An HPLC
column has a finite lifetime. It is recommended that you routinely monitor a
column's retention characteristic and performance using standards. If you plan
to store the column for a prolonged period of time, use a mobile phase that
inhibits microbial growth (a mobile phase containing sodium azide or high
concentrations of methanol, acetonitrile, etc.). For optimum performance and
column life, follow these recommendations.
Routinely monitor the column's
performance.
Switch only between miscible mobile
phases.
Avoid precipitating salts in the column.
Use filtered and degassed mobile phases.
Do not allow the column to dry out. Keep
it tightly capped when not in use.
For prolonged storage, use a mobile phase
that inhibits bacterial and mold growth.
Use the column in the direction indicated
on the label. An unusually high operating pressure may indicate a plugged inlet
frit and may be cleared by reversing flow through the column for 10-20 mL.
Use Hamilton
guard columns to remove particulate matter or impurities that may permanently
bind to the analytical column.
Restoring Column Performance The procedures listed below may restore the performance of a column that has become
fouled, but the procedures will not improve the performance of a column that is
damaged or beyond its useful life. Nor can all stationary phases be restored.
The unique supports in PRP-X500 and PRP-X600 columns make it impossible to restore
performance with a washing procedure. For technical assistance regarding these
columns, please Contact HPLC Support.
NOTE:
Before performing a restoration procedure, always make sure that the mobile
phases are miscible and that precipitation will not occur. If necessary, wash
the column with a suitable intermediate solvent before changing over to the new
mobile phase. A minimum of five column volumes of intermediate solvent should
be used.
Column Restoration Procedures Follow the appropriate restoration procedure listed below using a flow rate of 1.0 or 2.0
mL/min. unless otherwise specified. Evaluate column performance using the test
mix and conditions listed on the Performance Test Report shipped with each
column.
Column Restoration Procedures
Packing
Name
Restoration Method
PRP-1
PRP-3
PRP-h1
PRP-h5
PRP-Infinity
Run
a gradient of 100% water to 100% acetonitrile.
Repeat three times.
PRP-X100
Pump
approximately 50 mL of methanol with 1%
6 N nitric acid.
PRP-X200
PRP-X300
Inject
several times with 100 µL of 1 N nitric
acid.
PRP-X400
Inject
several times with 100 uL of 0.1 M potassium
EDTA.
RCX-10
Flush
with 50 mL 0.1 N sodium hydroxide.
RCX-30
Flush
with 150 mL 0.1 N sodium hydroxide.
Column Restoration Procedures
HC-40
Ca2+
Flush
with 1% calcium chloride at 0.1 mL/min overnight.
HC-75
Ca2+
Flush
with 1% calcium chloride at 0.1 mL/min overnight.
H+
Flush
with 0.1 N sulfuric acid at 0.1 mL/min overnight.
Pb2+
Flush
with 1% lead nitrate at 0.1 mL/min overnight.
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