Chlorophyll
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- Опубліковано 15 гру 2023
- In this video I spend some time extracting and running chemical tests on chlorophyll.
Specifically I show how chlorophyll can be extracted and purified using different solvents, centrifugation, and chromatography.
I also demonstrate the UV fluorescence of chlorophyll, and the mechanism of photosynthesis.
This was mostly done for fun, and the end goal for this project was itself.
#chemistry #science #hydrogen #gas #elements #fire #chemical #industury #color #green #light - Наука та технологія
I definetly could fall asleep at the fluorescent footage and music if it was only a few minute longer, amazing work, really loved the music.
Trust me when I say I had a lot of fun making that footage. I wasn't sure if it was smart to put a 5 minute block of colorful footage with no speaking in the middle of the video but I'm glad you liked it!
Nice colors! Cool photosynthesis demonstration too!
Thank you! 😁
great video. i learned i should water my plants with a baking soda solution.
Your light show was dope, and that photosynthesis demonstration was also, dope
Thank you 😁 the fluorescence was the real motivation for making the video but the photosynthesis demo did turn out to be a neat little proof of concept
Damn the fluorescence hits hard
Thanks! I had a lot of fun with it trust me
Great video - the simplicity and effectiveness of paper chromatography makes it so accessible. Really makes me miss my organic chem and IR labs I had in university.
Thank you! I actually am doing a much longer video on paper chromatography and all the other types of chromatography (would have it out already but column chromatography separation is tough to perfect without actual lab equipment).
Main motivation for the full chromatography vid was actually this one because even after the redo I wasn't really happy with how my paper chromatography turned out. Spots too big (obviously) and the separation could have been even better. In any case I agree it is a wonderful technique!
Next up. Chlorophyll dye laser!
That was an excellent demo of chromatography for solvent selection
Thanks man! I'm actually doing a whole video on several different types of chromatography and all the ways it can be used (since I really don't think that's been done on UA-cam yet at all). It would have been out already, but I really wanted to get a good seperation of Excedrin using column chromatography and it's been a lot tougher than I initially expected
Well done! Thanks! Paper or TLC is such a useful technique and so accessible, thanks for the great demo! Especially with visible chromophores. But how did the strips look under the UV? The photosynthesis demo was really cool.
Thank you so much! And honestly they were too dim under UV for the camera to pick them up. No idea why. They looked nice in person, only the chlorophyll-A and to a lesser extent chlorophyll-B were visible and they were both a very clean bright red color
When the sample is on a TLC plate you should put them under UV light while wet with solvent. When they dry sometimes fluorescence is quenched or diminished.
I wanna see a bioreactor powered by chlorophyll
Thank you a lot for this great video
I do my best :)
Fascinating! So you just demonstrate spirit in the beaker. 🙏♥️🙏
Ah yes... reminds me of my dorm days... ethanol extraction of herbaceous materials.
LMAO
Niceeeeee videooo :D
Thank you for making this video. I hope you get famous and very happy in life
Awe 😭😭 thank you so much. Need more comments like this lol
I regularly extract chlorophyll in lab, 80% acetone works well!
Hm. It works better diluted than pure? I hadn't considered that
The methanol separation is really cool. I used to love doing paper chromatography of pen ink when I was bored in the lab 😂 filter paper, pen, splash of acetone, just to see the colours 😍
Thank you so much! Weirdly enough I never got to do paper Chromatography in school, and I've only recently gotten into it. I agree though super fun I've been trying it on every different pen in my house 😅😅
Chlorophyll makes for a great UV Lava Lamp
Respect your hard work dude💐
Excellent! Like to see, you do some more biology videos involving chemistry. Perhaps you could isolate some enzymes and demonstrate the enzyme action via an assay. Thanks again.
Ah, avoiding chlorophyl is usually the target,
Interesting how your spinach experience is. If you can:
CO2 or Butane extract could be interesting too but there’s a risk to them.
Thank you, beautiful fluorescence, especially the droplets..
Thank you! I had a lot of fun with that part in particular
That fluorescence is wild; I guess I have some reading to do. 🤔
Yeah I had read about it a bit before this (tbh the fluorescence was the motivation for the video) and it just turned out way more vivid than I imagined
Watching the video to the end was worth it. Keep up the good videos.
Tysm 😁😁
I recently studied photosynthesis and it was really nice watching this!
I'm glad to hear it! It's crazy this was my longest video I've ever done and even still there was so much more I wanted to say. Insanely fascinating molecule
should have wilted the spinach first, you could easily have fit 12-18 bags of of spinach into that blender if you wilted it first : p
Very good point^^ I do think this all would have turned out better if it was wilted or dried and tbh I didn't only because I'm lazy
Would the heat of wilting change the chemistry?
I perfer my chlorophyl as alive as possible, drying would probably affect that aliveness. xD
@@DieKleinenSuchtis chlorophyll isn't alive. It's just a catalyst
@@mattschoolfield4776 You are probably right. My intention was a controversial way to remind humans: eat fresh greens and stay hydrated!
It is really pleasing to look at. Great work :)
Many thanks!
Put UV light to a bottle of absinthe and it did fluoresce red. Interesting.
Huh that is actually interesting, I wouldn't have thought to try that
Полезно.
Your videos are always nice. And your voice fits them perfectly. Please keep up the work. Sooner or later your channel is going to explode. Also thank you for all the effort you're putting in them.
Thank you so much, I love to hear that the effort doesn't go unnoticed. More than anything I just love sharing my interests, and more than anything else the growth of the channel makes me happy bc it's confirmation that other people find this stuff cool as well
Chlorophyll... more like borophyll 😂 sorry I had too.
It was a very good video.
LMAO I'm glad you thought so :)
You can use an oil or wax to sequester the plant lipids and oily compounds. Aggressively stir it in an aqueous extract then chill to solidify it and scrape it off the top.
huh I've never heard of that method.. sounds similar to winterization my buddy told me about that he uses for his cannabis extracts.
Do you think paraffin wax would work? I think thats the only wax I have lying around.
@@integral_chemistry it’s basically a winterization, I ran into it reading about morphine extraction from opium. From what I remember it was paraffin that they used.
Very cool visuals! But I have some concerns with the methodology here.
First, after your initial extraction, you comment that the chlorophyll is reasonably pure. But you hadn’t done a gel, or a column, or any other sort of assay to determine that.
Then I was pleased when you set out to do chromatography! Until my heart sank when I saw your technique. The spot size should be tiny, and above the level of the mobile phase in the developing chamber. Your spot size was hilariously massive, and immediately contaminated your mobile phase since half of it is below the solvent level! Haha, I would have been smacked if I did that in lab.
So a few adjustments maybe for next time, or maybe just ignore me. You’re up to some pretty cool stuff! Loved the video!
EDIT: Ok I see in round 2 the spots are at least above the solvent level, so that’s good. Still suuupppeeerrr huge spot size, so hard to determine the results. But at least the mobile phase stayed uncontaminated that time!
These are good points, but looking at it from another viewpoint, I think it goes to show how robust and accessible the technique can be, and it gives amateurs some confidence that they might also be successful with their own experiments even if they have limited resources or experience.
Thanks for the feedback 😁 I do agree with you 100%
I go into pretty much all of my projects completely blind, and since my background is in organic synthesis I can usually get away with going into my synthesis videos blind.
It usually doesn't always work out as well going into projects like this totally blind.
That said, all of my videos are a learning process for me more than anything, and I should probably make that more clear rather than making bold specious claims like "reasonably pure" when I really have no idea and I'm doing everything for the first time.
As a side note, I have started remaking projects and releasing them more as instructional pieces rather than experimental pieces. That's what my ether video was last week. My first ether attempt a year ago was as riddled with issues as this chlorophyll video. I actually might make two main playlists to separate experimental projects from instructional ones.
As for the Chromatography, I weirdly enough got through an advanced chemistry degree having never once done a single paper Chromatography (education these days is much more focused on instrumentation).
That said, when I do it myself I typically just do it as a qualitative test to see if there really is more than one compound present, and this is the first time I ever put it on camera. Because it was on camera I made the sample spots comically large to make them easier to see, I didn't realize that would make much of a difference but clearly it did.. I also didn't realize it was proper procedure to keep the sample out of the solvent but as you saw at the end I kinda figured out it made more sense logically lol.
Anyway I do genuinely deeply appreciate feedback like this from people with more expertise on different sub-fields of chemistry. I have the patience to watch videos or do as much reading as I should, so most of my learning comes from critique on my projects like this.
I absolutely agree! love your videos, but the chromatography was questionable :) I'm amazed that you made it through school without chromatography! I just finished organic chemistry this past semester and did it nearly weekly.
there are a couple tips I have to enhance the information you have from your chromatography output! for one, if you mark your spotting line and solvent front, you can track which compounds you've isolated. this would make it easier to note which chlorophylls, carotenoids, and phycobilins you've extracted. I'm not sure that was the point of your video, but it's really cool to see. Something else that might enhance your info would be to use silica-prepared plates for thin layer chromatography (tlc). It's definitely more investment making them (or buying them), but you can get more specific results. using this type of chromatography requires more specific placement (like dropping with a capillary tube), but seeing the spread or pigments separate by polarity is really fun!
Also, different pigments can be best extracted in different solvents. it might be interesting to do this again, using tlc, and comparing your findings with reagent chlorophylls to determine purity!
@@hooligansteriotype markers, inks, etc. from various products and art supplies can be interesting too
It should be noted that Photosynthesis does occur in a lot of bacteria as well that contain either or both Photosystem 1 and Photosystem 2, and that the main products of Photosynthesis are: a precursor molecule of either sugar or amino acid, and energy molecules such as NADPH and ATP. Not all photosynthetic cells produce oxygen either, even in some cells that do contain Photosystem 2 where, usually, water is broken down into O2.
Also, production of sugar is not limited/exclusive to Photosynthesis; almost all cells in life have the ability to produce sugar, because sugar is essential not just as a food source for heterotrophs, but also as a building block of other things like glycolipids and the cell wall.
I hope this channel grows. I love videos like this and the video is well made. Good luck for the future mister progammed cell death!
Thank you so much! I'm not really sure if it works to my benefit or to my detriment but the last time I really used UA-cam to watch videos was around 2014. I don't really like what UA-cam has become tonally with how over-the-top everything and everyone is and how videos are so long seemingly for the sake of selling adspace so I'm trying to do my part by making videos that are genuine to myself and something I'd actually want to watch lol
@@integral_chemistry Yeah youtube has been going down the drain in recent years. It's all about generic click bait crap that will attract attention rather than actual genuine content that is not only enjoyable to the viewer, but to the creator as well. Personally I grew up watching a fair bit of Cody's Lab so I love videos like this.
Do you have or can get a woods glass filter? It blocks out most of the visible spectrum but passes that red and UV.
I made a pair of glasses once from some. On a sunny day all the grass and living plants are that red while everything else is dark purple. Trippy sunglasses, lousy welding glasses.
I've actually never heard of such a thing, and it sounds pretty damn cool. I'm guessing you can just buy the sheet material online?
@@integral_chemistry small sheets, they don't make woods glass specificly anymore but there is a Kodak filter code for it and filters that do the same thing so it's not too hard to find.
Somewhere on my Facebook I got s picture of a bush in my front yard taken through one. Not a digital filter.
Wow, just wow!
BTW, there is medicine called Chlorophyllipt that can be used as a source of chlorophyll for experiments. Shouldn't be prescribed one.
Oh wow yeah I just looked that up and its only $5. Definitely need to pick that up. Thanks!
where did you buy that centrifuge?
Amazon actually. It took a long time for me to actually spring for it as a centrifuge is typically something you'd expect to spend $1000+ on and I didn't exactly trust a $350 model. Eventually it went on sale for $240 so I said screw it and it's actually turned out to be very solidly built. Highly recommend!
Very nice video! Thank you!
You could try next to remove the Mg and put other metals in there.
I was actually thinking something similar. No idea how to remove it without destroying the whole molecule, but it does have an uncanny resemblance to hemoglobin and phthalocyanine which is definitely intriguing
@@integral_chemistry I think it can be removed with sulfuric acid, but I don't know the conditions. EDTA could help chelate the Mg. Perhaps washing an organic solution of chlorophyll with an aqueous sulfuric acid and EDTA solution may work. It may take hours perhaps.
16:00 I assume you mean "put sodium bicarbonate in the water for the flower cuttings", *not* cutting the flower cuttings into little pieces, to make them stay alive longer. 😅
Oh whoops I didn't even catch that 😅😅 yeah your assumption is correct
what is the UV spectrum of your light? I would be interested to see the UV reaction of the chlorophyll with the carotenoids extracted
I'm wondering if methanol would have been a better choice?
I'm thinking it would have been, at least for quality over quantity. Honestly best is probably an 80:20 or so methanol:water solution as it seems like chlorophyll is pulled out most selectively by more polar solvents.
Do the purification with freezing cold solvent. Fats and lipids can be frozen out. Also may be a good excuse to run a chromatography column.
Can you make a video on synthetic chlorophyll a production
I'm not entirely convinced about the photosynthesis demo. The result could be explained as well by lysis and release of acids from vacuoles, etc. IE the leaf disks could have floated due the CO2 bubbles.
That's fair. You could test that by just putting one in the dark and the other in light yeah? I would like to expand on that one test
@@integral_chemistry Yes, that would convince me. At least I don't see how lysis could be dependent on light exposure here. Good thinking.
Question is what Chlorophyll it is? A? B? C? They have different solvents
It's a mix of A and B I believe. C shouldn't exist in spinach, and I saw mostly A with a little of what I believe is C in the Chromatography
Chlorophyll A: pratically insol in petrol ether, Chlorophyll B: sparingly sol in petrol ether, by C and D it dont say anything about(Merck Index)
Nice. Reminds me of my late childhood when I did the extraction of leave dyes first. 👍
If you do the paper chromatography with mineral spirit as a solvent you will see that you didn't extract pure chlorophyll, but also xanthophyll and carotin which colors the leaves autumnal when chlorophyll went back to the roots.
If you only need a small sample of extract you can simply grind a few leaf peaces with some sand and ethanol in a mortar and filter off the solid residues afterwards.
Btw: I tried extracting the dyes with acetone, which lead to a green extract, too, but the fluorescence test failed in contrast to the extraction with ethanol. 🤔 Any explanations?
I'm not sure if you got to the very end (its a rather long video so I would understand if you didn't) but I did go back and try a few more paper chromatography tests using hexane, DMSO, methanol, and acetone. The acetone just looked exactly like the ether and ethyl acetate, but both the methanol and DMSO resulted in very clear separation of beta-carotene and xanthophyll. The hexane interestingly enough also resulted in decent separation of the xanthophyll but not the beta-carotene.
As for your question I don't have a real answer but I have a hypothesis. Acetone 100% dissolves and strips out chlorophyll, but it also strips EVERYTHING.
Even though I used isopropyl in the initial extraction, which as you can see from the paper chromatography didn't dissolve the xanthophyll and lutein nearly as well as the less polar solvents (like acetone), it still dissolved enough that my final extract contained a TON.
That said, maybe acetone just stripped out so much other crap that it absorbed too much light to get a nice fluorescence.
In the fluorescence test I did here, I used a more than 100x dilution from the initial extract because the purer extract just absorbed too much light to get anything that looked nice.
You are right, I really didn't get it at the end. I am already too used to listen to your explanations so that I missed the small typed text... ☺️
The mentioned result was visible best with methanol which I have never used with leave dye so far. But you should really try it with mineral spirit for chromatography where all three components are separated and therefore clearly recognizable.
Your explanation for the use of acetone for extraction is very plausible. Thanks.
And yes, my sample maybe was simply too concentrated to give a chance for any light effect. With ethanol in a dilute solution there is always a perfect red fluorescence.
@@experimental_chemistry I'll definitely need to try that out. I've gotten so many good suggestions so far I think this video deserves a follow-up (despite it already being longer than I would typically make a video).
And yeah I was pretty surprised methanol gave the best separation, but as I mentioned this is not an area of chemistry I know much about.
@@experimental_chemistry Acetone extraction does yield fluorescence result. I just showed this to my son.
@@user255
Hm, then my solution probably was too concentrated. Or I used the wrong kind of leaves.
Does chlorophyll not dissolve in water? I thought it did.
Yeah weirdly enough it's completely insoluble in water. It does disperse in water to the point it looks like it's dissolved though
I have always asked myself, since the difference between chlorophyll and hemoglobin is the magnesium and iron atoms, can we convert chlorophyll to hemoglobin?
You could possibly switch out the magnesium for iron and make red chlorophyll which would be cool, but it would be tough because chlorophyll is a very fragile molecule. Also there are more differences than just the coordinate metal ion, they evolved for different functions and have several different functional groups you'd have to change as well, which would be a huge task. As a side note, did you know that B-12 is the most complex molecule that has ever been fully synthesized in lab? The process was insane, check it out!
The production of heme and chlorophyll from scratch in cells is more or less similar, until at a later step in the process - and chlorophyll has a few extra steps of production, particularly the addition of an isoprenoid tail, which makes it more lipid soluble.
I wonder what a magnesium heme would even do; would it just work like chlorophyll but with less of the lipid solubility?
Chlorophyll…. more like BORORPHYLL 😂😂
Were you the one who said the same thing on my chlorophyll video? xD
Came to the comments just to make sure someone had already said it
what dat song at 7:00
More like Bore-ophill..