Waters µStyragel Column - Manual (Page 3)

[ Care and Use ManUal ]
µStyragel Column 3
Follow the procedure below to connect your column:
1. Refer to the guidelines above for column sequence
requirements.
2. Remove the end plugs from your column and save them. Ensure
that solvent flow is in the direction shown by the arrows on the
column labels.
3. Thread the inlet and outlet fittings of each column until finger
tight, then tighten the fittings 1/4 to 1/2 turn.
Flow Rate and Back Pressure
For best resolution and maximum column life, do not allow the flow
rate to pressure exceed 1.0 mL/min or the back pressure to exceed
500 psi per column.
Note: The flow rate recorded on the Certificate of Analysis, included
with the column, may be higher than the guidelines provided. For
maximum column life in your lab, please follow the flow and back
pressure guidance we provide.
Note: When using the 100 Å and 500 Å columns, slowly increase the
flow rate, in 0.1 mL/min increments, to 1.0 mL/min. Increase the flow
rate over a period of at least 30 seconds per increment. Initially set
the pump at 0.0 mL/min. Then turn on the pump and ramp the flow
rate gradually.
Defective Columns
One or more defective columns in a series may cause the entire set
of columns to appear defective. One defective column can cause peak
spreading that cannot be overcome by any number of good columns.
d. Equilibrating the Column
Equilibrate the column when you install it and when you use it after
it has been stored. Follow the procedure below to equilibrate your
column:
1. Set the pump at 0.0 mL/min, then turn on the pump.
2. Increase the flow rate gradually in 0.1 mL/min increments to
1.0 mL/min.
3. Purge the column with the shipping solvent until you obtain a
stable baseline.
Initial Efficiency Tests
Waters recommends that you test your system and columns before
the first analysis. Run a test sample using the recommended
parameters for your system and columns, and record the results.
These results serve as a baseline to compare future performance. See
Section V, c, Efficiency Testing, for the procedures for determining
the efficiency of your system and columns.
Save the chromatograms from these tests. Also, for the calculated
column efficiency test, record the retention times, system settings,
and all experimental conditions so they may be reproduced exactly
for future comparison.
III. PRePaRIng solV ent anD samPles
a. Preparing the Solvent
The use of clean solvent is important for reproducible results and
maintenance-free operation. Use solvents of HPLC-grade or better,
filtered to remove micro particulate matter above 0.45 µm. Go to
www.waters.com to view filter offerings from Waters.
Solvent compatibility
It is important to use solvents compatible with your particular
column. Otherwise, your columns may experience gel shrinkage
and irreversible channeling. See Table 2 for column-solvent
compatibility.