraction could deactivate or degrade the heat-labile antioxidative and volatile components.

The data seems to be suggesting that a mushroom which is freshly harvested and immediately extracted, specifically with less damaging techniques (ie using cold water), is equally as affective as some other industry standards which use long extraction times and vigorous means of liberating fungi analytes from their cellular matrix. It even showed the antioxidizing content diminished over time in heated extracts. It appears that the most important factors in extraction is to move with precaution and preserve as best possible the volatile compounds. Freezing the mushrooms first has even led to better liberation of nutritional and other components from the fungal body.

This being said, at an attempt to preserve volatile compounds which can readily denature at higher temperatures that are not common in nature, we suggest performing cold water macerations on your mushroom material. Hermetically sealing it in a jar for a maximum of 14 days with daily shaking has shown to produce effective medicines. Do this with your mushroom.

When the 14 days is up, strain out the plant matter and press it of all aqueous content. Set aside your aqueous extraction

Ethanol

Next prepare your ethanolic maceration in your vessel of choice. Perform the extraction the same way and time as the aqueous extraction. When you are finished you will have two fractions, a water and and alcohol fraction both containing essences from you mushroom.

Reunification

Now that you have your two extracts, you need to recombine them in a way that does not spoil the unique medicinal chemistry of your extracts. Recombining your extracts in the improper ratios or sequence can potentially denigrate the quality of your medicine.

With this considered, we are going to follow the modern chemical law of adding acids into waters. With more volatile constituents, say a strong acid like hydrochloric acid, the concentration of base being added to the high concentration acid can cause violent exothermic entropic chemical reactions that can cause harmful by-products and even explosions. However, going in the opposite direction and adding the strong acid into the water, the small concentrations of acid are slowly neutralized by the water/base and violent and potentially hazardous chemical reactions are avoided.

This same principle is applied in reforming dual extractions into singular solutions. This is becasue likewise, adverse chemical reaction's can occur from improper unification that can potentially spoil our medicinal potential. The ethanolic portion of the extract represents the acidic portion. However you need to be careful here. You're not just going to dump your fractions together again, you need to proportion it right so that the chemical equilibrium between the two fractions remains within equilibrium and the dissolved medicinal constituents do not fall out (precipitate) from solution resulting in lower medicinal quality and waste of medicine (we don't waste medicine).

You will want to slowly add the ethanolic portion into the water portion of your extract, until you see the first signs of precipitation. If you add the entire ethanolic extract without signs of precipitation, then that just means the chemical constituency was mostly soluble in one solvent or the other. That is unlikely though. It is more likely you will begin to see precipitate form fairly quickly. Stop the addition the second you do.

Once you have recombined all you can without precipitate forming, you have completed your mushroom-dual extraction. If you did not surpass the recommended saturation threshold in reunification, then its likely you have three total extracts left over. One dual-extract, one water extract, and one alcohol extract. I recommend you hermetically seal them all away from light and store them in a cool place, even refrigerating them would not be a bad idea. Use the dual extract for maximum efficacy but the other two are powerful medicines as well. you can make more dual extractions be portioning out fractions of dual-extract with your remaining individual extracts and give them to your friends, or you can just keep them separate and take them as you need.

Let us know if this information helped at all!

Ratioing

A standard practice for liquid-solvent extractions is to use a 5:1 fluid ounce to ounces of herbal material ratio. IE if you have one ounce of mushroom use five ounces of solvent. If in case, you are like me, and suffer from laziness, like many other herbalists and medicine makers who standardize their medicine making practices across the board (not all, but many!) you can always simply recombine your separated extracts to a predefined ratio using a simple equation.

V1 * C1 = V2 * C2

Where, V = Volume and C = concentration
Use any unit (as long as they're the same)
Say for example you had a 500 mL of a 75% alcoholic extract and you wanted to dilute it to 30% final ratio for your dual extract. How much of the alcohol extract do you add to the water extract to make it 30% final concentration?

500 * .75 = .30 * X

Solve for X to find your total final volume of 30% dual-extract. Then subtract X from V1 to find how much water you need for the addition. (to see example equation worked out, watch part two video above).

A note on Vessels*

Mushrooms contain light-sensititve compounds. To retain the highest efficacy of your medicinal extract, wrap your vessels in tin foil. Dull side inwards and shiny side out.

Continuing Research

Much research could be done to advance the fields that work with and apply plant medicines. Accurate chemical profiles must be generated, comparative extraction methodologies on individual species must be reviewed, organic preservation techniques must be explored, affects of various solvents on chemical constituency, medicinal activation of naturally occurring medicines and the continual hunting of novel species must be explored. I encourage you all to do your own research and publish your own findings, experiences, ideas, and inspirations. So much research has been published into traditional preparation methodologies that have shown the ancient way of medicine making often produces significantly more effective medicines than the lab created medicines, standards, and experimental samples. This pharmacopeia makes an effort to discover and make available traditionally verified scientific formulas but more research into these traditional formulas and ethnobotanical recipes is necessary for the advancement of medicinal plant sciences and the total restoration of the world soul. The research is only just beginning and much work is to be done.


Sources

(1) Erica Sharpe, Aiden P. Farragher-Gnadt, Michael Igbanugo, Thomas Huber, John C. Michelotti, Adam Milenkowic, Sylvia Ludlam, Margaret Walker, Douglas Hanes, Ryan Bradley, Fadi Bou-Abdallah, Comparison of antioxidant activity and extraction techniques for commercially and laboratory prepared extracts from six mushroom species,Journal of Agriculture and Food Research,Volume 4,2021,100130,11SSN 2666-1543,https://doi.org/10.1016/j.jafr.2021.100130.

(2) Activity of Aqueous Extracts of Lion's Mane Mushroom Hericium erinaceus (Bull.: Fr.) Pers. (Aphyllophoromycetideae) on the Neural Cell Line NG108-15 Volume 9, Issue 1, 2007, pp. 57-65 DOI: 10.1615/IntJMedMushr.v9.i1.70 Get access Kah-Hui WongMushroom Research Centre, Institute of Biological Sciences, Faculty of Science, Universiti Malaya, 50603 Kuala Lumpur, Malaysia; Department of Anatomy, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia

(3) http://www.davidmoore.org.uk/Assets/Printed_documents/2002_Smith_etal_Medicinal_Mushrooms_complete.pdf

(4) The Pharmacological Potential of Mushrooms Ulrike Lindequist, Timo H. J. Niedermeyer, and Wolf-Dieter Jülich https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1193547/

(5)Art51.pdf (usamv.ro)

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