it is under a blacklight, and the purple spot (center) is the ergot alks; the yellow perimeter is just the solvent front
aniracetam
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Vesp
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What does this look like in normal light? To me this image only looks like how wet paper would look under a black light. If normal light revealed purple on white, that would be impressive.
Do you have any quantitative instead of qualitative results?
Do you have any quantitative instead of qualitative results?
aniracetam
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nope
it was a culture aliquot that had been sitting for a few days. the spot was dry, and checked it again under the UV
light this morning. it's definitely an indolic complex.
already adsorbed the culture on bentonite, next step is elution/extraction, conversion to a salt.
it was a culture aliquot that had been sitting for a few days. the spot was dry, and checked it again under the UV
light this morning. it's definitely an indolic complex.
already adsorbed the culture on bentonite, next step is elution/extraction, conversion to a salt.
aniracetam
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here
Vesp
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All right, the second picture is more helpful.
Good luck!
Looking very forward to the final write up, hopefully from start to finish.
Good luck!
Looking very forward to the final write up, hopefully from start to finish.
aniracetam
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raw alks from the filtrant (most are still in bentonite)
gonna test on MALDI, 2,5-DHB as the matrix
Tsathoggua
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Sick, sick sonofabitch!
aniracetam
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'ey y0...
reflectron, positive mode. 2,5-DHB matrix. m/z threshold cutoff at 200.
base peak looks like elymoclavine +4 (+ 0.021 m/z deviation)
..
this is just a raw alk sample from the filtrant, no chrom.
the real goods are in a brick of bentonite in the vac des.
reflectron, positive mode. 2,5-DHB matrix. m/z threshold cutoff at 200.
base peak looks like elymoclavine +4 (+ 0.021 m/z deviation)
..
this is just a raw alk sample from the filtrant, no chrom.
the real goods are in a brick of bentonite in the vac des.
xxxxx
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Wow aniracetam. This is really amazing work! This is the first time I have ever seen someone making such practical progress towards ergot alkaloids! I hope this works out for you and I know many people here would love a full writeup when you finish. BTW it might be a good idea to upload those images here rather than hotlinking them, they can give away members information and they can also get lost easier leading to broken images.
Anyways keep up the fantastic work aniracetam!
Anyways keep up the fantastic work aniracetam!
akcom
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Wow, I don't know whats more impressive: your results or your lab. If you don't mind my asking, are you doing this in a professional setting or are you just well funded?
aniracetam
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I just know how to shop for equipment.
the mass spec is something I work with, it'll be a little while before I have my own.
the mass spec is something I work with, it'll be a little while before I have my own.
akcom
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That makes me feel better haha, what is that big hunk of junk next to your rotovap?
Vesp
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Where did your graph picture that was in this post go?
http://127.0.0.1/talk/index.php/topic,1383.msg21357.html#msg21357
http://127.0.0.1/talk/index.php/topic,1383.msg21357.html#msg21357
Tsathoggua
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Aniracetam, have you got a writeup for the bentonite separation protocol? I have heard nothing but good things, but its one of the things I lack in my library.
I'll say it again...sick, sick fucking fuck Any idea yet what your strain is producing per liter, and what culture medium are you using?
I'll say it again...sick, sick fucking fuck Any idea yet what your strain is producing per liter, and what culture medium are you using?
aniracetam
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akcom, it is a UV-Vis detector with a capillary flowcell, syringe pump, and an ultrasonic power supply under them.
vesp, I took it down to edit it...i'm actually doing another run now, to improve resolution.
tsathoggua, it was a patent I looked at, for the process of separating raw ergot alkaloids. basically loosely followed two patents. the first was the tonolo et al. patent, which uses 5% mannitol, 1% sucrose, 2% succinic acid, 0.5% KH2PO4, 0.03% MgSO4; added 0.5% tween 80 for good measure.
latest batch follows the same formula, but I added 1 g l-tryp.
for this prior batch, I added 50 g/L bentonite, stirred for an hour, and vac filtered. put the cake in the oven at 80 C for about an hour and a half, then transferred to a vac des.
vesp, I took it down to edit it...i'm actually doing another run now, to improve resolution.
tsathoggua, it was a patent I looked at, for the process of separating raw ergot alkaloids. basically loosely followed two patents. the first was the tonolo et al. patent, which uses 5% mannitol, 1% sucrose, 2% succinic acid, 0.5% KH2PO4, 0.03% MgSO4; added 0.5% tween 80 for good measure.
latest batch follows the same formula, but I added 1 g l-tryp.
for this prior batch, I added 50 g/L bentonite, stirred for an hour, and vac filtered. put the cake in the oven at 80 C for about an hour and a half, then transferred to a vac des.
overunity33
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makes me happy to see this, things have come a long way since the lsa bioreactor post...
Look forward to getting more experimental details on the bentonite elution and further purification... youre a hero bro, this is a gamechanger..
Look forward to getting more experimental details on the bentonite elution and further purification... youre a hero bro, this is a gamechanger..
aniracetam
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it appears degradation of the the sample has occurred; relative abundance for 258 has dropped considerably, and 273 has increased, from the run yesterday and the run today.
did several sample runs.
thanks, overunity. been 15 years since I daydreamed about this; now I'm able and willing.
this a killer resource...The Handbook of Secondary Fungal Metabolites (Vol. 1 - 3), 52 MB.
hxxp://megaupload.com/?d=7PMP6KJM
did several sample runs.
thanks, overunity. been 15 years since I daydreamed about this; now I'm able and willing.
this a killer resource...The Handbook of Secondary Fungal Metabolites (Vol. 1 - 3), 52 MB.
hxxp://megaupload.com/?d=7PMP6KJM
overunity33
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Do you think that the degradation products will still hydrolize to LSA? Can you speculate as to what change occurred? i cant interpret spectra just yet..
Those daydreams man... its funny how fast capable people end up staring their daydreams in the face..
Those daydreams man... its funny how fast capable people end up staring their daydreams in the face..
aniracetam
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ok...after further examination, it seems 273 is background signal for matrix. 258 is part of the sample.
I loaded a couple spots on the plate with matrix only, shown in red, and matrix with sample, shown in dark blue.
the masses listed are for the blue peaks.
258 is likely chanoclavine (m.w. 256.343) +2.
pubchem CID 5281381. it has two proton acceptors
again, this is just from the filtered broth (did a base/acid extraction on it), i haven't eluted from the bentonite.
I loaded a couple spots on the plate with matrix only, shown in red, and matrix with sample, shown in dark blue.
the masses listed are for the blue peaks.
258 is likely chanoclavine (m.w. 256.343) +2.
pubchem CID 5281381. it has two proton acceptors
again, this is just from the filtered broth (did a base/acid extraction on it), i haven't eluted from the bentonite.
Tsathoggua
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You baked the bentonite in the oven at 80 C ?
that sounds like it would wreak havoc on the alkaloids absorbed.
that sounds like it would wreak havoc on the alkaloids absorbed.