Yes, as its gradient elution, so I packed the column with only ethanol as silica slurry and then slowly increased the polarity by adding water in ethanol as solvent system, which could have noticed in the video. Gradient elution will help to speed the process and save solvent loss. Thanks
Don't put sample .. atleast your mobile phase reach silica gel ...other wise you will not get 100 present purity...any way ..nice vedio ..thanks.. For your efforts... Take care...
If we are taking ethyl:hexane as Mobile phase in 1:2 ratio can i mix the solvents before wet the column .or can i use the hexane as how u used a water?
If u wish to do isocratic elution u can mix the solvents and elute with the same, if u are doing gradient elution then pack colum with hexane and gradually increase the polarity by adding ethyl acetate. Thanks for watching this video.
Thanks.if you have mixture a lot or small amount how can take silica amount in both case? I mean how many gram of silica can use? Is there standard depend on mixture ?
Depending on the number of spots and spot difference from TLC, you should opt the length of column and types of adsorbants for ex silica gel (60-120, 100-200 or 200-400 mesh), and depending on amount of ur sample (g, or mg) you should select the width of the column. As such there is no thumb rule, however u can to do it on trial and error and based on resolution obtained on TLC.
It depends on what kind of protein u wish to isolate. What u can do first develop the mobile phase using TLC, optimize the solvent system which will definitely help u to decide solvent system for column chromatography. If u wish to use HPLC Acetonitrile and TFA can be used in RP mode.
We can seperate many drug mixtures, purify intermediates during synthesis or extraction and many more. Its maintly and seperation tool. Thanks for watching this video. Regards
Sir pls I want seperation of inorhanic cation and anion seperation by column chromatography.. pls let me know what will be the mobile phase n how I should perform this
First of all my sincere apology for the delayed reply. Frankly speaking the selection of mobile phase totally depends on nature of captions or anions u r interested to separate, however u may try with aqueous HCl and ethyl alcohol, initially u may develop the TLC and then can run the column.
In an HPLC experiment 4-nitrophenol, acetophenone and benzophenone were analyzed. What is a good asymmetry value? And also if the values are below or above that value, what is a possible reason for that?
Ideally, to get a Gaussian shape peak we should get Asymmetry value 1. If the value is greater than 1 it shows tailing and if it is less than 1 it shows fronting. There are many possible reasons for not getting symmetrical peaks which u need to find out practically. Some of the possible reasons are aging of a column, contamination of HPLC system, exhausted guard column, improper flow rate, temperature, high ionic strength if RP is used, extreme pH conditions, improperly diluted/ conc. samples, solvent pH, etc.
You need to identify and optimize the solvent system first using TLC. Once u get separation of 6-gingerol on TLC u can try in column using developed solvent system. Probably u can try with Diethyl ether and n hexane with 7:3 ratio.
Yes, rough separation of crude oil is possible using column chromatography, u need to optimize the solvent system. If possible after column chromatography u can separate more purified fractions using GC or HPLC. U can also try using preparative TLC if it's on the microscale. Thanks for watching the video.
Plz watch the video at higher resolution setting with atleast greater than 360p. This video is recorded with resolution of 720p. Check in video setting of TH-cam. Thanks.
great source of knowledge for rural students and student who can not go big college. we all love u sir god bless you. Ganpati will take care of u.
Thank you so much Rajeshji for your compliments. Regards
Such as amazing video for students. Thank you so much.
Alots of respect from Pakistan to you Sir. 😊😊
Glad to receive compliments from our neighbors. Thanks u so much. Keep watching.
Very very excellent explanation..r i actual teacher sir ji
Glad to know u liked this video, will upload more videos on column chromatography in the coming days. Thanks
Very Crystal clear explanation with experiment.
Thank you very much sir
Thank u so much for ur encouraging words. Regards.
Sir.. U told the.. Mobile obsess slick and water,, 9:1...but u don't taken in video.... Why sir?
Yes, as its gradient elution, so I packed the column with only ethanol as silica slurry and then slowly increased the polarity by adding water in ethanol as solvent system, which could have noticed in the video. Gradient elution will help to speed the process and save solvent loss. Thanks
Nicely explained sir, please keeping making such helpful videos. 👏
Thank u so much for ur reply. Yes will be releasing another column chromatography video shortly. Keep watching.
@@dr.amolbkhade3003 thank you so much sir! 😃
Don't put sample .. atleast your mobile phase reach silica gel ...other wise you will not get 100 present purity...any way ..nice vedio ..thanks..
For your efforts...
Take care...
Thank u Vinodji for ur valuable input. Glad u liked it. Regards and tc.
My question is that ...mixture of ink red and blue is solid or liquid....?
Mixture of both inks were taken in liquid form and it was adsorbed/mixed on/with silica gel which is also used as stationary phase here.
If we are taking ethyl:hexane as Mobile phase in 1:2 ratio can i mix the solvents before wet the column .or can i use the hexane as how u used a water?
If u wish to do isocratic elution u can mix the solvents and elute with the same, if u are doing gradient elution then pack colum with hexane and gradually increase the polarity by adding ethyl acetate. Thanks for watching this video.
Sir.. U told add solvent.. Means which solvent
Solvent system is mentioned in the requirements i.e. ethanol and water in 9:1 proportion.
Thanks.if you have mixture a lot or small amount how can take silica amount in both case? I mean how many gram of silica can use? Is there standard depend on mixture ?
Depending on the number of spots and spot difference from TLC, you should opt the length of column and types of adsorbants for ex silica gel (60-120, 100-200 or 200-400 mesh), and depending on amount of ur sample (g, or mg) you should select the width of the column. As such there is no thumb rule, however u can to do it on trial and error and based on resolution obtained on TLC.
Very nice explanation
Thank u so much.
Good Explain Sir ji 🌹 God bless you.
Thank u so much, keep watching.
Pls, make the video in dry chromatography with suitable examples, and easy to us....
Yes surely will do that in a couple of days. Thanks for suggesting content and watching this video.
awesome understanding.... Thank you
Glad it was helpful! Keep watching. Thanks.
Sir which solvent has to be taken for purification of protein in mushroom
It depends on what kind of protein u wish to isolate. What u can do first develop the mobile phase using TLC, optimize the solvent system which will definitely help u to decide solvent system for column chromatography. If u wish to use HPLC Acetonitrile and TFA can be used in RP mode.
Nice video. its use, please? other than ink separation
We can seperate many drug mixtures, purify intermediates during synthesis or extraction and many more. Its maintly and seperation tool. Thanks for watching this video. Regards
Sir pls I want seperation of inorhanic cation and anion seperation by column chromatography.. pls let me know what will be the mobile phase n how I should perform this
First of all my sincere apology for the delayed reply. Frankly speaking the selection of mobile phase totally depends on nature of captions or anions u r interested to separate, however u may try with aqueous HCl and ethyl alcohol, initially u may develop the TLC and then can run the column.
In an HPLC experiment 4-nitrophenol, acetophenone and benzophenone were analyzed. What is a good asymmetry value? And also if the values are below or above that value, what is a possible reason for that?
Ideally, to get a Gaussian shape peak we should get Asymmetry value 1. If the value is greater than 1 it shows tailing and if it is less than 1 it shows fronting. There are many possible reasons for not getting symmetrical peaks which u need to find out practically. Some of the possible reasons are aging of a column, contamination of HPLC system, exhausted guard column, improper flow rate, temperature, high ionic strength if RP is used, extreme pH conditions, improperly diluted/ conc. samples, solvent pH, etc.
Tq sir
You are welcome
please sir tell me how can i separate [6]-gingerol from crude extract of ginger by organic layer seperation
You need to identify and optimize the solvent system first using TLC. Once u get separation of 6-gingerol on TLC u can try in column using developed solvent system. Probably u can try with Diethyl ether and n hexane with 7:3 ratio.
Ok sir thank you
Can we do it for oil also sir
Yes, rough separation of crude oil is possible using column chromatography, u need to optimize the solvent system. If possible after column chromatography u can separate more purified fractions using GC or HPLC. U can also try using preparative TLC if it's on the microscale. Thanks for watching the video.
@@dr.amolbkhade3003 sir can you please guide me how to extract lipids from cooking oil ? Is there any titration method can be followed ?
Screen isn't clear
Plz watch the video at higher resolution setting with atleast greater than 360p. This video is recorded with resolution of 720p. Check in video setting of TH-cam. Thanks.
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