ver.200603E
SHOWA DENKO K.K. (https://www.shodex.com/)
3
6.2 Column Installation
(1) Connect the column to LC system by following the “flow direction arrow” (
➡
) indicated on the adhesive label.
(2) Make sure to insert the tubing all the way to the end fitting and secure it with the male nut. It is important that
there is no extra space between the tubing and the column side of the end fitting. Presence of an extra space
will let the sample to spread out and may result in wide peaks.
(3) Set the initial flow rate at less than 0.35 mL/min and start the system. If using the column at an elevated
temperature, keep a low flow rate until the temperature of the column reaches to the set temperature, and
then gradually increase the flow rate to the desired.
(4) When using the column for the first time, before connecting the column outflow to a suppressor, wash the
column with the eluent at 0.35-mL/min flow rate for more than 30 minutes. This also applies to when reusing
a column with which the eluent was replaced with shipping solvent.
Caution!
· Verify that there is no solvent leak. It may cause electronic leakage, rust, and/or chemical
injury.
Attention!
· Make sure not to let air bubbles enter the column while installing the column. The air
bubbles may damage the column.
· When restarting the system after column installation or after holding the eluent flow, start
the system at less than 0.35 mL/min. A rapid increase in pressure can damage the column.
· If the column was used at an elevated temperature, lower the flow rate to less than 0.35
mL/min at the end of analysis. Then, turn off the column oven, and let the column
temperature return to room temperature before stopping the pump. If the pump was
stopped while the eluent inside the column was still hot, as the eluent temperature
decreases, its volume also decreases. This may result in creating an empty space in the
column and deteriorates the column.
6.3 Eluents
(1) Use freshly generated ultrapure water (
18.2 M
Ω
-cm
) to prepare the eluent.
(2) The use of an eluent generation system is highly recommend for the preparation of potassium hydroxide or
sodium hydroxide solution. If an eluent generation system is not available, dilute a reagent-special grade
potassium hydroxide or sodium hydroxide solution with ultrapure water.
(3) Degas the eluent fully to prevent the formation of air bubbles.
(4) Presence of small debris or insoluble substances may result in deterioration of the column and/or they appear
as noise on the chromatograms. Filter the eluent with a 0.45-
μ
m disposable filter to prevent the problems.
(5) Start the system at less than 0.35 mL/min when replacing the solvent.
Attention!
· Use HPLC grade organic solvent whenever possible. Solvents left in an opened bottle for
a long time should not be used. The content may have been changed, absorbed moisture,
or has been contaminated.
· Always use freshly prepared solvents. Solvents stored for a long time may have changed
their compositions and may influence elution patterns and/or damage the column.
· If using a manually prepared eluent, keep the eluent in a solvent reservoir with a carbonate
trap cartridge.
Note
· Use of on-line degasser is recommended.
· It is recommended to set the pump limiter to avoid exceeding the maximum pressure.