Waters Styragel Columns - Manual (Page 4)

[ Care and Use ManUal ]
Styragel Columns 4
Flow rate and backpressure for 4.6 mm
Solvent Efficient Columns
For best resolution and maximum column life, do not allow the flow rate to
exceed 0.3 mL/min or the backpressure to exceed 3.5 MPa (500 psi, 35 atm)
per column.
Please 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.
When using the HR 0.5, HR 1, and HR 2 columns, increase the flow rate in 0.1
mL/min increments at 30-second intervals until you reach 0.3 mL/min.
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. See Table 4, Table 5,
or Table 6 for column efficiency data.
Initial efficiency tests
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 on Efficiency Testing, for the procedures to determine the
efficiency of your system and columns.
Save the chromatograms from these tests. For the column-efficiency test,
record the retention times, system settings, and all experimental conditions
so they may be reproduced exactly for future comparison.
III. PreParIng solvent and samPles
This chapter describes:
• Preparing the solvent
• Changing solvents
• Converting columns to high temperature solvents
• Preparing the sample
a. Preparing the Solvent
Use clean solvent for reproducible results and maintenance-free operation. Use
solvents of LC-grade or better, filtered to remove micro particulate matter larger
than 0.45 µm. Refer to Waters catalog, filtration section for filter choice and
solvent compatibility chart.
b. Changing Solvents
Solvent compatibility
Waters Styragel columns are shipped in the solvent of your choice: toluene,
THF or DMF. Some applications require a different solvent. Changing solvents
works best between compatible solvents. Refer to the table below for solvent
compatibilities.
Note: The use of highly aqueous mobile phases may damage the resin and is
not recommended.
To use Waters Styragel columns for high-temperature chromatography in
solvents like trichlorobenzene (TCB) or orthodichlorobenzene (ODCB), you
must convert the column to the selected solvent at elevated temperature.
High-temperature conversion procedure
To convert the column bank to high-temperature operation:
1. Convert the system to the column shipping solvent at room
temperature.
2. Purge solvent efficient columns at 0.1 mL/min while gradually
increasing the temperature to 90 ˚C over a minimum of 3 hours.
Solvent Efficient Columns
(Purge solvent efficient columns at 0.1 mL/min while gradually
increasing the temperature to 90 ˚C over a minimum of 3 hours.)
Set the system to 55 ˚C when converting columns from THF to
HFIP or Y _ Butyrolactone.
To convert to:
Use columns
shipped in:
To convert to:
Use columns
shipped in:
o-Dichlorobenzene Toluene
Trichlorobenzene Toluene Hexafluoroisopropanol THF
Phenol/TCB Toluene N-Methyl pyrrolidone DMF
y-Butyrolactone THF m-Cresol DMF
Table 1. Solvent Conversion Table