Egg White

[Chris]

We have had a request from a client to directly inject egg white onto reverse phase HPLC with a 50/50 acetonitrile/water mobile phase. Has anyone had any experience of this as it doesn't sound like the best idea to me.

Thanks

[Noser222]

Umm...not that I have ever tried this, but my first thought is that you'd have significant protein precipitation. What are they interested in?

[Jumpshooter]

You need to tell that client to: 1) purchase/provide the analytical column and guard column and 2) purchase and provide tubing; and 3) purchase and provide a flow cell. These are precautions b/c by doing so you will shorely ruin your own column.

[Bryan Evans]

Assuming they want you to analyze the proteins in the egg white...
you can try it using reverse phase on Intrada WP-RP, 300A pore size
(but you will need to run a TFA reversed-phase gradient, not 50:50 ACN:Water).

Below are proteins found in egg white (albumin and lysozyme):
http://www.imtakt.com/TecInfo/TI289E.pdf

[Uwe Neue]

First thing to do: make sure that your client is willing to pay for the column.
Second thing to do: do it!

If the injection volume is low enough, you may get away with it - for a while. But don't expect to get a lot of injections out of the column in 50/50 MeCN/water.

If your injection is large enough, my bet is that you get a boiled egg on the column inlet.

It is difficult to pump through a boiled egg...

[Bryan Evans]

Still not sure what useful information they hope to obtain by
injecting egg white directly into an HPLC column but...

Below is a chromatogram for bovine serum (no protein precipitation)
injected directly on to the Intrada WP-RP column. For this example -
Imtakt used a 0.4 uL injection volume onto a 250x4.6mm, 3um column.

The chromatogram shows that the proteins elute off the column -
you'll want to use a very small injection volume
for the egg white as well (or larger i.d. column for a larger inj volume):

http://www.imtakt.com/TecInfo/TI274E.pdf

[Mark Tracy]

My experience with ovalbumin was that the MeCN concentration that will precipitate it is only slightly different than the concentration that will elute it from an RP column.