Showa Denko Shodex SUGAR SP0810 8C User manual

Operation Manual Shodex SUGAR SP0810 8C Version 211015E
Operation Manual
Shodex™ SUGAR™SP0810 8C
Europe, Middle East, Africa, Russia
www.shodex.de
Sales Office:
Showa Denko Europe GmbH
Shodex Business
Konrad-Zuse-Platz 3
81829 Munich
Germany
Technical Support:
[email protected]
+49 (0)89 / 93 99 62 37
Quotes and Orders:
[email protected]
+49 (0)89 / 93 99 62 34

Operation Manual Shodex SUGAR SP0810 8C Version 211015E
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Operation Manual
Shodex™ SUGAR™ SP0810 8C
(Please read this manual carefully to achieve the best and consistent column performance for a long time)
Important Handling Instructions
Caution! • Please consult the Safety Data Sheet (SDS) for the reagents and solvents used with
the columns and understand their proper handling methods to prevent potential
health hazard or death from occurring.
•Please wear appropriate personal protective equipment such as lab goggles and
gloves when handling organic solvents and acid and alkaline reagents. Avoid any direct
physical contact to prevent chemical injuries.
Before Using the Column
(1) Please visually inspect the package and outside of the column for any damage.
(2) Please check if product name and serial number written on the column package, column name tag,
and enclosed Certificate of Analysis (CoA) are matching and correct.
1. Introduction
Thank you for purchasing the Shodex™ product. Shodex™ SUGAR™ SP0810 8C is a high performance
chromatography column which is specifically designed for the separations and analysis of sugars. A
strong acid cation exchange type packing gel, prepared from a rigid resin of styrene divinylbenzene
copolymer provides a mixed separation mode. The analytes are separated by a combination of ligand
exchange and size exclusion modes. It is capable of rapidly separating and analyzing monosaccharides
or disaccharides from each other, and also sugar alcohols.
2. Column Components
3. Column Specifications
Product code
Product name
Column Size ID x
Length (mm)
Particle
Size (µm)
Theoretical Plate
Number (Per Column)
Exclusion
Limit*
F6378106
SUGAR SP0810 8C
8.0 x 100
7
≥ 3,000
1,000
F6700081
SUGAR SP-G 6B
6.0 x 50
10
(Guard Column)
-
* Reference value only / Measured with pullulan
Base Material: Spherical porous particles of styrene divinylbenzene copolymer
modified with sulfo group (counter ion: Pb2+)
Column Housing: SUS-316 (stainless steel)
Screw Type: Internally-threaded type No.10-32 UNF
Shipping Solvent: Water

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4. Usable conditions
Product Name
Flow Rate (mL/min)
Maximum Pressure
(MPa) (Per Column)
Temperature (°C)
Recommended
Maximum*1
Recommended*2
Maximum
SUGAR SP0810 8C
0.5 - 1.0
1.0
1.0
70-85
95
SUGAR SP-G 6B
-
-
-
70-85
95
*1 When analyzing under 40°C, set the flow rate at less than 0.5 mL/min to reduce the column load.
*2 Increased temperature slows down the formation of sugar anomers, and thus prevents peak splitting.
Usable solvents are listed below.
(1) The standard eluent is water.
(2) Up to 20 % (v/v) acetonitrile or ethanol is usable.
Caution! • The eluate may contain lead counter ion attached to the sulfo group. In particular,
presence of acids, bases, or inorganic salt ions in the eluent or sample makes it easier
for lead ion to desorb. Please be careful when disposing the eluate.
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.
•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-μm 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-μm filter to prevent the column from clogging or
deteriorating.
(3) Recommended sample injection volume is less than 10 μL per column.
(4) When analyzing an acidic or a basic sample, make sure to neutralize the sample prior to the
injection.

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(5) When a sample contains cations other than lead ions (the pretreated sample after neutralization),
remove them by using a cation exchange resin.
(6) When a sample contains protein or lipid, make sure to remove them prior to the injection. Proteins
may be removed by ultrafiltration or by adding acid or acetonitrile. If using acid to remove the protein,
neutralize the sample before injection. If using acetonitrile, make sure that the final acetonitrile
concentration is less than 20 % (v/v).
(7) When a sample contains a large amount of organic acids, use an OH-form anion exchange resin to
remove them.
(8) When a sample contains hydrophobic substances or surfactants, use a reversed-phase solid phase
extraction to remove them.
Note • Use of guard column is recommended to protect the analytical column.
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 introduce water.
Attention! • If the eluent left in the system is not compatible with the column to be used, it may
damage the column.
• A drastic change in the eluent compositions may remove substances adsorbed on
the system and they may enter and deteriorate the column.
7.2. Column Installation
(1) Connect the column to LC system by following the “flow direction arrow” () indicated on the
name tag.
(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 0.2 - 0.3 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) At the end of analysis, reduce the flow rate to 0.2 - 0.3 mL/min and turn the column oven off. Make
sure that the column oven temperature is at room temperature before turning off the pump.
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 0.2 - 0.3 mL/min. A rapid increase in pressure can damage the
column.

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• If the column was used at an elevated temperature, let the column temperature
return to room temperature before stopping the pump. If the pump was stoppedwhile
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.
Note • It is recommended to set the pump limiter to avoid exceeding the maximum pressure.
7.3. Solvent Exchange
When replacing the solvent, set the column temperature at 70 - 95°C and start the system at 0.2 - 0.3
mL/min. Recommended solvent volume to introduce at each step is 3 to 5 times of the column volume.
Attention! • Frequent eluent replacement may damage the column, and thus not recommended.
7.4. Column Cleaning (Regeneration)
Problems in peak shapes and elution time changes or elevated column pressure etc. are often caused
by insoluble or adsorbing components present in the eluent and reagents being deposited inside the
column. These problems may be resolved by cleaning and/or regenerating the column. When washing
and/or regenerate the column, let the washing solution flow from the column outlet go directly into
the waste container and not let the solution go through the detector.
<Cleaning Method>
Insoluble components that block the column inlet may be removed by reversing the flow direction,
i.e., introducing the eluent from the column outlet, with flow rate at less than half of the recommended
flow rate.
<Regeneration Method>
Presence of cations other than lead ion in the sample may replace the lead ion attached to the sulfo
functional group. This can change the separation patterns and peak shapes. Following regeneration
procedure reattaches the lead ions and may resume the column performance.
Set the column temperature between 70 - 95°C and introduce water at 0.5 mL/min flow rate.
Inject 50 μL of 0.2 M aqueous lead nitrate solution several times.
Caution! • Please be careful when disposing the eluate containing lead nitrate solution.
8. Column Storage
Remove the column from the system after replacing the in-column solvent with the shipping solvent.
Securely tighten the end caps and store the column at a location with stable temperature (a cool and
dark space is recommended). Refer to section 7.3 Solvent Exchange for how to replace the eluent.
Attention! • Never allow inside of the column to dry. It can damage the column.
9. Column Inspection
Inspection method is described in the Certificate of Analysis (CoA). Theoretical Plate Number (N) and
Asymmetry Factor (Fas) were calculated using the below equations.
(1) Theoretical Plate Number (N)

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(2) Asymmetry Factor (Fas)
10. Additional Warnings
(1) Do not remove end fittings.
(2) Do not make a strong impact on the column. Do not drop or hit the column on a hard surface.
(3) When disposing the column, please follow a proper waste disposal method specified by your local
regulations.
Refer to the Shodex website (www.shodex.de) for product details and their applications.
For additional assistance, contact the distributor from whom you purchased the column or contact
your regional Shodex support office.
11. Warranty (Version 4)
(1) Showa Denko K.K. warrants that the Shodex™product, at the time of delivery to the user, will conform to the
specification of the attached CERTIFICATE OF ANALYSIS, if the Shodex product is used in accordance with the
attached operating manual. The foregoing warranty is exclusive and is in lieu of all other warranties with respect
to the Shodex product, whether written, oral, implied, statutory or otherwise. No warranties by Showa Denko
K.K. are implied or otherwise created, including, but not limited to, the warranty of merchantability and fitness
for particular purposes.
(2) Any claim of inconformity to the specification must be notified to Showa Denko K.K. within ten (10) days after
delivery to the user. User’s exclusive remedy and Showa Denko K.K.‘s exclusive liability for such claim are limited
to the replacement of the Shodex product in question. In no event is Showa Denko K.K. liable for any indirect,
incidental or consequential damage arising out of in connection with the Shodex Column, whether or not such
damage is allegedly based on breach of warranty, negligence or otherwise.
(3) No warranty is made in any of the following cases:
3-1 If the Shodex product is not used in accordance with the operating manual
3-2 If the Shodex product is remodelled by anyone other than person or firm designated by Showa Denko K.K.
3-3 If the Shodex product is disposed of
3-4 If the Shodex product is resold by the user without giving prior written notice to Showa Denko K.K.
3-5 If the performance of the Shodex product is not conform to the specification of the attached CERTIFICATE OF
ANALYSIS due to any of the reasons below:
a. Computer virus
b. Impurities contained in the sample, reagent, gas air or cooling water provided by the user
c. Breakdown or malfunction of equipment, apparatus or component used in combination with the Shodex
product
d. Force majeure such as fire, earthquake, flood, other natural disaster, crime, riot, act of terrorism, war or
radioactive contamination
(4) In no event is Showa Denko K.K. liable for
4-1 the results of analyses or preparations using the Shodex product or any portion of the same, including, but
not limited to, the reliability, accuracy, efficacy and safety of said results, and
4-2 the occupational hazard in the use of the Shodex product, whether or not such use is made in accordance
with the attached Conditions for use.
(5) The Shodex Column is for laboratory use only. It must not be used for clinical diagnosis. Showa Denko K.K. is
not liable for any use of the Shodex product except laboratory use.