【4K】-- Column Chromatography (Purification)🌟
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- เผยแพร่เมื่อ 20 ต.ค. 2013
- #Column #Chromatography
Purifying mixtures of compounds using column chromatography.
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We are currently studying separatory techniques but they are just in textbooks, so I can't really understand how they happen. You know, it's hard to understand these things when I can't even visualize them. So I really thank you for demonstrating these. You're saving my grades.
You are MOST welcome! there are many more videos in this playlist and in my collection, free to subscribe and learn well! God Bless, Merry X mas.
explanation is excellent and Dr is helpful, this video is the best on youtube.
Thank you for posting this demonstration of flash chromatography.
The explanation was clear and detailed. Much appreciated !
thank you so much Kevin!
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Useful to visualise the concept of column chromatography for JEE/NEET students. Helps in remembering it for more longer period of time. Many thanks!
Most welcome!
Thank you so much for making a video on this! I watched your video on mestrenova before, both are really helpful and making my life so much easier!
you are kind! thanks for appreciating. share it with your peers.
Thank you so much Mr. Fun. I understood your explanation. Keep up the good work!
You are welcome!
Wow wow
It it just great. The detailed pros and cons 🌿. Thank you so much Fun Man ♥️
My pleasure!
This is really great. Well explained... Thanks
Glad you enjoyed it!
best explanation on TH-cam for #columnchromatography, useful for #chemists and #biologists 👌
Thanks a lot 😊
Thanks doc.
Most welcome!
Helpful indeed ✨
Glad it was helpful!
Thank you so much for giving English subtitle. It helps me sooo much.I request you that please add English subtitle in your all videos. Please 😊😊😊😊😊
Even better; Pay attention and learn to listen.
wow what a racist!
This is so cool
thank you for your support! merci! do subscribe and share our videos with your friends :-)
can you tell me how you prepare the sample chloropyll by using leaves ?
Why do people say this is so hard and complicated that it cant be done by an amateur chemist? Are there some compounds that are more complicated to run? Like for example lysergamides
Thank you doctor, please I need your help for this:
if my sample is solid, can I use any solvent to dissolve it?
Or I have to use only the same mobile phase as solvent to dissolved sample?
How to separate a compound where just above a little cap also presents in TLC. So, at the same polarity both are eluting.
it is helpful indeed , thank you Dr.
I have some question, please I need your help, because I am new in organic chemistry, and some information I can not get it in textbook.
1) how many gram from silica can use to sample? some talked 1 gram of sample to 20 gram of silica gel, i do not know about this.
Wow amazing vedio it is very helpful , I have questions about the solvent! after preparing the organic compound based on what we chose the solven? And what if the compound is solid how to add it as a solvent which solven we should use to desolve the compound?
Try to test various solvents to dissolve, such as acetone, acetonitrile, ethyl acetate, hexane , etc !
legend
+SixTough John?
+FunMan Fung hi no we dont know each other but grat videos man
+FunMan Fung Thanks, lad, from the Czech rep. ;)
Most Welcome, mate.
+SixTough LOL you didn't get it, but hey thanks!
If my sample is liquid, can I pack it direct in column on silica gel by pipette, or use rotary separator to completely dry, then dissolved in mobile phase after that
If I have sample which is mixture of components but it is a little bit such as 0.01 gram contains example three compounds, here:
Can I use any column example 50 Cm, regardless length ? or I have to use only narrow column and short one
i did this in my final semester of M.Sc biotechnology for a project work paper. i could revise all what i did and some additional information.thanks a lot. please continue this kind of video they are just too rare.
Thank you very much!
After preparation of column chromatography and it is already to start:
Is hexane is compulsory to use as first mobile phase, then after that use hexane : ethyl acetate and change polarity
Thank you for this video please can we use this method to separate dyes from plant
Yes we can
I have some inquiries about (column chromatography). please if you have time to answer, I think like these inquiries are important and I think the answers are very important for any student like me to start work in lab. I sent some question to some channel on youtube, but almost no reply, but here found your answers for almost comments, this is good, you are helpful and excellent doctor, thank you.
my question is :
If I used wet packing of silica gel to column (slurry),
here, can I use any solvent for silica gel until reached slurried, or must use only hexane
Okay so I plan on performing TLC and traditional column chromatography
and I need help. gravity isn't a strong enough force to elute obviously
and I don't have a license or funds to get a 5ft nitrogen/O2 tank, and I
was thinking an air compressor but those are also expensive. does
anyone have any experience with at home chromotography workup? please
let me know
Thank you so much .Very nicely explained. May I know if we use Amberlite XAD resins for column, can we store the resin after running column? If yes, how to store it so that we can use it again.
Why do you want to store?
@@chemfunman Thank you for the reply.. I am not sure, I need to purify enough amount, like around 20-30 litres of bacterial culture, so I was wondering if we can store the resin and reuse it. So is it possible?
sorry Dr. my questions were many. but I think were important for students or researcher, and I hope your answers will be reference to them and me or any visitor to your channel, for my case I could not get answers for these questions on youtube or textbook; I hope also your channel will be such as source the river in organic chemistry. sorry and thank you again.
thank you doctor for this nice video.
i have some question: if i did some experiments in the lab and my product was (liquid or oily) here how can run column, i mean add it direct over sand or also must add solvent to product, (this solvent is the same mobile phase or any solvent just until dissolve?)
plz doctor i need your answer,tq
No need
Hi friend you are very intelligent ❤️❤️ I am impressed , could u tell me if this method could be used to cbd/thc because I see u use hexane is this dangerous if I extract cbd/thc
I will use the same method to isolate CBD from a THC/CBD oil. Hexane is used as a solvent in this case. You can use isopropyl alcohol or ethanol for your experiment with CBD, that's common, and it works the same as hexane as a suitable solvent. Mention that isopropyl alcohol is as toxic as hexane. Please make sure to dissolve all solvent by heating it above 80°C. Good Luck!
@@jonasschweiger8938 No you can not simply exchange hexane with isopropyl alcohol, unless you move to reverse phase. Alcohols are very strong eluents, much stronger than esters. I would start with an ether first, but keep in mind, that it is somewhat more polar and an hydrogen bond acceptor, but not a donor, so the selectivity might be off. I think diethyl ether might be convenient.
You should never underestimate the toxicity of n-hexane. It is neurotoxic and for some complicated reason much more so than other light alkanes. Modern medical grade petroleum spirit is often explicitly hexane free because of this.
Good luck with your projects. And always keep in mind, that natural product separations are often on a different level than simple synthetic workup. This is the real deal for which chromatography was invented and not the harmless stuff encountered in undergraduate organic courses :3
Also one has to keep in mind, that solvents dissolve in your terpenoids (cbd and thc) very well, so they will not be removed by simply heating to their normal boiling point. To get entirely rid of them will need high vaccum and long time periods. Traces are really, really hard to remove even at high temperatures that would harm your products. I am not joking, this is a real problem and most laboratory workers ignore it, because in an organic laboratory, solvent traces do not matter that much. We learned it on an industry course about separation problems some months ago.
How much sand do you need for this chromatography column? and can I use water instead of hexane? Thank you
Fern Sutiyawan can't use water!! 0.3cm thick sand
thank you.......
can i know why you put sand??? and why stationary phase always suspend(covered) by mobile phase in Chromatography Columns
Sand will protect the silica layer from being disturbed, so that the solute mixture will move as a uniform band (doing its best to). If there are no sand, during the transfer of eluent into the column (while topping up), the silica may experience a 'dent' and not be levelled.
not to disturb the silica layers. sand is denser, it will provide a firmer support.
Thanks to the subtitle, the accent is a kind of difficult to recognize for a non-native speaker. But, anyway, it's a good tutorial vid for a beginner.
thank you for your kind comment. if you would like to support doing up a a subtitle in your native language, send to my email? so I can upload to benefit more people on earth. what do you think?
hi can anyone help me please i have some question about a recipe
the organic phase dried over anhydrous MgSO4, filtered, and the solvent removed under vacuum. The residue was chromatographed over alumina with elution employing a 3:1 C6H6/CHCl3 mixture, and the collected fraction stripped of solvent under hard vacuum to a constant weight. This free-base solid can be recrystallized
when i dried water from organic phase and getting fractions from column this gives me a solution but in the end of recipe said free-base solid how can i get solid from solution fraction with vacuum?
The product is probably a solid - so after you purify it by flash column, and you evaporate off all of the solvent (the hard vacuum step), you'll get a solid.
Ok, for flash purification it is suitable. But for thourogh purification it is unsuitable because of large number of silica cracks in the column (four white bands in the lower part). This happened when you abruptly changed the polarity of the eluent from hexane to hexane:ethyl acetate 1:1
It's my pleasure
Why did you dissolve your sample in dichloromethane and use hexane as your first mobile?
good solubility
What substances do the second test tube and the fourth test tube contain.??
Probably carotenoids
Dear sir, you are using a air pump for applying pressure on the column- can you tell me the model and company name of that pump? If you have any link for getting more details for purchasing that pump then please share here if possible. Please tell about column and connector dimensions.
Please give information about the air pump that you have used
Can the same method and steps be used for sephadex LH20?
try?
Это слишком хорошее деление само по себе и вещества окрашенные. Посмотрела бы я как он поделил таким способом что-то реально сложное и бесцветное
Sorry 😞 I don’t understand
How to calculate about what amount of silica gel to be added to column
Two spoonfuls
another question if my product solid, here can add it to column as solid ?or first disolve it in solvent?, (this solvent is the same mobile phase or any solvent just until disslove ?) tq
plz doctor i need your answer,tq
the solid would not dissolve sometimes, and it is OK! When I performed purification using column chromatography, i had many solid particles (insoluble) and did a successful separation any way!!
@@chemfunman thank you doctor for reply. sorry you mean just add the solid that insoluble direct above sand after that you got nice separation
@@chemfunman also i habe this question .if i did some experiments in the lab and my product was (liquid or oily) here how can run column,? i mean add it direct over sand or also must add solvent to product, (this solvent is the same mobile phase or any solvent just until dissolve?)
Can u explain me can i get the sand or how kin of that?i try without sand theres always any air
Buy fine sand from the lab?
can you plz mention the name model and brand of air pump used?
if feasible can you send mn link plz?
I will have to visit the lab and ask. Can u see from the video? at the beginning
Does eluent and mobile phase are same thing in chromatography?
yup
Hello, can i get the measurement of the chemicals used? Thank you.
Can you specific what chemicals you need advice on their quantity?
Fun Man FUNG the n-hexane, silica gel, sand and cotton wool.
@@erika_890 I thought i mentioned in the video. cotton wool, a small glob. silica gel, depends on your diameter of the column, n-hexane, prepare 250 mL
please dear Dr,
could you explain another video about Preparative Thin Layer Chromatography (PTLC), how can student or researcher prepare plate in lab( amount of silca gel to water), when can use this method, how can purification by using this method using mobile phases, thank you in advance
Yes, soon
@@chemfunman thank you Dear doctor
12:23 why would it feel warm when the firat compound is coming out?
positive interaction (bond formation) is an exothermic process. giving out heat
@@chemfunman hmmm... I have some Googling to do. Thanks for the quick reply/answer! Earned my subscription (WITH notifications!) Haha
@@chemfunman thanks for the reply
If we are using silica already as a stationary phase then what is the role of sand, please clarify
to block the silica from fleaking down.
@@chemfunman Is cotton not enough
@@chemicalsociety9094 Nein
What air pump did you use? Thanks!
NITROGEN / air
@@chemfunman May I ask what brand/model? Thanks again 😊
@@nocturnalrph1678 hah! That I am not sure because there's no brand I noticed:(
@@chemfunman Oh. It's okay. Thanks :)
Could you tell me the branch of small bump you use?
brand?
Fun Man Fung I mean a name that company uses for its products (trademark) , I want to buy a small pump like yours
@@henrythierry1778 now we under lockdown, no access to lab. we will find after we can go back to work
Fun Man Fung thanks a lot
why we choose Hexane ,. please
non polar solvents
The correct abbreviation of ethyl acetate is EtOAc! (12:42, 15:15)
thanks for the note, Yes! Absolutely right.
Which air pump should we use
normal air would do.
@@chemfunman thank you sir 🙏
Thanks for your support!
@@chemfunman sir, can I get your mail ID? . I have some quarries as I am doing column in my PhD.
@@shravankumbhare5785 let me think about it as I am very occupied with projects
what will happen if the solvent is stopped adding to the adsorbent?
Better do it! if not too many actions at the same time will be distracting. thank you! do share these videos and subscribe.
"Use hexane to rinse down your food pipe"
thank you! you are welcome - come back to this channel for more! like and subscribe :D
WHY ONLY LANGUAGE OPTION IS ENGLISH ???????
Help us translate please?
Using pressure pump is not the good practice 🤣