Making Fluorescein
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- Опубликовано: 9 сен 2024
- Today we will be making some fluorescein! I bought the resorcinol off ebay, and I used the phthalic anhydride that I made in a previous video.
The mechanism and procedure that I adapted mine from can be found here: sites.uci.edu/c...
Extraction from highlighter markers: • How to Make Fluorescei...
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Nile talks about lab safety: • Chemistry is dangerous.
Anyone else not sure about the chemistry but really enjoys most of his videos?
The newer videos are incredibly well made but I'm still able to watch his old stuff without feeling like its lacking.
42thsliker
Nigel's format has really not changed much over the years, only his recording equipment / software / labs. That's because he has a Formula That Works. He's also a chemist and follows directions to the letter, often, else explodes. I figure he's made the connection with RUclips being pretty similar 😅
Only the sound quality but i binge watch it all anyways
"i want to make it because i want to play it." the chief reason why anybody does science things.
YKTV
True
Biology: "No! Only medical!"
Me: 🙁
Truer words were never spoken🤣🤣🤣
1:48 Funny, that “stir bar” kind of looks like a paper clip...
It is
It's to stir the oil bath. In the actual flask is a proper stir bar lmao
I noticed that too. lol
You beat me to it....but only by a year! LoL
@@bdnugget I don't notice until it was brought up in the comments. It's a genius solution to needing to stir bars at the same time. If I ever take up chemistry myself I'm doing this, a lot. With both ice and oil baths.
DUUUUUUDE!!!!!!!! I use this stuff all the time with lidocaine to numb eyes!!!!
Makes it possible to view the cornea through the slit lamp on an eye exam with an indigo light that illustrates just like the beakers at the end of the video anywhere fluorescein has accumulated on any abrasions. The eye acts as the water in the beaker and the fluorescence remains unquenched illuminating the areas of higher fluorescein concentration as bright neon yellow/green. Exactly what it looks like at 12:29. You tha bomb.
Thanks for posting!!!!
why hasn't anyone asked you why you numb eyes in the past 3 years
@@nataliella97 Because the dude is obviously an optometrist or optometrist's assistant. It's strongly implied.
@@Daggeira Call him what you will, I ain't getting my eyes close to that guy
Pthalic anhydride just wants to give you a hug.
stonent lol
And then the resorcinol messed up its arm.
And you cut its one arm by making fluorescein
phallic anhydride wants to do more than that. 😏
Cool, if you got some bromine, you could make eosin, the brominated version of fluorescein. I don't have a protocol, but I'm sure you could find one easily.
I remember in my first undergrad lab, we use to perform a simple test to assess for the presence of bromide in a solution. By adding an oxidizing agent to the solution, you convert the bromide to bromine. You then heat that solution under a filter paper on which you've added a drop of fluorescein. If the spot went from yellow to pink, you had bromide in your solution!
+supersmashsam cool! I actually have a bit of bromine left. Maybe I could make some eosin. Ill look into it!
And with iodine you can make erythrosine. I made both back in the day when I was in high school. Eosin is tetrabromofluorescein, erythrosine is tetraiodofluorescein. There's also a dibromo-dinitrofluorescein AKA "imperial red"
a
Great video - particularly interesting to me because I have an eye condition and I've had many fluorescein angiograms done so the ophthalmologist can detect blood flow in the retina. Fun part about that is the stuff is unchanged by the body and flushes out in urine. I leave the rest to your imagination...
This stuff is great. I've used it as an indicator once
Nile red, i really like your videos. I loved the inclusion of the overall reaction along with the description of the mechanism, please do this in more of your videos. thanks
+Walter Meier I am glad you enjoyed it!
To separate pure Fluorescein, you can directly add some water ( sufficient amount to dissolve) into the crude product in the rb and boil the mixture at around boiling temp ( using a reflux, of course ), then all the other compounds present will be dissolved in the water ( like, pthalic acid and sulfuric acid or unreacted resorcinol etc ) but Fluorescein being insoluble will be precipitated on the side inner walls of the rb. Then you can vaccum filtrate the product by washing with water. we did this procedure and got nice 86% yield with absorption spectra peak at 488.5 nm.
Funny for someone who never had a chemistry set like myself to binge on this channel lol
I need one of those stirrers for my coffee :D
Absolutely incredible presentation. Very concise! Please don't change your format.
fluorescein and rhodamine are the two cheapest fluorescent dyes one can buy. They are like 10 quid for 100 grams on amazon, sold as tracers for leaks in plumbing.
What camera do you use? Your footage always looks very crisp.
+crazyboy0602 Canon 6D with a 24-70mm F2.8 lens!
You should see it now.
Love the paper clip in the oil bath.
Flourescine dye is a very common tool in an ophthalmologists office used almost every hour if not more frequently. It brilliantly delineates any abrasions/ulcers. It's also used for measuring intraocular pressure (applanation tonometry). Very cool!
Flow Cytometry, duude..
FITC, or fluorescein isothiocyanate is the basic fluorescent probe in leukemia and lymphoma diagnosis. Awesome video. Many thanks👍
you are awesome, keep on bringing it dude. 2 questions ; how long will the beaker glow if left alone, and how did you clean your equipment up? looks like it stained all the glass.
+One2rock It will stay forever! As long as your are exciting the fluorescein with blue light. It is fluorescing, not glowing. So the energy comes from light you put into it.
I cleaned everything with base solution. It makes the fluorescein water soluble and i can just wash it off then with tap water. My base bath is now glowing green though :p
I KNEW HIGHLITERS GLOWED! I KNEW IT!
Ok
I learned more info in just 3 videos than I did in chemistry class.
there is a huge difference between classes and this video. Videos like this dont go into full detail of the mechanisms that are happening and dont have a entire field to understand, just smaller ideas learned from said field. Yes you got a brief idea and application of chemistry but if you unironically think your learning more technical stuff in this video than in school, your actually a joke.
@@MrAwesomeHero1 dont care
Bit late to comment...and Im sure you know this...but dont forget the compound will dimerise etc. which causes a fluorescence concentration quenching effect. Hence the red colour when concentrated. Happens with Rhodamine 6G and others as well. Concentration vs. fluorescence will peak at a certain concentration.
Great videos by the way. Keep them coming! Just dont go "breaking bad"......
Oh balls....you already said. I hadnt watched until the end. Duuuuhhhhhhhh.
Hi,
always a nice experiment to demonstrate. However, when working with organic solvents, beware of the risk of fire. Solvent/air mixtures can even be explosive. When working with diethyl ether one should keep in mind that this chemical forms peroxides during storage. Especially when you evaporate etheric extracts to dryness as in this video, you can get a nasty explosion, with fragments of the flask flying around.
There is a procedure to make ether peroxide-free, but it is rather involved. Easier might be the use of tert. butyl methyl ether, which doesn't form peroxides for steric reasons. As a solvent it has similar properties as diethyl ether (except that it doesn't work in Gringard's reaction).
Fluorescein can also be purified by pouring the molten raw product into some dilute HCl (~ 1 M) and boiling. This extracts most of the contaminants without the use of organic solvents. Filter, discard the acid and dry the insoluble material, which is chromatographically pure fluorescein. Both the raw product and also commercial fluorescein have some brownish contaminant that forms a long tail during thin-layer chromatography.
Somebody mentioned eosin synthesis. This is quite simple: Disperse some fluorescein in absolute ethanol, stir and dropwise add a stoichiometric amount of bromine (which of course is a very corrosive and toxic liquid, so work carefully in a fume cupboard). Initially, the mixture will turn into a solution as ethanol-soluble dibromofluorescein is formed. Further addition of bromine leads to precipitation of tetrabromofluorescein, i.e., eosine. Simply filter and dry, if you started from pure fluorescein you do not need to further purify the eosin. Again, it is interesting to take samples at various stages of the reaction for TLC.
If you keep the eosine over a vessel of NH4OH over night, it reacts with the NH3 gas and you get the ammonium salt of eosine, which is more water-soluble than the free acid.
I have not tried this, but if the bromine is replaced by iodine, you should get erythrosin (eosine B for blueish), which has an even more intensive colour than eosine Y (for yellowish).
+ Engelbert Buxbaum vor 2 Monaten :
. - You should have mentioned it :
To prevent ether against *most dangerous peroxide* , there is pressed Na-wire into it, which catches the oxygen out of the diethyl ether peroxid 's . That often is used in laboratory 's, because ether - explosion / fire may destroy a laboratory , as it once happened in Hamburg / Germany , in the chemical university .
2017-04-11
I love how he's just like: "oh well." 1:15 😂 i love it, and even though i dont really understand whats going on, i really enjoy watching his videos 😙
Try shining a 488 nm wavelength light on the fluorescein. The glow is far brighter because fluorescein is particularly sensitive to this wavelength.
Great vid! But you forgot to include the highlighter extraction link in the description :)
+ZeroGames oh yes, ill add that now!
I once had a contact lens slip back on my eye where I couldn't find it on my own. I went to the eye doctor, and she used a few drops of a dilute solution of fluorescein in my eye. The fluorescein stained the contract, making it easier to locate when a UV light was shined around between my eye, and eye lid and socket.
I owe my continued sanity to fluorescein, because i was about to loose my mind trying to find that contact myself.
If you point a decent power 450 nm blue laser pointer (like 200mW) a dense soln of F will glow super-bright (much brighter than one can get with the UV) -- almost like sunlight.
I did that in uni. Nothing beats taking glowing green baths
If you dont have a black light buy some rgb LEDs and the blue light works just as well though ph affects what wavelengths are absorbed best
I am only in high school so I don't understand everything (yet) and I get classes in Dutch so I always gotta translate things. But I must say your channel has really gotten me into chemistry. My lazy teacher really left me underwhelmed with chemistry but now I am actually really wanting to take some kind of course in chemistry. Especially organic chemistry cause I really think carbon is pretty cool. And also. Awesome videos! Keep it up =)
+Tim Meermans I am very glad that I have sparked your interest in chemistry. I love to hear this, because this is one of my goals with the videos :)
Well it looks like me and your videos react quite well :P (wow that was a bad joke) But I can't wait to see what else you have planned. And I hope to see more great explanations. And one request. Could you do some carbon related stuff? I saw you did study organic chemistry after all
Also I just realized ethanol and methanol and those things are all carbon related. Whoopsie. Well never mind the request. Just keep being an awesome chemist
Thanks for getting me re interested in an old hobby I'd make my own resins and curing or hardening compounds I learn alot from you Thanks for the fun educational aspect
Thank you very much!
the year is 2021, this video is still awesome. I ended up here after watching the soap vid. I also watch nileblue
Fluorescein is used in drainage tracing dye. I remember using it several years ago and got some on my hands... for about three days afterwards my bodily fluids were a very bright yellow-green. It was quite funny in retrospect.
... is that a paper clip in the oil bath?
JimGee
Could be, if it is magnetic.
Yep. The stirring mechanism works in a magnetic way so a steel paperclip works just like a magnetic stir bar. I'm pretty sure a stir bar is just a steel bar covered in a non-reactive plastic.
They're samarium cobalt magnets coated in PTFE usually. You could just use a steel bar or whatever but it doesn't work very well to stir viscous stuff.
bro yeah i saw that too lol
This is so cool! I was diagnosed with may-thurner syndrome by use of fluorescence for diagnostics!
I use FITC in flow cytometry all the time and it can be excited by a blue laser(488nm)
today i was at a chemistry competition, where one of the the calculations was with fluorescein... they had an explanation about it, what it was, what it's used for, etc., and and as i was reading it i was so happy that i already knew what it was😁
We use this at work as an indicator for our water treatment in the boiler system. I have played with it many times when bored lol
This would be really cool to water cool a PC with
So so simple process! I'm gonna do it while I brush my theet
The thumbnail for this looks like it could be an album cover
What informs your choice in glassware? As in why use a beaker vs Erlenmeyer flask, vs round bottom flask vs test tube etc?
My Et2O-layer wont get this dark and my H2O-layer stays pretty dark. Can somebody tell me how i can change this. I added NaCl to the Mixture and hoped it will work but it doesnt.
Coming back and watching this is 2021 and seeing him use a paper clip as a stir bar gives me some sort of feels. I'm not sure what though.
I really like your videos hope you upload some inorganic chemistry videos soon...
This is the best stuff for finding leaks in your cars coolant and oil systems. Clean the engine, put a bit of this in the antifreeze and oil. Run the car for a day or two, then go out at night with a UV flashlight - any leak will be readily apparent.
I think ferric chloride solutions look a bit like this too, a dark red solution with greenish yellow edges. I have no idea why that would be though, it's driving me a little nuts!
this is so cool!
Resorcinol
The alcohol of a resource.
Hi¡ Is there any way to synthesize resorcinol in home?
+Nation Of Our Hobbies I am not sure if there is a very easy way. From what I see, what precursors to resorcinol could be even harder to get or annoying to make.
Yeah I imagined that, anyway..... thank you ¡¡¡
Also used as a marker in ocean search and rescue
I love this channel lol 👍☺
Saw your Uranium Glass video. Would it be possible to do something similar with fluorescein?
Powderize it an mix it in a silica mix?
I thought you were saying Phallic Anhydride
not sure how i got here but im just sitting here playing runescape letting your videos play in the back lol
9:48 You mean nilate?
Could MTBE be substituted for Ethyl Ether on the extraction?
Could someone explain me how does that equipment that mix/rotates liquid inside of the becker works? I mean, as far as I know a becker doesn't have any holes on its bottom
From what i can see, i think that capsule is a magnet. The machine below the beaker push the pole of this capsule and make it spin.
alienjantan Oh, I see! It makes sense, thank you
its a magnetic stir rod on a stir plate. the capsule is a magnet coated in teflon, and the plate has a big magnet that spins, whichs makes the rod rotate and stir the solution
When I was a kid, I was *fascinated* with that apparatus!
a team of tiny elves live inside of the little tube and they activate an electromagnet that causes it to spin with the magnet thingy spinning underneath
Please HF or carborane....and there reaction with glass...and please a how to make glass or chemical reactions while making glass.
youre like the new nurdrage cuz you upload more 😉:)
Hey, i rly enjoy your videos. I dont study chemistry yet but im planning to do so after school in about 2 years. Atm im just learning everything university level chemistry by myself with books or online lectures. Now my question is which level is this mechanism for the Fluorescein at ? Cause my teachers say that i might be B.Sc or even higher in Organic-level but tbh i would not be able to figure this mechanism out as clear as it is on the website. I would be nice if u could give me this information. :)
+Wiz Onio I would say that it is Org 1/2/3 which would be B.Sc level. The fischer esterification was guaranteed org 2 for me, but I don't remember when I covered Friedel-Crafts. I think it is first mentioned in 1, but covered in more detail in 3. It is a type of Electrophilic Aromatic Substitution.
Don't fear if the mechanism seems complicated...because it is. It is not super easy to see that those 3 reactions occurred in succession. I definitely didn't think it was obvious.
+Nile Red Thank you for your quick answer. I know all these mechanism in this synthesis but if i wouldn't see the real product/answer i probably get a slightly different product/answer.
Yeah, true. There are other reactions that occur though. There are always side reactions. So you might predict a side product, but that isn't wrong! Sometimes without running the experiment, you don't really know what will be the major product.
my favorite color
1:48 “I had a stir bar at the bottom”
That looks like a paper clip. I mean it works to stir a solution. But I’m pretty sure it’s not technically a magnetic stir bar
But would it be possible to use it with solar panels. So Box 10x 10 X10 and 5 panals around water and the fluorescent substance.
Does anyone have a link where I can straight up by the stuff. I want to do some experiments with it.
great video!
is the fluorescein still fluoresent in solid form?
No
Why the oil bath and 190-200 degree C is required. In our lab we did the reaction at around 50 degree C and by heating with a heat mantle. ?
How can sulfuric acid be used as an alternative catalyst to zinc chloride?
Could you rotovap the acetone and ether out rather than just boiling it?
+Mr.chang cooler because everyone has a rotary evaporator in their garage he could have also vacuum desiccated the product instead of putting it in the oven he showing us how he did it with his equipment.
+William Rice It's funny because I actually do have a rotovap...my vacuum is just too crap to run it with. It smokes after like a minute
u wot m8
Let's all just be friends!
Great video. I think there a little mistake in the color of the oxygen atom in the water generated as the blue oxygen water result from the double Friedel-Craft reaction (oxygen came from phthalic anhydride) and the red one is the resulting of the Fisher reation (came from the resorcinol moiety).
I am watching this after his liquid soap video and he said in that video that the flouroscein hurt a tiny bit but in here he said it was completely harmless so now im confused.
what...what majick is a stir bar? Magnets? How do they work?
Also, what's a Molecular Sieve :O
Sounds so cool.
What is it with the water that make it dissolve so much better in ether
What gives the deposit the red color??
great great video again
Can make a dye laser from that.
are you like a scientist or teacher or something or just a rich guy who knows about chemistry?
Twig 46
Why would he be rich?
because he can afford to buy all this lab equipment and chemicals and clean it up when hes done, so he must have some money to do that with. i mean, while this is educational this is basically a hobby for him.
Imperial Inquisition Ordo Hereticus chemicals are expensive. but yeah.
Defiantly not rich
Could just be a chemist. I mean this stuff isn't exactly a rocket
Very helpful, we have this topic on our lab and this video really help us to learn what will we gonna do. 🤣
Love your videos! Very clean and high quality footage, gladly subscribed!
I want a bunch of this to use as a laser target.
Btw if you have phthalic anhydride, you may prepare luminol, pretty interesting too but harder to prepare
+MethyleneVapour I am planning to make it. I actually just got some more, so I have a lot on hand. 100g+
Same sort of Green as automotive glycol antifreeze. IS this what is used as a colorant for antifreeze?
Put this into solid soap and make glowing soap.
NileRed: What kind of oil is used in the oil bath?
Great! Where's the video on how to make methamphetamine?
+ShroudedPanda Don't worry...it is coming. I am not even kidding. Well, I won't be making meth, but I do plan to make a video about meth production in general and the chemistry behind it.
Woo! Can't wait to do some real breaking bad stuff (don't actually have RV, knowledge, or drug dealers).
look forward to the education on that one nile, any chance of doing apiol type compounds extracting and isomerizing to the 3,4 ally benzene
2:48 I think you made a mistake in the source of H2O molecules. should F-C reaction generate no H2O? instead, it is the ether formation that gives one H2O. and I think there should be two H2Os produced in the reaction, not 3.
make a video on aurin synthesis please
Do you have a paper about this?
Is there another way to remove the ether without a vacuum pump?
Happy St Patricks day. Now you know how to dye your local river.
Hello, can I re-record some of your videos, but, in Portuguese? I like your experiments, so I want to reproduce them.
How is it not photobleaching like its reputation?
Why did the saturated salt solution make the ether layer clear?
+woofyams It pulls the water out of the ether layer!
maybe the water pulls the stuff apart and makes it easier for it to dissolve into the ether?
Missed opportunity to mention how you can actually see the concentration drop after it hits the water, you can see it stay red for a few seconds and from the outside inwards watch it turn to green.
Nice video ! =) More !! By the way, I'm really interested in fluorescence and especially how it work and what is happening in detail, like "why the light changes that way with this molecule and not others..." I read quite a few articles that explain that it's because of the shape of the molecule but I'd like to learn more about that or even about fluorescence in general :)
(-I'm a french fan, apologize if my spelling is wrong)
+SpinHalo76 Your spelling was good. I didn't even think engish wasnt your first language until you said it! Fluorescein and other fluorescent molecules are indeed very interesting. However, I unfortunately don't really have any resources to direct you to. :(
Ok, thanks for answering anyway and continue your videos there are great ! :)
I know this is a old video, if you are still active I was wondering if this molecule is in anyway related to those in certain rocks that are fluorescent under UV short wave lights ? Thanks
Not at all. I think you mean amber which is a tree sap that fossilised.
This can be found in eye drops to check for corneal abrasions, also super diluted in the green liquid in bubble levels.
This is not naturally occurring
hullo, love your videos! just one silly question, if you had the stir bar in the bottom oil bath how is it stirring the florocene? I don't get it...lol