Hydrogen Peroxide Bench-testing

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Hydrogen peroxide usage report
Jay Hemdal
April 3, 2021

Premise:
I use 7% hydrogen peroxide for my home water treatment system. Therefore, since I always have this material in the house, I wanted to explore other possible uses for it, including:
  • Disease control​
  • Water sterilization​
  • Bleaching of algae on surfaces​
  • Indirect production of dissolved oxygen (for overstocked tanks, etc.)​
Final conclusion:
After all of the tests were performed, the only additional use for this product that has merit was found to be “water sterilization”. It is easier to control hydrogen peroxide in this use than is sodium hypochlorite (bleach). All other possible uses either didn’t work well or had unwanted detrimental effects. Using peroxide as a dip to treat Amyloodinium marinum (velvet) has been reported in the literature. I had no cases of this disease to test, but will try it in the future if I run into that disease.

Overview:
With the chemical formula H2O2 this compound has some unexplored uses in aquariums. Hydrogen peroxide is available over the counter in pharmacies and supermarkets as a 3% solution. At this level, it is not reactive enough for most potential uses. Available through some home tap water treatment services, a 7% concentration solution of hydrogen peroxide is stronger and more useful. Do not use this product until you have read and fully understand the Safety Data Sheet (SDS) for it. Although even stronger concentration of H2O2 are available, they are very reactive and their use simply entails too much risk. In addition, these products have had stabilizers added to them which may not be suitable for use in and around aquariums. Perhaps the most immediate use for hydrogen peroxide is as a disinfectant / oxidizer. Filter elements and other aquarium items that have been fouled with organic materials can be soaked for 24 hours in a 7% solution of hydrogen peroxide. It is much easier to rinse off than bleach is, and does not leave a toxic residue like some other disinfectants might. Knowing the propensity for this compound to give off an oxygen atom in oxidizing reactions, some people have experimented with using it to raise the ORP level of aquariums, or to temporarily enhance the dissolved oxygen level in water. Remember, like ozone, too much hydrogen peroxide can pose a threat to the animals themselves. Some researchers have shown that protozoans can be killed by doses of hydrogen peroxide ranging from 12.5 to 25 ppm. Because this is in the range that some fish have shown sensitivity, more study is required. Testing for this compound can be done using a standard DPD chlorine test, at least in terms of presence or absence. There are now test strips available (see below) that can quantify peroxide concentrations from 0.05 ppm to 100 ppm. Additionally, an overdose of hydrogen peroxide can be neutralized with a standard aquarium dechlorinator. Fisheries biologists sometimes use hydrogen peroxide to treat bacterial gill diseases in freshwater fishes such as Columnaris, Flavobacterium columnare as a bath at a dose of 50 to 100 ppm for an hour each day for three days. It may also serve as a dip to treat marine fish for Amyloodinium. The typical dose for that is 20 ml of 3% hydrogen peroxide in one gallon of aquarium water for 30 to 45 minutes. A static test of 5 ppm daily additions for five days eradicated cyanobacteria in a freshwater aquarium, but also damaged the nitrifying bacteria, causing a rise in ammonia levels.

Hydrogen peroxide dosing:
Because hydrogen peroxide is in a solution, and not a 100% active material, calculating doses is not very straightforward.

A 7% solution is 70,000 ppm and a 3% solution is 30,000 ppm

Using a 7% solution, the following doses can be calculated:
1 ml in 14 l = 5 ppm​
.1 ml in .37 gallons = 5 ppm​
Static testing with freshwater fish:
A 12.5-gallon aquarium with a Betta and blue green slime algae was dosed with a hydrogen peroxide dose of 5 ppm daily. After three days, the algae growth was reduced. After eight days, the algae was eliminated, however the Ammonia tested at 0.31 ppm and the nitrite was at 0.027 ppm and still rising. This is borderline acceptable for Betta, so the peroxide additions were stopped. Seven days later, the algae began to return.

Static testing with marine aquarium:
Long term testing of my 16-gallon Bio cube at home showed that daily 25 ml additions of 3% hydrogen peroxide resulted in reduction of green hair algae and Valonia sp. However, bristle worms and shrimp were adversely affected. Despite copious water changes, when the peroxide additions were stopped, the GHA returned to its previous levels. Valonia was still present, but in fewer numbers. Additionally, heavy peroxide use (40 ml/day) eliminated the GHA, but it was replaced with a brighter green, turf species. Hand plucking and more water changes worked better.

Hydrogen peroxide drop-down test:
It is reported in the aquarium hobby literature that hydrogen peroxide, added to seawater breaks down within minutes if the solution is aerated. This was found to be incorrect. A 25-ppm solution of hydrogen peroxide in strongly aerated seawater still measured > 20 ppm after 72 hours. A similar test using deionized water showed even more stability – a 25 ppm solution was measured after five days at 10-20 ppm.

Platinum catalyst test:
Reports are that platinum acts as a catalyst for hydrogen peroxide, releasing oxygen in a nondestructive manner. A platinum coated titanium anode was acquired. The anode was added to a solution of deionized water to which 25 ppm of peroxide was added. Bubbles soon began to adhere to the anode, showing proof of concept. However, the peroxide was exhausted after 24 hours, and 25 ppm is too high of a concentration to use with fish.

Platinum anode catalyst.png

Platinum anode catalyst

Bleach / Hydrogen peroxide bleaching comparison:

Sections of plastic, overgrown with hydroids and brown algae were tested to see if hydrogen peroxide was an effective bleaching agent as compared to sodium hypochlorite (bleach).

After a six-hour exposure, brown coloration means less bleached..png

After a six-hour exposure, brown coloration means less bleached.

Bleaching of aquarium components is useful when completely renovating an aquarium. This of course cannot be done with animals present, but it is helpful when cleaning jellyfish exhibits. Bleach, however, is difficult to rinse off afterwards. Hydrogen peroxide is much easier to remove. However, this bench test showed that peroxide, even at a 1:2 dilution, did not have sufficient bleaching effect. The lowest concentration that achieved 100% bleaching was a 1:5 solution of bleach after six hours.

Hydrogen peroxide test on deionized water:
Chemical effects of hydrogen peroxide on deionized water were examined to see if there were any profound changes. A one-liter sample of deionized water had 50 ppm of hydrogen peroxide added, and then measurements were taken over time:

Prior to addition:
Dissolved oxygen = 8.29 mg/l @ 95% saturation​
pH = 3.67 Total dissolved solids = 3​
Oxidation reduction potential= 433.7​
After 30 minutes:
DO = 8.09 mg/l @ 93% saturation​
pH = 3.65​
TDS = 41​
ORP= 473​
After 45 minutes:
DO = 7.88 mg/l @ 90.3% saturation​
pH = 3.64​
TDS = 41​
ORP= 472​
After two hours:
DO = 7.28 mg/l @ 83.7% saturation​
pH = 4.21​
TDS = 42​
ORP= 477.2​
After three hours:
DO = 7.25 mg/l @ 83.3% saturation​
pH = 3.73​
TDS = 43​
ORP= 476​
Overall, there was a slight rise in ORP and a temporary rise in dissolved oxygen, but not many other changes following the 50-ppm addition.

A second challenge was made at 100 ppm hydrogen peroxide, but 5 grams of algae was added to serve as a biocatalyst:

Prior to algae addition:
DO = 7.22 mg/l @ 82.8% saturation​
pH = 3.65​
TDS = 43​
ORP= 474.4​
One hour after algae addition:
DO = 9.01 mg/l @ 103% saturation​
pH = 4.16​
TDS = 37​
ORP= 477​
Three hours after algae addition:
DO = 11.62 mg/l @ 136% saturation​
pH = 4.45​
TDS = 32​
ORP= 476​
Four hours after algae addition:
DO = 12.7 mg/l @ 147% saturation​
pH = 4.61​
TDS = 31​
ORP= 475.3​
Sixteen hours after algae addition:
DO = 13.5 mg/l @ 153% saturation​
pH = 4.65​
TDS = 29​
ORP= 482.7​
The conclusion is that hydrogen peroxide breaks down faster in the presence of organic material, and this releases oxygen. This is likely due to the presence of peroxidase in the algae cells.

High range peroxide test strips – 0.5 to 100 ppm.png

High range peroxide test strips – 0.5 to 100 ppm

Peroxide low range.jpg

Low range peroxide test strips – 0.05 to 4 ppm
 
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Randy Holmes-Farley

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Hi Jay,

A great many people have found that adding hydrogen peroxide to their reef aquaria causes a drop in ORP, not the expected rise. I have a couple of theories about why, but I was wondering that since you say a rise in DI water, if you happened to check it in marine aquarium water of some sort?

One theory is that it causes local killing and breaking open of bacteria, and the low ORP intracellular contents end up driving down ORP more than the peroxide initially raised it.

Second theory gets really deep in the weeds of what exactly is formed when hydrogen peroxide breaks down in seawater. Folks claiming it is only O2 (so must be safe lol) are not correct, as there are also other redox active species (e.g., reduction of Cu++ to Cu+) present that might somehow end up lowering ORP.


Third theory relates to stabilizers that might be in the peroxide that may be more redox active than the peroxide itself. Not sure how that would work though.
 
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Jay Hemdal

Jay Hemdal

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Hi Jay,

A great many people have found that adding hydrogen peroxide to their reef aquaria causes a drop in ORP, not the expected rise. I have a couple of theories about why, but I was wondering that since you say a rise in DI water, if you happened to check it in marine aquarium water of some sort?

One theory is that it causes local killing and breaking open of bacteria, and the low ORP intracellular contents end up driving down ORP more than the peroxide initially raised it.

Second theory gets really deep in the weeds of what exactly is formed when hydrogen peroxide breaks down in seawater. Folks claiming it is only O2 (so must be safe lol) are not correct, as there are also other redox active species (e.g., reduction of Cu++ to Cu+) present that might somehow end up lowering ORP.


Third theory relates to stabilizers that might be in the peroxide that may be more redox active than the peroxide itself. Not sure how that would work though.
Hi Randy,

Sorry, I did not test the ORP on operating marine tanks after H2O2 additions. We have an old Hanna probe, and really, the ORP values it gives are always suspect to me. I would agree with your hypothesis, as the ORP rise that I saw was really not very great, and it makes sense that bursting cells could change the direction of that value pretty easily.

Regarding different species of peroxide - I didn't think about that, but that sure makes sense, I saw that with some ozone work I did back in the late 1980's

The peroxide we use is the same that I use on my home well - Culligan 7%. I was told that it doesn't contain stannous stabilizers, but I have no way to confirm that.

Peroxide chemistry is not my forte! (well, no chemistry is really) I just ran this series of bench tests to try out some hypothesis regarding other possible uses for it at my facility (other than washing filter socks). My big issues is that I have a 11,000 gallon Malawi cichlid tank packed with fish and chronic low DO (75% saturation). One option would be to supplement with bottled O2. I was hoping that I could use H2O2 to boost the O2. Based on how I killed off the nitrifiers in the goldfish tank test, I'm loathe to try that on a system with 4000 cichlids in it (grin).

For me - R2R members should only have three "takeaways" from this "rushed" material -

1) There are hydrogen peroxide test strips available on Amazon that work pretty well.
2) H2O2 does NOT decompose "in minutes" as posted here in numerous places
3) It has some detrimental biological effects on beneficial bacteria.


Thanks,

Jay
 

taricha

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Thanks!

What do we make of 150+% of max Dissolved O2 (as the h2o2 was decomposed in the presence of algae)?
Is it actually possible to supersaturate water to that level temporarily, or did h2o2 interfere with the O2 measurement (chemistry kit or DO meter?)


3) It has some detrimental biological effects on beneficial bacteria.
fascinating.
Looks like in FW it knocked out enough nitrifiers to create measurable ammonia/no2.

But in SW, it didn't create ammonia/no2. My interpretation is it still could have harmed the nitrifiers, but the remaining algae etc in the system picked up the ammonia/no2 slack.
 
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Jay Hemdal

Jay Hemdal

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Thanks!

What do we make of 150+% of max Dissolved O2 (as the h2o2 was decomposed in the presence of algae)?
Is it actually possible to supersaturate water to that level temporarily, or did h2o2 interfere with the O2 measurement (chemistry kit or DO meter?)



fascinating.
Looks like in FW it knocked out enough nitrifiers to create measurable ammonia/no2.

But in SW, it didn't create ammonia/no2. My interpretation is it still could have harmed the nitrifiers, but the remaining algae etc in the system picked up the ammonia/no2 slack.
It was supersaturation of O2, when I aerated the sample it de-gassed really quickly.
Jay
 

Dan_P

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Interesting study that raises some questions. Here they are.

How did you come to the conclusion that H2O2 is easier to rinse out than bleach?

While toxic compounds can be produced by bleach? What is the evidence that residues are left on the equipment?

An increase in ammonia may not be an indication that H2O2 harms nitrifying bacteria. Ammonia could have increased from heterotrophic bacteria consuming organisms killed by H2O2.

By the way how did you measure NH3 in the Betta experiment? Did you actually measure total ammonia?

In the Betta experiment, the dose gave a 5 ppm H2O2 level. Was this measured or calculated? What was the H2O2 concentration after 24 hours? Same questions for the Biocube experiment.

What seawater sample did you use in the H2O2 stability test? Aquarium water rapidly consumes H2O2 for the very reason you state: the presence of peroxidase.

The pH was rather low in the test of hydrogen peroxide in DI water. Why was it so low? Would you have observed the same results at the pH the algae normally lives at?

Thanks!
 

brandon429

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I would like to see nitrifiers / nitrification verified killed or slowed in a reef tank this way:

a calibrated seneye is used (able to show thousandths ppm active rate on a typical reef initially)

or the hach lab nh3 gear

have a test reef, any bucket of currently matured reef rocks represents what we all encounter daily on a valid scale. Have some aged sand from a current reef (organics carryover matching model) + live rocks a few pounds, plenty of current, and input a few drops of ammonium chloride into the system and show the reading spike to hundredths then drop back to thousandths, calibration proof in action of the collective biofilter

dose one mil 3% common medical grade peroxide per ten gallons twice a day, zap with same ammonia a few times on the digital graph, show where cannot maintain thousandths at the end of some two a day sustained dosing runs. This is the working dilution nearly all reef tanks used, merely a random amount once began a decade ago and everyone ran with it. Highest target kill / non target preserve rate dose honed by the masses is one mil per ten gallons common off the shelf 3% oral antiseptic. I estimate it’s use in reef tanks to be half a million posted doses online now.
 

Dan_P

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Hi Jay,

A great many people have found that adding hydrogen peroxide to their reef aquaria causes a drop in ORP, not the expected rise. I have a couple of theories about why, but I was wondering that since you say a rise in DI water, if you happened to check it in marine aquarium water of some sort?

One theory is that it causes local killing and breaking open of bacteria, and the low ORP intracellular contents end up driving down ORP more than the peroxide initially raised it.

Second theory gets really deep in the weeds of what exactly is formed when hydrogen peroxide breaks down in seawater. Folks claiming it is only O2 (so must be safe lol) are not correct, as there are also other redox active species (e.g., reduction of Cu++ to Cu+) present that might somehow end up lowering ORP.


Third theory relates to stabilizers that might be in the peroxide that may be more redox active than the peroxide itself. Not sure how that would work though.
The ORP drop is also observed with the addition of H2O2 to a NaCl solution.
 

Malcontent

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Peroxide chemistry is not my forte! (well, no chemistry is really) I just ran this series of bench tests to try out some hypothesis regarding other possible uses for it at my facility (other than washing filter socks). My big issues is that I have a 11,000 gallon Malawi cichlid tank packed with fish and chronic low DO (75% saturation). One option would be to supplement with bottled O2. I was hoping that I could use H2O2 to boost the O2.

Have you considered air stones? Or an oxygen concentrator?

What do we make of 150+% of max Dissolved O2 (as the h2o2 was decomposed in the presence of algae)?
Is it actually possible to supersaturate water to that level temporarily, or did h2o2 interfere with the O2 measurement (chemistry kit or DO meter?)

It's certainly possible. I've read on other forums that people think it's impossible for some reason. I have a fish that apparently has some permanent gill damage (perhaps from chloramine exposure--took him to the vet twice). He has his own 40 gal tank and every day I turn on the oxygen concentrator for 15 minutes. That will take saturation to over 300% and it will still be >100% 48 hours later.
 

Dan_P

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Have you considered air stones? Or an oxygen concentrator?



It's certainly possible. I've read on other forums that people think it's impossible for some reason. I have a fish that apparently has some permanent gill damage (perhaps from chloramine exposure--took him to the vet twice). He has his own 40 gal tank and every day I turn on the oxygen concentrator for 15 minutes. That will take saturation to over 300% and it will still be >100% 48 hours later.
This study decomposed H2O2 with iodide to produce O2 concentration 100 times the equilibrium solubility.
 

taricha

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Dan_P

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Ill-tempered seabass:

Oh oh, this article is going to stimulate the H2O2 craze, but at least it brings a bunch of science. Addition of H2O2 via the skimmer appears to be the best dosing point.

@brandon429 , check this article. Biofilter impact by H2O2 tested. At higher waste levels, H2O2 might not bother it. In cleaner water it might. Something @taricha might answer in his nitrification study :)

Test on fish also provides good information.

@taricha , check out disinfection discussion. I am wondering how the boost in O2 level impacts nuisance microorganisms.
 
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brandon429

why did you put a reef in that
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Very happy: one of the best benchmarked seneyes out there is running the test in an established reef today, we should make predictions ahead of time: prediction = tank waste immediately offsets any harmful characters from the dose and no change in thousandths ppm stasis occurs. Will see initial test back in a few hrs

if the dose can move Jons meter into hundredths on that full reef it’s a confirmed insult to the bac
 
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piranhaman00

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I’m a microbiologist at a whey processing plant, we make protein powder among other things.

We started dosing H2O2 at the beginning of the process to control micro growth at approx 150ppm. I expected the H2O2 to be fully dissipated by the end of the process and for sure after it’s dried at 350F.

I have found the peroxide is still detectable at the end of the process even after 60 days. Take a full 90 days to fully dissipate. Really strange as I thought it would be gone after a few hours.

Works crazy well especially at such high concentrations :)
 
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Jay Hemdal

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Ill-tempered seabass:

Interesting paper. This was a seawater system, and I had issues with a FW biofilter. They also discussed having enough organics in the system to break down he peroxide before it enters the biofilter. That is really tricky to do as the organic loading changes with increased peroxide dosing.
Jay
 
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Jay Hemdal

Jay Hemdal

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Have you considered air stones? Or an oxygen concentrator?



It's certainly possible. I've read on other forums that people think it's impossible for some reason. I have a fish that apparently has some permanent gill damage (perhaps from chloramine exposure--took him to the vet twice). He has his own 40 gal tank and every day I turn on the oxygen concentrator for 15 minutes. That will take saturation to over 300% and it will still be >100% 48 hours later.
We’ve added tons of airstones. Water changes are limited to 15% at a time, but we change 3x week. The system is filtered with 3 1hp 48” sand filters, these are oxygen sinks as well. I hadn’t looked into oxygen concentrators, just bottled gas. The trouble is the tank is designed to be a “wall of fish” but the fish grow and reproduce. We are culling, but can’t keep up. The fish are fine so far, I’m just worried we’ll reach a tipping point.
Jay
 
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