Shodex SUGAR EP SC1011-7F User manual

Operation Manual Shodex SUGAR EP SC1011-7F Version 210415E
Operation Manual
Shodex™ SUGAR EP SC1011-7F
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 EP SC1011-7F Version 210415E
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Operation Manual
Shodex™ SUGAR EP SC1011-7F
(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) of reagents and solvents used with the
column and understand their proper handling methods to prevent potential health
hazards 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 EP SC1011-7F 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 chromatography modes. It is capable of separating and analyzing
monosaccharides or disaccharides from each other, and also sugar alcohols.
The column is suitable for mannitol analysis following JP, USP (L19, L22) and EP requirements. Only
water is required for the analysis of neutral sugars.
2. Column Components
3. Column Specifications
Product
Code
Product Name
Column Size (I.D.
x Length) (mm)
Particle
Size (µm)
Pore
Size (Å)
Theoretical Plate
Number (Per Column)
F6379300
SUGAR EP SC1011-7F
7.8 x 330
8
200
≥ 11,000
F6700090
SUGAR SC-G 6B
6.0 x 50
10
-
(guard column)

Operation Manual Shodex SUGAR EP SC1011-7F Version 210415E
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Base Material: Spherical porous particles of styrene divinylbenzene copolymer modified with
sulfo group (counter ion: Ca2+)
Column Housing: SUS-316 (stainless steel)
Screw Type: Internally-threaded type No. 10-32 UNF
Shipping Solvent: Water
4. Usable Conditions
Product Name
Flow Rate (mL/min)
Maximum Pressure
(MPa) (Per Column)
Temperature (°C)
Recommended
Maximum
Recommended
Maximum
EP SC1011-7F
0.5
1.2
2.5
70 –85*
95
SUGAR SC-G 6B
-
-
-
* 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) Aqueous solutions of calcium nitrate or calcium sulfate salts can be used instead of water. Keep the
pH of solution between 3 and 7. For the analysis of samples containing large amount of heavy metals,
addition of 10 - 50 μg/mL Ca-EDTA will prevent the deterioration of column performance.
(3) Up to 20 % (v/v) acetonitrile or ethanol is usable.
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.
• Do not use salts other than calcium salts. Use of other salts may replace the calcium
ion attached to the sulfo functional group with other cation and may deteriorate the
column.
• When using a mixture of salt solution 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 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.

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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 20 μL per column.
(4) When analyzing an acidic or a basic sample, make sure to neutralize the sample prior to the
injection.
(5) When a sample contains cations other than calcium 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. If guard column is used, position the guard column in front (before the inlet) of the analytical
column.
(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.

Operation Manual Shodex SUGAR EP SC1011-7F Version 210415E
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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.
•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 50°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.
If a guard column is used with the analytical column, first remove the guard column and check the
performance of the analytical column alone. If the problem is solved, most likely the cause is from the
guard column. In this case, clean and/or regenerate the guard column.
If the problem is not solved by removing the guard column, clean and/or regenerate both guard and
analytical columns. Make sure to clean and/or regenerate the guard and the analytical columns
separately. 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.

Operation Manual Shodex SUGAR EP SC1011-7F Version 210415E
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<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 calcium ion in the sample may replace the calcium ion attached to the
sulfo functional group. This can change the separation patterns and peak shapes. Following
regeneration procedure reattaches the calcium ions and may resume the column performance:
Set the column temperature at 50°C and the flow rateat 0.5 mL/min. Introduce 50 mL of 0.1 M aqueous
calcium 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)
(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.
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.

Operation Manual Shodex SUGAR EP SC1011-7F Version 210415E
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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.