Testing of Chemiclean

[MnFish1]

I care too. @MnFish1 - your involvement in the whole Vibrant thing started out poorly, continued to be poor and still is poor, that I can tell. This is like opening the same playbook. You can stay out of this one if you don't care or don't intend to help - I will catch you on the next one where you contribution can be good when you care or even care about other people caring. Everybody else should just ignore what is posted. This is all that I am going to add unless some skin is added to the game... and I mean actual skin and not just words.

You must have missed the link I posted on a method to determine erythromycin. Whether it is helpful or not - I don't know. However - I'll try to help and let me clarify my point.

I don't care if its erythromycin specifically, I care if its ANY antibiotic at all - as its my philosophy to not use antibiotics unless treating a known pathogen to a fish. So - all of the research in the world on erythromycin won't help if in fact it's tetracycline or some other antibiotic.

So one way you could experiment would be to obtain some agar plates, a gram-stain kit, a bacterial culture, and see whether chemiclean acts like an antibiotic. Now - common sense would suggest that it is an antibiotic (since it removes cyanobacteria). You can even grow cyanobacteria on agar (likely requiring a specific type)https://pubmed.ncbi.nlm.nih.gov/2513249/

 

[MnFish1]

Blink twice if someone is forcing you to read and take part in this thread...

Actually - this forum was designed for people to propose experiments - and then have a discussion about them. No one is forcing me. IMHO - there are issues with the experimental plan. Taricha, though - has provided the most significant indirect information that erythromycin is not present.l

 

[nereefpat]

Sophisticated chemistry lab testing (HPLC, in this case) seems like the way to identify EM undisputedly.

Apart from that, a microbiology lab might be some use and would likely be fairly cheap/easy. For example, a few plates containing known EM in the agar and a few plates containing Chemi Clean. Compare what species of bacteria grow or are inhibited by both.

Just a thought. I know it wouldn't be a smoking gun like the NMR in the Vibrant study.

 

[MnFish1]

Sophisticated chemistry lab testing (HPLC, in this case) seems like the way to identify EM undisputedly.

Apart from that, a microbiology lab might be some use and would likely be fairly cheap/easy. For example, a few plates containing known EM in the agar and a few plates containing Chemi Clean. Compare what species of bacteria grow or are inhibited by both.

Just a thought. I know it wouldn't be a smoking gun like the NMR in the Vibrant study.

This is how they identified bacteria in the past - gram stain and which plates they would grow on. Now it's mostly DNA based - but agree this would be a way to suggest it or not.

 

[nereefpat]

This is how they identified bacteria in the past - gram stain and which plates they would grow on. Now it's mostly DNA based - but agree this would be a way to suggest it or not.

Right. The challenge is this would be backwards. Nailing down the exact Abx will be difficult, although you might be able to rule certain ones out. You would also show that it is an antibiotic (it is).

 

[Hypnotize]

Funny you can buy chemiclean on amazon.de (35 Euros approx)

Check where it’s coming from not available „in“ Europe

 

[MnFish1]

Check where it’s coming from not available „in“ Europe

Isn't amazon.de in Germany?Do they not have distribution centers in Europe? Whenever I order items from AMZN German site - I pay local (or no) shipping. If I click on Chemiclean on the site there is a big red warning that 'this item cannot be shipped to your location (in the US)'. It asks for a German postal code - suggesting that its being sold in Germany - I may be wrong - I heard this court issue in the EU was settled some years ago?

 

[Hypnotize]

Isn't amazon.de in Germany?Do they not have distribution centers in Europe? Whenever I order items from AMZN German site - I pay local (or no) shipping. If I click on Chemiclean on the site there is a big red warning that 'this item cannot be shipped to your location (in the US)'. It asks for a German postal code - suggesting that its being sold in Germany - I may be wrong - I heard this court issue in the EU was settled some years ago?

It’s shipped from Amazon US and since it’s clearly declared what ingredients it has it may be shipped.

Same with Flux RX. We can order it through Amazon but it’s not actually legal since you would only be able to get it from a pharmacy with a subscription from a doctor.

and the company Colombo had something similar to Chemiclean called Mycosidol on the shelf’s in LFS all over Europe.
They often stated that no antibiotics where in the product. Some aquarists but money together and sent it to different labs and all came to the conclusion that there were in fact antibiotics so they were forced to take it if the market.

they have a new product with the same name now but haven’t heard anything good from it.

 

[MnFish1]

It’s shipped from Amazon US and since it’s clearly declared what ingredients it has it may be shipped.

Same with Flux RX. We can order it through Amazon but it’s not actually legal since you would only be able to get it from a pharmacy with a subscription from a doctor.

and the company Colombo had something similar to Chemiclean called Mycosidol on the shelf’s in LFS all over Europe.
They often stated that no antibiotics where in the product. Some aquarists but money together and sent it to different labs and all came to the conclusion that there were in fact antibiotics so they were forced to take it if the market.

they have a new product with the same name now but haven’t heard anything good from it.

You are correct - when I changed the 'zip code' - I could see this"
Versand
Amazon US
Verkäufer
Amazon US"

However, what I don't understand is how an 'illegal substance' can be sent through customs?

 

[Hypnotize]

You are correct - when I changed the 'zip code' - I could see this"
Versand
Amazon US
Verkäufer
Amazon US"

However, what I don't understand is how an 'illegal substance' can be sent through customs?

Because of luck that’s all.

 

[MnFish1]

Because of luck that’s all.

Not to get off subject - but it's also illegal for amazon to ship it (from the US). So it surprises me that they allow it. Perhaps they don't realize it's an antibiotic.

 

[taricha]

Here's some reasons that it seems to me (totally inexperienced with medications, properties, etc) that the earlier suggested tylosin tartrate might be a good candidate along with EM for lab comparison to chemiclean.

1) some posted LD50 toxicity descriptions match as pointed out in this thread

Analytical chemist here. When certain "scarce" terms are plucked from the MSDS for the product and put into Google, I find some interesting similarities:

"LD50 Oral Rat > 6200 mg/kg" H317 H319

Specifically, there are some striking similarities between the product's SDS and that for sulfamethoxazole. However, I do not get any good hits with "LD50 Oral Rat > 6200 mg/kg" + erythromycin.

And now that I dig some more, tylosin tartrate also matches, and is a salt, unlike sulfamethoxazole.

https://chemi-pure.com/wp-content/uploads/2019/08/ChemiClean_SDS_US_122815_FINAL.pdf
https://northamerica.covetrus.com/content/sds/056666.pdf

2) solubility in water
Erythromycin 1-2 mg/mL
Sulfamethoxazole <1 mg/mL
Tylosin tartrate - various: 200 mg/mL at 25C, "freely"
(as I said earlier - a full 50mg scoop of chemiclean dissolves in 0.1ml of water.)

3) high toxicity to cyanobacteria
"For A. flos-aquae, we obtained a 96 h EC50 of 0.06 µmol L−1, which is within an order of magnitude of a published EC50 of 0.037 µmol L−1, which was reported for another cyanobacterial species, Microcystis aeruginosa, after 72-h exposure to tylosin"

0.06umol/L for tylosin is around ~0.06 mg/L so the ~1 mg/L dose of the chemiclean powder is plenty - and many report that a 1/2 dose of chemiclean also kills cyano.

(though there is inconsistency in the literature - some other papers have tylosin as toxic to cyano only at much higher concentrations.)

4) safe for nitrifiers
Paper showing tylosin needed to be at 50+ppm to have any effect on nitrifiers.

5) Melting point 126C
I put it in my heating block and between 117 and 126C the powder got darker/clumped and had clearly melted by 131C.

(left tube was chemiclean, right was the API E.M. erythromycin - it's mostly salt so it's not going to do much at those temps.)

6) pH between 5-7.2
on my pH test strips, dissolved in 1mL of distilled water, it was above 5 and below 7pH.

7) it's apparently an agricultural antibiotic for chickens and pigs (and bees somehow), so seems like it'd be quite cheap to acquire.
@jda looks like you could get it on chewy for $40 as a comparison to chemiclean and treat as many chickens as you'll ever need.

 

[MnFish1]

Tylosin (brand name: Tylan®) is an antibiotic in the same family as erythromycin. It is primarily used in cats, dogs, and small mammals to treat diarrhea and inflammation of the gastrointestinal tract.

Tylosin | VCA Animal Hospitals

Tylosin is a macrolide antibiotic similar to erythromycin. It is active against gram-positive and gram-negative bacteria, spirochetes, Chlamydophila and Mycoplasma organisms. Tylosin is usually used to treat certain types of diarrhea in dogs and cats. It may also be used to treat bronchitis...

vcahospitals.com

It seems it's clear that chemiclean is an antibiotic.

Tylosin - Wikipedia

en.wikipedia.org

 

[osprey101]

Here's some reasons that it seems to me (totally inexperienced with medications, properties, etc) that the earlier suggested tylosin tartrate might be a good candidate along with EM for lab comparison to chemiclean.

1) some posted LD50 toxicity descriptions match as pointed out in this thread
you'll ever need.

I was pretty surprised how clean the signal was; I finally found the time, ran it through Q-TOF to figure out what it was. Easier than I thought it would be.

 

[taricha]

I was pretty surprised how clean the signal was; I finally found the time, ran it through Q-TOF to figure out what it was. Easier than I thought it would be.

!!

Had to look that one up.


analytical chemists have the craziest toys.

 

[taricha]

I was pretty surprised how clean the signal was; I finally found the time, ran it through Q-TOF to figure out what it was. Easier than I thought it would be.

For those of us who aren't chem people but enjoy seeing how this stuff works, would you mind telling us what you saw in the QToF mass spectrum?

As I understand it, what you get is the mass of the whole compound (194 for caffeine example below) plus detections for the masses of different pieces possible if the collisions break it apart (109, 82 etc)

(example from this lecture ppt)

 

[MyFirstCar]

If anyone has access to a 96 well plate reader, an ELISA (an immunoassay) would give a pretty definitive answer about whether it's erythromycin. https://www.elabscience.com/p-ery_erythromycin_elisa_kit-205713.html . If anyone want to do one, I'd get someone who's experienced with doing them though, they're pretty finicky.

 

[EeyoreIsMySpiritAnimal]

.

 

[Randy Holmes-Farley]

For those of us who aren't chem people but enjoy seeing how this stuff works, would you mind telling us what you saw in the QToF mass spectrum?

As I understand it, what you get is the mass of the whole compound (194 for caffeine example below) plus detections for the masses of different pieces possible if the collisions break it apart (109, 82 etc)

Yes, that's basically how a mass spec of any sort is interpreted. Details of how they are done help determine the pieces that get detected and also how intense the pieces are. Fortunately, for identifying a molecule such as erythromycin, one would not need to try to predict the breakdown pattern since it is known already and/or can be run as a standard.

Here is erythromycin run one particular way. Note that the whole molecular ion at about 734 g/mole is not necessarily the largest peak.

Erythromycin Mass Spectrum

This MassBank Record with Accession MSBNK-IPB_Halle-PB005803 contains the MS2 mass spectrum of 'Erythromycin' with the InChIKey 'ULGZDMOVFRHVEP-RWJQBGPGSA-N'.

massbank.eu

 

[taricha]

Thanks, Randy.
I think @osprey101 was implying that the mass spec they got was a match for what they guessed it was earlier - tylosin tartrate, a macrolide antibiotic, but a different one than erythromycin.
(Much larger mass than EM, so should be clearly distinguishable between those two.)