Showa Denko Shodex IC Y-521 User manual

HPLC Columns
MANUAL
IC Y-521
Shodex HPLC Columns
Europe, Middle East, Africa, Russia
SHOWA DENKO EUROPE GmbH
Shodex Business
Konrad-Zuse-Platz 3
81829 Munich, Germany
E-mail: [email protected]
Phone: [phone]
www.shodex.de
For technical support please use
contact details shown below:
Columns manufactured by Showa Denko K.K Japan
Made in Japan

IC Y-521 Ver. 640
Manual
2
Operation Manual
Shodex™IC Y-521
(Please read this manual carefully before using the column to ensure performance and life.)
1. Introduction
Shodex IC Y-521 is designed for exclusive use in ion chromatography for cationic separation.
Packed with a polystyrene-based strong cation exchange resin prepared specially for ion
chromatography, the column is suited for use in separation of alkaline metals, alkaline earth
metals and amines.
2. Specification
1) Size: ID, 4.6 mm; length, 150 mm.
2) Material: SUS 316
3) End fittings: Swagelok-type, No. 10-32 UNF
4) Packing: Polystyrene-based cation exchange resin with low exchange capacity
5) Plate number: 3,000/15 cm
Note: The plate number was calculated on the following conditions.
i) Test specimen: 20-ppm Na+.
ii) Injection volume: 50 µl.
iii) Mobile phase: Aqueous 4-mM nitric acid.
iv) Flow rate: 1.5 ml/min.
v) Column temperature: 40°C.
3. Mobile phase
1) For separation of alkaline metals and amines
Aqueous nitric acid in a range of 2 to 15 mM is used as the mobile phase. No organic
solvents other than methanol can be added to it and the concentration of such methanol
must be 1096 maximum.
CAUTION! Water for use in separation of alkaline metals must be ion exchange water with
electrical conductance of 1 µS/cm maximum that has passed through a 0.22-µm-mesh filter.

IC Y-521 Ver. 640
Manual
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2) For separation of alkaline earth metals
A solution of ethylene diamine and tartaric acid is the mobile phase.
NOTE: Recipe for mobile phase of 2-mM ethylene diamine/4-mM tartaric acid
i) Dissolve 6.01 g of ethylene diamine and 30.02 g of tartaric acid in ion exchange
water that has passed through a 0.45-µm-mesh filter to obtain one liter of the
solution.
ii) Dilute the solution 50 times and pass it through a 0.22-µm-mesh filter to prepare the
mobile phase.
3) Degassing
Upon heating the mobile phase to 60 to 70 C, degas it by means of an ultrasonic bath and an
aspirator.
4. Preinjection treatment of test specimen
1) Be sure to pass the test specimen through a 0.45-µm-mesh filter.
NOTE: Disposable filter, Shodex DT ED-13 or ED-03, is recommended for this purpose.
2) Be sure to remove protein, if contained in the specimen.
3) Remove any metal ions other than those of alkaline metals from the specimen that is to
undergo alkaline metal separation.
Injection of test specimens containing alkaline earth metals or heavy metal ions will temporarily
deteriorate the column performance for separation of alkaline metals.
5. Procedure to follow before column installation
1) Purge the chromatograph thoroughly with the mobile phase.
2) Remove pressure gages and dampers, if any, from the chromatograph, to wash them with the
mobile phase, and restore.
Pressure gages and dampers or a now-through type are recommended for your use.
CAUTION! When a Bourdon tube gage is used, be sure to install line filter column IC Y-
521L between the gage and the injector.
3) In case alkaline metals are to be separated with a chromatograph equipped with a stainless
steel piping, be sure to passivate the inside surface of the pipes.

IC Y-521 Ver. 640
Manual
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CAUTION! Although aqueous nitric acid can be used with such a chromatograph, the
inside surface of the pipes is likely to partially dissolve out until it has been completely
passivated.
Since the substances so dissolved in the aqueous nitric acid deteriorate the performance
of the column, if passed through it, take the following procedure to purge the
chromatograph before mounting the column on it.
i) Keep passing aqueous 20-mM nitric acid through the chromatograph at a low
rate or 0.5 to 1.0 ml/min or a few hours.
ii) Stop the pump and leave the chromatograph as is overnight.
iii) Keep passing aqueous 20-mM nitric acid through it for two hours.
iv) Replace the aqueous nitric acid with the mobile phase for use in separation
and keep passing the latter through the chromatograph for one hour.
6. Preheating of column
Preheat the column for better reproducibility, but not higher than 70 °C.
The temperature of 40 to 50°C is recommended for normal operation.
7. Start-up
Be sure to set the now rate at the specified level before starting the pump.
CAUTION! 1) Do not allow the flow rate to exceed 3.0 ml/min.
2) Do not build the pumping pressure/column above 50 kg/cm².
3) Do not abruptly change the pumping pressure or the flow rate during the
separation.
8. Column dismounting and storage
1) In case the column was kept heated during ·the operation, reduce the flow rate to 1.0 ml/min
maximum; turn off the column heater and keep the mobile phase flowing until the column cools
down to room temperature.
2) Stop the pump and dismount the column.

IC Y-521 Ver. 640
Manual
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3) Cap both ends and store the column in a place where no direct sunlight reaches and ambient
temperature fluctuates little.
CAUTION!
i) Do not store the column with aqueous nitric acid or 10 mM or more contained in it.
ii) In case the column is to be stored longer than one month, pass aqueous 0.1-M sodium
chloride through it at a flow rate of 0.5 ml/min to change the ion exchanger to an Na+
type and then pass aqueous 0.1% sodium azide before capping both ends.
ii) Do not loosen the column ends under any circumstances or the column
performance will deteriorate.
9. Deterioration of column performance and corrective actions
The causes given in the table below temporarily deteriorate the column performance,
expediting elution of the constituent substances or the test specimen to unsatisfactory
separation. A special precaution, therefore, must be exercised in separating alkaline metals.
Cause
Corrective Action
Reactivation
1) Contaminants in
mobile phase, e.g.,
those from
chromatograph.
1) Use a precolumni).
2) Use a line filter
columnii).
1) Inject 100µl of
aqueous 1-N nitric acid
4 to 6 times.
2) Pass aqueous 0.1-M
sodium tartrate and
0.1-M nitric acid in
that orderiii).
2) Metal ions in test
specimen.
1) Use a precolumni).
1) Same as 1) and 2)
above.
3) Proteins and nitrogen
compounds in test
specimen.
1) Remove proteins.
2) Use a precolumni).
1) Pass aqueous 0.5-N
sodium hydroxide and
0.1-N nitric acid in that
orderiv).
4) Use for separation of
alkaline metals of the
column that was used
for alkaline earch
metals separation
1) Use a column designed
exclusively for the
specific purpose.
1) Inject 100 l of aqueous
1-N nitric acid 4 to 6
times.

IC Y-521 Ver. 640
Manual
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Notes: i) Shodex IC Y-521P is recommended.
ii) 10 Shodex IC Y-521L is recommended.
Since It Is filled with ion exchange water by the manufacturer, be sure before use to pass
ca. 30 ml or the mobile phase through it.
The line filter column can be reactivated by passing through it 50 ml each or aqueous
0.5-N sodium hydroxide, 0.1-M tartaric acid and 0.5-N nitric acid in that order.
iii) Flow rate, 0.5 ml/min; volume, 30 ml each.
iv) Flow rate, 0.5 ml/min; volume, 30 ml each. There are some cases where the
column cannot be reactivated because or adsorption of the test specimen on the
packing.