Alcohol Extraction

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Graywolf

Graywolf

Judging by the links you assume that the water dissolves or weakens the attachment of grains of chlorophyll within the chloroplast?
Unfortunately, these articles are not readily available to my understanding. My familiarity with the anatomy of the cell is limited only by -


I was interested in the location of the chlorophyll in the cell when suddenly got an emerald-green butane extract, washed from dry sepals peeled from fem seeds.
Green puzzled me, because the chlorophyll does not dissolve in nonpolar butane.

Then I decided that this strange behavior of chlorophyll depends on how the grains of chlorophyll are attached to the tissues of plants.
If they are held by some grease or wax, which are soluble in butane, after the dissolution of these compounds are released and the chlorophyll grains fill butane solution in a suspended state, but not dissolved.

With the naked eye, we cannot distinguish individual objects smaller than 50 microns and visually perceive suspension as a solution. Chl grains are 2-10 microns.
But even in these articles they write that the membranes are hydrophobic and water-soluble proteins are rare.

All chlorophyll (Chl)-binding proteins involved in photosynthesis of higher plants are hydrophobic membrane proteins integrated into the thylakoids. However, a different category of Chl-binding proteins, the so-called water-soluble Chl proteins (WSCPs), was found in members of the Brassicaceae, Polygonaceae, Chenopodiaceae, and Amaranthaceae families.



I used this polar->nonpolar gateway, transferring the desired resin from the green solution in ethanol to non-polar petroleum ether, it was the British lighter fluid "Newport."
The procedure was carried out in several stages.
I added small portions of petroleum ether, shaking vigorously, waiting for the separation of layers, took a yellowed layer of ether with a syringe and added a fresh portion instead of removed.
Of course, each successive portion was painted paler than the previous one.

Only in the last step I added water. Without water, the layers are separated much faster and a very inconvenient boundary layer of emulsion does not form.

Chlorophyll, as it is insoluble in a nonpolar solvent, always remains in alcohol and after adding water, chlorophyll remains in alcohol, diluted with water, but not in water.

In my opinion, this ancient technology from the era before the butane in no way indicates the solubility of chlorophyll in the water.

Unfortunately Water cannot wash away Chlorophyll.
 
Graywolf

Graywolf

Judging by the links you assume that the water dissolves or weakens the attachment of grains of chlorophyll within the chloroplast?
I was tossing Water Soluble Chlorophyll Binding Protein on the table as the possible elusive water soluble substance binding the chlorophyll in the plants and allowing water washing to exhibit some of the properties it does.



They are proteins that are soluble in water, have some solubility in alcohol, and are insoluble in non polar solvents.



Broken loose, the chlorophyll could form micelle, by clumping with their polar ends out and their hydrophobic tails in, which would allow them to be transported by the water.



Freezing would make them less available to the water, and with them in place, the chlorophyll would be protected from the alcohol.



What that doesn't explain, is the light green hue BHO picks up without the presence of alcohol.



That brings us to the definition of insoluble, which gets lightly tossed about as an absolute, when in fact it is a relative term. The alkane butane, considered insoluble in water, is not totally and will hold up to 32ml/L water.



Since there are no simple oxygenated alkane alcohols present, what is responsible for extracting the green?



I submit that polar chlorophyll is also slightly miscible with polar water, because it has both polar hydrophilic surfaces and hydrophobic surfaces.



Unfortunately, these articles are not readily available to my understanding. My familiarity with the anatomy of the cell is limited only by -

I was interested in the location of the chlorophyll in the cell when suddenly got an emerald-green butane extract, washed from dry sepals peeled from fem seeds.

Green puzzled me, because the chlorophyll does not dissolve in nonpolar butane.

Then I decided that this strange behavior of chlorophyll depends on how the grains of chlorophyll are attached to the tissues of plants.

If they are held by some grease or wax, which are soluble in butane, after the dissolution of these compounds are released and the chlorophyll grains fill butane solution in a suspended state, but not dissolved.

With the naked eye, we cannot distinguish individual objects smaller than 50 microns and visually perceive suspension as a solution. Chl grains are 2-10 microns.

But even in these articles they write that the membranes are hydrophobic and water-soluble proteins are rare.

All chlorophyll (Chl)-binding proteins involved in photosynthesis of higher plants are hydrophobic membrane proteins integrated into the thylakoids. However, a different category of Chl-binding proteins, the so-called water-soluble Chl proteins (WSCPs), was found in members of the Brassicaceae, Polygonaceae, Chenopodiaceae, and Amaranthaceae families.

I used this polar->nonpolar gateway, transferring the desired resin from the green solution in ethanol to non-polar petroleum ether, it was the British lighter fluid "Newport."

The procedure was carried out in several stages.

I added small portions of petroleum ether, shaking vigorously, waiting for the separation of layers, took a yellowed layer of ether with a syringe and added a fresh portion instead of removed.

Of course, each successive portion was painted paler than the previous one.

Only in the last step I added water. Without water, the layers are separated much faster and a very inconvenient boundary layer of emulsion does not form.



Have you tried using saturated brine, instead of plain water?



Chlorophyll, as it is insoluble in a nonpolar solvent, always remains in alcohol and after adding water, chlorophyll remains in alcohol, diluted with water, but not in water.

In my opinion, this ancient technology from the era before the butane in no way indicates the solubility of chlorophyll in the water.



Unfortunately Water cannot wash away Chlorophyll.

Ummm, and yet salt water does regularly in our lab.

Plain water also washes green out of plant material in our bubble washing machine, so you can see why I'm not just jumping on the total insolubility/miscibility thang?

The bubble machine water looks like a colloidal suspension, but it is still being washed away by the water. How else did it get there, even as a collodial suspension, from its position locked in the plant material?

Do you have an alternate theory leaving chlorophyll totally insoluble, and yet explaining the observable?
 
jump

jump

Plain water also washes green out of plant material in our bubble washing machine, so you can see why I'm not just jumping on the total insolubility/miscibility thang?

The bubble machine water looks like a colloidal suspension, but it is still being washed away by the water. How else did it get there, even as a collodial suspension, from its position locked in the plant material?

Do you have an alternate theory leaving chlorophyll totally insoluble, and yet explaining the observable?

I know or maybe I believe that water is H2O, and ethanol is C2H5OH.

You can consider me a dogmatist, but for me there is the concept of an undisputed scientific fact.
Moreover, I think it is absolutely necessary tool in the learning process. Their value cannot be neglected.

For almost a hundred years scientists have seriously investigate Chlorophyll in various aspects.
I see no reason to doubt the validity of their statements about the solubility properties of chlorophyll.

If the explanation for the observed phenomenon contradicts the scientific facts, then most likely it is wrong and some other explanation must be found.

I think that green water is a suspension, hydrosol, colloidal solution formed by the grains of chlorophyll released from the chloroplast as a result of chemical or mechanical destruction of the membrane that holds them in a thylakoid cassette inside the plant cell. I think that green butane solution has the same nature.

I do not see anything surprising in Healthy Kitchen-Green Smoothies ... My manic friends drink them for eternal youth.
--------
Salt ... No, I did not add salt in water, it was long ago and since then I have not had any problems with the chlorophyll.
Do I understand correct that salt is used in combination with the two solvents, when the first was polar? Or primary hexane can also pick up Chlorophyll?
 
jump

jump

It is interesting to know whether it is possible through a microscope to distinguish
the solution of Сhl + iso
from
the emulsion Сhl + iso + H2O
and from
the suspension of Chl + H2O?
 
Graywolf

Graywolf

Not ignoring you brother Jump, just having problems downloading Joe's detailed response, which unfortunately is in a later MS Word version than mine.

I predict we will solve that issue shortly and that it will cut to the chase on the issue, as he understands it.

Peace!
 
jump

jump

Thank you Gray Wolf !

I would venture to suggest that the document will set out the beginning of colloid chemistry.

The theory of colloidal particles is so complicated thing

that in discussing the physics of the micelles and the double electron layer,

we risk easily stop the interest of readers to the extraction of alcohol in general.
 
Graywolf

Graywolf

Ok, here is Joe's analysis without his attached pictures:

Chlorophyll Info
Chlorophyll is an intra-membrane chemical within a thylakoid. A thylakoid is a membrane-bound compartment inside chloroplasts.
The thylakoid membranes of higher plants are composed primarily of phospholipids and galactolipids that are asymmetrically arranged along and across the membranes
Chlorophyll is shown as photosystem I and II in this illustration.

Both phospholipids and galatolipids have hydrophilic (water loving) heads and hydrophobic (water fearing) tails.
In biology this theme is used in almost all life forms to compartmentalize for energy storage, isolate invaders or encase their genome to protect it and many others reasons.
Solubility is a term that has more than one definition. In inorganic chemistry it refers to waters (or another compound) ability to break covalent and ionic bonds of most compounds, dependent on time temperature and pH. The “solubilized” atoms are then bonded to their polar opposite ion H3 (+) or OH (-) and are in solution.
In biology however, it is used in the first tense but, it is also used to describe the ability of an organic molecule or complex to form an association with water and be in solution but not be “solubilized” by it. Proteins and other organic molecules use charged ions such as phosphate (PO4) and Sodium (Na) to form micelles. Micelles are little balls of hydrophobic molecules surrounded by a charged ion. Just like a cells membrane bilayer. Sometimes micelles are formed by complexes of proteins surrounding a small molecule for transport through water.
Chlorophyll specifically, is only able to form complexes with other molecules to stay in solution at biological pH (7.4). Its natural environment is at a pH of around 4 not 7.4. At this pH it has a net charge of -2 so that it can form a chemo-gradient for electron transport during photosynthesis.
So since pH =-log [OH-/ H+] when at pH 4 the [H+] concentration is higher thus giving an environment that is more likely to associate with the (-) charged area of the chlorophyll molecule. Hence the low solubility of unbound chlorophyll in water, the large hydrophobic areas compresses together and presents their hydrophilic areas to exclude water from the center becoming a mass that will sediment in water.
The point of this is to illustrate that while purified chlorophyll is not likely to stay in solution in pure water; we don’t extract pure chlorophyll and we don’t use water. We use brine to keep the charge on the phospholipid bilayer (Na+ with PO4-3) and no detergents. The alcohol (ROH+) wants nothing to do with Na+ while in its protonated (H+) state. Non polar solvents for obvious reasons won’t either and also won’t form much of an emulsion with alcohols in their bent state because; the alcohol is denser and forms a micelle like layer to protect itself from the charged Na+. If there is an excess of alcohol it will start forming an emulsion layer at the upper interface.
The phospholipid bilayer of the chloroplasts and of the thylakoid being intact or mostly so prevent the chlorophyll from being disassociated with the Na+ water and are able to be excluded from alcohol or non-polar solvents . Alcohol is able to associate with chlorophyll and proteins in its native conformation but not when bent by Na+ because the electron pool concentrated at the (O-) repels the (-) region of the chlorophyll molecule and the PO4-3 of the membrane bilayer. If the membranes have been broken up by a detergent or solubilized by enzymes then the only way to exclude chlorophyll from a non-polar solvent or alcohol is with lots of Na+ and water. Because of the large non polar area of the chlorophyll molecule it can more easily form a hydrophobic interface and shield the charge in the center. The salt exposes the charge (because ionic is a stronger bond than Van Der Wals forces) and precipitates the chlorophyll into water.
So in conclusion, chlorophyll is not wholly soluble in water, but in its biological complex is able to associate with it. By manipulating charge/charge interactions chlorophyll can be forced into solution with water and away from polar organic and non-polar solvents. While it doesn’t meet the inorganic chemistry definition of solubility, it will form micelle complexes with an ionic solution and it can be precipitated from that solution under the right conditions. From a biological perspective it can also be solubilized by any solvent under the right conditions.
 
T

Turtle Man

9
3
Interesting read, thanks Joe and Graywolf!

Starting off with 99% iso, during the purifying steps (brine wash etc) is there a significant advantage of using hexane instead of acetone?
 
sox

sox

Yes. Acetone isn't non polar.
look at this weird shit i made from acetone...is it safe to smoke?? i wouldnt think sooo_O
It smells SOOO GOOD though! its trippy.
i think im gunna try and wash it with 99% and then evap that...i cant find hexane for shit!
2012-06-29_18-51-41_576.jpg
 
deep buddy

deep buddy

look at this weird shit i made from acetone...is it safe to smoke?? i wouldnt think sooo_O
It smells SOOO GOOD though! its trippy.
i think im gunna try and wash it with 99% and then evap that...i cant find hexane for shit! View attachment 230238

you can smoke it. i would vac purge it for a bit first. well for a while, but you can smoke it. im sure its the bomb
what method did you use with the acetone?
 
sox

sox

you can smoke it. i would vac purge it for a bit first. well for a while, but you can smoke it. im sure its the bomb
what method did you use with the acetone?

pretty much i just ran it straight over, evap then froze it...only time it ever turned out that way..
 
S

ShiftyThumb

Boo! Lol. So 99% is at Randall's, Safeway, blah blah chain. Little bottles but what can ya do. Trying out some different ISO making variations and wash times. The GDP batch that started this thread was my finest batch I've ever made though. I'm sure the material used was a big factor. That was fiyah. Stay safe n smokin
 
squiggly

squiggly

I see that was an old post by graywolf, but just a little addition to it--you can think of the effect not so much being about whether or not chlorophyll is "water soluble" its more about it being "more soluble" in the water than a different solvent under a set of conditions.

Things aren't usually "soluble" or "insoluble" there are degrees of solubility. That is to say things are generally more or less soluble, not soluble or insoluble. As Graywolf points out you can do manipulation to the kinetics in your solution to make certain molecules be "more soluble" in a preferred layer.

In chemistry, we do this all the time. It generally comes at the end of a reaction, and we call it "working up" a reaction.
 
S

ShiftyThumb

So I didn't read any of what had been discussed before posting last, and after doing so, I'm so glad that this thread went from basic ISO extraction to a very detailed and complex debate about the finer aspects of the chemistry and physics involved in the process. I love learning from people who have a deeper understanding or a differing view of a subject than me. Always creates an environment for learning and a bettering of the craft.
 
S

squarepusher

im pretty clueless at chemistry, but what if you did something like this. Say you have 3 liquids, say hexane (NP), ethanol and water. Hexane and water are not miscible, but ethanol is miscible in both. Based on this, is there a way to isolate/remove cholorphyl?

like i said I am a chem noob just wondering if someone can explain to me why I am wrong.
 
Graywolf

Graywolf

Chlorophyll is mostly insoluble in a non polar solvent, so if you extract it with alcohol and then mix that alcohol with equal parts salt water and hexane, the cannabinoids will migrate to the Hexane, and leave the chlorophyll with the water and alcohol.
 
S

squarepusher

ok here's a question, since Methanol and Hexane are not miscible, if you simply washed hexane with methanol, would that separate the chlorophyll/cannabinoids as well?
 
Graywolf

Graywolf

ok here's a question, since Methanol and Hexane are not miscible, if you simply washed hexane with methanol, would that separate the chlorophyll/cannabinoids as well?

A Hexane extraction is unlikely to have much chlorophyll in it in the first place, but if it did, the cannabinoids would go to the hexane and the polar elements like chlorophyll to the Methanol. If you also used brine, you could take out the water solubles. http://skunkpharmresearch.com/getting-the-green-and-waxes-out-afterwards/
 
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