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OfflineP.Menace
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Re: Salicylic Acid Agar [Re: Psuper]
    #8727402 - 08/05/08 01:31 AM (15 years, 7 months ago)

Well, After doing a lot of reading and research, I found that Salicylic acid belongs to the Dihydroxybenzoic acids group.
These acids are 1 of that are used as additives in Antifreeze for Gasoline and Diesel Engines.
Being a mechanic I do know that the addition of Antifreeze (ethelyne-glycol) to water effectively raises the boiling point of the water, based on %, yet never exceeding a raise of more than about 15oF.
And adding Pressure to water also effectively raises the boiling point of water by 3oF for every 1 PSI gained.

So, I am to assume that since the Acid is used as an additive in Automotive Antifreeze, there would theoretically be no degradation of the substance under normal Sterilizing procedures, since after all water boils @212oF and if 15psi is added the boiling point of the water would be 252oF, and the range of most commercial antifreeze once mixed with water is -24oF to +272oF


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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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OfflineMycoAu
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Re: Salicylic Acid Agar [Re: P.Menace]
    #8728686 - 08/05/08 11:59 AM (15 years, 7 months ago)

You should be fine with that substance up to much higher temps than you'll probably ever see in substrate sterilization and probably even in dry heat sterilization temperatures for equipment.

most of the simpler benzoic acids are very stable at higher temperatures.

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OfflineP.Menace
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Re: Salicylic Acid Agar [Re: MycoAu]
    #8728783 - 08/05/08 12:26 PM (15 years, 7 months ago)

I know right. Its because of the Organic salts contained within the acid that allows it to reach such high temps without allowing for any degradation.

See what some simple off site reading can do for ya:thumbup:

I am PCing a 2.5% and a 5% acid/agar solution right now.
I will pour it and take some pics for documentation purposes.


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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Re: Salicylic Acid Agar [Re: P.Menace]
    #8732149 - 08/06/08 01:40 AM (15 years, 7 months ago)

Nicotinic acid should also be heat stable. Its melting point is 236.6 °C, it doesn't have a boiling point though because it decomposes before it boils.


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Re: Salicylic Acid Agar [Re: BlimeyGrimey]
    #8732191 - 08/06/08 02:03 AM (15 years, 7 months ago)

Well. As for the first experiment.

!. 2% Salicylic Acid Concentration:
Acid added before PC cycle
Ph. 6.8
Failed
2. 5% Salicylic Acid Concentration:
Acid added before PC cycle
Ph. 6.4
Failed

The Agar never re-constituted during cool down, also has a darker Brown coloration than normal.

On to Experiment 2:
Adding the Acid to the Agar after the PC cycle, in front of flowhood.

Make observations:
Color before and after PC cycle and the addition of the Acid
Re-Constitution Time vs. Normal MEA without Acid additive


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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Re: Salicylic Acid Agar [Re: P.Menace]
    #8733261 - 08/06/08 11:27 AM (15 years, 7 months ago)

Why don't you try rising the pH a little bit? In acidic conditions agar doesn't solidified as it should.

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Re: Salicylic Acid Agar [Re: Cindor]
    #8733303 - 08/06/08 11:35 AM (15 years, 7 months ago)

The Ph of the agar without the acid additive is 6.9... Im not that far off.
I will try that if adding the acid to the agar after the PC cycle fails. (experiment underway)

I have also take my Failed agar attempts, tested Ph today and see that it is at 6.8-6.9, so the Ph stabilized itself over 12 hours, yet the agar never set up.
What I might do is try an LC with like 30% Acid Agar solution to Water, and at least test its toxicity to Myc. If there is growth then it could potentially show that the solution isnt toxic to myc. and only impedes the agar itself.

I wonder if Salicylic Acid acts as a surfactant?


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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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OfflineMycoAu
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Re: Salicylic Acid Agar [Re: P.Menace]
    #8733395 - 08/06/08 12:03 PM (15 years, 7 months ago)

Here's an idea for you to try.  Take your agar, make it per usual procedure, but leave out enough volume to allow for addition of the acid solution after autoclaving your agar.  The acid solution should be made up  heated to near boiling (probably 80-90*C) and sterilized while still hot by using a syringe filter and added directly to the agar solution (that is still hot) with stirring (available for a lot of different places).

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Re: Salicylic Acid Agar [Re: MycoAu]
    #8733418 - 08/06/08 12:09 PM (15 years, 7 months ago)

Quote:

MycoAu said:
Here's an idea for you to try.  Take your agar, make it per usual procedure, but leave out enough volume to allow for addition of the acid solution after autoclaving your agar.  The acid solution should be made up  heated to near boiling (probably 80-90*C) and sterilized while still hot by using a syringe filter and added directly to the agar solution (that is still hot) with stirring (available for a lot of different places).




Im all on that already. The Acid solution is in a pyrex tube so i can boil it. I used up some of the agar I made yesterday at the same time as the Acid agar, re-heated it and Im about to go in and finish up this business, and see if the agar sets up this time.
If not what i will try is pouring plates of basic agar and dropping a drop or two of the solution on the plates after the agar sets up, then moving the dishes around and spreading out the solution... on a second thought, why do that tho, Im really only going to need the acid in the center of the trays where I will be placing the Myc. samples, the genetics will hopefully exchange there and then move on out beyond that point on the tray, where IMO the acid may be useless.
I dunno, does that seem right? cause its almost like the best Idea IMO. may give that a shot if this next experiment fails.


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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Re: Salicylic Acid Agar [Re: P.Menace]
    #8733504 - 08/06/08 12:29 PM (15 years, 7 months ago)

The idea about using a syringe filter to add a drop to the center of the plate is great.  The only true problem I foresee (at the moment) is being able to control the effective pH and concentration at the site of the drop. 

This may not change the effectiveness of the technique though- it is definitely worth a try.

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Re: Salicylic Acid Agar [Re: MycoAu]
    #8733606 - 08/06/08 12:48 PM (15 years, 7 months ago)

I *could* always test the Ph of the Acid in water, see where it baselines and perhaps adjust that solution.
But i did happen to notice yesterday that the Agar was a bit low and today it was almost neutral again. this could mean one of two things.
1. the agar, malt, dex and whatnot neutralized the acidic nature of the beast.
2. the Salicylic Acid itself broke down in 12 short hours.

Ideas? thoughts?
I could always try a test and see if it returns to a more neutral Ph after 12-24 hours in just water, try neutralizing it some.
I could even give er a go by placing 2 seperate strains in the center of an agar plate, allow them to foothold and create the obvious territories and then put a drop of sterile Salicylic acid solution right on the myc. itself... technically that should work too right?


--------------------

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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Re: Salicylic Acid Agar [Re: P.Menace]
    #8734979 - 08/06/08 05:11 PM (15 years, 7 months ago)

The boiling point of salicylic acid is  211 °C.  Typical autoclave temperature is 131 °C.  You should be fine.


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OfflineP.Menace
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Re: Salicylic Acid Agar [Re: Seuss]
    #8735031 - 08/06/08 05:20 PM (15 years, 7 months ago)

Yeah, the temp isnt the proving problem... its the fact that it does something to the agar itself.


--------------------

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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Re: Salicylic Acid Agar [Re: P.Menace]
    #8735273 - 08/06/08 06:04 PM (15 years, 7 months ago)

Be sure to check the pH after a 24 hour period.  If it is self-adjusting with time, then you've got a winner.  Of course, growing the mycelium out and then placing the acid on top.  You could also place a line of acid solution on the plate, let it set until the water is evaporated or absorbed and then use the plate for growth of the two strains.  This way, any mycelium crossing that line is sure to have gained the benefit of the acid.

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Re: Salicylic Acid Agar [Re: MycoAu]
    #8735301 - 08/06/08 06:12 PM (15 years, 7 months ago)

The ph will be tested later on tonight after im sure 24 hours have passes... but its not SOLID the acid destroyed the "agar" properties of the AGAR. Turns out that salts can do that when added to agar, and salicylic acid is a salt.

About the acid on the trays, or myc. That was already discussed earlier on in this thread. the best idea yet was mine to put a portion in the center of the tray and place my new transer material on top in the center of the acid... there are still problems with this tho. Where to transfer from AFTER the acid plate, to ensure that im choosing the best possibility for a crossed set of genetics, thats why i suggested allowing it to grow a bit first and perhaps placing the acid on the line (visible line) between the 2 strains, and see if they Merge, if they do i can almost be sure that the set of genetics at/after the "merge" are crossed.
sound theasable?


--------------------

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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Re: Salicylic Acid Agar [Re: P.Menace]
    #8735419 - 08/06/08 06:47 PM (15 years, 7 months ago)

Ok, first I want to clarify something.  Salicyclic acid is NOT a salt.  It is an organic acid.  Because of certain properties, it dissociates easily in water or aqueous based mediums (ie agar).  These properties, along with the acids ability to catalyze hydrolysis reactions are what destabilize the agar-agar gelling action and destroy the molecules that make up agar.  I don't care to go into any more detailed explanation than that.  (I'm only pointing out some facts that needed to be clarified, please don't take it for more than that.)

Second, other than drawing tissue samples from some area inside the known acid-treated agar, I don't know how you could best isolate or choose a region of crossed-genetic material.  I'm sure that to some degree it will be a crap-shoot.  Use a needle with some water, loosen some of the tissue in the acid-treated zone, suck it into the syringe and transfer to multiple plates.  Then, grow it out and check for fruiting strains.  Isolate strains from tissue of the fruits from here on out.  Be sure to do many, many initial transfers of the acid-treated tissues to help ensure your chances of getting crossed strain that fruits.

(I hope that "answers" your question - or at least is covering information you were most intereted in.)

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Re: Salicylic Acid Agar [Re: MycoAu]
    #8735620 - 08/06/08 07:26 PM (15 years, 7 months ago)

Taken from:
http://www.freepatentsonline.com/7387748.html
Sorry if the off site link gets me in trouble, but I feel its good information here.
This is an article on Antifreeze, read if your interested... I was.
Quote:

DESCRIPTION OF THE PREFERRED EMBODIMENTS

In accordance with the present invention, it has been found that the high temperature stability of glycol-based antifreeze coolant compositions can be enhanced by the addition of certain additives comprising organic compounds with a carboxylic acid moiety and a hydroxyl moiety, and also tricarballylic acid. Examples of such additives include alkali salts of the mono-, di- and trihydroxy benzoic acids and their derivatives. These include but are not limited to the salts of salicylic acid, acetylsalicylic acid, the resorcylic acids and gallic acid.

Monohydroxy benzoic acids include salicylic acid (2-hydroxybenzoic acid), 3-hydroxybenzoic acid, and 4-hydroxybenzoic acid.

Dihydroxybenzoic acids include 2,3-dihydroxybenzoic acid, 2,4-dihydroxybenzoic acid, 2,5-dihydroxybenzoic acid, 2,6-dihydroxybenzoic acid, 3,4-dihydroxybenzoic acid, and 3,5-dihydroxybenzoic acid.

Trihydroxybenzoic acids include gallic acid (3,4,5-trihydroxybenzoic acid) and 2,4,6-trihydroxybenzoic acid.

Benzenediols include 1,2-benzenediol (pyrocatechol), 1,3-benzenediol (resorcinol), and 1,4-benzenediol (p-hyroquinone).

Especially effective are the alkali salts of tricarboxylic acids such as citric, isocitric and tricarballylic acids.




And totally on the multiple inital plates and transfers, I was already intending on somewhere in the realm of 40 trays (2 sleeves) treating them somehow, probably after the agar has set up in the trays... that seems the best idea after all of the evidence stated against treating the agar itself


--------------------

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roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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Re: Salicylic Acid Agar [Re: P.Menace]
    #8736617 - 08/06/08 10:48 PM (15 years, 7 months ago)

I see where you picked up on the "salt" thing now. 

That does bring up an interesting idea that I can't believe I overlooked earlier.  Get some salicylic acid and then choose a salt of that acid (ie- sodium salicylate or preferably the calcium salt).  The use of both the acid and a salt of that acid in the same mix will create a buffer system that, depending on your choice of salt, will buffer your agar at deferent pHs. 

Do a little research into the formation of buffer systems (quick google search) and you'll quickly figure out what I'm refering to.  Then, you can prepare your stuff in the PC and likely have it set up (gel) properly afterwards as long as the acid/salt concentrations are not too high.

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Re: Salicylic Acid Agar [Re: MycoAu]
    #8753969 - 08/10/08 04:16 PM (15 years, 7 months ago)

Well. after some small experimentation, I found that Salicylic Acid does not dissolve in ambient temperature water, the temperature must be brought up to around 200of before the acid dissolves, so in a small brash of experiments here are some results:

1. Sterilized water, ambient temp. salicylic acid added 1 gram to 100ml water
Observations made: Does not appear to dissolve any, even after vigorous agitation, so the solution was brought to near boiling with small torch, then acid began to dissolve into water, Ph tested. 6.6. water was allowed to cool once all acid was dissolved, 24 hours acid stays suspended in water after this. ph tested again 6.7. small to no change
2. Boiling Acid solution, Small amounts of lime added to raise ph, clouded water badly didnt personally care for the result. lime settled out of solution after 24 hours
3. Boiling acid solution, small amounts of "target 7.0" added until ph raised to 7.0. water clouded at first then cleared up as it cooled:thumbup: Ph tested after 24 hours 7.0 (good product)

Now that all of my trays have been poured with a slightly "light" MEA mix, cured and sat for a few days now with no contaminates appearing its time to give a few trays a shot of my Salicylic Acid Solution and watch for results, decomposition of agar or whatnot, many random outcomes we may be able to expect of this. it could potentially break down the agar even after it has set. if so i will need to try a smaller ratio of solution.:shrug:


--------------------

http://www.sporeworksgallery.com/Cubensae/Psilocybe_cubensis_Menace


roby000 said: thats true a shotgun is almost like a college degree in a sense that if you show it to the right person at the right time you could make a lot of money.

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