FlytrapCare Carnivorous Plant Forums

Use 1 part distilled white vinegar (5%) to 4 parts hydrogen peroxide (3%). Place the 1 part white vinegar in the microwave until it boils. Then add the hydrogen peroxide (3%). Let it sit in the fridge until it cools back down to room temperature.

A good soak time to start with on Dionaea leaves is 2 to 2.5 minutes. Plate the leaves without rinsing.

Hmm, should one wait until the vinegar cools before adding the H2O2?

hb

Hackerberry,

No, it's important that You add it while it is very hot. I guess it helps with the initial reaction, so I'm told.

gary

Thanks Gary!

hb

Peracetic acids are unstable in hot temperatures.

Reaction of acetic acid and hydrogen peroxide develops water,

CH3COOH + H2O2--> CH3COOOH + H2O

Reaction has low equilibrum constans, and if you use so much dilluted solutions, i think that in that mixture concentration of peracetic acid is very low, and real sterylising agent is oxygen developing during decomposition od H2O2 in acidic pH.
Normally it is prepared from anhydrous acetic acid and H2O2 30%, not 3%.

http://www.jproeng.com/qikan/manage/wenzhang/207239.pdf -interesting paper, something about kinetic model od reaction.

I really don't think that concentration of PAA in this mixture is high....

sativ wrote:Peracetic acids are unstable in hot temperatures.

Reaction of acetic acid and hydrogen peroxide develops water,

CH3COOH + H2O2--> CH3COOOH + H2O

Reaction has low equilibrum constans, and if you use so much dilluted solutions, i think that in that mixture concentration of peracetic acid is very low, and real sterylising agent is oxygen developing during decomposition od H2O2 in acidic pH.
Normally it is prepared from anhydrous acetic acid and H2O2 30%, not 3%.

http://www.jproeng.com/qikan/manage/wenzhang/207239.pdf -interesting paper, something about kinetic model od reaction.

I really don't think that concentration of PAA in this mixture is high....

Hi, Sativ!

You are right, It isn't high; We aren't making rocket fuel oxidizer.
You could be right on about the oxygen doing the work, as well. That is very interesting.

We call it PAA, for a lack of better term, and is easier to type than CH3COOOH + H2O, as it does contain some, whether 3% or whatever.

You add the H202 to the Very Hot Vinegar, this brings the solution of now forming Peracetic acid to slightly above room temperature and is always used at room temperature. It is never re-heated.

This is a very weak solution 'containing' Peracetic acid, that is great for being very aggressive on contaminates, (as evident of the formation of oxygen bubbles), but preserving clean, healthy tissue. (lack of bubbles).

But the keyword here is. . . SIMPLICITY. That is what everyone strives for, especially Kitchen enthusiasts. I have successfully formed 3 callous' on 3 Dionaea sections in the same batch! Not to mention the others (go to Peracetic acid in Sterilization thread).
This is with no Alcohol dip, no sterile rinses afterwards, no Peroxide soak, etc. . .
That is the main attractiveness to this methodology; the hardest part was the last 3 months of developing this protocol, with its timeline, experimenting, testing; waiting, waiting, waiting. . . as My hacked up Collection will attest to, but it has been worth it.

The chlorox method works great, especially with much more forgiving tissue like the African Violet, etc., but Dionaea leaves have always been hard, much harder to propagate, than alot of other species.

This alternative experimental method vastly limits the time the tissue is in the open air, going from jar to jar, sterile rinse to sterile rinse, (which has to be prepared fresh) which in itself, limits risk of contamination for those without a $3,000 laminar flowhood, and the benefit of getting very fresh, un-beat up tissue on the media quick so it can do it's thing. The timer is only used once.

Now with some much more experienced TC'rs jumping aboard and giving it a try, I think We can nail this thing down with further refinement, and I think We can bring up the number of strikes per batch with this stuff, as I was learning TC at the very same time I was experimenting with PAA. Made many mistakes. That is why I think someone with experience will be successful MUCH faster than I was.

See pics on page 10 of 5 week old Dionaea leaves, on the Peracetic Acid in sterilization thread. They look almost as fresh as the Day I plated them.

This solution has a great surplus of H202 (and water) adding oxygen to the weak PAA as it is consumed or gassed off; and will do so for a very long time; maturing at 1 month to full strength, if left at room temperature, (I've done some reading, too). I burned My explants to a crisp using some PAA at 2 weeks old at the same time line. This is (whatever it is) a very powerful oxidizing agent that is at a safe level for Us, but is still MORE than strong enough for what We do, hence the short soak times. People of the Yahoo list serv. even use it as an alternative to the 10% chlorox to clean their boxes, and use for a tool soak and sterilization, and are doing so successfully. I, too, use it as a tool soak, but it can rust out Your tools if forgotten; even stainless.

I'll let You go there and talk to Gregorio about the Chemistry, I'm sure He will enjoy that.


Gary

I'm doing one experiment and will let you guys know in a couple of days. I was doing Ceph leaves with PAA but vinegar was not heated and added to h2o2 right away. I will see some contamination if this did not go properly. I did 3 minutes soak with no rinse. Also, I cleaned the leaves first in running tap water.

hb

Thanks for the participation, Hackerberry!

Are Ceph leaves pretty hardy? Are they inherently pretty contaminated to start with?

Gary

Do not forget on the other forum they are using 100ml 5% vinegar solution. This was not mentioned until just recently (unless I missed an earlier post). I had already made up a solution of 100ml of 100% vinegar + 400ml H202 3%.
I have a client I have been working with for the last 2 years on doing bamboos they want 100,000 per year so it is worth while pursuing. I used 43 bamboo nodes from nursery grown plants. I did 43 different tests and had 43 contaminations in 24 hrs. I figure the 100% acetic acid neutralized the H202. I then made up a solution of about 5% (20ml 100%) vinegar + 420ml H202 3% and redid the tests. It is now over 24hrs. I have had a couple of contaminations but it is looking very good. The material I am using is one of the dirtiest plants I have tried to clean up (except for nepenthes). In the past I have used everything from antibiotics, bleach, H202, etc, etc, etc. I was so desperate I even used Mecuric Chloride - yes I do know how dangerous the stuff is. Most treatments failed to sterilize them or over sterilized them. If I obtain 30% success I will consider it a massive success.
Oh I nearly forgot I do not heat up the vinegar. I doubt it is necessary.
Geoff

Multiclone wrote:I then made up a solution of about 5% (20ml 100%) vinegar + 420ml H202 3% and redid the tests. It is now over 24hrs. I have had a couple of contaminations but it is looking very good.

Great! I'm excited to hear the contamination count after 5 days or so.

Multiclone wrote:Oh I nearly forgot I do not heat up the vinegar. I doubt it is necessary.

I haven't had chemistry since I was in college and I am in no way a chemist, but I think I've heard others say that they doubt it is necessary and I tend to agree with them and you Geoff.

Do not forget on the other forum they are using 100ml 5% vinegar solution.

Yes, it is grocery store strength in the USA. 5%

Oh I nearly forgot I do not heat up the vinegar. I doubt it is necessary.
Geoff

I've heard One other Person say that as well, but since I'm no Chemist, so I do like Monkey, and heat.
Gregorio insists on boiling the Vinegar first.

Good Luck and thanks for participating, this will be VERY INTERESTING!

Gary

goldslinger wrote:Yes, it is grocery store strength in the USA. 5%

Here in Australia off the supermarket shelf, 2ltrs for $1.09 100%.
Geoff

I use 12% acetic acid and it seems to work too. So far.
Also put some stuff in jars last week and everything looks great.

Now we need to make a thread about what PAA can do . When I read in the mailinglist the people seem to love it and use it for everything like clean the house....

Multiclone wrote: Here in Australia off the supermarket shelf, 2ltrs for $1.09 100%.
Geoff

I know you Aussies are a tough bunch, but that can't be right. Taking a good whiff of 100% acetic acid will burn your sinuses and make your nose run for a week as if you have a cold. I doubt eating it on a salad would be very pleasant either.