Extracting bioactive compounds from mushrooms for production purposes involves a combination of cutting, grinding or crushing the mushrooms, ultrasonically-assisted solvent extraction, and subsequent isolation of the bioactive compounds such as beta-glucans. Here is a practical instruction of ultrasonic mushroom extraction in large-scale batch production using larger barrels, eg. a 120L extraction vessel.

Instructions for Large-Scale Mushroom Extraction using Probe-Type Sonication

You can use the following step-by-step instructions for the ultrasonic extraction of any mushroom species including medicinal, culinary and psychodelic mushrooms. Use this protocol to easily extract beta-glucan and other bioactive molecules from Chaga (Inonotus obliquus), Lion’s Mane (Hericium erinaceus), Reishi (Ganoderma lucidum), Cordyceps (Cordyceps sinensis), Maitake (Grifola frondosa), Turkey Tail (Trametes versicolor), Shiitake (Lentinula edodes), Agaricus blazei (Agaricus subrufescens) etc.

Materials and Equipment

• Mushrooms
• Solvent: Distilled water or aqueous ethanol (60% or higher). Choose your solvent depending on your target compounds.
• Blender or grinder
• Probe-type sonicator UIP2000hdT
• Mechanical stirrer
• Food-grade container or batch (e.g. stainless steel, plastic)
• Filter cloth and/or a vacuum filtration setup
• Rotary evaporator
• Storage containers

Sonication-Assisted Mushroom Extraction Protocol

Preparation:

1. Ensure that all equipment is clean and sterilized.
2. Measure and prepare the desired amount of mushroom material for extraction.
3. Mount the stirrer and sonicator onto the extraction vessel.

 
Extraction Procedure:

1. Chop or grind the your dried or fresh mushrooms into a coarse particles of approx. 1 to 5 millimeters. For our example, we use 10kg of mushroom particles.
2. Then add the mushroom particles to the extraction vessel.
3. Next, add 100L of the solvent of your choice to the beaker containing the mushroom material to be extracted. We use here a 1:10 ratio of mushroom to solvent. The mushroom-to-solvent ratio might vary depending on the specific mushroom species, your target compounds and the particle size.
4. Turn the stirrer on in order to mix the slurry. After approx. 2 minutes, the mushroom-solvent slurry is pre-mixed.
5. Keep the stirrer running. Set the UIP2000hdT sonicator to 100% amplitude and switch the sonicator on. Sonicate the mushroom slurry for approx. 45 to 120 minutes using the UIP2000hdT. In order to prevent the thermal degradation of heat-sensitive bioactive compounds, we recommend to maintain an extraction temperature below 60ºC.
6. Temperature Control: Use the temperature sensor of your sonicator to observe the process temperature. Use the temperature control function of your sonicator setting a maximum temperature limit. When the upper temperature limit is reached, the sonicator pauses automatically and continues when the temperature is within the set temperature zone. Additionally, you can use pre-cooled solvent or cool the extraction slurry during sonication.
7. After the sonication process, filter the sonicated mixture through filter cloth or use vacuum filtration to separate the aqueous ethanol containing the extracted bioactive compounds (such as beta-glucans) and other bioactive compounds from the solid mushroom residue.
8. Then use distillation, evaporation or rotary-evaporation to isolate the mushroom compounds from the solvent.
9. After that, you can purify the extract further to obtain specific compounds or fill your extract into storage containers.

Scale-Up: From Ultrasonic Batch to Inline Extraction

If you plan to scale up your mushroom extract production from batch to continuous inline extraction, we will recommend you a suitable ultrasonic flow cell for your industrial probe-type sonicator. Often, you can keep the solid-to-liquid ratios and all other parameters constant allowing for a linear scale-up to higher extract production. Extracting larger quantities of mushroom compounds using an ultrasonic flow-through system is a sophisticated method to produce large extract volumes. Hielscher Ultrasonics offers flow cell reactors designed for the extraction of bioactive molecules from fungi and plants.
If you are interested in processing large mushroom streams, we encourage you to reach out to us for more in-depth information. Our technical team and sonication experts will be glad to assist in determining the most suitable setup for your specific needs. We offer various industrial extraction setups for food- and pharma-grade extraction of mushroom compounds. The image below shows a large flow cell reactor with a UIP4000hdT ultrasonic homogenizer for the extraction of mushrooms at approximately 50 to 200 liters of mushroom-solvent slurry per hour.

Design, Manufacturing and Consulting – Quality Made in Germany

Hielscher ultrasonicators are well-known for their highest quality and design standards. Robustness and easy operation allow the smooth integration of our ultrasonicators into industrial facilities. Rough conditions and demanding environments are easily handled by Hielscher ultrasonicators.

Hielscher Ultrasonics is an ISO certified company and put special emphasis on high-performance ultrasonicators featuring state-of-the-art technology and user-friendliness. Of course, Hielscher ultrasonicators are CE compliant and meet the requirements of UL, CSA and RoHs.