Jump to content
The Corroboree

Fractalhead

Members2
 View Profile  See their activity

  • Content count

    374

  • Joined

  • Last visited

Posts posted by Fractalhead

  1. Posted

    Here's an interesting article. I wonder if anyone has tried eating these...

    Biochim Biophys Acta. 1971 Aug 19;244(2):322-8. Related Articles, Links

    Urinary excretion of tertiary amino methoxy methylenedioxy propiophenones as metabolites of myristicin in the rat and guinea pig.

    Oswald EO, Fishbein L, Corbett BJ, Walker MP.

    I've got a few more references to put up including the one in which amphetamine derivatives were reported...

  3. Posted

    What these guys have done is isolate from cannabis the protein (enzyme) that catalyses the final step in the biosynthesis of THCA (the stuff that gets converted to THC in the pot smoking process). Because they had the pure enzyme, they were able to determine its amino acid sequence. This allowed them to find and copy the corresponding gene from cannabis into other organisms including tobacco root cells. When they fed the transgenic tobacco root cell culture with cannabigerolic-acid (the biosynthetic precursor for THCA) the tobacco cells turned it into THCA.

    If the gene for THCA synthase were introduced into a plant that makes its own cannabigerolic-acid (i don't know of any other than cannabis spp.) then you might have a new THCA producing plant.

  4. Posted

    That's one down... now we just need to clone the rest of the THCA biosynthesis genes You can get this journal article for free from the Journal of Biological Chemistry website.

    : J Biol Chem. 2004 Jun 9 [Epub ahead of print] Related Articles, Links

    The gene controlling marijuana psychoactivity molecular cloning and heterologous expression of Delta1-tetrahydrocannabinolic-acid synthase from Cannabis sativa.

    Sirikantaramas S, Morimoto S, Shoyama Y, Ishikawa Y, Wada Y, Shoyama Y, Taura F.

    Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Fukuoka 812-8582.

    Delta(1)-Tetrahydrocannabinolic-acid (THCA) synthase is the enzyme that catalyzes oxidative cyclization of cannabigerolic-acid into THCA, the precursor of Delta(1)-tetrahydrocannabinol. We cloned a novel cDNA (Genbank accession number AB057805) encoding THCA synthase by reverse transcription and polymerase chain reactions from rapidly expanding leaves of Cannabis sativa. This gene consists of a 1635-nucleotide open reading frame, encoding a 545-amino acid polypeptide of which the first 28 amino acid residues constitute the signal peptide. The predicted molecular weight of the 517-amino acid mature polypeptide is 58,597 Da. Interestingly, the deduced amino acid sequence exhibited high homology to berberine bridge enzyme from Eschscholtzia californica, which is involved in alkaloid biosynthesis. The liquid culture of transgenic tobacco hairy roots harboring the cDNA produced THCA upon feeding of cannabigerolic-acid, demonstrating unequivocally that this gene encodes an active THCA synthase. Overexpression of the recombinant THCA synthase was achieved using a baculovirus-insect expression system. The purified recombinant enzyme contained covalently attached FAD cofactor at a molar ratio of FAD to protein of 1:1. The mutant enzyme constructed by changing His-114 of the wild-type enzyme to Ala-114 exhibited neither absorption characteristics of flavoproteins nor THCA synthase activity. Thus we concluded that the FAD binding residue is His-114, and that the THCA synthase reaction is FAD-dependent. This is the first report on molecular characterization of an enzyme specific to cannabinoid biosynthesis.

  7. Posted

    Was having a discussion the other day about establishing homozygous gene knockout plants and this led to the possibility of going via a haploid intermediate. This got me thinking again about a paper i read ages ago on anther culture of tobacco to give haploid plants that will happily grow vegetatively but can't produce gametes unless diploidy is restored by intervention with something like colchicine. The process seems to be readily adaptable to new species. Suddenly it occured to me just how readily this process could be applied to our special plants.

    The wonderful thing about it is that virtually every haploid plant you produce by anther culture will have a different combination of alleles. Some of these alleles will be recessive in the diploid plant and hence the possibly weird and wonderful characteristics they might impart are not observed in the diploid. Once in the haploid plant, these recessive genes (which might just include genes of secondary metabolism) have a chance to work their magic and show us what they're made of. This has the potential to speed up our plant breeding efforts enormously and yield some weird-arse or particularly kick-arse new chemotypes!

    For a rundown on anther culture by androgenesis induction see Wang et al. (2000) Plant Physiology vol 124, pp. 523-530

  9. Posted

     

    quote:
    I think the warnings re oxalic acid in Aizoaceae really apply mostly to smoking and snorting.

     

    Torsten, is that because you don't think regular oral dosing at the typical oral doses for aizoaceous materials would lead to an ingestion rate of oxalate in excess of the body's oxalate excretion rate?

  11. Posted

    SHIT! I'm so sorry! I misunderstood the status of this information. Its just so bizzare. Fuck. I should have just assumed it should have been left to the researcher to bring it up. I'm way out of line here. BIG apologies to all involved parties. I realise I've probably made too much of a contribution already, but if i can help with details of my extraction or repeat it and provide a sample pls let me know.

  13. Posted

    I don't think mesembrine has the long systems of conjugation required for fluorescence. Its got one aryl ring but the rest has no conjugated systems. Certainly looks like it would react with the right detection reagent on tlc - not that it really matters anyway because it is yellow. Nuciferine has a chloroform soluble hydrochloride and it was this property that led to the first successful isolation of nuciferine. I transcribed selected parts of the paper onto the forum some time ago. You should find it if you search for nuciferine and nelumbo nucifera. The guy who isolated it was from a south eastern university from memory.

    BTW, this is way off topic, but do you know anything about the 'Ed' Sceletium sp. being tested negative for alkaloids by GC/MS?

  14. Posted

    but i heard two types were involved: one in the form of bark chunks and one powdered and boiled. i heard the bark chunks were effective on their own prior to the brew. or was this the same material? i also heard it was in general very mild.

    [ 25. June 2004, 22:07: Message edited by: Fractalhead ]

  15. Posted

    I can't remember... have there been any positive activity reports from teh red caapi powder? If so, then the dull fluorescence of the aqueous extract may be due to the absence of harmaline. The green fluorescence of harmaline is particularly strong and even very dilute solutions of harmaline glow bright green. In peganum, the blue of harmine is responsible for the aqua colour of the mixture. Perhaps your red caapi bark is devoid of the potent fluorophore harmaline but loaded with the less intense but nice and active tetrahydroharmine (pale blue). It might be easy to miss this glow next to a smashed-up-glowstick-like harmaline rich sample (especially if the tetrahydroharmine one is diluted considerably).

    john, i guess its the interference of the solvents/buffers that was the problem preventing us from knowing what the ionisation state of the alkaloids really is in vivo. Still, your experiment would be worth a shot as it would be extremely unusual if an apolar solvent extracted the alkaloids from non-basified plant material. I really don't think the protonation state of the alkaloids in vivo or the extraction efficiency of the neutral water is the cause anyway.

  16. Posted

    The reports I've read indicate that:

    Mild but distinct and beautiful open eye phosphenic visuals can be obtained from 2-3 fractal seeds *fresh off the vine* and this probably goes for any other top-quality seed. 10+ seeds that fresh and things can get a bit 'gaa-gaa' and constant open eye fractals can overwhelm one along with intense feelings of euphoria and e-like empathogenicity. 20+ seeds would be a heroic effort at this potency.

    However, even after several months you can munch 10-20 seeds and not get any visuals. You might have a nice trip but nothing like the places you can reach on the fresh ones. The major downside is that the compounds responsible for the negative somatic effects appear to be much stabler than the goodies. so the bang for your buck factor goes down steadily reaching almost pointlessness by about 1-1.5 yrs.

    Luckliy we have two hemispheres and hence two major HBWR seasons so the freshest seed around is going to be at most 6 months old at any point in the year... depending on import restrictions where you live

  17. Posted

    I'm pretty certain the common ravey blacklights are generally made using the same materials and rely on the same physics to emit the characteristic spectrum inherent in that setup. They emit mainly between 350 nm and 370 nm a little bit in the short wavelength visible region and peak at 365nm. This is commonly referred to as longwave UV. This wavelength is low enough to make all the b-carbs glow.

    I'm still puzzling over the possible mechanism(s) behind the effect of the formic...

    * Is the intensity of fluorescence of the beta carbolines pH dependent or dependent on the nature of the salt?

    I guess the way to test that would be to observe the fluorescence of an aqueous solution of isolated b-carbs while gradually adjusting pH. Repeat with different acids and compare.

    * Are the alkaloids really fully protonated in the tissue?

    Don't know how you can really find this out.

    * Are there possibly other very weakly basic compounds present in the materials that are only fluorescent at extreme pH?

    The thing that caught my eye was some of the strange colours coming from your materials wetted with formic acid. So is there a colour change upon enhancement of fluorescence with formic acid? The fluorescence from peganum seeds is usually distinctly aqua blue-green more than yellow but i guess it can look yellowish under some conditions... :confused: From memory (and i will have to check this), there is no yellow fluorescing alkaloid reported in HBWR (except maybe tryptamine?? - i wish i had my notes here ) although i have a sneaking suspicion....

    * Does the formic enhance release of the fluorescent compounds from the tissue?

    Maybe try extracting with a little water and then observing the change in fluorescence upon addition of the formic acid to a certain concentration versus extraction with formic acid solution of that known concentration for the same amount of time. The fact that an aqueous extract of rev's powdered red caapi bark didn't glow much would be a concern to me since if there is any appreciable amount of alkaloid present in it, what the result implies is that neutral water does not extract them while formic acid does. Why would water extract alkaloids from one strain of caapi bark just fine but not the other one unless the second one doesn't have much in it? I mean this stuff was *powdered* wasn't it??

    Good work guys! I'm glad to see someone has set aside time time to do these things. Its important that we get to the bottom of this so we can understand what the acid-enhanced tests really tell us. I mean, if you are srictly looking for 'useable' concentrations of things in plant material then maybe if it doesn't show up without acid, the concentration is too low to bother with.

    [ 24. June 2004, 21:30: Message edited by: Fractalhead ]

  18. Posted

     

    quote:
    That Saliva and wetting of plant material results in fluorescence of varying degrees from medium to low, whereas the addition of Formic acid results in a level of fluorescence much much greater than moistened material

    woops! i can't believe i missed that point. :confused: This is very interesting. Will have to try this out. I checked my Elaeagnus angustifolia the other day with only saliva and got no noticeable glow. Will try with acid tonight.

    Rev, I have only ever seen dull fluorescence coming from the *veins* of caapi leaf - fresh or dry.

    [ 23. June 2004, 21:57: Message edited by: Fractalhead ]

  19. Posted

    The number one rule any gnome should be aware of when dabbling in the HBWR is NO ALCOHOL! The vast majority of negative reports i hear about are where the gnome has been drinking or drinks in the early stages of the trip before it really kicks in. I have seen this time and time again and i have a very close gnome friend who has even experienced the effect of alcohol on HBWR himself so I am sure this is not a myth. The number two rule would be the fresher the better. El Duderino, I highly recommend your gnome try again without alcohol. The THC... no probs It would be a shame to overlook this wonderful plant because you didn't realise the alcohol contraindication.

    [ 22. June 2004, 21:27: Message edited by: Fractalhead ]

  20. Posted

    The b-carbs will not fluoresce unless they are solvated salts. The freebases don't fluoresce and neither do dry crystals of the salts. Hence, all you need to see full fluorescence of the b-carb containing plant material is a little water to rehydrate it if it is very dry. A little saliva also does the trick. I'm not sure whether dry alcohols work as well as water but that would make an interesting experiment. If masticating samples, beware not to cross contaminate. Its amazing how far the fluorescence from a single rue seed will spread Its interesting that you made a similar observation to myself in the confinement of the fluorescence to the veins/petiole in the caapi leaf.

  22. Posted

    I think I found the document you were looking for Rev. Looks like a good paper.

    Phytochemistry. 2000 Mar;53(6):623-37. Related Articles, Links

    Chemotaxonomy and geographical distribution of tropane alkaloids.

    Griffin WJ, Lin GD.

    School of Pharmacy, University of Queensland, St. Lucia, Australia.

    Its got a map showing duboisia myoporoides alkaloid profiles in different parts of queensland. Interestingly, while the ones up in north qld do both have nicotine, they also have scopolamine, hyoscyamine and norhyoscyamine. Perhaps more interesting is that the only region on the map not having tropanes listed as present is down in south east qld at 1000m altitude on the Acacia Plateau.

  24. Posted

    Ok. I'd be really interested to read about that. I still can't think what the big difficulty would be. Just three more questions:

    1. What exactly were they trying to do using HPLC and GC/MS?

    2. Surely they found *something* in their samples so what turned up??

    3. What's this about asparagus?

    [ 17. June 2004, 20:13: Message edited by: Fractalhead ]

  25. Posted

    Well i searched erowid for LSD + HPLC and LSD + PURITY and LSD + PURITY + HPLC and couldn't find any references to any HPLC analyses of street acid. I only had a quick skim so i may have missed the article you read. I did find a reference to the use of HPLC/TLC/MS to identify LSD in urine and other bodily fluids. You can apparently detect half a microgram of LSD with Van Urk's reagent on a TLC plate so I figure that's sensitive enough to detect LSD and any other possible indole contaminants in a 100 microgram trip. I'm not implying that contaminants are neccesarily indoles. In any case, TLC has the sensitivity to detect trace contaminants and once you find any suspect spots, you can scrape them off and have them identified by MS. Sounds pretty straight forward to me. Can you show me who couldn't identify compounds present in street acid??

×