Waters µStyragel Column - Manual (Page 5)

[ Care and Use ManUal ]
µStyragel Column 5
Very High Molecular Weight Polymers
The same behavior is typical for most very high molecular weight
polymers, although the effect may not be as easily observed as it is
with narrow molecular weight standards. Very high molecular weight
polyolefin fractions are subject to possible shearing and incipient
precipitation. Use especially dilute solutions (for example, 0.02%) to
minimize precipitation.
IV. UsIng the ColUmn
a. Chromatography Guidelines
Gel permeation chromatography columns have a finite lifetime
directly related to their care and use. Column life is reduced
by contamination from samples and eluents, frequent solvent
changeover, and improper handling and storage.
Guidelines for Column Use
When using µStyragel columns, observe the following guidelines:
• For best resolution and maximum column life, use a flow rate of
1 mL/min or a back pressure not to exceed 500 psi (3 MPa or
30 bars) per column.
• Protect the column from vibration and mechanical shock.
• Protect the column from rapid changes in pressure, which can
result from rapidly changing the composition of the solvent.
• When changing to a solvent with a different viscosity, it may
be necessary to adjust the flow rate to maintain the operating
back pressure specification of 500 psi (3 MPa or 30 bars) per
column.
• Avoid precipitation. Dissolve samples in the mobile phase.
• Always use high-quality HPLC solvents.
• Dedicate each column or series of columns to a specific applica-
tion. Frequent switching of samples and solvents accelerates
column deterioration and loss of resolution.
b. Calibrating the Column
Whenever you replace a single column or a complete column bank,
generate a new calibration curve to ensure the reproducibility of your
application. Figure 5 shows typical calibration curves by column fam-
ily. The calibration curves were obtained with polystyrene standards.
Figure 5: Calibration Curves for µStyragel
4 6 8 10 12
107
6
5
4
3
2
10
10
10
10
10
MW
Elution Volume [mL]
100Е
500Е
10^3Е
10^4Е
10^5Е
10^6Е