Derek,
I have spent several years working with lipoproteins and separations. In my experience, the lipids must be removed prior to using RP-HPLC as the lipids are strongly retained by the RP column and under normal protein elution conditions will not be removed. The use of very strong solvents can remove the lipids from the columns; however, a simple method of delipidation prior to using HPLC can eliminate this. The following is a reference for a new delipidation method using solid phase extraction.
L.K. Watkins, P.V. Bondarenko, D.C. Barbacci, S. Song, S.L. Cockrill, D.H. Russell, and R.D. Macfarlane, Fast C18 solid-phase desalting/delipidation of the human serum apolipoproteins for matrix-assisted laser desorption ionization and electrospray ionization mass spectroctrometric analysis, J. Chromatog. A, 840, (1999), 183-193.
Only once was there a problem with protein precipitation using this method and it was a very large (MW 550,000), extremely hydrophobic protein that always precipitates in the absence of lipids. If quantitative recovery of the protein is necessary, a modification of this method works better and I can supply you with the details. If you have any questions at all, please feel free to contact me at LKW7389@acs.tamu.edu.
Regards,
Layle K. Barbacci
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