Waters Glyco-Pak DEAE Columns - Manual (Page 6)

[ Care and Use ManUal ]
Glyco-Pak DEAE Columns 6
To aid in diagnosing potential column problems:
• Equilibrate the column in 15 mM sodium acetate adjusted to a pH of 5.0
with acetic acid at a flow rate of 0.8 ml/min for 1 and 1/2 hours
• Inject 10 µl of a 10 mg/ml solution of N-acetylneuraminic acid in
15 mlVl sodium acetate buffer at pH 5.0
• Set detector to 220 nm.
Retention time will vary with absolute concentration of the buffer as well as
instrument configuration, however, the peak shape and retention should be
similar to that seen in the chromatogram below.
Figure 8: Test Chromatogram, N-Acetylneuraminic Acid
Performing this test prior to discussing the problem with your Waters repre-
sentative will help in diagnosing the problem.
a. Column Storage
• Store columns in distilled water and refrigerate at 4 - 6 °C. DO NOT
FREEZE.
• DO NOT allow buffers or other potentially harmful materials to remain
in the system overnight when not being used. Run buffers with a slow
flow rate (0.1 ml/min) overnight if necessary.
• Return the column to its box with the end caps firmly in place for stor-
age. Allowing columns to dry out can result in poor chromatographic
performance.
b. Troubleshooting
Use Table 2 to troubleshoot problems with your column.
Table 2: Column Problems and Solutions
V. warranty and serVIce InformatIon
a. Service Information
Waters Corporation staff of experienced service specialists provide mainte-
nance assistance on both preventative and/or corrective levels. For complete
information and assistance, please call Waters Service Department at 1-800-
252-HPLC. For solutions to particular applications questions Waters team of
technical support personnel are available to help you with specialized support.
They may be contacted at 1-800-252-HPLC in Milford, MA.
Waters Corporation staff of experienced service specialists provide mainte-
nance assistance on both preventative and/or corrective levels. For complete
information and assistance, please call Waters Service Department at 1-800-
252-HPLC. For solutions to particular applications questions Waters team of
technical support personnel are available to help you with specialized support.
They may be contacted at 1-800-252-HPLC in Milford.
Problem Cause Solution
Excess pressure
buildup
• Filters plugged with
Particulates
• Sample precipitates
on column (sample
not soluble in
mobile phase)
• Clean in an ultrasonic bath or
replace.
• Always alter mobile phases
and samples.
• Slowly purge with a strong
mobile phase that is both
appropriate to dissolve the
contaminate and compatible
with the column.
Loss of resolution,
broad peaks, low
plate counts
• Filters partially
plugged
• Replace or clean inlet and
outlet filters in an ultrasonic
bath.