How can we measure void volume after packing the column? What are the main steps of packing the column? What is the protocol to use silica gel column chromatography?
Great questions: To answer them, please refer the manuals and guides provided by the support manufactures and/or your teachers (read them). If your question relates to Low pressure (LP) columns using glass tubes/columns packed with Silica Gel (i.e. Silica Gel 60) and gravity fill or with the aid of a low pressure pump, then pack the columns using a slurry solution of the support (example guide: https://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_for_the_Lab_Column_Packing.html ).
Column Void volume will be determined by selecting and running a std through the column that is not retained by the support and elutes out quickly (you need to know the elution Time and the Flow Rate to calculate it). The information needed can be found in the manuals and guides provided by the various support manufactures of the supports used. "Packing" involves making a simple slurry of the beads in solvent, then carefully pouring or "pumping" the slurry into the cylinder (with glass wool or frits installed) allowing the bed to form on top. Use degassed liquid for the solution and try to minimize voids or air bubbles when creating the column. Practice is key. Please refer to the support manuals and guides for the specifications of the supports, including their volume and best gravity or low pressure packing techniques. For tips on packing LP columns, try a keyword search on the web (i.e. GOOGLE) for hundreds of examples.
https://collegedunia.com/exams/column-chromatography-principle-procedure-applications-chemistry-articleid-709
http://fac.e-learningcenter.tanta.edu.eg/pluginfile.php/27894/mod_resource/content/1/Column%20Chromatography%202.pdf
Good luck.
Dear Fnu Harshvardhan for an instructive tutorial video about the use of silica gel column chromatography pleas have a look e.g. at the following YouTube video:
Silica Column Chromatography
https://www.youtube.com/watch?v=PE3J-vNi55Y
Also please see the following manual in which the basic steps are described in detail:
Column Chromatography Procedures
https://orgchemboulder.com/Technique/Procedures/Columnchrom/Procedure.shtml
Good luck with your work!
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