Shodex SUGAR KS-800 Series Manuel utilisateur
Operation Manual Shodex SUGAR KS-800 series Version 2020
Operation Manual
Shodex™ SUGAR™KS-800 series
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:
+49 (0)89 / 93 99 62 37
Quotes and Orders:
+49 (0)89 / 93 99 62 34
Operation Manual Shodex SUGAR KS-800 series Version 2020
2
Operation Manual
Shodex™ SUGAR™ KS-800 series
1. Introduction
Shodex SUGAR KS-800 series are packed with ion exchange resin gels of sulfonated styrene-
divinylbenzene copolymer. They are designed for high-speed and high-performance liquid
chromatography. They are capable of separating carbohydrates such as monosaccharides,
disaccharides, oligosaccharides and sugar alcohols, as well as non-ionic water-soluble substance, with
distilled water or 0.001 M sodium hydroxide aqueous solution used as the mobile phase.
The columns demonstrate an excellent performance, particularly, in high-speed analysis of saccharides
present in living bodies, food, pharmaceuticals and industrial wastewater.
The columns fulfill USP L22 and L58 requirements.
2. Specifications
Product
code
Product name
Counter
ion
Particle
size (µm)
Exclusion limit
(Pullulan)
Plate
number
(TP/per
column)
Size (mm)
Shipping
solvent
F6378010
SUGAR KS-801
Sulfo (Na+)
6
1,000
≥17,000
8.0 x 300
H2O
F6378020
SUGAR KS-802
Sulfo (Na+)
6
10,000
≥17,000
8.0 x 300
H2O
F6378025
SUGAR KS-803
Sulfo (Na+)
6
50,000
≥17,000
8.0 x 300
H2O
F6378035
SUGAR KS-804
Sulfo (Na+)
7
400,000
≥17,000
8.0 x 300
H2O
F6378050
SUGAR KS-805*
Sulfo (Na+)
17
5,000,000
≥9,000
8.0 x 300
H2O
F6378060
SUGAR KS-806*
Sulfo (Na+)
17
*(50,000,000)
≥9,000
8.0 x 300
H2O
F6700020
SUGAR KS-G 6B
Sulfo (Na+)
10
-
(guard
column)
6.0 x 50
H2O
* columns marked with asterisk are available until end of 2021
Packing material: Styrene divinylbenzene copolymer
Sulfonated strong cation exchange resin gel with sodium ions
Column material: SUS 316 (stainless steel)
Screw type: Internally-threaded type No. 10-32 UNF
*() Estimated value
Usable conditions
Mobile phase
Column temperature
Flow rate
Column pressure
H2O or 0.001 M NaOH aq.
Max. 20% ethanol
20 - 29°C
0.5 mL/min
Max. 5 MPa
30 - 49°C
1.0 mL/min
50 - max. 85°C
Max. 1.5 mL/min
The columns can also be used in row.
Operation Manual Shodex SUGAR KS-800 series Version 2020
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3. Mobile phase
1) Use distilled water or 0.001 M sodium hydroxide aqueous solution as the mobile phase.
2) Remove extraneous substances and insoluble from the mobile phase by passing it through a 0.45 µm
filter into a bottle and thoroughly degas it by, for instance, heating it about 60°C in a hot water bath
and shaking in an ultrasonic washing vessel with the ambient air pressure being simultaneously
reduced by an aspirator. Use of solvent degassing device will facilitate the degassing work.
Note! Adsorbed sometimes by the packing, molecules of highly hydrophobic specimens are too slow
to flow out of the column. In such a case, add ethanol in an amount of 20% maximum to the mobile
phase to reduce the absorptivity.
4. Mounting and start-up
1) Before mounting the column on the liquid chromatograph, replace the solventin the chromatograph
with the mobile phase.
Caution! In replacing a water-insoluble organic solvent, replace it first with a water-soluble organic
solvent, such as acetone or ethanol, and then replace the water-soluble solvent with the mobile phase.
2) Set the flow rate at 1.0 mL/min.
3) Connect the column to the chromatograph in such a way that the arrow mark on the column will
point to the direction of the flow.
Caution! Do not let the air into the column during the connection.
4) Heat the column to 40 - 50°C and start the pump.
Caution!
i) Keep the column pressure below 5 MPa per column; otherwise, the column will be ruined.
ii) The maximum working flow rate depends on the working temperature, as follows:
Working temperature Maximum working flow rate
50 - 85°C 1.5 mL/min
30 - 49°C 1.0 mL/min
29°C or below 0.5 mL/min
iii) Do not abruptly change the column pressure or flow rate.
iv) Do not heat the column higher than 85°C.
Precautions to be taken when using KS-806 and KS-805 TE080820
When Shodex SUGAR KS-806 or KS-805 is used for the first time or after they have been stored for a
long period of time, there are cases in which white cloudy liquid comes out from the column.
This does not have an effect in the performance (characteristics) of the column, but it could cause the
contamination of the detector. Therefore, use the column in accordance with the following
procedures:
1) Remove the plugs from both ends of the column and connect it to the chromatograph. However, do
not connect the column to the detector!
2) Pass 20 to 30 mL of the eluent in use through the column at 1.0 mL/min. The liquid discharged from
the column should be disposed of.
3) Connect the column to the detector.
4) Start measurement when the baseline has stabilized.
Operation Manual Shodex SUGAR KS-800 series Version 2020
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5. Pre-treatment of specimen
1) When the specimen is solid, dissolve it, using some of the degassed mobile phase.
Note: Do not use any other solvent; otherwise, the solute sometimes settles in the mobile phase to
plug the column.
2) When the specimen is liquid and contains an organicsolvent such as alcohol, dilute it with the mobile
phase so as to reduce concentration of the organic solvent to 20% maximum; otherwise, the column
performance will deteriorate.
3) Pass the specimen through a 0.45 µm filter to remove extraneous substances and insoluble. Use of
the disposable filter unit is recommended.
4) When the specimen contains proteins, add 2 or 3 drops of distilled water in which sulfosalicylic acid
is dissolved at the rate of 20g per deciliter, to 5 mL of the specimen, stir and pass it through a 0.45 µm
filter to remove the proteins.
6. Dismounting and storage
1) Set the flow rate at 0.2 mL/min, turn off the heater and let the column stand as is until it cools down
to room temperature.
2) Stop the pump and dismount the column.
3) Cap both ends of the column, place it back in the same case in which it was delivered from the
manufacturer, and store the case in a place where the temperature does not vary more than ±5°C from
the ambient temperature.
Caution!
i) Do not remove the end fittings of the column; otherwise, its performance will deteriorate, or the
column may be rendered unserviceable.
ii) Do not let the in-column solvent freeze during storage; otherwise, the column will swell and become
unserviceable.
iii) Storage at a normal temperature; otherwise, high temperature will also cause the column to swell
and become unserviceable.
7. Band broadening
Adsorption of organics and metal ions by the packing sometimes causes the bands of fructose and
glucose to be broadened.
If such adsorption occurs, pass 0.2 M NaOH aqueous solution through the column at a rate of
0.5 mL/min to desorb them and completely replace the solution with distilled water. If, however, the
band broadening is moderate, injection of 40 µL of 1 M NaOH aqueous solution into the column will
suffice to carry out the desorption.
8. Guard column
Use of pre-column, SUGAR KS-G, is recommended immediately before the column to protect the
packing from contamination with pollutants or readily absorbable substances, such as metal ions and
amino compounds.
Operation Manual Shodex SUGAR KS-800 series Version 2020
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9. Performance test
The performance of the column can be checked by calculating the number of theoretical plates under
the following conditions:
1) Specimen and injection amount: KS-801 1.0% ethylene glycol aqueous solution 5 µL
KS-802 2.5% ethylene glycol aqueous solution 5µL
KS-803 0.5% ethylene glycol aqueous solution 15 µL
KS-804 0.8% glucose aqueous solution 5 µL
KS-805 0.4% glucose aqueous solution 5 µL
KS-806 0.8% glucose aqueous solution 5 µL
2) Mobile phase: distilled water
3) Flow rate: 1.0 mL/min
4) Column temperature: 50°C
5) Detector: High-sensitivity refractive index detector such as Shodex RI
6) Calculation formula: NTP = 5.54 x (Rt/W)2
where NTP: Number of theoretical plate, Rt: Retention time, W: Peak half width
Operation Manual Shodex SUGAR KS-800 series Version 2020
6
10. Warranty (Version 4)
1) Showa Denko K.K. warrants that the ShodexTM 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 product, 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 analysis 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 product 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.
Ce manuel convient aux modèles suivants
14
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