Analytical HPLC-columns

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Chris.L
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Analytical HPLC-columns

Postby Chris.L » Wed Apr 16, 2014 11:51 am

Hello,
please excuse my poor englisch.
I want to make my own analytical HPLC column (150 x 4.6 mm, C18, 5µm) but i can´t find any information how to produce one.
So, have sombody an idea where i can find it ?

Thank you for your help.

Greetings.

daniswan
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Postby daniswan » Wed Apr 16, 2014 2:37 pm

The UWPR has a lot of useful information on this: http://proteomicsresource.washington.edu/protocols05/protocols05_LC_MS.php

Chris.L
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Postby Chris.L » Wed Apr 16, 2014 8:46 pm

Thank you for your help. But i think their are only informations about making capillary columns. Is it possible to transfer this informations about paking capillary or preparative columns to analytical columns with an ID about 4.6 mm ? Because i try to transfer the mechanics to get a preparative column to my analytial column and it workt not so good.

tsbatth
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Postby tsbatth » Tue Apr 22, 2014 1:53 pm

Out of curiosity why not just buy one? I mean there is a lot of variety in analytical columns with dimensions and sizes you're interested in.

Chris.L
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Postby Chris.L » Wed Apr 23, 2014 1:47 am

It its just to expensive to buy everytime a new column because we need a lot of them.
So its all about the money :)

JDRCHEM
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Postby JDRCHEM » Wed Apr 23, 2014 5:18 am

Unfortunately, you will will most likely be very disappointed with self-packed, standard-size HPLC columns for applications above ~ 2000 psi. Millions upon millions of R&D/engineering dollars have been spent to fine tune packing of HPLC columns to make them as reproducible as they are from column-to-column. The details of how these columns are packed by commercial vendors are closely guarded secrets. There is a reason these columns cost as much as they do, in part, it is an effort by vendors to recoup the cost of developing a reproducible column packing process.

Can columns be self packed? Absolutely, but you will most likely have voids that will both affect the reproducibility and reduce the number of theoretical plates relative to a commercial column. If you figure out how to pack your own columns such that their quality is as good as commercial columns, you would have a very marketable/profitable skill that few people have.

Chris.L
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Postby Chris.L » Wed Apr 23, 2014 10:26 am

Hi Jdrchem
that is one of my main problems. The reproducibility is not so bad but the columns have far less theoretical plates than commercial columns.

OliviaRBanks
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Postby OliviaRBanks » Fri Jul 11, 2014 7:26 am

Good morning,

My name is Olivia Banks I am learning how to pack my own capillary columns now for our Easy- nLC 1000 and I am having some trouble with the whole packing process. I am using a 75 um ID silica and the packing material is Hamilton PRP- C18 100 Angstrom 5um resin. I use a micro-torch to pull the thin glass tip for my columns. It’s taking me about 2-3 hours to pack about 15 cm of column at 800 psi. Is there any way this time can be cut down? I also inserted a packed column into the Easy-nLC and ran isocratic flow and all of the packing material came out through the glass tip. I do not use frits when packing my columns because of the glass tip. The packing material that I was using before Michrom Magic C18 200 Angstrom 5um, but this packing material no longer exists. I did research and the Hamilton packing material is the most similar and cost effective so we went with that one.

Any information or help will be greatly appreciated. I am using the protocol on the UWPR website because it gives a great explanation on how to pack columns and frits.

Infinity
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Postby Infinity » Mon Jul 14, 2014 10:38 am

Typically HPLC column should last long and you can use it for hundreds of separations. If it is not the case I would say it could be more useful to invest time into cleaning your sample/optimizing separation conditions to extend lifetime of the column. There should be a reason why commercially available column doesn't last for long. If your sample contains something that interfere with separation and you are not able to remove it you could try to buy a precolumns (works like a filter, contains the same material as analytical column and installed just before the column) but they are also expensive (but much cheaper compared to analytical columns).

Christopher
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Postby Christopher » Fri Oct 31, 2014 8:53 pm

I guess the tip ID just isnt small enough and the beads are coming out. You could try putting a conventional Kasil frit in there before you pull the tip. Just put the frit liquid in, and let the capillary sit vertically for 10-20 minutes and the frit liquid will drift down the tube. Toss it in an oven and let it solidify, then pull your tip in the remaining capillary after the frit. Just try to keep the amount of liquid for the frit to a minimum, and you shouldnt really have any post-column peak broadening troubles.


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