Author Topic: Ergot Culture Medium for Alkaloid Production (C. Purpurea & Paspali) (Read 1616 times)

aniracetam · « **Reply #140 on:** May 05, 2011, 12:28:31 PM »

it doesn't.
the patent describes drying at 80 C for at least half an hour on a fluid drying bed.
this variety of montmorillonite has a platelet structure, the alks are intercalated and stable.

aniracetam · « **Reply #141 on:** May 06, 2011, 10:16:44 PM »

from a bentonite extract (conc. ammonia in MeOH, extracted with dcm. rotavaped, and tartaric acid in MeOH was added)
all sample (no chromatography conducted, no matrix added).

Vesp · « **Reply #142 on:** May 07, 2011, 12:20:14 AM »

So what does this all mean?

aniracetam · « **Reply #143 on:** May 07, 2011, 12:38:22 AM »

this means the products (which are likely terminal N-substituted ergolines) following bentonite elution may be hydrolyzed, then separated on a column.
the bentonite seems to preserve the alks, as ou suggested in another thread. also seems to clean up the broth
this analysis was done on MALDI-TOF..I'll have to do more precise characterization, using LC/ESI-MS, on monday.

next run, i'll skip the bentonite step and just hydrolyze the alks.

Shake · « **Reply #144 on:** May 07, 2011, 09:03:52 AM »

hey aniracetam, id like your opinion on this http://www.youtube.com/watch?v=Sqi_qBlPwc4&feature=related

coming from someone with experience, and being written by someone who obviously had not done it himself

aniracetam · « **Reply #145 on:** May 07, 2011, 02:16:02 PM »

i've heard a lot of criticism of the book, mostly from prick-wavers from the hive; honestly, I've never read the book, so I can't make any conclusions. however, I will search for it, and reply back.

taking the conventional scientific approach, I find the book LSD by Otto Snow to be the most comprehensive
book on the art. It's an excellent read, very informative; particularly the chromatographic separations section.
the only vagueness I find in the book is mention of applying inert gas. there should be more detailed research paper-oriented methods to describe which steps involve inert gas, and a suggested flowrate

Shake · « **Reply #146 on:** May 07, 2011, 02:56:22 PM »

i know what you mean i bought otto snows love drugs.. great read other than i could not understand 1 word of it.. i wish i hadnt thrown it out now!

the books falsely advertise as though they are noob friendly when they are not!

aniracetam · « **Reply #147 on:** May 07, 2011, 05:23:12 PM »

yea, forgot to mention...they are rather esoteric

this Uncle Fester's book...definitely an easy read for people not familiar with the esoteric side of
chemistry. I found a couple issues: while the idea of farming ergot is nice, I did not see any mention of saprophytic c.purpurea cultures lacking the ability to produce alks in submerged culture. This is an important point, as it took, what, something like 40 years? and thousands of plates to isolate the right mutated purpurea strain to produce said alks. someone please correct me if I'm wrong about this, I've seen it mentioned several times

the other issue: the author suggests using conc. ammonia in methanol, and adding dcm for extraction, claiming that it will form separable layers. it will not. methanol and dcm form an azeotrope, I recently witnessed this; even in a -20 C freezer, the liquids will not separate.

Shake · « **Reply #148 on:** May 07, 2011, 05:48:48 PM »

i agree with the azeotrope that thing is a fucker, i got it not to long ago, killed me

let me edit..

i guess im trying to say, how hard is it compared to how he makes it sound?

i mean do you just laugh it off? or is there some hope for someone following his book?

tbh i suspect no, no clanny has a chance

aniracetam · « **Reply #149 on:** May 07, 2011, 05:59:56 PM »

well, like with anything, it takes practice; but you don't need a PhD to make acid.
and if you spend over $5000 on equipment, you've spent too much (unless it's a mass spectrometer)

with enough preparation, planning, and practice, any competent person can pull it off. it's not impossible.
that book is a decent guide, and it provides a lot of references, which I highly recommend seeking

I say...bee optimistic, and patient.

The Lone Stranger · « **Reply #150 on:** May 07, 2011, 06:01:15 PM »

And now a short break for some adverts .......

The Ark i`m touting contains 3 books by strike , 3 by fester and 5 by snow 2 of wich cover LSD ...... plus others ......

And now back to the main program........

Shake · « **Reply #151 on:** May 07, 2011, 06:27:18 PM »

^^^^^
this guy is nuts

lol

a mas specrometer uses light refractions to measure the density of it.. the mass..

have you ever thought that one could be home made? all it is is light and a spectrum.. the rest is the reading no?

aniracetam · « **Reply #152 on:** May 07, 2011, 07:03:25 PM »

lol

that is not a mass spectrometer, that is a spectrophotometer.

if you could make a mass spectrometer in your home, that would be very impressive.
it typically requires high vacuum (diffusion pump, or turbomolecular pump), backed by a roughing pump;
RF signal generators, an electron multiplier, and various controllers and sensors, not to mention extensive programming
to monitor said sensors.
you can find a spectrophotometer on a popular auction site for $50 - 300. a complete mass spec is usually >$5000

Shake · « **Reply #153 on:** May 07, 2011, 07:07:08 PM »

tomato tomarto

mas spectrum i swear i am right here... what is it then?

i mean, we both know im not.. but still

aniracetam · « **Reply #154 on:** May 07, 2011, 07:08:01 PM »

an absorbance spec is a less information-rich method of analysis than mass spec. the mass spec does not use UV or visible light. it uses RF and an electron multiplier instead of a photomultiplier tube. they operate on similar principles (amplification), but have different detector mechanics.

mass spec is standard for ID'ing compounds, along with NMR (which requires mg of sample, vs ng and ug for mass spec)
and tandem mass spec gives structural information

for all practical purposes here, the addition of a reagent, like Van Urk's, is useful for the identifying the different alks; in this case
a spectrophotomer would be useful to differentiate, AND determine the concentration of alks in solution using Beer's law.
this is probably what you had in mind

but yea...a spectrophotometer and a spectrometer are not the same thing
NMR and mass spec are both spectrometers

Tsathoggua · « **Reply #155 on:** May 08, 2011, 02:42:51 AM »

I was under the impression that at least, in the visual range, van urk/erlich's was quantitative only?

overunity33 · « **Reply #156 on:** May 08, 2011, 02:45:00 AM »

Quote from: Tsathoggua on May 08, 2011, 02:42:51 AM

I was under the impression that at least, in the visual range, van urk/erlich's was quantitative only?

*qualitative?

It shows the presence of indole alkaoids, by reading the absorbance you can use beers law to determine the concentration... of course you need some standards or some really good educated guessing

aniracetam · « **Reply #157 on:** May 08, 2011, 02:51:43 AM »

true.

each compound has an extinction coefficent and max abs at a given wavelength. but ergot alk UV absorbances are pretty
close to each other: hxxp://sciencemag.org/content/83/2146/166.extract

for this reason, the spectrometers give more info. tandem TOF or tandem MS will give structural info
of the mass fragments; but, it's also time consuming and complex

LC-MS/MS ftw. instead of a new car, get the bank to finance one of those.
this one's $34,995 on a certain popular auction site.

I'd love to have a bare-bones LTQ, but it's just the mass spec, and a used one costs as much as a new maserati gran turismo
the LTQ Orbitrap Velos is even cooler

Tsathoggua · « **Reply #158 on:** May 08, 2011, 03:33:25 AM »

I meant, by quantitative, being able to determine rough concentration of alkaloid by fluorescence intensity, either alone, or in tandem with a counterstain of fluorescein.

Qualitative in a way, but limited to boolean 0-1.

Aniracetam, you seem to know a fair lot about spectrometry, I know next to nothing about the practical workings of the tools, and which are appropriate for which purposes. I am thinking of getting myself a cheap, old handmedown spectrophotometer, for the analytical work, and I could really do with some advice as to what to choose, and why. You can leave mass specs of any kind out, likewise NMR, my bank will be funding nothing, as I'm unemployed, and have been, not for want of trying, trying and trying again, since I left school at 16. Chances of mr.fatcat reaching into his(or rather everybody else's) pockets to fund a mass spec, or HPLC-MS for someone whos only income is benefits, and the fruits of his own creativity and resourcefulness, is about the same as waking up tomorrow, to find out I am actually a fruitbat.

aniracetam · « **Reply #159 on:** May 08, 2011, 03:41:13 AM »

I hear ya

I'm gonna have to finagle one. been working with them for about two years, and know what to look for in one.
was able to differentiate between THH and harmine (seen in the attachment) in a sample using an LCQ, which I've seen go for 2800 - 7500 on 3bay.
already bought a harvard apparatus 22 series LC syringe pump for $130

Author Topic: Ergot Culture Medium for Alkaloid Production (C. Purpurea & Paspali) (Read 1616 times)

aniracetam

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aniracetam

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Vesp

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aniracetam

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Shake

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aniracetam

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Shake

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aniracetam

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Shake

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aniracetam

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The Lone Stranger

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Shake

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Shake

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aniracetam

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Tsathoggua

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overunity33

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aniracetam

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Tsathoggua

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aniracetam

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