Waters Glyco-Pak DEAE Columns - Manual (Page 4)

[ Care and Use ManUal ]
Glyco-Pak DEAE Columns 4
II. InstallatIon
a. Column Installation
Remove the end-cap fittings from your Glyco-Pak DEAE column with a
5/16-inch wrench. Save the end-cap fittings to recap the column when it is
removed from the system.
Mobile phase flow direction for the Glyco-Pak DEAE column is indicated by an
arrow on the column label.
Note: When changing columns, always replace the ferrules. Carefully reseat
the ferrule to avoid creating dead volume.
Follow the next four steps of this procedure to cut tubing to connect a new
steel column, or to improve the end connections on your existing fittings.
1. Using a three-cornered file with a cutting edge, scribe the circumfer-
ence of the tubing at the desired break.
2. Grasp the tubing on both sides of the scribe mark with cloth-covered
pliers (to prevent marring the tube surface) and gently work the tube
back and forth until it separates.
3. Slide the compression fitting, followed by the ferrule (large end of the
taperr first) over the tube. Be certain to bottom the tube in the fitting
seat to assure a leak-free connection.
4. Use a 5/16-inch wrench to install the column in the system.
Note: Attach a union in place of the column and flush the lines before install-
ing the column. This flushes previous, possibly incompatible mobile phases
from the system.
Figure 5: Ferrule and Compression Assembly
b. Mobile Phase and Sample Preparation
Refer to the following list for mobile phase and sample considerations:
• Use LC grade salts filtered through a Durapore ® membrane, with a
Waters Solvent Clarification Kit (P/N 85113) or other suitable membrane
to remove microparticulate matter above 0.45 µm. A particulate-free
mobile phase reduces the problem of plugged filters and column beds
and preserves column life.
• Use vacuum filtration or sonication to remove dissolved gasses which
could affect your mobile phase delivery system.
• Filter particulate matter from samples to prevent excessive pressure
buildup. Use Millex
® -HV Filters (P/N 83996) or the Waters Sample
Clarification Kit (P/N 26865) to filter samples.
• Use a Waters In-line Precolumn Filter (P/N 84560) to obtain maximum
column life.
• Anionic species, such as protein denaturing agents, should be removed
from samples prior to fractionation.
• Buffers typically used with other DEAE columns can be used with the
Glyco-Pak DEAE column. These include sodium, potasium, amonium
and iris salts of phosphate, acetate and chloride.