Understanding Vibrant: Algaefix, Polixetonium Chloride / Busan 77

[rmorris_14]

Hello fellow MTRC member.

They have been tageed numerous times and are supposedly waiting for results of their own testing.

Right now it is "hurry up and wait".

 

[Randy Holmes-Farley]

Thank you. I tried to get people to help figure this stuff out many months ago - https://www.reef2reef.com/threads/vibrant-what-is-it-actually.820541/

Following

Has anyone tagged the owers of vibrant yet? Seems as if they have been lying.

UWC is doing some tests (such as NMR) and should have the answers shortly. I confirmed this with them yesterday.

 

[a.t.t.r]

UWC is doing some tests (such as NMR) and should have the answers shortly. I confirmed this with them yesterday.

You really think they sent in vibrant unmodified off the shelf? I am fully expecting a totally different chart then what we saw at this point.

what we really need is a trustworthy member to allow users to ship them several unopened bottles and then send them off for a true independent test with no manipulation.

 

[rtparty]

You really think they sent in vibrant unmodified off the shelf? I am fully expecting a totally different chart then what we saw at this point.

what we really need is a trustworthy member to allow users to ship them several unopened bottles and then send them off for a true independent test with no manipulation.

There is NO chance they show a test with the same results we see here. Unless their play is a 3rd party screwed them over and that is a poor excuse at best.

 

[Eagle_Steve]

There is NO chance they show a test with the same results we see here. Unless their play is a 3rd party screwed them over and that is a poor excuse at best.

If the results do not match, can it no be refuted by typical reefers buying a bottle, not opening it, sending it to someone trust worthy (not sure who that would be) and then running the same tests on a ton of bottles?

I am sure there are plenty of people who would go buy one and mail it to a respectable person and has to be a way to ensure all bottles are sealed, etc. etc.

 

[Atrumblood]

If the results do not match, can it no be refuted by typical reefers buying a bottle, not opening it, sending it to someone trust worthy (not sure who that would be) and then running the same tests on a ton of bottles?

I am sure there are plenty of people who would go buy one and mail it to a respectable person and has to be a way to ensure all bottles are sealed, etc. etc.

I still have 80% of a bottle I recently purchased. I stopped using it after reading these posts though. I'd be happy to send in a sample to have it tested by an independent third party. I actually might know a guy that has access to NMR and FTIR equipment. I will see if I can get him to run a sample.

 

[elysics]

There is NO chance they show a test with the same results we see here. Unless their play is a 3rd party screwed them over and that is a poor excuse at best.

A poor excuse that works isn't poor. I figure continuing to sell large amounts of vibrant isn't their priority, keeping the rest of their business running is.

I figure "we are just a small local company, betrayal and mistakes can happen to anyone, please have sympathy, we hope to do better in the future blablabla" or something similar might work perfectly well.

 

[rtparty]

If the results do not match, can it no be refuted by typical reefers buying a bottle, not opening it, sending it to someone trust worthy (not sure who that would be) and then running the same tests on a ton of bottles?

I am sure there are plenty of people who would go buy one and mail it to a respectable person and has to be a way to ensure all bottles are sealed, etc. etc.

Absolutely, we should look at doing this no matter what. BRS, SWA, and any other respectable vendor that carries Vibrant should do it as well.

Is anyone really going to trust whatever UWC releases if it doesn't match here? UWC would have to PROVE that taricha was on a witch hunt for some reason and show how everything was purposely skewed.

 

[Randy Holmes-Farley]

Absolutely, we should look at doing this no matter what. BRS, SWA, and any other respectable vendor that carries Vibrant should do it as well.

Is anyone really going to trust whatever UWC releases if it doesn't match here? UWC would have to PROVE that taricha was on a witch hunt for some reason and show how everything was purposely skewed.

At least one other person has run an NMR and it matches the proton NMR taricha got. I don't want to out them so I'll leave it to them to post where and when (and if) they want to.

 

[Malcontent]

I'm totally not buying small bottles of Prime every few months to keep an "archive" that spans several batches.

 

[jda]

Some background, in case anybody wants to do this themselves...

There are places around you with NMR and IR instruments and most will analyze things for you for a fee. There are three public universities within an hour from me and all three have chemistry departments that can and will do this. Call around and ask. Some universities have NMR technicians on their websites that you can reach out to directly. The staff at these Universities have real jobs too, so just be a bit patient with them. I had to register as a vendor and set up a payment account, so expect a few hoops, which anything legit is going to have.

Here is what you need to know if you wish to do this. Call/email them and ask for 400mhz 13C and 1H NMR. You can also ask for FTIR if you want. Tell them that they need to dry out the samples completely, preferably with vacuum, before they bring it up to D2O - they will know what this means. You can use higher mhz NMR instruments, but the tests are more expensive and there is no need. I could have paid more to use a 500mhz instrument, but choose not to even though it would have worked just fine.

The bottle of Vibrant has a seal, so drop off or mail a sealed bottle and ask the tech to remember or pay attention that they broke the seal on the bottle, just in case. Just bring the bottle, pay and let them work as independent technicians.

Lastly, tell them that the substance is a EPA pesticide so that they can dispose of the extra safely. They will know how to do this too.

Edit 03/11 (forgot to add this): You will need somebody with access to special software to make a graph/chart in the ranges that you see - the ranges of the X axis on the charts already provided. The wider range makes it too hard to see where the spikes/peaks are. Again, they will know how to do this.

Expect to pay about $600-1000 for this... at least in my area. It might be less if you want less tests done, like just the 13C or the 1H and not both. The IR is pretty cheap. NMR is much more expensive.

I did this over the last few weeks. I went and purchase a bottle at a local shop, took it to Colorado State University unopened and had their staff start to analyze it.

The tests are being run, but the 1H Hydrogen came back already - graph below. It is the same substance that is in the first post. I don't know how there can be any doubt to the work that @taricha did. I have the raw data from the machine in a .zip file if any real scientist wants to look at or analyze it... PM me. The numbers are shifted a bit, which is common and happens when the zero point is calibrated different from instrument to instrument - what is important is that the graphs are identical when you shift them together. Transparency: the black box on the graph is the name on the directory of the machine where the files were stored, which IDs a person, so I took it out.

I will post the Carbon and IR when I get them back. I think that we all know what to expect... they will match the work done on page one.

1H 400mhz NMR:

Receipt for recent purchase. No markings on the bottle with a batch number or anything. Guy working there had no recollection when this stock came into the store:

 

[taricha]

And here's what it looks like when you do an overlay of the data I received and what @jda received... You can see at the bottom, the x-axes (chemical shift - ppm) are aligned.

Talking to technical people, this is what it looks like when the same signals are read on different machines with different calibrations.

 

[LRT]

Incredible contribution to the community and thank you to the folks putting money where the mouth is.

 

[Christoph]

Fantastic to have so much NMR data here on R2R.

Im synthetic organic chemist by training and thus have had many NMRs to measure and to interpret. There is no doubt the substances in all 3 measurements (the two initially submitted by taricha and the recent one from jda) are matching, meaning all samples had the same organic chemical composition.

Also the NMR data is matching what would be expected from a chemical structure like Busan 77.

Best regards,
Christoph

 

[Atrumblood]

Fantastic to have so much NMR data here on R2R.

Im synthetic organic chemist by training and thus have had many NMRs to measure and to interpret. There is no doubt the substances in all 3 measurements (the two initially submitted by taricha and the recent one from jda) are matching, meaning all samples had the same organic chemical composition.

Also the NMR data is matching what would be expected from a chemical structure like Busan 77.

Best regards,
Christoph

100% agree with you. Having this data available to the community is huge!

 

[a.t.t.r]

So we now have multiple accounts peer reviews and independent testing. RIP @UWC. I hope @Bulk Reef Supply stops all business with them as they have shown to be malicious at worst and incompetent at best.

 

[JCOLE]

Also, I was one of the members that sent their bottle to @taricha for initial testing. This was the same bottle I used right before my crash.

 

[avidhexagrammid]

You're telling me, some guy in his garage has a prep-grade UPLC column and can sell liters of this extract for dollars? Why is he selling to aquarists? He should be selling the miracle elixir to people managing waste water streams.

This is such a bizarre line of reasoning to me. The most likely explanation is that it was developed by a biotech firm contracted by UWC. I personally don't expect scientific/technical explanations from the owner because the people "in charge" don't always know the nuances of what they're talking about when it comes to process. However, I did land on the topic of cyanophycin because of this comment.

Please, if you could just make some relevant arguments to these points that would make your case more and less like off-topic whataboutisms and wild guesses that require a miracle of miracles involving esoteric biosynthetic pathways.

If you'll notice, I haven't actually made a whole lot of arguments, I've been asking for clarity because I prefer to make informed arguments. Dismissing a reasonable suggestion because you've decided it's complicated, or too specialized, or you don't have an inherent understanding of it is such a weird thing to do and doesn't actually rule it out as a possibility. Seems cyanophycin is fairly well-studied and could be easily engineered for this purpose.

Simply just address these points people have repeatedly raised to you:

1. Why are we assuming it's crude extract and that no purification steps were taken before packaging?
2. As above.
3. Once again, this is why I am asking how sample preparation and conditions during analysis could influence losses, reactions, and lower sensitivity to certain compounds in an unknown mixture.
4. I never suggested that the NMR spectra didn't indicate the presence of QAC, just the IR. In fact, I think the C NMR is the most compelling aspect of all of this, but I've still found a potential alternative explanation that doesn't involve polixetonium chloride. I'm not blind, I can see the spectra are uncanny, but you're comparing two unknowns and haven't characterized anything from them. I'm not a big fan of inductive reasoning when it comes to making such steep accusations. Everything here has only been presented as confirmatory for polixetonium chloride, without actually characterizing it and without any mention of what other factors could explain these results (which is a necessary part of analysis). Pretty much all of the mini assays conducted have some glaring confounding variables that make the results inconclusive yet they go wholly unacknowledged. Some quick examples:
- bromophenol blue also stains enzymes
- glucose is a reducing agent and will decolorize methylene blue
- classic COD tests exclude oxidation of ammonia from quantification

It would be courteous to at least acknowledge people's counter points before spouting out conjecture without any evidence whatsoever.

Speaking of not acknowledging counterpoints, are you (or anyone else) able to at least address why the cholinium ion isn't a possibility here? Seems like a good candidate given it's utility for algal cell wall disruption, flocculation, and reduction of planktonic bacteria. None of the results rule it out as far as I am aware.

 

[Malcontent]

 

[avidhexagrammid]

I have no idea what a metabolite line of questioning is.

I saw comments from UWC that 1) indicated the use of bacterial metabolites (i.e., cell free) from cyanobacteria and select Bacillus spp. that involve the production of polyaspartic acid; 2) suggested that the quat-like response is normal; and 3) that the lab who developed it is slated to present this research at an upcoming conference for the American Society of Microbiologists. I'm assuming it's a biotech firm and the 'bacteria blend' has remained a proprietary secret because they're working on a patent and/or publication. That's pretty normal. I've checked and the book of abstracts for that conference is not out yet but it seems plausible to me due to how easily it can be verified.

Even if you somehow believe that EVERY organic molecule in this "bacterial" mixture, EXCEPT the polixetonium was masked from view, it still contains a ton of polixetonium, a wholly unnatural algaecide. It really doesn't matter if there are other molecules present or not, the ALGAECIDE IS THERE.

I don't somehow believe this, I am asking for clarity. If certain compounds are present at low enough concentrations and would normally have some peak overlap to begin with, could their signals have poor resolution compared to something like a QAC that's present in larger amounts? Also, it contains a QAC, are we sure it's polixetonium and not something like cholinium?

Certainly, some peaks in NMR and IR move around as pH changes protonation of structures like carboxylic acids, non quaternary amines, thiols, etc..

Thank you! I was asking for specifically this reason, not QACs.

Edit: fixed a hyperlink.