it will not be an issue. i did a lot of work with trehalose back at the univeristy. super cool stuff.
i can figure out a ratio. it might take a little dicking around, but it should be easy enough to find.
as for the price of chlorin e6, that is cheap. welcome to the wonderful world of chemical supply. i actually think i can get it cheaper tho. i'll keep y'all updated.
it's a natural product though. it shouldn't cost that much. see this is where i normally say fuck buying it and figure out the extraction process but for something im putting in my eye i may bite the bullet. ill browse around as well.
Just found a patent describing the synthesis and extraction of chlorin e6. it's actually super easy. you just extract chlorophyll a, treat with HCl to remove the magnesium then add NaOH to remove the tail and voila, cheap easy chlorin e6. just purify the product which isn't hard and you're good to go.
I had a thought. if you look at the structure of the chlroin e6 it looks very similarto dinofalgellate luciferian. i wonder if other pigments could be used to give other colors visibility.
the cost in time and resources to make the yield you want at a purity safe to put in your eye is far beyond the cost of buying it from a bulk chem supplier.
This is the same reason that we ended up buying the A2 at $1k for 50mg (and that was a discount). Making it from scratch would have required building a lab to make it which cost more than just buying it. This is why we ended doing this whole lab building thing, but we still don't have some of the needed tools to do this now...
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$40 for 100mg is very reasonable for lab grade chemicals.
you gotta stop saying "it's super easy" to do things. Define how one purifies the product with the given tool set that most people have? How is purifying a product not hard? i've spent days purifying a product using lab grade chemicals and a proper tools. I'm all about DIY bio and chem but you still gotta decide what is important in terms of tools and cost. Where to draw the line at doing every step oneself and what corners you are willing to cut. I am reading this patent, and based on the wording, they are glossing over the casual tools that you have in an industry lab (one skilled in the art) that are really expensive.You have to remember, you are considering you are doing this to make something that you put in your eye. Using ish from home depot as your reagents might not cut it...
Beyond that, the fact that this process isn't as time intensive as the previous reference means that it only takes, from my guess, about 3 days, once you have gotten the protocol down. So probably a month of dedicated work to make sure that you can do this without fucking it up.
It's like the whole extracting lidocaine things that came up earlier. I mean, you could, but why would you do that? Why make your life hard?
That being said, with proper funding, SfM would be happy to be the chem builder for the community. A full rotvap costs about 1k. An HPLC costs about 35k. We can make these things. We will make these things for you. But we (and you all) shouldn't make bathtub 65% yield crap with shit tools
I say it's super easy in that it's two simple reactions. also if you can get chlorophyll a in pure form cheaper than the chlorin e6 than it could be converted with less impurities than trying to extract it. Yes of course toolage is expensive (although you can build a rotovap easily enough) and hplc cannot really be made but the sysnthesis is interesting enough to be worth trying it even if it's not going to end up in your eye. I say thing's like "it's super easy" only because i have a way about things that is not what you would call typical. when someone tells me something is doable i'll find a way to do it if not out of spite but out of curiosity. Ive done plenty of things people say aren't feasible. also im a student on a student budget. mind you if all goes to plan all of this will be meaningless and i'll be able to afford as much of the lab grade stuff without having to both synthesizing it. im just curious to see if there is some alterante pathway to a product. again all i want is to explore the option sheerly out of curiosity. I want to learn how it's done and learn why it's not viable. there are few thigns i hate more than being told I can't do something or its impossible. I'd rather understand the science and see that it's impossible than just be told. Remembering, we're working on things that most people think are insane or impossible.
The following procedure was reported in Oncology Reports, 2009, 22(5). 1085-1091 by Korean scientists Yeon-Hee Moon et al. Live chlorella (Chlorella ellipsoidea) 100 g (dried weight) was sequentially washed with 500 ml of water and 300 ml of 50% ethanol in water to remove polar materials and the residue was extracted twice with 500 ml of 100% ethanol to obtain chlorophyll a rich fraction (extraction yield 4.3%). Stirring the combined ethanol solution of chlorophyll a in 1 N HCl (pH 2.5) for 3 h at room temperature afforded pheophytin in the form of precipitates. The precipitate was dissolved in dichloromethane washed with distilled water, dried with anhydrous sodium sulfate, and rotary-evaporated to dryness. The residue was purified by a chromatography using neutral alumina (Aldrich, Brockmann, ~150 mesh) with a gradient elution from 30% dichloromethane in n-hexane to 100% dichloromethane. The main green band was collected and evaporated to dryness. The crystalline powder was dissolved in acetone, adjusted pH 12.0 with 1 N NaOH, and stirred for 12 h. The precipitated Ce6 was filtered, washed with acetone and dissolved in 100 ml of water, and filtered to remove insoluble impurity. After lyophilization of the filtered water solution, a fine black powder of Ce6 was obtained. The purity of Ce6 is 93-98% (yield of Ce6: 1% from dried weight of chlorella).
again what immediately jumps out is the chromatography which is the expensive part if you use hplc. but that being said i've now settled my curiosity in how it's made.
It's great that you are interested in how these things work. I am always happy to discuss this. I guess I got stuck on the practical aspects of making the drops and get worried that either the conversation will dissolve into what ifs, or worse, that someone will decide that they are going to give it a shot with what they have in their kitchen and end up hurting themselves.
I agree that the protocol is fairly straightforward. Sadly, any decent chromotography setup is going to really cost a pretty penny. especially with 1% yield :(
It's definitely on my list of things to get for the lab, but we need some serious cash before that happens.
The amount of Ce6 needed for 1 ml is 20mg. This means with 100mg you can make 5ml. That's 100 drops of solution. The method calls for application every 4 hours while awake (3 times) for 4-7 days. Even at the full 7 days, this still comes out to less than 100 drops.
I have sent out quote requests for larger amounts and availability.
ya at 3 drops per eye assuming you use it once a day you'd get a month of use out of it. i don't know why you'd be in the dark for more than 3 hours everynight anyway
are we talking equipped to make them from scratch as the conversation has kinda veered towards or just 'we have the ability to source and mix these cause we're a lab"
from scratch doesn't stand to talking about. still a "buying core lab tools" situation. way too much. i can give you numbers, but its just depressing.
now, making 100ml from purchased materials, we are talking about roughly 1k. not counting the tools that we already have, shipping, etc.
as an initial investment, i shy away from making 100ml of something when you haven't tested 5ml.
however, as a struggling scientist who wants to make cool things for the community, i will happily take your money if you want a bunch of this ;)
for everyone interested, i am getting quotes now on materials and will be making a few small batches of this for people to try (including myself and Cassox). if you are interested, or you want to support this work, please pm me.
idea. what if other pigements are used? or what if just chlorophyll was used? who knows maybe one of them could let us see fun new colors XD it's jsut speculation but i'll dig into a bit see if i dig anything up. if not maybe it's worth a try if we can get pure enough products.
i am goingto guess that before publishing, they ruled out the other chlorophyll. You dont get to publishing point with something that specific without testing a lot of other stuff
fair point. or they got really lucky but i doubt that. That said there's a pretty wide variety of pigments. and maybe chlorophyll just didn't work AS well so they didn't bother working with it. I dunno it was just a thought but you're probably right.
Congrats on the results, guys! I can't wait to see the data when you publish it. Regarding the chlorin e6, would we be able to organise a group buy of an eye drop preparation for anyone who's interested? And from reading the article (and most of the patent, jesus that was tedious) I'd say that it does work with other pigments, since they said some flies use a similar method to see in the UV range. The difficult part is to know which pigments will safely be incorporated into the correct cells and interact with this pathway, which is something that requires a lot of research to find out.
Buffer? From what Im reading, they used sodium bicarb to buffer it. The insulin is being used as an uptake mechanism with the addition of glucose. The patent also discusses the use of a slew of other other substances such as ketamine, iodine, and topical anesthetics. This patent appears to be an attempt to cover as much area as possible. It's non-specific as to what insulin type, basically saying short, medium, and long term. I've seen no documentation regarding trials and results. Until I've researched this further I will have no involvement in this project.
After looking into it more Im in. I still have a few concerns though. One: safety. Dude, this stuff is used as a sensitiver to kill cancer cells with light. Has anyone looked up its Ld50 even? There are also some solubility issues.it is soluble in water at certain ph at low density but works better in am oil. I do want to move forward, but as of right now i wouldn't put it in my eye. I need to do some serious research here first. I'd love to be able to get my hands on some to experiment with, but im leery to promise a product of any kind.
I am a bit late to this so sorry for that I am a new member, in one of the first posts where this is explained you mentioned monitors and the really bright colors.
Have you tried to change the colors that are prominent in the settings to see if it appears "Normal" at a set range?
Sorry just another question, would this work as well if I used this on contact lenses IE bought some that do not refract light the ones that change your eye color without the removing of melanin.
Comments
The following procedure was reported in Oncology Reports, 2009, 22(5). 1085-1091 by Korean scientists Yeon-Hee Moon et al. Live chlorella (Chlorella ellipsoidea) 100 g (dried weight) was sequentially washed with 500 ml of water and 300 ml of 50% ethanol in water to remove polar materials and the residue was extracted twice with 500 ml of 100% ethanol to obtain chlorophyll a rich fraction (extraction yield 4.3%). Stirring the combined ethanol solution of chlorophyll a in 1 N HCl (pH 2.5) for 3 h at room temperature afforded pheophytin in the form of precipitates. The precipitate was dissolved in dichloromethane washed with distilled water, dried with anhydrous sodium sulfate, and rotary-evaporated to dryness. The residue was purified by a chromatography using neutral alumina (Aldrich, Brockmann, ~150 mesh) with a gradient elution from 30% dichloromethane in n-hexane to 100% dichloromethane. The main green band was collected and evaporated to dryness. The crystalline powder was dissolved in acetone, adjusted pH 12.0 with 1 N NaOH, and stirred for 12 h. The precipitated Ce6 was filtered, washed with acetone and dissolved in 100 ml of water, and filtered to remove insoluble impurity. After lyophilization of the filtered water solution, a fine black powder of Ce6 was obtained. The purity of Ce6 is 93-98% (yield of Ce6: 1% from dried weight of chlorella).
again what immediately jumps out is the chromatography which is the expensive part if you use hplc. but that being said i've now settled my curiosity in how it's made.
I'm interested, I'll shoot you a PM.
Regarding the chlorin e6, would we be able to organise a group buy of an eye drop preparation for anyone who's interested?
And from reading the article (and most of the patent, jesus that was
tedious) I'd say that it does work with other pigments, since they said
some flies use a similar method to see in the UV range. The difficult
part is to know which pigments will safely be incorporated into the
correct cells and interact with this pathway, which is something that
requires a lot of research to find out.
Have you tried to change the colors that are prominent in the settings to see if it appears "Normal" at a set range?