ver.211215E
SHOWA DENKO K.K. (https://www.shodex.com/)
2
4.
Usable Conditions
Usable solvents are listed below.
(1) A mixture of water and acetonitrile or water and ethanol of any ratio can be used. Up to 100 % water,
acetonitrile, or ethanol can be used. Use of organic solvents other than acetonitrile or ethanol is not
warrantied.
(2) Aqueous and buffer solutions can be used instead of water. They include acidic aqueous solutions such as
phosphoric and acetic acids; alkaline aqueous solutions such as ammonia and sodium hydroxides; aqueous
salt solutions of sodium chloride, potassium chloride, or sodium sulfate; and buffers of phosphates or
acetates.
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.
· When using highly corrosive salts such as sodium chloride, wash out the salts at the end
of analysis. The metal parts of the devices and/or the columns may rust.
·
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.
· Do not use hydrochloric acid as it corrodes stainless steel.
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.
e.g. When using a mixture of water (buffer) and over 50 % acetonitrile for the eluent: If the sample does not
dissolve in the eluent well, first dissolve the sample with water (or buffer), and then add acetonitrile to
make the final acetonitrile concentration in the sample more than 50 % (v/v).
(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 5
μ
L per column.
(4) When a sample contains protein or lipid, make sure to remove them prior to the injection. Presence of
those substances may damage the column.
Product Name
Flow Rate (mL/min)
Maximum
Pressure (MPa)
(Per Column)
pH Range
Temperature
Range (°C)
Recommended Maximum
Asahipak NH2P-40 2E
0.1 - 0.2
0.4
20
2 - 13
15 - 50