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Blue Juice: A new method of extraction using Ice by Paul Stamets


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#61 cg3p0

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Posted 08 February 2019 - 09:33 PM

Hyphaetion

 

I had been wondering about how to juice out mushrooms to get a concentrated liquid, looks like you figured it out.  I thought that was a meat grinder at first, but that wheatgrass juicer would probably be about the best machine for juicing them you could find.  From what we've uncovered, you could have dropped some kind of antioxidant in there to keep it from degrading turning that dark blue,probably just toss in a crushed vitamin C pill would work.  I thought it would be cool to make a concentrated liquid and get some blotter paper and soak it up and make mushroom acid tabs.



#62 cg3p0

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Posted 08 February 2019 - 10:24 PM

pharmer

 

I think soliver already did that test for us, his 40% blued and his  90% didnt, although your right about structuring the test better.  I would assume the 50% light blue, 75% barely blue, and 100%no blue.  Seriously any chemist could probably explain this in 2 seconds but i guess we dont got any here.  I know alcohol is used to preserve lots of stuff so its not suprising it appears to be preserving the psilocin.

 

 

I got a bunch of tests that SWIM wants to prefrom but it will be a few weeks.  Im more interested in trying to degrade the chemicals and then put them back together.  From psilocybin -> psilocin  -> blue and then back from blue -> psilocin -> psilocybin.  I think maybe this thread is getting derailed so i may post it elsewhere if there are any results.


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#63 hyphaenation

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Posted 09 February 2019 - 12:01 PM

Also ..if you agitate the mycelium of certain strains of cubes in water it will turn quite blue.

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#64 hyphaenation

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Posted 09 February 2019 - 12:50 PM

Yeah the wheatgrass juicer seems to be the right tool to squeeze the goods out. It would be good to mitigate the degrading of the alkaloids through light/oxidation etc. I hadn't heard of using citric acid...

The thought I had at the time was to try and create a liquid-M type solution that could deliver a drip & trip experience.

#65 cg3p0

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Posted 09 February 2019 - 03:01 PM

If you ever get around to trying to juice them again you might try rehydrating the pressed mushrooms with fresh water, about the same volume as you pressed out, and then juicing them a second and maybe third time as it looked like you still had lots of stuff left behind in the pressed mushrooms.  I had been wondering about that too, whats the best way to make sure you get it all out of the mushrooms.  I would think large volumes of liquid, like this cold water tek, or alcohol extraction, or making tea  would probably do a better job of getting all the goods out on the first go compared to juicing them.  However, if you wanted to concentrate the liquid, im not sure you wouldnt lose some to evaporation or something else, so the juicing would maybe have the advantage for concentrating it.  Im sure others must have tried concentrating thier extracts im just not aware of how well it turns out.  

 

Im guessing some of those strains had active mycelium, thats interesting!  You might be able to just make a bunch of liquid culture and just drink that, wouldnt have to go through the whole process of actually growing the mushrooms.  That would be awesome.  Probably worth further testing if you still got them around.



#66 RutgerHauer

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Posted 09 February 2019 - 04:27 PM

Interesting extraction! Simple.

Has anyone an idea why you would do this extraction in stead of for example making a tea?

 

RH


Edited by RutgerHauer, 09 February 2019 - 04:27 PM.


#67 PistolPete13

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Posted 09 February 2019 - 05:01 PM

pharmer

 

I think soliver already did that test for us, his 40% blued and his  90% didnt, although your right about structuring the test better.  I would assume the 50% light blue, 75% barely blue, and 100%no blue.  Seriously any chemist could probably explain this in 2 seconds but i guess we dont got any here.  I know alcohol is used to preserve lots of stuff so its not suprising it appears to be preserving the psilocin.

 

 

I got a bunch of tests that SWIM wants to prefrom but it will be a few weeks.  Im more interested in trying to degrade the chemicals and then put them back together.  From psilocybin -> psilocin  -> blue and then back from blue -> psilocin -> psilocybin.  I think maybe this thread is getting derailed so i may post it elsewhere if there are any results.

 

 

Hey sorry guys I have been soo busy lately, I had seen this post earlier and had a quick read but I did not know it had become more interesting......

 

It has been pretty widely documented in the literature that the bluing reaction is for the most part enzymatic, and an acid phosphatase is responsible. Psilocybin is not oxidized easily and is a lot more stable in aqueous solution than psilocin which oxidizes quite readily in aqueous solution. So the bluing reaction you are seeing as psilocybin is dephosphorylated by these enzymes which then leaves the psilocin is vulnerable to further oxidation (some researchers believe the psilocin forms dimers or trimers which basically means two or three molecules stick together forming these colored products) these enzymes are being extracted along with the actives and is not good for the shelf life of the actives in solution. Not a problem if you want to consume it there and then, it will actually feel stronger as the psilocybin will be for the most part dephosphorylated into psilocin which is available straight away.

 

So a gentle warming of shroom tea people (like Mikey once pointed out) produces a much stronger bluing than a fierce heat and boil which denatures the enzymes and stops the reaction.

 

A high strength alcohol will denature the enzymes and stop the reaction.

An aqueous alcohol will not denature the enzymes and stop the reaction.

 

I will be back! (when I get a minute).....


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#68 cg3p0

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Posted 09 February 2019 - 05:35 PM

PistolPete13

 

That is the kind of info i was looking for, thanks!  So its mainly enzymes that break down psilocybin.  Does drying mushrooms kill the enzymes?   Are you onboard with the info Cubit found about antioxidants preserving psilocin from oxidizing?  What are your thoughts on adding antioxidants into an already blued solution, which makes the blue go away?  Could this be putting the oxidized psilocin back to regular psilocin?  Some simple tests crushing the hell out of fresh mush's and letting them fully ozidize and testing a sample with and without antioxidants could shed some light on this im thinking.  

 

Another question.  If one were wanting to get the most bang out of thier mushrooms, turning all the psilocybin into psilocin, would it be best to do something like this:  Blend up whole  fresh mushrooms into a solution with antioxidants.  Leave the mushrooms in there so there are plenty of enzymes to break down the psilocybin into psilocin, and the antioxidant would preserve all the psilocin from breaking down and keep it stable.  Some gentle heat to speed up the reaction. Then after some time when the enzymes broke down all the psilocybin, filter the mushrooms out of the solution.

 

Sorry lots of questions


Edited by cg3p0, 09 February 2019 - 05:42 PM.


#69 PistolPete13

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Posted 09 February 2019 - 06:59 PM

 

Incubation of psilocybin or psilocin with a crude enzyme extract resulted in the formation of a.blue color usually after 5-15 minutes incubation and gave a linear rated over 180-minute-period (Figure 10). The addition of NaCN was effective in delaying the blue color formation, but did not influence the phosphatase activity'.

 

The dephosphorylation of PDP by a crude enzyme extract proceeded at a rapid rate as shown the liberation of phenolphthalein (30-120 seconds for enzyme extract A and 5-25 minutes for enzyme extract B). The dephosphorylation of psilocybin was rapid for both enzyme extracts. The blueing phenomena occurred in 3-5 minutes incubation in enzyme extract A and after 15 minutes in enzyme B. The liberation of psilocin reached a maximum just before the reaction mixture turned blue (Figure 11). When the psilocin was used as substrate, its concentration remained
unchanged before it turned blue.

https://open.library...831/1.0094541/2

 


The use of citric acid could possibly work as a preservative, and could be easily tested(if it does not turn blue).

 

 

What are your thoughts on adding antioxidants into an already blued solution, which makes the blue go away?

 

It is not quite as simple as that, the anti-oxidants are useful before the reaction as they can prevent the it from happening in the first place. Once it has been done, it is not as simple to put the toothpaste back into the tube so to speak. There has been a little research into the blue product, and they have not had much success finding out exactly what is was. There are theories to what it is, but at the end of the they are just theories......

 

 

Your last question I am sure you will be glad you asked;

 

In 1985 in the Journal of Forensic Science some researchers realized that because of the phosphoryloxy group that psilocybin has it makes it very poorly soluble in non-polar solvents (like ether, xylene ect). So they converted their samples into psilocin and used a classic A/B extraction and got great yields of very clean psilocin.

 

So to answer your first question;

 

A representative sample of 2 to 10g of dried mushrooms is ground to a fine powder by mortar and pestle. The powder is mixed with 100 mL of dilute acetic acid in a 250-mL beaker. The pH is readjusted to pH 4 with glacial acetic acid. After standing 1 h, the beaker is placed in a boiling water bath for 8 to 10 min or until the internal temperature of the acid mixture reaches 70°C. The beaker is removed and cooled to room temperature under running water. The acid mixture is separated from the mushroom powder by suction filtration using glass wool.

 

Basically putting dry sample into vinegar making sure pH is 4, standing for 1 hour, place in a boiling water bath(saucepan with boiling water in it) until it gets to 70*C. Cool and filter. Then.....

 

The filtrate is brought to pH 8 with concentrated ammonium hydroxide and quickly extracted with two 50-mL portions of diethyl ether. Gentle mixing instead of shaking should be used to prevent an emulsion. The ether is dried over sodium sulfate, filtered, and evaporated under nitrogen with no applied heat.

 

Crude psilocin will appear as a greenish residue. Recrystallization from chloroform/heptane (1:3) yields white crystals. The resulting powder can then be submitted to infrared and mass spectral analyses.

 

I have a paper that uses (I think, I have to double check this) bicarb soda to adjust the pH to 8 and said it was much gentler on the psilocin molecule which is vulnerable at higher pH.

 

Hope this helps....



#70 PistolPete13

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Posted 09 February 2019 - 07:00 PM

Ohh how rude......I am sorry.

 

Here is the paper I just referenced;

https://erowid.org/a....extraction.pdf

 

Just think I should also add, that psilocin is pretty unstable. For example you maybe would not want to store the above extract in your drawer for any length of time. In the paper above you can see when they are evaporating the solvent off they are piping in nitrogen gas so the psilocin at no stage comes into contact with the oxygen in the air. The salt would be more stable and would need to be stored correctly.


Edited by PistolPete13, 09 February 2019 - 07:32 PM.


#71 cg3p0

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Posted 09 February 2019 - 07:39 PM

So in that process of converting the psilocybin to psilocin, is it just the acid bath that is responsible for converting the psilocybin to psilocin?  And then the next part with the base only necessary if your wanting to dry it into crystals?  If that is true, wouldnt just the acid bath (using the lemon tek or vinegar or similar) be all thats nessessary for the home brewer to convert all the psilocybin into psilocin?  It would just be left diluted in the solution instead of concentrated into crystals?  thanks for the info!


Edited by cg3p0, 09 February 2019 - 07:41 PM.


#72 hyphaenation

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Posted 09 February 2019 - 08:04 PM

 

 

Has anyone an idea why you would do this extraction in stead of for example making a tea?

One reason would be the amount of liquid you potentially need to get a buzz... The idea is to take a dropper-full of Liquid-M or even a few drops With potent woodloving species such as cyanescens the extract would only be needed in very small quantity.

Tea is great. No doubt ... This is just something potentially interesting and different.


Edited by hyphaenation, 09 February 2019 - 08:20 PM.


#73 cg3p0

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Posted 09 February 2019 - 08:22 PM

Nevermind i found my answer, should have read your paper first :blush:

 

 

Psilocybin is completly dephosphorylated to psilocin by heating the acid extract.

 

 

Thats good to know.  The paper warns ,and i see you edited you post to warn too, that the psilocin will degrade quickly if left in the solution.  But this information was from 1984.  Did you check out what Quboid dug up. Its form 2019

 

 

 

  • Adding an antioxidant (e.g., ascorbic acid and/or sodium ascorbate) prevents bluingwithin a suspension of mushroom material in water.

https://psychedelicr...luing-reaction/

 

So maybe a simple solution to not have to worry about the degredation of psilocin would be to choose ascorbic acid as the acid used for the extraction instead of acetic acid.



#74 PistolPete13

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Posted 09 February 2019 - 11:59 PM


 

So maybe a simple solution to not have to worry about the degredation of psilocin would be to choose ascorbic acid as the acid used for the extraction instead of acetic acid.

 

The acetic acid is mainly to help with the extraction by aiding in pulling the actives out of the biomass, it protonates the amino group which changes it from its natural zwitterionic form to a salt(acetate in this case) that is much more soluble in the water.

 

Vitamin C does not stop the enzymatic dephosphorylation of psilocybin or help with the extraction, it acts as an antioxidant that slows the oxidation of psilocin. It can protect vulnerable parts of the molecule and react with any free oxygen that is present, by reacting with the oxygen first the psilocin does not get a chance to be oxidized. But with the law of averages and the amount of oxygen present in our environment this effect not 100% perfect.

 

In fact light has been shown to have a much more detrimental effect on the molecule than anything else, it catalyses the oxidation(quickly)..



#75 PistolPete13

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Posted 10 February 2019 - 12:39 AM

A Rapid Extraction and GC/MS Methodology for the Identification of Psilocyn in Mushroom/Chocolate Concoctions

 

The presented acetic acid facilitated extraction of psilocyn from mushrooms is more rapid and convenient versus
traditional methanolic extraction procedures, which require long time frames or potentially destructive heating. In
addition, the use of sodium bicarbonate as a neutralization agent keeps the pH below 8.5, thereby avoiding base-
facilitated destruction of psilocyn. The Total Ion Chromatogram (TIC) of the mushroom only sample shows a
clean psilocyn peak (Figure1). Analysis by TLC also shows only psilocyn. No psilocybin was detected - this is
perhaps due to the activity of the phosphatase enzymes present in the mushrooms, which can dephosphorylate
psilocybin to psilocyn in aqueous medium (19).

 

 

 

Found it!!

 

 

Procedure for Pure Mushrooms:

  1. About 0.2 to 0.5 gram of mushrooms are transferred into a mortar.
  2. The mushrooms are covered with 10 % acetic acid, and ground with the help of a pestle.
  3. Another 5 mL of deionized water are added and the mixture is ground into a fine slurry.
  4. The slurry is then transferred into a test tube and centrifuged for about 3 minutes.
  5. The supernatant is transferred into a small beaker
  6. The supernatant is neutralized by adding small amounts of sodium bicarbonate (neutralization is judged to be complete when the foamy effervescence stops). A little excess bicarbonate is then added.
  7. The resulting solution is transferred into a test tube and extracted with an equal amount of chloroform.
  8. The biphasic solution is centrifuged, and the chloroform layer collected in a shell vial.
  9. The chloroform extract is concentrated under air, transferred to a micro vial, and analyzed on the GC/MS.

Total extraction takes around 15 minutes. The results are shown in Figure 1.

 

[Direct Link]


Edited by PistolPete13, 10 February 2019 - 12:55 AM.


#76 PistolPete13

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Posted 10 February 2019 - 03:13 AM

Nevermind i found my answer, should have read your paper first :blush:

 

 

Psilocybin is completly dephosphorylated to psilocin by heating the acid extract.

 

 

Thats good to know.  The paper warns ,and i see you edited you post to warn too, that the psilocin will degrade quickly if left in the solution.  But this information was from 1984.  Did you check out what Quboid dug up. Its form 2019

 

 

 

  • Adding an antioxidant (e.g., ascorbic acid and/or sodium ascorbate) prevents bluingwithin a suspension of mushroom material in water.

https://psychedelicr...luing-reaction/

 

So maybe a simple solution to not have to worry about the degredation of psilocin would be to choose ascorbic acid as the acid used for the extraction instead of acetic acid.

 

Sorry I kind of rushed through and I think I misunderstood your question a bit, you were not talking about ascorbic acids antioxidant properties or the enzyme that starts it all off which I was referring to. But you were referring to one of the downstream enzymes that actually causes the bluing reaction, and ascorbic acids chelating properties. And its ability to stop the bluing reaction.....

 

The problem as stated above is not actually the bluing reaction(it definitely does not help, and seems it may speed things up further), but as soon as that first enzyme creates psilocin...the writing is on the wall. Unless you take measures to preserve it.

 

In a link I will attach, they study the stability of psilocin and psilocybin in aqueous solutions. And it is not does not last long, and does not turn the water blue when it degrades when it is not in presence of these metal ions!

 

Hopefully this helps!

Attached Files


Edited by PistolPete13, 10 February 2019 - 03:15 AM.


#77 cg3p0

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Posted 10 February 2019 - 12:32 PM

Well thanks for all the info.  But that sucks.  So just because the antioxidants keep it from turning blue, that doesnt mean it preserved the psilocin.  And unless your really set up with a lab it would be difficult to extract and preserve correctly.  So im assuming when drying mushrooms your really just preserving the psilocybin and all the psilocin that was in there is lost.  Sucks for the species high in psilocin like pans, seems like your wasting half the potential potency.  I guess the best thing most of us can do would be to pick them early before most of the psilocybin converts to psilocin from the enzymes?

 

 

psichart1.jpg

 



#78 pharmer

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Posted 10 February 2019 - 02:11 PM

The acetic acid is mainly to help with the extraction by aiding in pulling the actives out of the biomass, it protonates the amino group which changes it from its natural zwitterionic form to a salt(acetate in this case) that is much more soluble in the water.

 

If it's in salt form why not just drive off the solvent(s) and store as crystals? Is that possible?



#79 PistolPete13

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Posted 10 February 2019 - 03:12 PM

Sorry that sounded so pessimistic....I think you will find ascorbic acid still does have a preservative effect, as I said above;

 

 

Vitamin C does not stop the enzymatic dephosphorylation of psilocybin or help with the extraction, it acts as an antioxidant that slows the oxidation of psilocin. It can protect vulnerable parts of the molecule and react with any free oxygen that is present, by reacting with the oxygen first the psilocin does not get a chance to be oxidized. But with the law of averages and the amount of oxygen present in our environment this effect not 100% perfect.

 

So if added it should still protect the molecule to some extent and scavenge free oxygen and have an overall preservative effect. I was just trying to point out that stopping the blue reaction which is only degradation in the presence of these enzymes or as it may turn out the metal ions they posses does not stop the oxidation in aqueous solution(just ties up the metal ions so they cant react and produce those colors in the process). It should extend shelf life but it will have its limits (it is not a panacea).

 

 

Procedure for Pure Mushrooms:

  1. About 0.2 to 0.5 gram of mushrooms are transferred into a mortar.
  2. The mushrooms are covered with 10 % acetic acid, and ground with the help of a pestle.
  3. Another 5 mL of deionized water are added and the mixture is ground into a fine slurry.
  4. The slurry is then transferred into a test tube and centrifuged for about 3 minutes.
  5. The supernatant is transferred into a small beaker
  6. The supernatant is neutralized by adding small amounts of sodium bicarbonate (neutralization is judged to be complete when the foamy effervescence stops). A little excess bicarbonate is then added.
  7. The resulting solution is transferred into a test tube and extracted with an equal amount of chloroform.
  8. The biphasic solution is centrifuged, and the chloroform layer collected in a shell vial.
  9. The chloroform extract is concentrated under air, transferred to a micro vial, and analyzed on the GC/MS.

Total extraction takes around 15 minutes. The results are shown in Figure 1.

 

And that procedure I posted above using dilute acetic acid(vinegar), bicarb soda and a non-polar solvent. Evaporated under air, and takes 15 minutes and by no means needs a lab.

 

And psilocin is preserved in dried mushrooms, you just need to get it out of solution quickly to preserve as much as possible. It cannot be safely stored for any real length of time in aqueous solution. But once it is out of solution and completely dry (especially as a salt) and stored properly it is a lot more stable. And stored properly should keep for over a year.

 

You can see from this quote. If you remove all of the water quickly(freeze drying also retains nearly all the actives) and store them correctly shrooms can retain their activity(and psilocin) for up to two years. And indefinitely when freeze-dried and stored below -5*C......

 

Freezedried samples showed no detectable loss of psilocybin or psilocin when
stored at -5 “C or at -60 “C, but some freeze-dried samples lost both psilo-
cybin and psilocin over periods of one to two years when stored at room
temperature. Methanolic extracts were stable for over a year at -5 “C, but
within six months lost all psilocin and some psilocybin when stored at room
temperature.

https://www.ncbi.nlm.../pubmed/7201053

 

The methanol they use is reagent grade so there is no water at all present(hence the longer than expect shelf life for methanolic extracts).

 

So you just need to get the actives out solution as soon as possible after picking, one way or the other(and store them correctly). Or dry them bone dry and store them correctly and just take what you need and extract on a batch by batch basis......

 

I should have really been clear, that I was warning against the dangers of blue juice (or trying to keep aqueous solutions) not trying to discourage extractions!!! :dry:


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#80 PistolPete13

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Posted 10 February 2019 - 05:19 PM

Also tried literally juicing cyans ... in a wheatgrass juicer. The liquid came out clear and then turned blue , bluer and finally black. I was to chicken to dose on it but a friend did some and said it was potent (although details were scarce). I would definitely try it again in the future and try dosing the liquid. I think it could be juiced in to a bowl of alcohol and then put in a dark bottle and dosed though a dropper.
 

post-104353-0-05114100-1477531025_thumb.

 

 

post-104353-0-77135400-1477531010_thumb.

 

post-104353-0-37477000-1477535800_thumb.

I forgot that I filmed the maiden juicing ! 

https://mycotopia.ne...90#entry1292388

 

Just wanted to add one more thing, Hypaenations great idea to juice the mushrooms does away with the need for acetic acid to aid in pulling the actives out of the biomass. Because he did it mechanically, in fact in the procedure I posted above you could theoretically go straight to step 6. with his juice.

 

And a quote from a paper that I linked in my second post above;

 

The liberation of psilocin reached a maximum just before the reaction mixture turned blue (Figure 11). When the psilocin was used as substrate, its concentration remained
unchanged before it turned blue.

 

It seems like once that blue color appears there is not going to be much psilocybin left, maybe time to do steps 6 and 7?

 

  1. The supernatant(hyph's shroom juice) is neutralized by adding small amounts of sodium bicarbonate (neutralization is judged to be complete when the foamy effervescence stops). A little excess bicarbonate is then added.
  2. The resulting solution is transferred into a test tube and extracted with an equal amount of chloroform.
  3. The chloroform extract is concentrated under air

 

Hopefully there is not too much water present in the juice to make it difficult?

There would be no effervescence as no acetic was added, you would just have to adjust pH to 8 with bicarb or just adding an excess may suffice?

And I remember reading about having to be careful of emulsions forming when doing the non-polar pull, maybe if the juicer produced crystal clear juice it would help a bit also?

Also chloroform could be substituted for another suitable non-polar....


Edited by PistolPete13, 10 February 2019 - 05:39 PM.





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