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

[ Care and Use ManUal ]
Glyco-Pak N Columns 4
II. InstallatIon
CAUTION: Excessive pressure will damage the Glyco-Pak N Column. Do not
exceed a flow rate of 1.0 m1/min or backpressure of 500 psi.
a. Column Installation
Remove the end-cap fittings from your Glyco-Pak N 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 N column is indicated by an
arrow on the column label.
Note: When changing columns, always replace the ferrule. Carefully reseat
the ferrule to avoid creating dead volume. Improper seating of the ferule will
result in reduced resolution on the Glyco-Pak N column.
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
taper 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. Solvent and Sample Preparation
Refer to the following list for mobile phase and sample considerations:
• Use LC grade solvents filtered through a Durapore ® membrane on 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.
• Mobile phase solvents should be prepared by weight for maximum
reproducibility.
• Use a Waters In-line Precolumn Filter (P/N84560) to obtain maximum
column life.
• 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.
• Samples should be disolved in acetonitrile/water (50:50) for optimal
chromography.
III. operatIon
CAUTION: Improper equilibration of this column may result in decreased
column performance. Please read this section before attempting to use this
column.
a. Column Equilibration
To equilibrate the column, set up a linear gradient from 100% water to
65 - 75% acetonitrile in water (depending on desired retention) at a rate
of 0.5 ml/min for one hour. Then flow premixed (acetonitrile/water) mobile
phase at final conditions for one additional hour at 1 ml/min.