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Mushroom Testing Case Study

Terpene and Terpenoid Testing (HPLC)

First, we obtained Lion’s Mane (Hericium erinaceus) mushroom and performed our own extraction. Our proprietary process was used to extract the bioactive small compounds (terpenes) in the fruiting body. Next, we performed a similar extraction to all four of our extracted powder samples purchased from Amazon. All solutions were then placed on our HPLC instrument and analyzed under optimal conditions.

In the above figure we include a blank solvent (with no mushroom sample), the four samples purchased from Amazon and our own extracted mushroom. From this overlay you can see that LM-002 and LM-003 are similar to the solvent blank, exhibiting no peaks (compounds). Samples LM-001 and LM-004 are comparable to the mushroom we extracted. We can qualitatively compare the peaks and their relative intensities (amounts) in the samples to our mushroom extract. Think of this as more of an identification test, determining if the product is real.

We believe this test shows that LM-001 and LM-004 are powders made from Lion’s Mane mushrooms, while products LM-002 and LM-003 do not contain Lion’s Mane. However, let’s compare the Beta-Glucan and Heavy Metals testing for additional confirmation.

Beta-Glucan Testing

The four Amazon samples were tested for their Beta-Glucan content. The samples were prepared for analysis as instructed by the Megazyme test kit. Two of the samples, LM-002 and LM-003, were visually distinct from the other samples during this lengthy testing procedure. The results are in the table below.

The test kit also includes a Control sample with a known value. The Control is tested with the samples and has a Certificate of Analysis (COA) Beta-Glucan value of 49%. Megazyme’s documentation states that the specification of the Control is +/- 5%, meaning the test itself should give results that are within 5% of the listed value. We meet this specification, giving confidence to our sample results.

Samples LM-001 and LM-004 are once again quite similar, with Beta-Glucan values of 42-43%. These samples are within the expected range of actual Lion’s Mane extracted powders. Samples LM-002 and LM-003 yield some confusing results. LM-003 is only 16.5%, which is much lower than most listed extracted Lion’s Mane powder values that we were able to find online. However, the interesting results is for LM-002 where a negative number is obtained. What is going on?

First, the test for Beta-Glucans is not directly a test for Beta-Glucans. It is a test for Total and Alpha-Glucans content, then the Beta-Glucans is mathematically determined.  Total Glucan = Alpha-Glucan + Beta-Glucan. If Beta-Glucan is not in the sample or is at an extremely low level, the calculated value will not be correct. Second, the test kit and its procedures are designed to determine glucan content in mushrooms. If the sample is not from a mushroom, then the results will most certainly vary.

From this data it is shown that LM-001 and LM-004 are both consistent with each other and our expectations as a mushroom extracted powder. Samples LM-002 and LM-003 are once again giving erroneous results, further adding to our questioning their validity.

Heavy Metals Testing

The last test performed on these four Amazon purchased samples of Lion’s Mane Extracted Powders was Heavy Metals. We investigated Lead (Pb), Mercury (Hg), Arsenic (As), and Cadmium (Cd). See the results in the table below.

The FDA has not established limits for heavy metals in dietary supplements, so at this time it’s difficult to comment on the levels we see in our samples. It’s at least assuring that samples LM-001 and LM-004 have similar values for the four metals we investigated. Samples LM-002 and LM-003 have slightly higher Arsenic values, but keep in mind these are all very small amounts. The values shown here are the average of duplicate determinations and they should be rounded when reported, but we wanted to show the minor differences.


Scout Scientific set out to demonstrate our ability to analyze mushroom extracts. We ordered Lion’s Mane Extract Powders from four different vendors selling their products on Amazon. We anticipated that we would be looking for minor differences between the vendor samples, but instead found that two of the four samples were not actual Lion’s Mane mushroom powders.  They may possibily be mycelium on grain products. The HPLC work clearly shows that not only do samples LM-002 and LM-003 not contain the target compounds, but they also do not contain any detectable (UV active) compounds. Once we reviewed the data we collected, we were curious as to what was in these products. The lack of compounds in the UV spectrum of the chromatogram leads us towards it being a simple grain or starch, but we can only guess as these tests are designed to test for Lion’s Mane compounds.

The FDA does not require specific analytical methods for compliance with cGMPs for dietary supplements, which is how extracted mushrooms are categorized. However, because these products claim to be Lion’s Mane, the FDA would consider these products (LM-002 and LM-003) to be misbranded, thus potentially adulterated.

What about the vendors? Scout Scientific will reach out to all vendors of this case study to let them know what we found. It is our belief that the vendors of LM-002 and LM-003 do not know that they are selling fake products. It does not make it okay, and they are certainly responsible for what they sell, but we feel they may have been misled by those at the wholesale level of the mushroom extract market. If there are any updates to our interactions with these vendors, we will post them to this page.

What’s Next?

There is much more work to be done on not only Lion’s Mane, but other species of medicinal mushrooms. We are currently working on the following projects:

Medicinal Mushrooms in Other Products

Just like some of the currently sought after cannabinoids (CBD, Delta 9, etc.), medicinal mushrooms are increasing in popularity in the functional product marketplace. Lion’s Mane mushroom is now being introduced into gummies, candies, chocolate bars, hot chocolate drinks, coffee, sports drinks, cosmetics, skin care lotions and more as it continues to expand across the industry. However, these vendors may not be receiving an actual mushroom extract to add to their product. Our methods provide vendors with the confidence that their starting material is authentic or that a third-party has formulated their products correctly.  Scout Scientific could also confirm that the terpenes are not degraded by the manufacturing process of the target products.

Extraction Optimization

We would like to work with mushroom vendors, including individual foragers, small businesses, and large industries to optimize their extraction processes. We are interested in investigating not only the best methods to perform the extraction, but also the optimal conditions for maximizing specific bioactive compounds. Does a double extraction with hot or cold water make a difference? How about various percentages of ethanol? We can help answer these questions.

Compound Quantitation

A goal of Scout Scientific is to isolate and purify terpene compounds in Lion’s Mane fruiting body. Using LC-MS we believe we have identified most of (if not all) the compounds in our HPLC analysis. We have been working towards the isolation of these compounds. On the small scale we have been very successful, as shown in the following chromatograms. Once we have begun to use more advanced manufacturing techniques it is our hope to be able to provide quantitation of Lion’s Mane Extracts and make these materials commerically available allowing others to do the same in their own laboratories.

All mushroom testing is performed in a non-GMP, non-ISO laboratory.