Pictures. Saturated saline solution for Nomans tek.

[Kerberos]

After reading the salty threads and receiving some pointers from ChemisTryptaMan and cyb i am having a go at this.

i am following my normal extraction method except i am starting with a saturated saline solution and also i will be cooking the mixture for 24 hours.
Step 1
I have a 2ltr kilner jar which is filled with 1900ml of de-ionised water. This is placed in a 1.5ltr crock pot (£10 from a store that sounds like bilko)which i am using as a water bath. To this i have added initially one tub of rock salt 362g. I have used rock salt as all the other salt contained anti caking agents. Pic attached.

i Heated this in my water bath up to the same temperature as the mixture will be cooking at. I then kept adding salt until no more would dissolve, this was a total of 450g. I then on ChemisTryptaMan advice let this cool down to room temperature and remove any salt left in the bottom of the jar, approx 5g.

Step 2
i added lye at the rate of 1g per 15ml of water as per Nomans tek a total of 127g. the solution turned milky white and stayed this way. i have taken a small sample to see if it clears over time. I have taken a ph reading from this sample and it is 14. (Pic attached as "ph reading") As of writing it still is milky white several hours later. UPDATE there appears to be a denser layer separating out on the bottom of the sample. i initially thought it was salt crashing out but it is liquid. i will post pic when i get new batteries for the camera.

Step 3a
Added 40g of powdered bark, this was produced with a dry spice grinder. All the fibers where removed and kept with all the other fibers from previous grindings. It is my intention to do an extraction just with these fibers to see what the percentage of DMT is contained in them.

Step 3b
As i added the bark, which i always do slowly, the mixture turned a very strange (scary)dark green colour. I didn't take a pic because my hands where full at the time. The mixture ended up a dark chocolate colour and looks thin if that makes any sense.

Inner coating
The jar looks like it is coated with oil on the inside and makes the jar look smoaky. pic attached.

The mixture is now in my newly acquired water bath and will be cooking until this time tomorrow. It will be allowed to cool to room temperature before being pulled with hot solvent as per the advice of ChemisTryptaMan.
I will be doing three pulls initially as this is the way i normal extract, 40ml, 30ml and 30ml before freeze precipitating. I will then take suggestions on how to proceed after we see if this has worked or not!

All observations gratefully received.

Speak to you tomorrow.
Kerberos
Edit. just had a smell of this and it does not smell of the usual lye/caustic smell. I smells more like a hit of DMT tastes if you know what i mean.:shock:

 

[ChemisTryptaMan]

The container will likely have a layer of oil in it tomorrow, the salinity is probably too great for the oils to stay in solution for as long as they normally do. I'm awaiting your results eagerly.

edit: and the milky color may mean that the lye addition caused some NACL to come out of solution and is suspended, but this should also settle and not effect the extraction. The salinity will prevent the aqueous layer from mixing with the NPS, at least in theory, we will see soon enough.

 

[Kerberos]

The container will likely have a layer of oil in it tomorrow,

The jar has a layer already CTM, in fact as soon as i started to add the bark you could see an oily film on the surface of the solution.

I have just edited my first post to say that the mixture does not smell like normal either. It smells very active, so much so that i had a touch of pre flight anxiety! The mixture seems to be at temperature of 135f, do you think this is OK CTM? or is it to low?

 

[ChemisTryptaMan]

135 is perfect. If you are going to let it go overnight then that is the perfect temp to work at. Once we see what you get here we will know there is or isnt an upper limit to the effectiveness of ionic strength. Great work!

I expect to see something abnormal tomorrow, but this will likely be a three layered system, plant oils will not likely fully integrate into either layer, but slowly will integrate with the upper layer, this is why later pulls tend to be more yellow. You may have an orange/yellow layer that after removing will ensure what is left is going to be really clean, but post a picture in the morning so we can see what it looks like.

 

[ChemisTryptaMan]

If I don't get back to you soon enough tomorrow and there is a significant oil layer, remove it prior to adding any naphtha as this would be a defat without having to do anything but remove the separated oil. That would really kick ass. Then the partition coefficient would be maximally high and a clean product would still be achieved. My biggest worry was that too many fats would get pushed into the NPS, but if they separate on there own that would be a huge blessing.

 

[Legit]

Really looking forward to seeing how this turns out. Also looking forward to seeing how deadhor5's upcoming extraction goes. Damn, I wish I was in a position to extract. The more people we have running extractions, the better.

 

[Kerberos]

I have never done a cook of the mixture before and would like to know what it should smell like?

I am asking because as previously stated this mixture of salt, bark and lye smells more like my used solvent with that DMT smell it builds up over time has. Is this normal?

 

[ChemisTryptaMan]

I have never done a cook of the mixture before and would like to know what it should smell like?

I am asking because as previously stated this mixture of salt, bark and lye smells more like my used solvent with that DMT smell it builds up over time has. Is this normal?

I wouldnt let this concern you, the smell is probably due to the fact the that the DMT wants out of the water as well. Keep the oil layer you pull out in case a significant amount of spice migrates into the oil. At least if this happens you will still be able to recover that spice.

 

[Kerberos]

Thanks for that CTM.

On the oil layer front; it has not materialized as of yet. You do notice a layer clinging to the inside of the jar when you move the mixture around though. This is going to be in the water bath for at least 24 hours as i am working and will not have the time until then. If there is an oil layer i will take this off. i will post pictures of the mixture when i take it out and any subsequent oils that maybe present.

Do you think the oils will be more like to form a layer as the mixture cools back down to room temp?

 

[ChemisTryptaMan]

The oil is more miscible with the water layer at higher temps so this should be the case.

However, I was really expecting you to do cyb's tek with the acid wash in the beginning being the heated part and here's why.

While base and acid will both catalyze the cleavage of the amide bonds in the protein, basic conditions make the spice soluble in oily solvents over aqueous ones. I expect that if an oil layer does form it will tend to hold on to any spice in contacts and will lower the yield from your pulls. If an acid cook is done the spice is far more comfortable in the water layer while the proteins are being digested. Once the long heated acid bath is over, then base can be added, layer or not, and the spice will become insoluble in the water at this point, right before you add the NPS, ensuring not enough time has passed for the spice to integrate into the oils. the "layer" may also just be attached to the walls of the container as well, it may not settle at either the bottom or the top. Eventually it will, but this could take a very long time. Also, did you lyse the cell walls with a freeze/thaw before you started this process. I'll explain how to do this optimally in another post where we were already discussing it but I have to remember which one it was first.

 

[ChemisTryptaMan]

Also, if the oily layer stays stuck to the jar, just pour the aqueous contents into a new container before you basify the solution. This will keep the pulls cleaner.

 

[Infundibulum]

The oil is more miscible with the water layer at higher temps so this should be the case.

However, I was really expecting you to do cyb's tek with the acid wash in the beginning being the heated part and here's why.

While base and acid will both catalyze the cleavage of the amide bonds in the protein, basic conditions make the spice soluble in oilly solvents over aqueous ones. I expect that if an oil layer does form it will tend to hold on to any spice in contacts and will lower the yield from your pulls. If an acid cook is done the spice is far more comfortable in the water layer while the proteins are being digested. Once the loong heated acid bath is over, then base can be added, layer or not, and the spice will become insoluble in the water at this point, right before you add the NPS, ensuring not enough time has passed for the spice to integrate into the oils. the "layer" may also just be attached to the walls of the container as well, it may not settle at either the bottom or the top. Also, did you lyse the cell walls with a freeze/thaw before you started this process. I'll explain how to do this optimally in another post where we were already discussing it but I have to remember which one it was first.

Chemistrypaman,

Most of the plant oils saponify very quickly in an STB and will thus blend nicely with the rest of the aqueous soup. I'd be very surprised if oils form a distinct layer as you theorise.

Also, proteins hydrolysis does not happen as easily as you suggest, and I am not convinced that you can predict the impact of any free amino acids in the extraction system.

I believe that you have nice ideas and I enjoy educated posts. I also like that you trouble-shoot extractions. However, I do think that you theorise a bit too much and you need to cut that down a bit. You will never know all the fine details of what is going on in an extraction. Examples given:

1. you failed to mention potential saponification of oils and that this would make them very miscible in water and went on to predict that they'd separat

2. you assumed protein hydrolysis and the effect that may have without actually giving a demonstration of whether proteins actually degrade in the above given conditions or why this is of any significance to begin with.

3. your predictions also fail to mention dmt chelation of potential clathrate structures. etc

The list of factors that one can include in any theoretical predictions can really go on forever. Please remember that extractions are quite complex systems, not a simple aqueous solutions. We truly appreciate knowledge of said factors but let's not over-predict things!

 

[Kerberos]

No to acid wash and no to lyse.

it was always my intention to compare adding salt with my usual way of extracting for a personal comparison. However some more bark has been ordered and i will follow cybs tek but using a saturated solution (if no one else has done it by the time it turns up).

One consolation is that we will know more about how much DMT does get locked in to the oils (and the fun in getting them out again)

 

[ChemisTryptaMan]

infindibulum, I theorize a layer may form as I have seen it happen myself in highly salinated solutions in the lab many times. I'm very experienced with organic synthesis as I have worked in research labs as a graduate student and have practiced everything I've talked about in a real life setting. I don't think every oil will separate, only those of low enough polarity to be fully immiscible in that high of an ionic strength. I have done acid base extractions using more solvents and conditions than I can count as this is usually part of the purification process when recovering products after a reaction. I failed to mention saponification because I intended the heated bath to be done under acidic condition where it would not occur. If anyone wants to research acid or base catalyzed hydrolysis of proteins to see what conditions are necessary for the reaction the information is very available as this is a widely covered topic in biochemistry. Acid/Base chemistry is the basis for most properties of biomolecules.

Sorry if I am being too defensive, but I really do know what I'm talking about, and for the speculation part of this it is a collaborative effort between several members to find the optimum conditions. I am theorizing about what might happen and I make that clear, I see this extraction as an experiment to give us a greater understanding of what happens during these extractions just as Kerboros does. I apologize again if I seem too defensive, I just don't want to be viewed as someone who states theories as fact. When I'm speculating I say so, when I am spreading knowledge I have gained through years of study and years of experience, I don't call it speculation.

edit: also, not all oils saponify, this is only true for fatty acids attached to glycerol, which are just one type of oil in the mix.

 

[Infundibulum]

Thank you for your reply,

Of course there is no need to sound defensive nor you need to put your credentials up front. We far prefer to talk with demonstrations, rather than taking ab initio for granted that if someone has credentials then he's bound to be correct.

Re the acid or base protein hydrolysis, from protocols I found through google you heed to do it in either very concentrated acid or very concentrated base at over 100 Celsius for many hours. Neither conditions are matched in a typical A/B or STB. So if you want to to claim that you have protein hydrolysis in the typical A/B or STB teks then you have to demonstrate that it happens. I am not discussing the latter point, I am straight telling you that should you need to make a point about protein hydrolysis as you do, you will be requested to provide proof (either yours or other's).

Re the oils and saponification, of course not all oils saponify, and not at the same rate either. But in a product like mhrb which is inherently oil-poor material, you would expect to find primarily tryglycerides (as constituents of plant membranes), which are quite saponifiable.

As a final note, please try to avoid ad hocs as well; in the long run, inadequate interpretations like this:

The oil is more miscible with the water layer at higher temps so this should be the case.

and

but I really do know what I'm talking about,

..do not click together nicely.

 

[ChemisTryptaMan]

Re the acid or base protein hydrolysis, from protocols I found through google you heed to do it in either very concentrated acid or very concentrated base at over 100 Celsius for many hours. Neither conditions are matched in a typical A/B or STB. So if you want to to claim that you have protein hydrolysis in the typical A/B or STB teks then you have to demonstrate that it happens. I am not discussing the latter point, I am straight telling you that should you need to make a point about protein hydrolysis as you do, you will be requested to provide proof (either yours or other's).

Re the oils and saponification, of course not all oils saponify, and not at the same rate either. But in a product like mhrb which is inherently oil-poor material, you would expect to find primarily tryglycerides (as constituents of plant membranes), which are quite saponifiable.

As a final note, please try to avoid ad hocs as well; in the long run, inadequate interpretations like this:

The oil is more miscible with the water layer at higher temps so this should be the case.

and

but I really do know what I'm talking about,

..do not click together nicely.

Protocols do call for reflux, but the reaction is carried out at lower temps regularly when working with molecules unstable at 100C, the reaction just takes longer. This is a fact that shouldn't even need to be debated. Obviously if the temperature is too low the rate becomes negligible and the hydrolysis could take years to complete.

I have found some very heavy oils/waxes in mimosa, especially when using a salt tek. Everything from Dark red/brown wax to orange wax have shown up after recrystallizing.

I don't see my first statement there as adhoc, as it is also fact, though i do not know the exact miscibility nor its exact relation to temperature, but adding energy in the form of heat will overcome some of the hydrophobic interactions and allow for greater miscibility.

I apologize for throwing out my credentials as I can see that it may be interpreted as boasting but my intent was simply to confirm that I don't just pull this information out of thin air. I have been a chemist for the better part of a decade, admittedly not the longest time, but I do know this stuff and I felt like you were attacking my attempts to contribute to this thread, as I believe it was me who inspired the experiment to begin with.

I will try to do the experiments that some of us are collaborating on in the PM's for now. Until we have the data we should probably not post anything about our work, as it seems my brainstorming with kerboros about what he might expect caused some misinterpretation of what was being said.

 

[benzyme]

I have been a chemist for the better part of a decade

that's it?
you call that credentials?

sorry, didn't mean to sound rude. I've been a chemist for almost two decades (17 years), before there was a hive. I still learn new ways of solving old problems.

 

[ChemisTryptaMan]

I feel like if you stop learning new ideas you have effectively stopped living. I love chemistry with a passion and research has been my life for so long. I love spending time searching the literature for whats happening in all areas of chemistry. I recently lost my access to sci-finder as my old employer finally took me off their license, but I'm continuing to learn something new and exciting everyday. I Initially went into grad school intending on studying metallo-organic chemistry and ended up finding myself doing natural product synthesis. You never know what might spark your interest next. As a chemist, you know just how much we learn in grad school and just how intensely we study every aspect of what we do. I don't post ideas based on fantasy, and when I do speculate, I try to make it clear that's what I'm doing. I hope that at some point we can work together on solving some problem. The amount of coordination several of us are putting into optimizing yields right now is a statement for the power of this forum. A few minds come together and what results is beautiful. Cyb's tek alone has made 2% the standard yield for many people working with MHRB. I think I may have taken infin's comments too much to heart, but I honestly felt like I was being made to feel stupid or something for talking about things I do have a solid understanding of to say the least. I love this forum and everyone on it, so please be sure any ideas I talk about are things that I feel will benefit the community in some way. This thread is an experiment to test the effects of complete saturation on yield, so that we have a value that we know is coming from above optimum salinity, and prove that one must exist.

Love and Light
CTM

 

[benzyme]

don't limit your learning experience to grad school, or institutionalized learning. you'll learn way more outside of school, with "real world" experience. theories are good for coffee banter.

 

[ChemisTryptaMan]

I think I learned more from my last job than from the end of grad school. I was making toner for a printer company and learned a lot about production on industrial levels, which put a lot more perspective on things for me. Meetings with the chemistry team were downright hostile at times when there was a disagreement, which I never really understood. I worked before that developing assays for biomolecules to be used in medical machinery, learned a lot about manufacturing and applying chemistry to actual real life situations. I refer to grad school more cause that was where I learned how to learn. This is a field where you really have to know how to teach yourself and solve problems on your own. The third mystery school of ancient Egypt was apparently life itself, but I have to double check my sources on that info. Most facts start out as theories as well, so I love to discuss theory nearly as much as fact. Who knows, some of the strangest theories prove to be true sometimes.

I hope that my ideas are beneficial to this community, If I put forth an idea and it proves to be incorrect, I will be sure to say that it is not yet tested as I always do for the most part. That's really what we are trying to do is test some theories to see if we can improve yields for everyone as our sources become more difficult to acquire.