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[ CARE AND USE MANUAL ]
[ CARE AND USE MANUAL ]
IV. TROUBLESHOOTING
To achieve reliable performance of the method, ensuring
sensitivity and peak shapes are acceptable, contamination
of the LC system should be eliminated. For cleaning of the
Waters LC system and other troubleshooting please refer
to the startup guide.
Please contact your local Waters representative if you need
more guidance and advice on troubleshooting or cleaning your
ACQUITY liquid chromatography system.
For non-Waters LC systems, please contact your supplier for
specific information on liquid chromatography system cleaning.
V. COLUMN CLEANING, EFFECTS OF
ADDITIVES, AGING, AND STORAGE
Changes in peak shape, peak splitting, shoulders on the
peak, shifts in retention, change in resolution, or increasing
backpressure may indicate contamination of the column.
Flushing the column while taking care not to precipitate
buffers, is usually sufficient to remove the contaminant.
Using a two step flushing procedure, divert the flow to waste
and flush the column with an appropriate volume of 20:80
acetonitrile:water, and gradually return to 100% acetronitrile.
Be sure to use a flow rate that does not over pressurize the
system or column. If the flushing procedure does not solve the
problem, purge the column using the following cleaning and
regeneration procedures.
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©2019 Waters Corporation. Produced in the U.S.A. September 2019 720006666EN Rev A IH-PDF
a. Cleaning
Use the cleaning routine that matches the properties of the
samples and/or what you believe is contaminating the column.
Flush the Anionic Polar Pesticide Columns with 50:50
acetonitrile:water to remove polar contaminants. If this
flushing procedure does not solve the problem, purge
the column with 5:95 acetonitrile:water. To clean polar
contaminants from the Anionic Polar Pesticide Column, run
a 10-minute gradient from 0–100% water and return to the
initial gradient method condition. Please note that as aqueous
concentration increases, backpressure will rapidly increase
as well. Reduce flow rate when operating at greater than 60%
aqueous. Repeat if necessary.
Retest the column after using the cleaning procedure to
determine if the specific problem has been fixed. If so, continue
using the column, avoiding samples and solvents that may
clog the column inlet. If the column performance is poor after
cleaning and regenerating, call your local Waters office for
additional support.
b. Storage
Purging out any additives with water/acetonitrile (50:50) and
storing in 100% acetonitrile is recommended. Alternate storage
solvents can be used but may impact initial column equilibration.
Waters Corporation
34 Maple Street
Milford, MA 01757 U.S.A.
T: 1 508 478 2000
F: 1 508 872 1990
www.waters.com
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