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Author: Subject: Alternatives to Nitrogen.
NEMO-Chemistry
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[*] posted on 13-2-2017 at 17:09
Alternatives to Nitrogen.


I cant easily get Nitrogen (no jokes please), I need to transfer and sample some microbes. This is methane related, so I wanted to know if Hydrogen would work instead of Nitrogen?

I understand the small risk (small amounts involved), but as Hydrogen can easily be made on demand and on a small scale, can anyone see any reason not to use it to exclude Oxygen?

As its such a stupid question I will expand it slightly, I am pretty positive it would work, but you know how sometimes you think about ways around a problem, then you begin to doubt your right.

Thats kind of the situation, yes I could simply try it. Except I have had a really hard time getting this culture going, if I mess it up now I would throw the towel in with it.

Anyway thank you in advance. And sorry if its really stupid to ask!
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[*] posted on 13-2-2017 at 17:13


Just as a side note.................

I had always Assumed that cows fart methane, so obvious place to looks for bugs is cow shit. Months of failure looking for the bug I wanted made me turn to vet books, as I am sure you lot know cows actually belch more Methane than they fart.

Significance? Well the bug I was after is at the opposite end, so its taken ages to get a culture that wasnt dead.
And no this isnt actually about making Methane, its about a particular bug thats all (One i cant spell!).
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JJay
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[*] posted on 13-2-2017 at 17:17


I've wondered about using hydrogen as the atmosphere for doing metal hydride reductions before. I think it would work, but hydrogen is actually a pretty dangerous gas to work with. It's extremely flammable and tends to gather in high places like along the ceiling. Also, when you generate hydrogen, you'll need to purify it.

You can also generate nitrogen. From what I remember, it can be generated in extremely pure form from azides, and you can also generate it from sulfamic acid and sodium nitrite, although it will require some purification.




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[*] posted on 13-2-2017 at 18:05


Quote: Originally posted by JJay  
I've wondered about using hydrogen as the atmosphere for doing metal hydride reductions before. I think it would work, but hydrogen is actually a pretty dangerous gas to work with. It's extremely flammable and tends to gather in high places like along the ceiling. Also, when you generate hydrogen, you'll need to purify it.

You can also generate nitrogen. From what I remember, it can be generated in extremely pure form from azides, and you can also generate it from sulfamic acid and sodium nitrite, although it will require some purification.
Thanks for that.

I did consider making Nitrogen, but getting hold of sulfamic acid is likely a mare in the UK, I just thought Hydrogen was very cheap to make in small amounts, very easy and obtainable.

Quantity wise we are looking at a couple of 500ml conical flask type containers. Its kind of 'jam jar' scale, so i didnt think the amount would be an issue.

Its an attractive idea on a small scale, my main problem is I have half convinced myself it wont work! I guess I could use Propane?? Or maybe its Butane in lighter refill cans?

I got the starter culture from a slaughter house, but its been a real PITA and taken ages to get to this point. Dont get me wrong I am not doing anything that interesting (except to me), but you know how sometimes you put so much effort in...........I would be gutted to mess it up now :D.

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JJay
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[*] posted on 13-2-2017 at 18:10


Probably best to just buy it if you need it for something critical



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[*] posted on 13-2-2017 at 18:42


Quote: Originally posted by JJay  
Probably best to just buy it if you need it for something critical


The problem is getting hold of it in small amounts. Here it costs a fair bit for a cylinder.

Sorry my last post seems to read a bit rude?? Not sure why, rudeness isnt intended. Quantity wise we are probably looking at 1-2 balloons full (party type not people carrying type :D).

You would be surprised just how hard it is to get simple things around here, no decent sized towns for a long way.

Well I cant let it sit there forever, its declining as it is. So bite the bullet and try it I guess. As long as there is no Oxygen, I dont think it matters too much what else is in it (now watch that statement come back and bite me! lol).

Actually Butane might be better, its cheap and the small cans for lighters are cheap and pretty pure.... Sod it I will try it Friday! I should know within a few days if its worked or not.


Thanks for the comments, BTW much appreciated.
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[*] posted on 13-2-2017 at 23:27


Heat copper powder in air and it will remove the O2, leaving you mostly N2 and Ar. To get rod of CO2, use NaOH solution.



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[*] posted on 13-2-2017 at 23:53


Nemo:

Sulfamic acid is readily available in the UK from ebay and Amazon at about £10/500g
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[*] posted on 14-2-2017 at 02:13


Quote: Originally posted by vmelkon  
Heat copper powder in air and it will remove the O2, leaving you mostly N2 and Ar. To get rod of CO2, use NaOH solution.


You beat me to it. I was going to suggest, well, that we ARE surrounded by a 79% mix of it. If one doesn't want to heat copper powder, then one way that I remove oxygen from air is with acidic cuprous chloride. It would still be necessary to wash with NaOH, and dry.

Having said that, even if air is washed through a few bottles of CuCl solution, it's likely still necessary to scrub the remaining bits of oxygen with a heated copper catalyst with maybe a 2% mix of hydrogen in nitrogen. This is followed by a desiccant bed to remove moisture. It all depends on how sensitive your microbes are to oxygen.




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[*] posted on 14-2-2017 at 03:50


Some have built pressure swing adsorption system to get a steady feed of decent quality nitrogen. It'd be a lenghty build though...
Alternatively there is this reaction:

NH4Cl + NaNO2 → N2 + NaCl + 2 H2O

From here http://pubs.acs.org/doi/abs/10.1021/ed044p475
I'd guess you'd need a wet scrubber to ensure the evolving gas is free of contaminants, but still relatively simple.




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[*] posted on 14-2-2017 at 04:13


Easier would be to use CO2 from dry ice.
You can buy small quantities at ice cream stores.
Or use compressed CO2 that is sold in small containers for different uses.
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[*] posted on 14-2-2017 at 06:26


I was going to suggest CO<sub>2</sub> also. If your only aim is to exclude oxygen, it's heavier than air and easy to make or buy.
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[*] posted on 14-2-2017 at 07:46


CO2 is a bad idea for cell cultures. It will rapidly make everything acidic and probably wipe it out. You truly need an inert gas like nitrogen, but I'm sure argon would also work. Argon is sold cheaply at welding shops everywhere. But, I suppose if you could afford to buy an argon bottle, you could probably afford a nitrogen bottle as well.

Iron powder is great at eating up oxygen when treated with certain salts. It is the basis of operation for many disposable hand warmers. You might try opening up a bunch of those and packing a column with that material, and use a fish tank compressor to blow air through it.





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[*] posted on 14-2-2017 at 08:18


Quote: Originally posted by NEMO-Chemistry  
I cant easily get Nitrogen (no jokes please),


What country are you in? I bought a large cylinder of it for about £20 from B.O.C. last year. It is one of the cheapest and easiest to buy. If you live in the UK just ring them and place an order.




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[*] posted on 14-2-2017 at 10:51


Could you just use propane as your inert gas? Sure, it's flammable, but it's not like we don't know what precautions to take when dealing with a flammable gas. The ethyl mercaptan or whatever they put in gas to make it smell would be at an extremely low concentration. Plus, it has the benefit of being able to smell it if there are leaks.

edit: just realized that's what OP was suggesting and nobody seems to have answered him. I've considered using propane as an inert gas for reactions myself, at least when the scale is small enough that it poses little if any danger. If you're not using fire around it, I see no reason why it shouldn't work.

As for removing oxygen from air, one convenient way to do it is with those "Hot Hands" single-use hand warmers. They contain reduced iron powder, and some moisture and salt in order to catalyze the iron oxidation. If you wanted to remove the oxygen faster without generating CO2, you could ignite steel wool or magnesium. Eventually the metal would stop burning when the oxygen was gone.

[Edited on 2/14/17 by Melgar]
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[*] posted on 14-2-2017 at 13:32


Quote: Originally posted by DrP  
Quote: Originally posted by NEMO-Chemistry  
I cant easily get Nitrogen (no jokes please),


What country are you in? I bought a large cylinder of it for about £20 from B.O.C. last year. It is one of the cheapest and easiest to buy. If you live in the UK just ring them and place an order.


What size was this? I did ring them a while back and was told you had to hire cylinders, maybe i asked for the wrong thing?? They wanted £20+ a month rental and around £85 for the gauge, then the gas on top.

Someone else mentioned buying a cylinder from them, so I must be ringing the wrong division or depot.

If I can buy it for that kind of money then its the way I will go, the reason I havnt done it was the rental costs as I dont need much.

thanks for all the suggestions, CO2 does indeed seem to kill most methane cultures off. I thought of hydrogen because it was simple, and the quantity i need is small.

I will have another go at calling BOC and see if I can buy instead of rent a cylinder.

Thanks alot guys you have given me alot of options.

I have got myself a bit muddled with this now, its taken so long to get this far (far far longer than it should have), I cant see the wood for the trees now lol.
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[*] posted on 14-2-2017 at 15:01


H2 should be fine.
CO2 too.

Maybe use methane, ethane, propane, butane or ethylen, propylen, butadien or acetylen.
Alternatively Helium or Argon.

H2S may be too crictical but is often present next to CH4 into anaerobic riots.




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[*] posted on 14-2-2017 at 15:40


Many hardware stores around the world sell small (around 500mL) cylinders of argon for reasonably cheap (not cheap compared to a larger cylinder, of course, but perhaps cheap enough for your purpose). They're similar to the small propane cylinders for hand-held blowtorches or camping stoves.
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[*] posted on 14-2-2017 at 16:16


Another source for argon is wine shops. Cans of Ar are sold to fill the head space of opened bottles of wine so that they last longer.
https://www.danmurphys.com.au/product/DM_365521/winesave-som...

It is an expensive source on a mass basis but the cans are really convenient and a cheap option if all you need is a small amount.
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[*] posted on 14-2-2017 at 19:24


Great suggestions guys and thanks alot, I know its difficult for many to understand this, but I honestly live a long way from anything resembling a hardware shop.

The closest we get is farm supply shops, most farms have welding and cutting gear, but they get large cylinders delivered and they are rentals. I do occasionally go into the city but thats 4-5 hours round trip, ebay was good and so was Amazon. but the postage these days up here is a fair bit.

I am really glad I asked though, hearing the suggestions makes sense now. But I got to the point where I was unsure. Lighter gas came to mind, but it seemed too simple a solution, maybe because Nitrogen is mentioned in most books.

One reaction above that interests me is the Copper one. I know it says powder but would fine copper wires work? If it has to be powder, what kind of heat are we looking at?

My thinking is heating a container (no idea which yet), containing copper powder, using say nichrome wire to heat the container and therefore the copper.

I doubt its much use for this application, but I can see applications where passing air over something like that could be handy. I dont post much these days, dont know if it was Blogfast getting some trouble or what. But anyone else starting to feel like they are doing something 'wrong' every time they mix Bicard and vinegar?

Sorry I am going off topic :D, just all feels like a witch hunt going on.

Not sure if my project counts as chemistry, and Methane production (small scale) isnt exactly exciting, but if I get this final culture working, I will post it up if anyone is interested.
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[*] posted on 14-2-2017 at 19:26


Quote: Originally posted by j_sum1  
Another source for argon is wine shops. Cans of Ar are sold to fill the head space of opened bottles of wine so that they last longer.
https://www.danmurphys.com.au/product/DM_365521/winesave-som...

It is an expensive source on a mass basis but the cans are really convenient and a cheap option if all you need is a small amount.


I didnt know that! ironically we do have a home brew shop in town.......Might be a good place to mooch for other Chems as well :D.
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[*] posted on 15-2-2017 at 08:52


Again, I'm not sure how sensitive your microbes are to oxygen, nor do I have the experience to make that determination. Possibly, a small can of argon, or propane, might work for you, even if it isn't ultra high purity or something like that.

As to the question about copper, the reason for it to be in powder form is simply that of surface area. Its reaction with oxygen will be faster and more complete at a lower temperature (250-350°C). Copper wire would probably never react fully through its own bulk, except at very high temperatures and long periods of time. Keep in mind that isolating an inert gas can end up being just as much of a project as the one you're already working on, if not more so.

Anyway, another place to find argon is in light bulbs. If you have old burned-out incandescent bulbs, one could break the bulbs underwater (by placing a flat-head screwdriver at the base of the bulb, and then wacking it sharply with a hammer) and harvest the gasses in a bag. I wouldn't recommend using brand-new bulbs for that, but this project might be an excuse to replace some burned-out ones. Generally, the bulbs that have tight, coiled-up filaments contain argon; the ones with a long-straight, wire filament, contain a vacuum.

If you want to go fancy with isolating nitrogen, this can be scaled up:

https://www.sciencemadness.org/whisper/viewthread.php?tid=55...

The catalyst beads can be reduced to copper with hydrogen or alcohol vapors, and then the gas stream switched to air to remove its oxygen. Enough catalyst beads would need to be used to give you enough nitrogen to work with, more than what I used in the link. Or, the same depleted air can be passed multiple times through the repeatedly regenerated catalyst bed, until no color change occurs in the catalyst (from oxidation).

[Edited on 2-15-2017 by WGTR]




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[*] posted on 15-2-2017 at 10:40


If you break a lightbulb or two inside a plastic bag or bottle filled with water, you can collect a mixture of nitrogen and argon at the top and with a little rudimentary engineering, force it into a system. Of course you won't end up with completely oxygen-free air this way, but something tells me that 1 or 2 percent isn't going to make too big of a difference to most living things.
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[*] posted on 15-2-2017 at 12:42


The transfer is from liquid to liquid, so the simple solution is to just force out some of the liquid with the gas. the airline has a one way aquarium type valve, so I dont have to worry about any oxygen in the container itself.

I found some cheap argon cylinders from the wine shop. No idea what I thought was inside light bulbs (a vacuum I suppose), I didnt know it was/could be Nitrogen.

Some oxygen for most Methogens isnt the end of the world, however for some it is. I need to look the name up again, but one of the ones I am trying to isolate produces Hydrogen Sulphide in some conditions, but methane in others (different pathways).

In the bottom of lakes and other various places, it lives very happily and has no problems, but trying to isolate it!!!! utter nightmare. Its been alot of work, especially for something that is nothing more than fun/curiosity.

I appreciate the answers, I had kind of lost confidence in what I was doing. I am going to split the cultures into 4 and try a different gas with each.

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[*] posted on 15-2-2017 at 13:59


Liquid to liquid...then simply avoid any gas....only use liquid boiled water for example (almost O2 and N2 free...and sterilized)...use a seringe filled with that water to displace the fluid in front of it to the other location and on the other side use another empty seringe and pump the liquid...simple liquid transfert.

[Edited on 15-2-2017 by PHILOU Zrealone]




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