Ultrasonic Cold Water Extraction of Psilocybin

Isolating psilocybin from fresh and dried magic mushrooms, e.g. psilocybe species, is known to be time-consuming and often quite inefficient. With ultrasonic extraction psilocybin, psilocin, baeocystin and norbaeocystin are rapidly extracted from fresh and dried psilocibe mushrooms. Ultrasonic extraction breaks the cell walls of the mushroom so that the bioactive substances are completely released, so that a highly efficient and rapid extraction process is achieved.

Ultrasonic Psilocybin Extraction

Ultrasonic psilocybin extraction is highly efficient and outcompetes alternative extraction methods by high yields, short extraction time, as well as simple and safe operation. Being a highly efficient, rapid, reliable, and environmental-friendly extraction method, ultrasonic extraction is used to extract numerous bioactive compounds from plants and fungi. Hielscher ultrasonicators are already widely used to extract bioactive compounds from magic mushrooms including hallucinogens such as psilocybin, psilocin, baeocystin and norbaeocystin. The ultrasonic extraction of psychedelic compounds gives high yields of superior extract quality within a turbo-extraction procedure. Ultrasonic extraction is compatible with any solvent and allows for the production of full-spectrum extracts.

Ultrasonication is a fast and mild extraction method to produce high-quality mushroom extract. In the video, a UP400St is used for shitake extraktion.

Cold Mushroom Extraction using UP400St with 22mm probe

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Ultrasonic extractor UP100H, a compact 100 watts powerful ultrasound probe, is used for the extraction of hallucinogens from magic mushrooms of the psilocibe species.

Portable ultrasonicator UP100H for psilocybin extraction.

Ultrasonic Extraction – Gentle Cold Extraction of Mushroom Substances

Many bioactive compounds are heat-sensitive, which means they degrade and decompose under high temperatures so that they loose their effect and potency. Therefore, most procedures for isolation of secondary metabolites from botanicals and fungi benefit from a non-thermal extraction method, which is not based on heat as basic working principle. Ultrasonic extraction is a purely mechanical procedure, whose working principle is based on the generation of acoustic cavitation.

What is Ultrasonic Cavitation and How does it Impact Extraction?

When a liquid is sonicated with an ultrasound probe at 20kHz, 20,000 vibrations per second are transmitted into the medium. The ultrasonic waves travel through the liquid, where they create alternating high-pressure (compression) / low-pressure (rarefaction or expansion) cycles. During the low-pressure vacuum cycle, minute vacuum bubbles or cavities occur in the liquid, which grow over several pressure cycles. During the compression phase of the liquid and bubbles, the pressure is positive, while the rarefaction phase produces a vacuum (negative pressure.) During the compression-expansion cycles, the cavities in the liquid grow until they reach a size, at which they cannot absorb more energy. At this point, they implode violently. The implosion of those cavities results in various highly energetic sonomechanical effects, which are known as the phenomenon of acoustic / ultrasonic cavitation. During the implosion of ultrasonic cavitation bubbles, extreme energy dense forces such as high shear, liquid streaming and turbulences are created in the sonicated liquid. These cavitational forces disrupt cell walls of plants and release the intracellular compounds, including bioactive substances such as the hallucinogens psilocybin, psilocin, baeocystin and norbaeocystin from mushroom species. The alternating high-pressure/low-pressure cycles also promote intense mass transfer rates, which results in the superior extraction yield obtained by sonication. Via ultrasonic extraction, almost the complete amount of active substances present in botanicals and mushrooms can be isolated.

Ultrasonic Psilocybin Extraction: Step-by-Step Protocol

  • You can extract psilocybin and other bioactive substances from mushrooms using water, ethanol or 200 proof ethyl alcohol as solvents.
  • Chop the fresh or dried psilocybin mushrooms into small bits and pieces. You can use caps and stems. Both contain the psychoactive compounds, however the caps consistently are richer in psilocybin. The spores do not contain psilocybin.
  • Place the chopped mushrooms in a glass beaker and fill it with cold water.
    Optionally, you might place the glass beaker in an ice bath to keep the process temperature as cold as possible.
  • Sonicate the mixture of mushroom and solvent with an ultrasonic probe. Depending on the volume, the UP100H or UP400St will be a good choice. Our technical staff will recommend you the ideal ultrasonicator for your extraction process and share with you process parameters such as amplitude setting and sonication time.
  • The sonicated slurry will turn blue. The blue water containing extracted psilocybin from the mushroom fruiting bodies.
  • Separate the mushroom pieces and debris by straining or filtering. Optionally, keep the mushroom material from filtration for a second extraction run using either water or ethanol.
  • Use an evaporation system to remove the solvent. 1 to 2 grams of dried magic mushrooms should yield in approx. 1cc of final extract. As mushrooms are a natural product, the content of psychedelic substances can vary.
  • By an optional crystallization step, the shelf-life of the psilocybin extract can be prolonged further.
  • Fill the final extract into a dark glass container or vial and store in the freezer.

 

Ultrasonic mushroom extract can be filtered and concentrated using a rotary evaporator.

Ultrasonically extracted bioactive compounds can be filtered and further concentrated using a rotary evaporator. The picture shows an ultrasonic chaga mushroom extract.

Ultrasonic extractor UP400St (400 watts, 24kHz) for the extraction of psilocybin, psilocin, baeocystin and norbaeocystin from mushrooms

UP400St – a 400 watts powerful ultrasonic probe-type extractor for the extraction of psilocybin from mushrooms

Advantages of Ultrasonic Extraction
 

  • More complete extraction
  • Higher yields
  • Rapid process
  • Cold / non-thermal process
  • Compatible with any solvent
  • Easy and safe to operate
  • Low maintenance

Why is a Probe-Type Ultrasonicator Best for Mushroom Extraction?

Ultrasonication is a process in which high-frequency sound waves are used to create cavitation bubbles in a liquid. When these bubbles collapse, they create intense localized shear forces that can break down cells and release the contents of the cells into the liquid.
In mushroom extraction, ultrasonication can be used to break down the cell walls of the mushrooms and release their bioactive compounds into a solvent. There are two types of ultrasonicators: bath-type and probe-type.
A bath-type ultrasonicator is a device in which the sample is placed in a container filled with the solvent, and relatively weak ultrasonic waves are applied to the entire container. This method is known as quite ineffective, as as ultrasonic bath distributes ultrasonic energy unevenly and with a low intensity. As in an ultrasonic bath the mushroom sample is sonicated indirectly, the ultrasound cannot penetrate deep into the sample. The ultrasound waves have to go through the walls of vessel before they hit the mushroom material. Thereby, the already low intensity ultrasound waves of the ultrasonic tank are even further diminished.
On the other hand, a probe-type ultrasonicator is equipped with a tip – the so-called sonotode or probe – that can be inserted directly into the sample, allowing for more focused and localized application of ultrasonic energy. This results in a significantly more efficient cell disruption and extraction of bioactive compounds, especially in dense or hard-to-reach areas of the sample.

Chitin is a polysaccharide that is a major component of the cell walls of many fungi, including mushrooms. Chitin is a tough and rigid material, which makes it difficult to break down using conventional extraction methods.
Ultrasonication can be effective in breaking down chitin and releasing the bioactive compounds that are trapped within the cell walls of the mushrooms. However, the high-intensity of probe-type sonication is necessary because the chitin in the cell walls is a relatively tough material. The focused and localized application of ultrasonic energy provided by the probe-type ultrasonicator ensures that the chitin is subjected to a sufficient amount of energy to be broken down.
Additionally, the probe can be moved around to different areas of the sample creating additional macro-mixing to ensure that all parts of the mushroom are adequately sonicated. This is particularly important for mushrooms with thick cell walls or dense structures, where it can be challenging to ensure complete extraction using other methods.
In summary, the high-intensity of probe-type sonication is necessary to break down the chitin in the mushroom cell walls and release the bioactive compounds. The focused and localized application of ultrasonic energy provided by the probe-type ultrasonicator ensures that the chitin is adequately sonicated, resulting in more efficient and thorough extraction of bioactive compounds from the mushrooms.
A probe-type ultrasonicator is generally considered to be more efficient for mushroom extraction, as it can provide more uniform and thorough extraction of bioactive compounds compared to a bath-type ultrasonicator.
Read more about the differences between processing liquids with a probe-type ultrasonicator and an ultrasonic bath!

Psilocybin-Containing Mushrooms

The most well-known psilocybin-containing mushroom is the species Psilocybe cubensis. Ultrasonic extraction is used to extract full-spectrum extracts from mushrooms.Psilocybin can be found in more than 200 species of mushrooms of the four fungus genera Conocybe including the species Panaeolus, Psilocybe, Stropharia, which are all suitable for extracting the active compound psilocybin ultrasonically. In general, mushrooms that contain psilocybin exhibit the following characteristics: They have dark spores, are lamella / gills fungi, grow mostly in humus-rich soil in meadows and woods of tropical and sub-tropical regions and exhibit blue bruises which occurs due to an oxidative reaction, when the mushroom is handled or damaged.
The most well-known species of psilocybin-containing mushroom is Psilocybe cubensis, which contains two major psycho-active compounds, psilocybin and psilocin. Commonly known as shrooms, magic mushrooms, golden tops, cubes, or gold caps, Psilocybe cubensis belongs to the fungus family Hymenogastraceae, and was formerly known as Stropharia cubensis. As it is widely distributed and easy to cultivate, Psilocybe cubensis is the best-known pscilocybin-containing mushroom species.
Psilocybe cubensis is mainly famous for its hallucinogenic compounds, which trigger psychedelic experiences after ingestion. These major psychoactive compounds are:

  • Psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine)
  • Psilocin (4-hydroxy-N,N-dimethyltryptamine)
  • Baeocystin (4-phosphoryloxy-N-methyltryptamine)
  • Norbaeocystin (4-phosphoryloxytryptamine)

Advantages of Ultrasonic Extraction Systems

The advantages of ultrasonic extraction are the high extraction yield of psychedelic compounds (i.e. psilocybin, psilocin, baeocystin, norbaeocystin), the free choice of solvent (e.g. water, ethanol, water-alcohol mixture etc.), as well as the simple and safe operation. Due to the intensive mechanical forces of sonication, ecological and mild solvents such as water, ethanol etc. are usually sufficient to achieve an extraordinary extraction rate and yield. As a result, ultrasonic extraction shortens the extraction time and enables a reduced use of solvents or the use of milder, gentler solvents. This means that ultrasonic extraction allows for both, higher extraction rates and extracts of superior quality (e.g. cold water extracts). Since the process temperature can be precisely controlled during the sonication, thermal decomposition of the extracts due to excessively high temperatures as well as evaporation of the substances are avoided.

Sophisticated Ultrasonicators for Mushroom Extraction

Hielscher Ultrasonics product portfolio covers the full range of high-performance ultrasonic extractors from small to large scale. Additional accessories allow for the easy assembly of the most suitable ultrasonic extractor configuration for your process. The optimal sonication setup depends on the envisaged capacity, volume, raw material, batch or inline process and timeline. Our long-experiences, well-trained staff will recommend you the most suitable ultrasonic extractor and will assist you during the establishing phase of your mushroom extraction process.

Ultrasonic botanical extraction gives higher yields. The Hielscher UIP2000hdT, 2000 watts homogenizer is powerful enough to extract batches from 10 liters to 120 liters easily.

Ultrasonic Extraction of Botanicals - 30 Liter / 8 Gallon Batch

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You can buy Hielscher ultrasonic processor in any different size and exactly configured to your process requirements of mushroom extraction. From treating fungi in a small lab beaker to the continuous flow-through mixing of mushroom slurries on industrial level, Hielscher Ultrasonics offers a suitable ultrasonicator for you! Please contact us – we are glad to recommend you the ideal ultrasonic setup!
The table below gives you an indication of the approximate processing capacity of our ultrasonicators:

Batch Volume Flow Rate Recommended Devices
1 to 500mL 10 to 200mL/min UP100H
10 to 2000mL 20 to 400mL/min UP200Ht, UP400St
0.1 to 20L 0.2 to 4L/min UIP2000hdT
10 to 100L 2 to 10L/min UIP4000hdT
n.a. 10 to 100L/min UIP16000
n.a. larger cluster of UIP16000

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Precisely Controllable Amplitudes for Optimum Results

Hielscher's industrial processors of the hdT series can be comfortable and user-friendly operated via browser remote control.All Hielscher ultrasonicators are precisely controllable and thereby reliable work horses in production. The amplitude is one of the crucial process parameters that influence the efficiency and effectiveness of ultrasonic extraction of bioactive phyto-nutrients and hallucinogenic compounds from botanicals. Raw material such as soft fresh mushrooms require milder sonication and lower amplitude settings, whilst dried mushrooms might require sonication at higher amplitudes to release the intracellular content. All Hielscher ultrasonicators allow for the precise setting of the amplitude. Sonotrodes and booster horns are accessories that allow to modify the amplitude in an even wider range. Hielscher’s industrial ultrasonic processors can deliver very high amplitudes and deliver the required ultrasonic intensity for demanding applications. Amplitudes of up to 200µm can be easily continuously run in 24/7 operation.
Precise amplitude settings and the permanent monitoring of the ultrasonic process parameters via smart software give you the possibility to produce your mushroom extract under the most effective ultrasonic conditions. Optimal sonication for best extraction results!
The robustness of Hielscher’s ultrasonic equipment allows for 24/7 operation at heavy duty and in demanding environments. This makes Hielscher ultrasonic equipment a reliable work tool that fulfils your extraction requirements.

Easy, Risk-free Testing

Ultrasonic processes can be completely linear scaled. This means every result that you have achieved using a lab or bench-top ultrasonicator, can be scaled to exactly the same output using the exactly same process parameters. This makes ultrasonication ideal for risk-free feasibility testing, process optimization and subsequent implementation into commercial manufacturing. Contact us to learn how sonication can increase your mushroom extract yield and quality.

Highest Quality – Designed and Manufactured in Germany

As a family-owned and family-run business, Hielscher prioritizes highest quality standards for its ultrasonic processors. All ultrasonicators are designed, manufactured and thoroughly tested in our headquarter in Teltow near Berlin, Germany. Robustness and reliability of Hielscher’s ultrasonic equipment make it a work horse in your production. 24/7 operation under full load and in demanding environments is a natural characteristic of Hielscher high-performance ultrasonicators.

Ultrasonic high-shear homogenizers are used in lab, bench-top, pilot and industrial processing.

Hielscher Ultrasonics manufactures high-performance ultrasonic homogenizers for mixing applications, dispersion, emulsification and extraction on lab, pilot and industrial scale.



Literature / References

  • Mason, N.L; Mischler, E.; Uthaug, M.V.; Kuypers, K.P.C. (2019) Sub-Acute Effects of Psilocybin on Empathy, Creative Thinking, and Subjective Well-Being, Journal of Psychoactive Drugs 2019.
  • Pedersen-Bjergaard, S.; Sannes, E.; Rasmussen, K.E.; Tønnesen, F. (1997): Determination of psilocybin in Psilocybe semilanceata by capillary zone electrophoresis. Journal of Chromatography B, 694; 1997. 375–381.
  • Stamets, P. (1996): Psilocybin Mushrooms of the World. 1996

Facts Worth Knowing

Magic Mushrooms

Mushrooms, which contain psychoactive / psychedelic substances such as psilocybin, psilocin, and baeocystin, are known as psychedelic mushrooms, magic mushrooms, shrooms, magic truffles, and mush. More than 180 species of mushrooms containing psilocybin or its derivative psilocin are known.
Psilocybin mushrooms are illegal in many countries, e.g. USA, Canada, Europe, India. In the USA, they are classified as a Schedule I controlled substance, though recently, the FDA and DEA have allowed some small, highly controlled human studies on their potential for use in medical and psychiatric drug administration. The effects of psilocybin and other psychedelics have been researched for their effects and potential treatment of disorders such as cluster headaches, mood disorders, and addiction.

Species

Psilocybe cubensis is one of the most popular magic mushrooms offering a medium psilocybin concentration of approx. 0.63% psilocybin and 0.60% psilocin in dried wild mushrooms. Indoor cultivated mushrooms mostly have higher concentrations of their active compounds.

Psilocibe Species % PSILOCYBIN % PSILOCIN % BAEOCYSTIN
P. azurenscens 1.78 0.38 0.35
P. bohemica 1.34 0.11 0.02
P. semilanceata 0.98 0.02 0.36
P. baeocystis 0.85 0.59 0.10
P. cyanescens 0.85 0.36 0.03
P. tampanensis 0.68 0.32 n.a.
P. cubensis 0.63 0.60 0.025
P. weilii 0.61 0.27 0.05
P. hoogshagenii 0.60 0.10 n.a.
P. stuntzii 0.36 0.12 0.02
P. cyanofibrillosa 0.21 0.04 n.a.
P. liniformans 0.16 n.a. 0.005

Psilocybin

Psilocybin is an psychoactive, hallucinogenic substance found in mushrooms of the genera of Psilocybe, Panaeolina, Panaeolus, Copelandia, Conocybe, Gymnopilus, Stropharia and Pluteus and is considered as a narcotic drug.
Besides psilocybin, psilocin and baeocystin are two further active substances found in magic mushrooms.
The potency of mushrooms can vary greatly since mushrooms are a natural product, which develops in response to its environment. The various parts of mushrooms differ in potency, too (e.g. cap and stem may have a different concentrations of the psychoactive compounds). Due of these natural variations, the maceration, homogenization and ultrasonic extraction might help to reduce the risk of unexpectedly strong (or weak) doses of psilocybin.
Use and Effects: After administration of psilocybin, a wide range of subjective effects have been reported which range from feelings of disorientation, lethargy, giddiness, euphoria, joy, and depression. Approx. a third of users experience feelings of anxiety or paranoia. Low doses of the drug can induce hallucinatory effects, closed-eye hallucinations, and/or synesthesia (the experience of tactile sensations when viewing colors). At higher doses, psilocybin can induce the intensification of affective responses, enhanced ability for introspection, regression to primitive and childlike thinking, and activation of vivid memory traces. Open-eye visual hallucinations are common and can be very detailed although rarely confused with reality.
Anecdotal reports as well as recent studies suggest that a single administration of psilocybin can enhance reative thinking and empathy and could therefore be a potential treatment for cognitive and social behavior. Results of the study “Creative thinking and empathy” (Mason et al. 2019) indicate that psilocybin enhanced divergent thinking and emotional empathy the morning after use. Enhancements in convergent thinking, valence-specific emotional empathy, and well-being persisted seven days after use. Sub-acute changes in empathy correlated with changes in well-being. The study demonstrates that a single administration of psilocybin in a social setting may be associated with sub-acute enhancement of creative thinking, empathy, and subjective well-being.

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