Prime Does Not Remove Ammonia

SMSREEF

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I can easily adapt guppies to saltwater fairly quickly. They would work - I still expect an ethical complaint from people here - I'm receiving my Seachem multi-tests tomorrow. I'm going to approach it a little differently that @Dan_P
I don’t see any other way to know for sure. I wish there were another way. But using feeder fish seems the most ethical path.

I am very interested in these results. And like you, I have no clue whether prime actually detoxifies ammonia.
 

Rick Mathew

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I can easily adapt guppies to saltwater fairly quickly. They would work - I still expect an ethical complaint from people here - I'm receiving my Seachem multi-tests tomorrow. I'm going to approach it a little differently that @Dan_P
Would you care to share your experimental design....It would be very interesting to me to see how you are going to approach this. It would seem to me to be quite a complex data acquisition using live animals response as one of the data points, at least from my limited experience. Would be very interesting to see what you are thinking...In my view it will be quite an accomplishment to pull this off...or am I thinking about this all wrong..:oops:
 

Brian_68

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I don't follow that. Since NH3 is the primary toxic form, and Dan showed it did not change before and after, how does that not refute a claim to detoxify ammonia by converting it into a nontoxic form? That is the exact Seachem claim.
Would that not have to assume the measurement system is accurate, which in this case is uncertain at best. Sure it did not change color but is it even working in the first place, and how much does it change vs. concentrations applied. If you are going to prove something you need to ensure your measurement system is accurate and repeatable.
 

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I don’t see any other way to know for sure. I wish there were another way. But using feeder fish seems the most ethical path.

Well, you could use Daphnia which are commonly used in aquatic toxicity testing. For saltwater you'd have to find an equivalent critter.
 

MnFish1

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Well, you could use Daphnia which are commonly used in aquatic toxicity testing. For saltwater you'd have to find an equivalent critter.
Yes - @Dan_P tried that with (I cant remember) - but it required an extremely high pH - and high ammonia level like 8 ppm - which was outside the parameters that the Seneye measures accurately
 

MnFish1

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Would you care to share your experimental design....It would be very interesting to me to see how you are going to approach this. It would seem to me to be quite a complex data acquisition using live animals response as one of the data points, at least from my limited experience. Would be very interesting to see what you are thinking...In my view it will be quite an accomplishment to pull this off...or am I thinking about this all wrong..:oops:
I was thinking of 2 tanks. One pre-dosed with Prime one not (otherwise identical 5 gallon tanks) - with let say 15 guppies each - that have been acclimated to SW.

The water (pH, temp, filtration, etc would be equal. I.e. there would be no filtration.

The ammonia would be measured at the start - and then each tank adjusted to (total) ammonia - 0.25 ppm - then gradually increased until there was distress in either tank. The tank with Prime would be blinded (i.e. the person watching the fish would not know which tank had prime).

Directions on the Prime bottle would be followed (ie As ammonia levels increased Prime would be increased according to directions)

Distressed fish would be removed and euthanized immediately - per preset criteria (suggestions?)

Endpoints would be time of first fish removal, time of 50 percent fish removal and time of 100% fish removal.

Comparison would be between the 2 tanks.
 

MnFish1

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Yes - @Dan_P tried that with (I cant remember) - but it required an extremely high pH - and high ammonia level like 8 ppm - which was outside the parameters that the Seneye measures accurately
I think it was amphipods
 

SMSREEF

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Well, you could use Daphnia which are commonly used in aquatic toxicity testing. For saltwater you'd have to find an equivalent critter.
I don’t know that I would trust a test that was not done on fish to translate and be applicable to fish. Maybe it would… But testing on fish seems like it would remove that variable.
 

Malcontent

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Distressed fish would be removed and euthanized immediately - per preset criteria (suggestions?)

You don't have to euthanize them. Just move them to clean water.

Yes - @Dan_P tried that with (I cant remember) - but it required an extremely high pH - and high ammonia level like 8 ppm - which was outside the parameters that the Seneye measures accurately

What happens if the same thing happens in your experiment (TAN going over 5 mg/L)?

In a bit of irony, I'm dealing with a moderate ammonia spike in my freshwater tank right now.
 

Randy Holmes-Farley

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Would that not have to assume the measurement system is accurate, which in this case is uncertain at best. Sure it did not change color but is it even working in the first place, and how much does it change vs. concentrations applied. If you are going to prove something you need to ensure your measurement system is accurate and repeatable.

I'm quite confident that was exactly what Dan did, but I'll let him speak for himself.

This is not hard. Take water with no ammonia, and demonstrate the tests show no ammonia. Then add ammonia and see that the tests show ammonia. Then test the same ammonia plus Prime and see what the test shows.

If the Prime plus ammonia shows little or no ammonia, the Prime reduced ammonia. If it shows the same as without Prime, it did not.
 

Rick Mathew

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I was thinking of 2 tanks. One pre-dosed with Prime one not (otherwise identical 5 gallon tanks) - with let say 15 guppies each - that have been acclimated to SW.

The water (pH, temp, filtration, etc would be equal. I.e. there would be no filtration.

The ammonia would be measured at the start - and then each tank adjusted to (total) ammonia - 0.25 ppm - then gradually increased until there was distress in either tank. The tank with Prime would be blinded (i.e. the person watching the fish would not know which tank had prime).

Directions on the Prime bottle would be followed (ie As ammonia levels increased Prime would be increased according to directions)

Distressed fish would be removed and euthanized immediately - per preset criteria (suggestions?)

Endpoints would be time of first fish removal, time of 50 percent fish removal and time of 100% fish removal.

Comparison would be between the 2 tanks.
Thank you...This should be interesting...Looking forward to your report....By the way who gets the "night shift" watching the tanks :)
 

MnFish1

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You don't have to euthanize them. Just move them to clean water.



What happens if the same thing happens in your experiment (TAN going over 5 mg/L)?

In a bit of irony, I'm dealing with a moderate ammonia spike in my freshwater tank right now.
They are being bought for feeding either way.

The problem with the other study (again to my recollection) - is that amphipods are much more resistant to ammonia than fish. Thus I believe that 8 ppm ammonia and pH 8.6 was used. That should not be required here.
 
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Dan_P

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Why disregard Seachem alert badge result?

Maybe Dan can post the procedure used with it and the results. :)
How Seachem Alert Detected No Ammonia Removal By Prime

Seachem Alert ammonia sensor detects free ammonia. It is semi-permeable membrane embedded with dye molecules that becomes colored when ammonia is present, the color becoming darker with increasing concentration of ammonia. Full color development requires about five minutes. If Prime removes all the ammonia from solution, the Seachem Alert will not change color when exposed to the solution. To ensure objective observations of color change of the sensor, the intensity of the light reflected from the sensor was measured with a spectrometer. The wavelength selected (603 nm) coincides with the absorbance of the dye when it reacts with ammonia. The graph below demonstrates the response of the Seachem Alert to a 0.48 ppm total ammonia solution in Instant Ocean.

4DDAEE42-1231-409B-BA16-B0C28DF42AB4.png


The objective I had was to observe many exposures of the Alert sensor to ammonia in Instant Ocean or ammonia in Instant Ocean with five times the recommended dose of Prime. The protocol was to alternatively expose the sensor for a short fixed time to either the ammonia solution or the ammonia plus Prime solution. The Prime was allowed to react two hours with the ammonia before testing. If Prime removed ammonia from solution, the sensor absorbance would not change when exposed to the Prime containing solution but would change when exposed to the ammonia solution. The graph below shows the results from the experiment. Each sensor data point is labeled with the solution the sensor was exposed to. Despite the variation in absorbance data, the response curve to the mixed solution exposure resembles the response curve for the Alert sensor exposed to just ammonia (first plot).

5CE54927-9D42-4062-9FD9-7C2E59EA0735.png


The table below summarizes the results. There is little difference between the first pair of absorbances or the sum of absorbances for the entire series of exposures. The sensor response to the ammonia solution dosed with Prime is nearly identical to that of the ammonia solution.

27025A86-356A-48E7-990A-DC14EB6D2AE4.png


Moreover, the absorbance at 150 seconds in the Prime experiment was accurately predicted from the 150 second time point of the ammonia only solution experiment in the first plot. This seems pretty conclusive that Prime has no effect on ammonia concentration.

28285F54-5740-4576-A373-3D31CB79DE5F.png


Subsequent tests performed with two other ammonia sensors produced similar results. There is simply no evidence that Prime removes ammonia.
 

MnFish1

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How Seachem Alert Detected No Ammonia Removal By Prime

Seachem Alert ammonia sensor detects free ammonia. It is semi-permeable membrane embedded with dye molecules that becomes colored when ammonia is present, the color becoming darker with increasing concentration of ammonia. Full color development requires about five minutes. If Prime removes all the ammonia from solution, the Seachem Alert will not change color when exposed to the solution. To ensure objective observations of color change of the sensor, the intensity of the light reflected from the sensor was measured with a spectrometer. The wavelength selected (603 nm) coincides with the absorbance of the dye when it reacts with ammonia. The graph below demonstrates the response of the Seachem Alert to a 0.48 ppm total ammonia solution in Instant Ocean.

4DDAEE42-1231-409B-BA16-B0C28DF42AB4.png


The objective I had was to observe many exposures of the Alert sensor to ammonia in Instant Ocean or ammonia in Instant Ocean with five times the recommended dose of Prime. The protocol was to alternatively expose the sensor for a short fixed time to either the ammonia solution or the ammonia plus Prime solution. The Prime was allowed to react two hours with the ammonia before testing. If Prime removed ammonia from solution, the sensor absorbance would not change when exposed to the Prime containing solution but would change when exposed to the ammonia solution. The graph below shows the results from the experiment. Each sensor data point is labeled with the solution the sensor was exposed to. Despite the variation in absorbance data, the response curve to the mixed solution exposure resembles the response curve for the Alert sensor exposed to just ammonia (first plot).

5CE54927-9D42-4062-9FD9-7C2E59EA0735.png


The table below summarizes the results. There is little difference between the first pair of absorbances or the sum of absorbances for the entire series of exposures. The sensor response to the ammonia solution dosed with Prime is nearly identical to that of the ammonia solution.

27025A86-356A-48E7-990A-DC14EB6D2AE4.png


Moreover, the absorbance at 150 seconds in the Prime experiment was accurately predicted from the 150 second time point of the ammonia only solution experiment in the first plot. This seems pretty conclusive that Prime has no effect on ammonia concentration.

28285F54-5740-4576-A373-3D31CB79DE5F.png


Subsequent tests performed with two other ammonia sensors produced similar results. There is simply no evidence that Prime removes ammonia.
Thanks @Dan_P -This is interesting. Becasue this may help how I'm going to do my experiment. Just want to understand - you took a solution of .48 ppm total ammonia. - and added Prime to one solution and No Prime to another. Assuming pH 8 and Temp 78 - that would equate to a free ammonia level of 0.0226 ppm. If the pH were lower (or higher) - the level of free ammonia will also change.

According to Seachem, Safe is < .02 ppm and Alert is .05 ppm. My question - when you visually looked at the disk - what color was it? It must have been very close to 'safe'. i.e just barely above yellow. I just wonder - In your write up above you state that it shows that Prime does not 'remove all (free) ammonia' - but even if it only removed 1/2 - it would be in the 'safe' zone. I wonder if your test is sensitive enough to detect such small quantities?

I'm asking because I'm trying to decide how much ammonia to use in my seachem multi-test (which is supposed to be coming today). Also - did you soak the seachem alert for 24-48 hours before using?
 

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It was a colorimeter interpretation, mechanical assistance better than the human eye guess as I read it

I bet without that magnification we'll need seneye doing the analysis for us at those minute levels?
 

MnFish1

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It was a colorimeter interpretation, mechanical assistance better than the human eye guess as I read it

I bet without that magnification we'll need seneye doing the analysis for us at those minute levels?
I don't know Brandon - what the sensitivity of the spectrophotometer used is/was. But - it could be that any change in ammonia (free) - was too small to be seen.

I agree that the seachem instructions are extremely difficult to wade through. On one hand says that prime should nearly immediately 'detoxify' free ammonia. However, the Seachem alert states that once ammonia is present, it can take a couple hours to show any change in color.

Then in the dosing instructions it says the product should dosed based on 'ammonia'. It does not say 'total ammonia', or 'free ammonia'. -but I'm assuming they are talking about 'total ammonia'.
 
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Dan_P

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Thanks @Dan_P -This is interesting. Becasue this may help how I'm going to do my experiment. Just want to understand - you took a solution of .48 ppm total ammonia. - and added Prime to one solution and No Prime to another. Assuming pH 8 and Temp 78 - that would equate to a free ammonia level of 0.0226 ppm. If the pH were lower (or higher) - the level of free ammonia will also change.

According to Seachem, Safe is < .02 ppm and Alert is .05 ppm. My question - when you visually looked at the disk - what color was it? It must have been very close to 'safe'. i.e just barely above yellow. I just wonder - In your write up above you state that it shows that Prime does not 'remove all (free) ammonia' - but even if it only removed 1/2 - it would be in the 'safe' zone. I wonder if your test is sensitive enough to detect such small quantities?

I'm asking because I'm trying to decide how much ammonia to use in my seachem multi-test (which is supposed to be coming today). Also - did you soak the seachem alert for 24-48 hours before using?
I would say that within the variation of the data of the above data, Prime did not remove half of the ammonia. Even at higher concentrations of ammonia with other sensors, no reduction occurred with Prime. Ditto for Cloramx with 38 times the recommended dose. I hope someone takes the experiment to the next level with an ISE. Then small reductions in ammonia (<25%) might be detected.

I did soak the sensor per instructions.

In a separate experiment with a different Alert sensor, I ran up the ammonia to hit most of the colors on the Alert card. I was surprised how closely the sensor color matched the reference color patch. When I hit 0.05 ppm free ammonia, that was the color I visually saw. Ditto for the next highest color level. 0.02 ppm barely colored the sensor and maybe if it was a blinded observation, I might have called it <0.02 ppm.
 
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Dan_P

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It was a colorimeter interpretation, mechanical assistance better than the human eye guess as I read it

I bet without that magnification we'll need seneye doing the analysis for us at those minute levels?
Definitely will need something other than the human eye for sensitivity AND objectivity. This situation is no different than the error prone method of visually comparing the color of test solutions vs color chart.

I used the Seneye sensor recently, not the device itself, and measured the response with my spectrometer. It behaved well with Prime and Cloramx. Having worked with the Seneye device, it seems to be pretty good overall. It still needs calibration though. No short cuts doing good science.
 

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