Why doesn't Nootropics Depot and Real Mushrooms provide terpene % for lion's mane?

Okay, I am going to respond to /u/Kostya93's word salad he made on the other thread here, because I don't want to drive any traffic to a subreddit that I feel promotes propaganda. Sorry OP, this post is not directed at you, but at Kostya93.

It is an interesting subject and almost identical to a thread I started myself on r/Nootropicsdepot but which was deleted instantly without any explanation.

Predictable... Immediately jumping to conspiracy theories about censorship again. That thread was removed before I could even see it, much less respond. Had I seen it I might have responded, but I have also been very busy. It's also very exhausting arguing with people not operating in good faith, or with complete misunderstandings of science. Regardless, there is no conspiracy here. I did not even see that post before it was removed.

u/misteryouaresodumb is the owner of Nootropics Depot. Judging from his reply in the original post he appears to be blinded by his ambitions. Like Paul Stamets. I see a lengthy reply with little actual content or facts. It comes across as a marketing speech to be honest.

Again, predictable. Character attacks and misdirection in an attempt to influence the reader's perception of my statements. That's propaganda 101. Let me write some more "marketing speech" for the readers here. You all can decide who is operating in good faith.

Here's the gist of his reply (correct me if I'm wrong): - Testing for erinacines / hericenones cannot be done due to a lack of standards and references. - Testing for total terpenes makes no sense because nobody knows which terpenes will be included that number. - Then he's linking to a blog about cannabis terpenes to make his point again - HPLC is unsuitable to test for terpenes according to him.

Yes, you are wrong. It's called an analogy. I brought up the fact that I would not even test for common terpenes in cannabis using HPLC. Then I linked a quick blog on why that is. I was not saying the blog spoke exactly about diterpenes in lion's mane. However, let me go into more detail. HPLC-UV cannot properly detect things without a chromophore. Many terpenes don't have chromophores, so they are not able to be accurately detected on an HPLC-UV. Let me go into even more detail, as I have a feeling you have no idea how any of this works. HPLCs consist of two main parts: the chromatography part and the detection part. The chromatography is handled by a column in an HPLC. That column separates compounds from each other. Then you have the detector, or the thing that actually detects what comes out of the column. Most HPLCs use a UV detector. That's why you see it referred to as HPLC-UV. So for HPLC-UV to be a valid methodology for a specific test, you need to be able to accurately separate the compounds you are looking to quantify with the column and solvents you choose, AND you need to be able to see their absorbance on a UV spectrum. If what you are looking for does not have a chromophore, then you cannot accurately detect it when it comes out of the column. This is especially true as the concentrations get lower. You can set your detector at a certain wavelength, and see anything that comes out of the column that has a density difference. However, that is a really shitty way of doing it, as you're mostly just measuring the liquid refraction rather than true absorption, and you certainly aren't going to get peak purity information. So without a chromophore in the compounds you are trying to quantify, you are not going to get accurate data. Hericenones do have chromophores, because they have a benzene ring. So at least you could detect hericenones on an HPLC-UV detector. The numbers might not be accurate, but you can at least see them. Most of the erinacines don't. I guess you can see Erinacine-A or H on a UV detector, but not B, C, D, E, F, G, or I. That's where mass spectroscopy comes in.

A mass spec can then detect the compounds coming out of the HPLC and get their masses. You can also use NMR for that as well. A mass spec uses a quadrupole to separate different masses, and a detector to give you the data. Then there are triple quad MS systems, which I spoke about in my other comment. That is essentially two mass specs run in tandem, with a 3rd quadrupole between them, which gives you much more accuracy and sensitivity for separating compounds of similar mass and structure. An NMR uses a strong magnetic field to actually detect the number of protons in a compound. This allows you to interpret the data and identify what compound something is. For things like terpenes, which can be very close to each other in both structure and mass, you need these things to be accurate. Then you have GC-MS/MS, which separates compounds using a gas mobile phase, rather than a liquid. This is good for volatile compounds like terpenes, but I won't get into all that right now. The moral of the story is that HPLC-UV is not an accurate methodology for the purpose we are speaking about.

That's incorrect and shows ignorance.

Cute! It certainly shows someone's ignorance...

For starters, musechem.com has a few isolated erinacines available as reference standards.

Never heard of musechem. Are they even reliable? I have already had my team reach out to them regarding the erinacines. We'll see what they say. We have dealt with this before, though. Every time we reach out to someplace that lists they have a standard, they tell us they actually don't. Finding a link in Google, and actually getting valid and reliable standards in-hand, are two totally different things.

See this research paper which includes the procedure to test for diterpenes using HPLC in Lion's Mane (page 18). All erinacines are diterpenes and all diterpenes are erinacines in the case of Lion's Mane. This is according to research. So, testing for diterpenes is actually testing for erinacines as a group in the case of Lion's mane. Even if it is not 100% accurate (but what is ? analytical tests always have a margin of error) it will provide a pretty good indication of what is there I think.

LOL! Did you even read the damn study you linked... That's a QUALITATIVE method for detecting erinacines, not QUANTITATIVE. Furthermore, the very study you linked discusses the limitations of HPLC-UV for quantitative analysis of diterpenoids! They even go through the exact same things I spoke about on my other comments!!!

Absorption of UV light is a phenomenon that is broadly used in the detection and identifiation of diterpenes. Although their UV spectra are rather nonspecific and have maxima in the low wavelength values (λ =  190–230 nm) [6,91], UV detectors or diode array detectors (DADs) are still the most popular and accessible in the HPLC kits. In the case of UV detection problems, the application of an evaporative light scattering detector (ELSD) [6,61] or refractive index (RI) detector [17] is a solution. Another interesting method of detection applied in the analysis of diterpenes is ELISA, which is quite complicated (time-consuming, and one has to have appropriate antibodies for the compounds analyzed) but highly selective and drastically lowers the limits of detection and quantitation. Therefore, the ELISA method can successfully be used for trace analysis in complicated natural matrices. ELISA analysis of gibberellins (besides with cytokinins) in oil palm is an example of the application of this method [100].

So the very study you linked says UV detectors in HPLC have limitations in detecting terpenes! They talk about possibly using the less common ELSD detectors, or evaporative light scattering detectors, but very few people are using those. Then they talk about a very time consuming and difficult enzymatic technique called ELISA, or enzyme-linked immunosorbent assay, but that's totally separate from HPLC. Then they even go on further...

Coupled techniques in HPLC analysis relate to identifiation and detection problems. The most widespread “coupled technique” is the connection of HPLC separation with spectral methods (MS, rarely nuclear magnetic resonance [NMR]). The main advantage of such a connection is the improvement of identifiation of compounds in complicated mixtures: without the need of timeconsuming isolation procedures and in low concentrations (when we do not have enough sample), even if we do not obtain suffiient separation of compounds analyzed. Obviously, LC-MS and LC-NMR have some technical disadvantages (time-consumption, costs, problems related to use of several eluents, such as evaporation, or potential ionization of analytes), but they are still indispensable in the search of new compounds, mainly pharmacologically active metabolites. For example, for the identifiation of ginkgolides using LC–sonic spray ionization (SSI), ion trap MS was applied [17]. ESI coupled with time-of-flght (TOF)–MS or ion trap MS is more popular in the identifiation of natural compounds and broadly applied in diterpene analysis [37,62,68,72]. Figure 24.14 shows an example of HPLC–ESI–ion trap MS analysis of diterpenes and flvonoids in an herb of Isodon rubescens, compared with the corresponding HPLC-DAD analysis [71].

Hmmm, LC/MS or LC/NMR. I wonder where I heard that before!

There are also examples of application of coupled ultra performance liquid chromatography (UPLC)–ESI–MS techniques used for the identifiation of metabolites in plants—for example, the identifiation of 47 taxanes from other compounds as metabolic profiing of Taxus hair roots [136].

Ohhhhh, UPLC. Never heard of that... LOL

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Have to split my comment up, due to character limits on Reddit.