Back in Iowa State University we used Waters HPLC for protein purification during many years without noticeable damage to the stainless steel tubings. But Dan was right about the pumps, someone in the lab forgot to flush the high salt pump with water after its use and damaged the pump...
Alex On May 29, 2012, at 5:14 PM, Daniel Anderson wrote: > Hi, Ho, > > Your question has a lot of variables. > > "HPLC" columns should not be used on the Akta within my field of view > because the Akta within my field of view does not have gradual pump > acceleration and deceleration. "HPLC" columns can be damaged by sudden > changes in pressure or composition. > > The "HPLC" within my field of view has wetted stainless steel surfaces > and the mobile phase should not contain chloride ion or reductant. > Chloride ion would accelerate corrosion of the stainless steel (and > result in metal ions in the protein). Reductant would strip off the > "passivation" (during maintenance I soak the stainless parts in nitric > acid to keep them stainless) later resulting in corrosion. > > The Waters sales representative once told me that the pumps have to be > salt-free and methanol-flushed at the end of every working day. Good > luck implementing that policy. > > -Dan > > Ho Leung Ng wrote: >> Hello, >> >> My Akta Purifier is being repaired, and I'm thinking about >> borrowing a colleague's HPLC in the interim. What makes the Aktas >> different from HPLCs? I've used HPLCs for purifying small molecules >> and peptides but not proteins. Anything I should be careful about >> regarding keeping the machines, columns, and proteins happy? >> >> >> Thank you, >> Ho >> >> Ho Leung Ng >> University of Hawaii at Manoa >> Assistant Professor, Department of Chemistry >> h...@hawaii.edu <mailto:h...@hawaii.edu>
