ver.211214E
SHOWA DENKO K.K. (https://www.shodex.com/)
2
4.
Usable Conditions
*
In general, lower temperature setting is more suitable for the separations of optical isomers.
Usable solvents are listed below.
(1) A mixture of water and an organic solvent, such as acetonitrile, of any ratio can be used. Up to 100 % water
and organic solvents can be used.
(2) Ethanol, 2-propanol (IPA), and tetrahydrofuran (THF) can be used.
(3)
Various buffer and aqueous salt solutions can be used instead of water. They include phosphate, or acetate
buffers, and sodium chloride solution. Please keep total concentration of salt under 1 M.
(4)
Acetic acid can also be added. Please keep its concentration under 6.0 % (w/v).
Attention!
· Use the column within above stated flow rate, pressure, and temperature ranges. Using
the column outside the given range may damage the column and lower its performance.
· When using a mixture of buffer (or aqueous solution of salt) and organic solvent, make
sure there is no precipitation of salt.
·
Column pressure is influenced by the eluent composition, flow rate, and column
temperature. When changing the eluent compositions, adjust the flow rate and column
temperature so that the column pressure remains below the usable maximum pressure.
5. Eluent Preparation
(1) Degas the eluent fully to prevent the formation of air bubbles.
(2) 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.
Attention!
· Whenever water is required, use ultra-pure water freshly generated by a water purification
system or water from a newly opened HPLC grade distilled water bottle. 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.
Note
· Use of on-line degasser is recommended.
6. Sample Preparation
(1) If possible, use the eluent for analysis to dissolve or dilute samples. If this is difficult, use a solvent which
has a composition that is as close as possible to the eluent's composition, but which fully dissolves or dilutes
the sample. When gradient elution is used, it is recommended to use the initial eluent to prepare the sample.
(2) Filter the sample solution using disposable 0.45-
μ
m filter to prevent the column from clogging or deteriorating.
(3) Recommended sample injection volume is less than 20
μ
L per column.
(4)
For the samples containing proteins, remove the proteins prior to the sample injection.
7. Column Usage Procedure
7.1 HPLC System Preparation
Wash entire LC system prior to the column installation, including all flow-lines and sample loop by switching the
valve, and then replace the washing solution with the eluent to be used. If desired new eluent has low
miscibility/solubility to the eluent of previous analysis, first use the eluent that is miscible/soluble to both eluents,
and then replace it with the desired eluent.
e.g. When replacing chloroform to water first run methanol and then replace it with water.
e.g. When replacing highly concentrated buffer solution or salt solution to water/acetonitrile, first run water and
then replace it with water/acetonitrile.
Product Name
Flow Rate (mL/min)
Maximum
Pressure (MPa)
(Per Column)
pH
Range
Temperature (°C)
Recommended Maximum Recommended
*
Range
ORpak CDBS-453
0.3 - 0.8
1.2
20
2 - 7
10 - 30
5 - 60