Author Topic: ergot and agar  (Read 13240 times)

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formula54

  • Guest
Re: ergot and agar
« Reply #20 on: September 21, 2001, 08:17:00 PM »
word, thanks agent. any info is good.
btw, what is that country hest?

spric

  • Guest
Re: ergot and agar
« Reply #21 on: September 26, 2001, 06:34:00 AM »
A 5 acre size petri dish otta' work for a enough to dose your hometown.  You could always just pH your media before use with KOH.  Also, methinks those sulfates don't hurt.

4:20

Bozakium

  • Guest
Re: ergot and agar
« Reply #22 on: September 27, 2001, 12:34:00 AM »
For a good starter's guide on ergot farming(and subsequent isolation, synth & workup), try "Practical LSD Manufacture" by Uncle Fester, available from Loompanics Unlimited, Port Townsend Washington.  Its a great springboard, and includes many useful references. I havent heard of any suceccful cultures in a large scale in the lab, it seems one must grow a crop of rye and infest it with Claviceps Purpuria intentionally and thoroughly. Happy not heard of anyone sucessfully cultivating the fungus in Farming.

formula54

  • Guest
Re: ergot and agar
« Reply #23 on: September 27, 2001, 05:20:00 PM »
yeah, but were talking about ergot for those of us that arent farmers. fester is useless for us.

formula54

  • Guest
Re: ergot and agar
« Reply #24 on: September 28, 2001, 08:38:00 PM »
"sometimes i feel like a dont have a partner"
im going to cook this chemical if its the last thing i do.

cilliersb

  • Guest
Re: ergot and agar
« Reply #25 on: October 01, 2001, 01:24:00 PM »
Well, at least there's some light on the horizon.

Some dude I've never heard of went surfing the net for no more than 15mins. He found a site that actually distributes Claviceps Paspali. Hint: they are based in Germany

They suggested this agar medium for CP:

Medium 129: POTATO DEXTROSE AGAR 

 Infusion from potatoes (see below)         1000.0     ml
 Glucose                                      20.0      g
 Agar                                         15.0      g

Potato infusion:
Boil 200 g scrubbed and sliced potatoes in 1000 ml water for 1 hour. Pass through fine sieve. Avoid
new potatoes.

And that from the dudes that grow it commercially, how nice!
 

Trenchcoat

  • Guest
Re: ergot and agar
« Reply #26 on: October 02, 2001, 07:41:00 AM »
Did fester ever actually make LSD?

Better loving through chemistry.

Trenchcoat

  • Guest
Re: ergot and agar
« Reply #27 on: October 02, 2001, 07:43:00 AM »
It'd probably be less expensive and less time-consuming to learn a foreign language and go to eastern europe in search of pure ET. Besides from what I've read this is the only way to make real pure LSD.

Better loving through chemistry.

formula54

  • Guest
Re: ergot and agar
« Reply #28 on: October 02, 2001, 10:36:00 PM »
i think its funny that so many cooks make it seem like lsd synthesis is impossible. it seems very viable, especially if we figure a way to mass grow claviceps, (besides farmer festers idea of owning a wheat field.) it is the basic eqivalent of sayin you need to own a washington forest to grow shrooms.

formula54

  • Guest
Re: ergot and agar
« Reply #29 on: October 11, 2001, 03:16:00 AM »
ALOT of you must have real bad migraines, if you know what i mean. i cant believe that theres not more discussion on claviceps cultivation.

IudexK2

  • Guest
Re: ergot and agar
« Reply #30 on: October 13, 2001, 12:56:00 PM »
SWIM has lots of antibacterial MEA... if someone can tell me if it is worth trying to grow ergot on it, and how best to approach it, SWIM would bee happy to try it.

Could the ergot possibly just bee grown large-scale on agar and extracted from there?

cyril

  • Guest
Re: ergot and agar
« Reply #31 on: October 14, 2001, 02:11:00 AM »
Got this from Psycadellic Chemisty Book:

Make up a culture medium by combining the following ingredients in about 500 milliliters of distilled water in a 2 liter, small-neck flask:

Sucrose 100 grams
Chick pea meal 50 grams
Calcium nitrate 1 gram
Monopotassium phosphate 0.25 grams
Magnesium sulphate 0.25 grams
Potassium chloride 0.125 grams
Ferrous sulphate heptahydrate 8.34 milligrams
Zinc sulphate heptahydrate 3.44 milligrams
Add water to make up one liter, adjust pH 4 with ammonia solution and citric acid. Sterile by autoclaving.

Inoculate the sterilized medium with Claviceps purpurea under sterile conditions, stopper with sterilized cotton and incubate for two weeks periodically testing and maintaining pH 4. After two weeks a surface culture will be seen on the medium. Large-scale production of the fungus can now begin.

Obtain several ordinary 1 gallon jugs. Place a two-hole stopper in the necks of the jugs. Fit a short (6 inch) glass tube in one hole, leaving 2 inches above the stopper. Fit a short rubber tube to this. Fill a small (500 milliliter) Erlenmeyer flask with a dilute solution of sodium hypochlorite, and extend a glass tube from the rubber tube so the end is immersed in the hypochlorite. Fit a long, glass tube in the other stopper hole. It must reach near the bottom of the jug and have about two inches showing above the stopper. Attach a rubber tube to the glass tube as short or as long as desired, and fit a short glass tube to the end of the rubber tube. Fill a large, glass tube (1 inch x 6 inches) with sterile cotton and fit 1-hole stoppers in the ends. Fit the small, glass tube in end of the rubber tube into 1 stopper of the large tube. Fit another small glass tube in the other stopper. A rubber tube is connected to this and attached to a small air pump obtained from a tropical fish supply store. You now have a set-up for pumping air from the pump, through the cotton filter, down the long glass tube in the jug, through the solution to the air space in the top of the jug, through the short glass tube, down to the bottom of the Erlenmeyer flask and up through the sodium hypochlorite solution into the atmosphere. With this aeration equipment you can assure a supply of clean air to the Claviceps purpurea fungus while maintaining a sterile atmosphere inside the solution.

Dismantle the aerators. Place all the glass tubes, rubber tubes, stoppers and cotton in a paper bag, seal tight with wire staples and sterilize in an autoclave.

Fill the 1-gallon jugs 2/3 to 3/4 full with the culture medium and autoclave.

While these things are being sterilized, homogenize in a blender the culture already obtained and use it to inoculate the media in the gallon jugs. The blender must be sterile. Everything must be sterile.

Assemble the aerators. Start the pumps. A slow bubbling in each jug will provide enough oxygen to the cultures. A single pump can, of course, be connected to several filters.

Let everything sit a room temperature (25 C) in a fairly dark place (never expose ergot alkaloids to bright light - they decompose) for a period of ten days.

After ten days adjust the culture to 1% ethanol using 95% ethanol under sterile conditions. Maintain growth for another two weeks.

After total of 24 days growth period the culture should be considered mature. Make the culture acidic with tartaric acid and homogenize in a blender for one hour.

Adjust to pH 9 with ammonium hydroxide and extract with benzene or chloroform/iso-butanol mixture.

Extract again with alcoholic tartaric acid and evaporate in a vacuum to dryness. The dry material in the salt (i.e., the tartaric acid salt, the tartrate) of the ergot alkaloids, and is stored in this form because the free basic material is too unstable and decomposes readily in the presence of light, heat, moisture and air.

To recover the free base for extraction of the amide of synthesis to LSD, make the tartrate basic with ammonia to pH 9, extract with chloroform and evaporate in vacuo.

If no source of pure Claviceps purpurea fungus can be found, it may be necessary to make a field trip to obtain the ergot growths from rye or other cereal grasses. Rye grass is by far the best choice. The ergot will appear as a blackish growth on the tops of the rye where the seeds are and are referred to as "heads of ergot." From these heads of ergot sprout the Claviceps purpurea fungi. They have long steams with bulbous heads when seen under a strong glass or microscope. It is these that must be removed from the ergot, free from contamination, and used to inoculate the culture media. The need for absolute sterility cannot be overstressed. Consult any elementary text on bacteriology for the correct equipment and procedures. Avoid prolonged contact with ergot compounds, as they are poisonous and can be fatal.

Thats the way I'd do it,
Cyril

PS: You can buy infected grain for $250-$400/Lb, if you look hard enough.



So you need a precursor, to a precursor, just to make a precursor?

IudexK2

  • Guest
Re: ergot and agar
« Reply #32 on: October 14, 2001, 02:57:00 AM »
Yes, that extract is on Erowid, we have all seen it.

cyril

  • Guest
Re: ergot and agar
« Reply #33 on: October 14, 2001, 03:19:00 AM »
In that case why not just use it, it's simple as all hell, otherwsise maybe some antibacterial TSA actually as per the method above TSB would seem more appropriate. (lol)  BTW that isn't only an extraction it's a growth media. 

When infected grain is so inexpensive to buy, and you have a method that apparently "we have all seen"  then why not just do it instead of waste time talking about other ways to do it. . .

The supplies to grow it are easily found at any biosupply house, (heck my neighborhood chemsupply store is also a biosupply store), and unless you skipped microbiology Lab for an extra Chem class the method is simple as can be.

Anyhow maybe the real lesson to be learned is to apply the knowledge that you have read before to the current situation.

Anyhow I guess I can't get too mad, probably just wasting my time trying to explain myself to an idiot.

Cyril  



So you need a precursor, to a precursor, just to make a precursor?

IudexK2

  • Guest
Re: ergot and agar
« Reply #34 on: October 14, 2001, 12:58:00 PM »
Coz I like growing stuff on agar.


Anyhow I guess I can't get too mad, probably just wasting my time trying to explain myself to an idiot.




Probably  ;)


cyril

  • Guest
Re: ergot and agar
« Reply #35 on: October 15, 2001, 07:18:00 PM »
Okay if you like to grow stuff on agar so much then I would assume you have prepared agar media before.  The above refernced growth media could easily be made into a growth media for growth on agar.  Seems pretty simple to me.  I would perhaps put something in to inhibit bacterial growth, and perhaps use a soy base rather then the chick peas, but still add the other components. I bet you'd get growth on a plate.  Sorry about assuming that it would be an easy extrapolation from one to the other. 

Cyril


So you need a precursor, to a precursor, just to make a precursor?

formula54

  • Guest
Re: ergot and agar
« Reply #36 on: October 15, 2001, 09:49:00 PM »
what im wondering is what each of those chemicals do in this cultivation writeup...im guessing some are nutrients, what are the rest for?

cyril

  • Guest
Re: ergot and agar
« Reply #37 on: October 20, 2001, 12:49:00 AM »
Here is the method I would use:

Ingredients:

6.5 g Potato Dextrose Agar
5.0 g Bacto Agar
500 ml distilled water
15 mg Rifampicin in 10 ml Methanol
15 mg Penicillin G in 10 ml 70% Ethanol

Mix the first 3 ingredients, autoclave for 20 min. and cool to room temperature. Add the antibiotics and pour into sterile petri dishes.

Plate out the fungi using a 10-4 dilution starting with 5 g dry weight of compost in 45 ml of the autoclaved phosphate buffer. Put this first dilution in a blender at high speed for 40 sec.

Perform serials dilutions to 10-4 and add 0.l ml of the final dilution to each plate.

Incubate the plates at 28C for 3 days.

Take counts and samples of fungal colonies at 3 days.

Sorry it took so long,
Cyril


So you need a precursor, to a precursor, just to make a precursor?

formula54

  • Guest
Re: ergot and agar
« Reply #38 on: October 20, 2001, 12:50:00 AM »
yes. thank you cyril, thats what we were all waiting for.

bujinkan

  • Guest
Re: ergot and agar
« Reply #39 on: December 10, 2001, 10:49:00 PM »
Does anyone know about Claviceps paspali vs. claviceps pupurea for ergotamine yeilds?
also, here is a page i found for all you bees trying to find ergot in the wild.

http://www.oda.state.or.us/Plant/ppd/Ergotmanual2.pdf