Monosaccharides and Oligosaccharides
Elution volumes of saccharides for seven different columns, SUGAR SP0810, SC1011, KS-801, SZ5532, SC1211, RSpak DC-613, Asahipak NH2P-50 4E and GS-220 HQ are shown in the table. As only the peak top position can be obtained from the table, if you want to to see the chromatogram with actual peak width, please contact us.
| Substances | Elution Volume (mL) | ||||||||
|---|---|---|---|---|---|---|---|---|---|
| SUGAR | RSpak | Asahipak | |||||||
| SP0810 | SC1011 | KS-801 | SZ5532 | SC1211 | DC-613 | NH2P-50 4E | GS-220 HQ x 2 | ||
| Pb2+ | Ca2+ | Na+ | Zn2+ | Ca2+ | Na+ | ||||
| N-Acetyl-alpha -D-glucosamine |
8.86 | 7.75 | 6.68 | 2peaks | 4.10 | 2peaks | 6.66 | ||
| D(+)-Arabinose | 10.42 | 8.91 | 8.21 | 5.11 | 5.56 | 5.75 | 6.18 | ||
| D-Arabitol | 15.86 | 11.33 | 7.63 | 7.27 | 8.16 | 5.81 | 6.29 | ||
| Aspartame | – | – | – | – | 34.02 | – | – | ||
| 2-Deoxy-D-glucose | 8.83 | 7.58 | 7.15 | 4.34 | 4.02 | 4.43 | 6.02 | ||
| Difructose anhydride III | 7.07 | 6.30 | 5.81 | 4.30 | * | 4.51 | 7.77 | ||
| Dulcitol | 20.18 | 12.76 | 7.40 | 9.46 | 11.28 | 7.33 | 7.45 | ||
| meso-Erythritol | 12.70 | 10.09 | 7.86 | 5.73 | 6.27 | 4.84 | 5.43 | 17.38 | |
| Ethanol | 11.13 | 11.33 | 10.09 | – | 4.27 | – | – | ||
| 1-Fructofuranosyl -D-nystose |
6.05 | 5.27 | 4.76 | 31.43 | * | – | – | 14.24 | |
| D(-)-Fructose | 11.05 | 8.85 | 7.71 | 5.37 | 5.90 | 6.19 | 6.75 | 17.05 | |
| D(+)-Fucose | 10.48 | 8.84 | 8.09 | 4.50 | 4.96 | 4.81 | 5.43 | ||
| D(+)-Galactose | 9.74 | 7.98 | 7.58 | 6.46 | 4.98 | 7.28 | 8.10 | 16.60 | |
| 4′-Galactosyllactose | 7.42 | 6.02 | 5.40 | 19.02 | * | 21.24 | 21.66 | ||
| alpha-D-Galacturonic acid | – | 6.28 | 4.36 | – | 5.63 | – | – | ||
| Gentiobiose | 7.22 | 6.08 | 5.75 | 10.50 | * | 13.18 | 16.36 | ||
| Glucose | 8.63 | 7.30 | 7.17 | 5.87 | 4.76 | 6.83 | 8.61 | 16.75 | |
| Glycerol | 4.26 | 17.83 | |||||||
| Glycyrrhizin | – | – | – | – | 2.71 | 2.00 | – | ||
| myo-Inositol | 12.77 | 8.86 | 7.99 | 12.63 | 7.87 | 15.80 | 9.96 | ||
| Isomaltose | 7.68 | 6.26 | 5.95 | 10.57 | * | 13.82 | 15.18 | 15.57 | |
| Isomaltotriose | 7.09 | 5.75 | 5.34 | 21.17 | * | 32.02 | 27.55 | 14.72 | |
| 1-Kestose | 6.79 | 5.75 | 5.26 | 13.09 | * | – | 20.11 | 15.31 | |
| Kojibiose | 7.56 | 6.21 | 5.88 | 9.65 | * | 11.47 | 14.82 | ||
| Lactitol | 13.27 | 8.09 | 6.13 | 16.35 | 6.67 | 14.04 | 11.82 | 15.50 | |
| Lactose | 8.05 | 6.51 | 5.99 | 10.12 | 4.07 | 11.69 | 13.27 | 15.71 | |
| Lactosylfructoside | 7.12 | 5.88 | 5.29 | 14.69 | * | 15.75 | 18.98 | ||
| Lactulose | 9.13 | 6.99 | 6.19 | 9.16 | 4.65 | 10.80 | 10.72 | ||
| Maltitol | 12.23 | 8.26 | 6.03 | 13.04 | 6.77 | 11.81 | 11.82 | 15.82 | |
| Maltoheptaose | – | 14.82 | |||||||
| Maltohexaose | – | 14.56 | |||||||
| Maltopentaose | – | 14.31 | |||||||
| Maltose | 7.85 | 6.34 | 5.94 | 8.67 | * | 10.61 | 14.24 | 16.11 | |
| Maltotriose | 7.48 | 5.89 | 5.38 | 13.79 | * | 17.88 | 24.96 | 15.56 | |
| Mannitol | 15.80 | 11.10 | 7.23 | 8.75 | 9.03 | 6.84 | 7.39 | ||
| D-Mannose | 10.72 | 8.17 | 7.64 | 5.83 | 5.01 | 6.72 | 7.84 | ||
| D(+)-Melezitose | 6.94 | 5.79 | 5.24 | 13.60 | * | 14.68 | 19.27 | ||
| Melibiose | 8.16 | 6.45 | 5.98 | 11.69 | 4.23 | 14.83 | 14.70 | ||
| Methyl-alpha -D-mannopyranoside |
11.13 | 8.87 | 7.78 | 3.99 | 4.39 | 4.15 | 4.71 | ||
| Nystose | 6.38 | 5.45 | 4.93 | 20.05 | * | – | 31.90 | 14.72 | |
| Palatinit | 2peaks | 2peaks | 5.90 | 2peaks | 2peaks | 2peaks | 12.73 | ||
| Palatinose | 7.84 | 6.45 | 5.89 | 8.08 | 3.99 | 9.81 | 12.12 | ||
| Panose | 7.14 | 5.78 | 5.32 | 16.87 | * | 23.14 | 25.60 | ||
| L-Phenylalanine | – | – | – | – | 31.58 | – | – | ||
| D(+)-Raffinose | 7.14 | 5.78 | 5.29 | 16.36 | * | 19.11 | 20.25 | 15.08 | |
| D(+)-Rhamnose | 9.77 | 8.23 | 7.37 | 3.93 | 4.43 | 4.09 | 5.52 | ||
| D(-)-Ribose | 19.35 | 13.66 | 9.04 | 4.82 | 8.64 | 5.30 | 5.45 | ||
| Rutinose | 7.81 | 6.49 | 5.80 | 6.65 | * | 7.21 | 10.87 | ||
| Saccharin sodium | 6.96 | 5.44 | 4.33 | 6.77 | * | 2.04 | 5.68 | ||
| D(-)-Sorbitol | 21.61 | 13.31 | 7.42 | 9.79 | 11.88 | 7.27 | 7.09 | 16.60 | |
| D(+)-Sorbose | 9.67 | 8.03 | 7.38 | 5.12 | 4.92 | 5.91 | 7.35 | ||
| Stachyose | 6.82 | 5.57 | 4.97 | – | * | – | 36.22 | 14.33 | |
| Stevioside | – | – | – | 4.14 | * | 4.44 | 6.07 | ||
| Sucrose | 7.54 | 6.29 | 5.87 | 7.91 | * | 8.68 | 11.87 | 16.26 | |
| alpha-D-Talose | 21.33 | 12.59 | 8.76 | 5.69 | 8.51 | 6.32 | 6.47 | ||
| Theanderose | 2peaks | 2peaks | 2peaks | 2peaks | * | 2peaks | |||
| Trehalose | 7.62 | 6.27 | 5.78 | 10.85 | * | 11.49 | 13.25 | ||
| Trehalulose | 8.92 | 6.95 | 6.10 | 9.54 | 4.78 | 11.38 | 11.68 | ||
| Xylitol | 19.87 | 13.14 | 7.94 | 7.77 | 10.16 | 6.19 | 6.10 | 16.88 | |
| Xylobiose | 8.16 | 6.68 | 6.40 | 5.65 | * | 6.71 | 9.05 | ||
| D(+)-Xylose | 9.21 | 7.90 | 7.71 | 4.55 | 4.48 | 5.21 | 6.58 | 17.46 | |
| D-Xylulose | 10.64 | 9.02 | 8.04 | 4.06 | 5.07 | 4.56 | 5.41 | ||
(-)–>cannot be detected
(*)–>cannot be separated from solvent peak
Columns : Shodex SUGAR SP0810, SC1011, KS-801 (8.0mmID*300mm each) Eluent : H2O Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 80deg-C Column : Shodex SUGAR SZ5532 (6.0mmID*150mm) Eluent : H2O/CH3CN=25/75 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 60deg-C Column : Shodex SUGAR SC1211 (6.0mmID*250mm) Eluent : H2O/CH3CN=65/35 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 70deg-C Column : Shodex RSpak DC-613 (6.0mmID*150mm) Eluent : H2O/CH3CN=25/75 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 70deg-C Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : H2O/CH3CN=25/75 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C Column : Shodex Asahipak GS-220 HQ (7.5mmID*300mm) x 2 Eluent : H2O Flow rate : 0.5mL/min Detector : Shodex RI Column temp. : 60deg-C
After reduction of the sugar and saccharide, the terminal groups may have various types of cyclic forms in addition to the open-chain form. In such cases, these saccharides exist as two diastereomers because the carbon atom of the terminal reducing group is asymmetric; these diastereomers are called anomers (alpha- and beta-anomers respectively). Under certain conditions where the rate of conversion of such diastereomers is low, alpha-anomer and beta-anomer are separated as they pass through the column. This causes a undesirable splitting or broadening of the peak. For the analysis of sugars, therefore, it is necessary to prevent the anomer separation. Below are possible methods for controlling this separation:
1) It has been reported that the anomer separation does not take place at high temperature. Therefore, when using SUGAR series (SP0810, SC1011, SC1211 or KS-800), set up the column temperature at 70 to 80deg-C.
2) It has been reported that the anomer separation does not take place under strong alkaline conditions. Polymer-base columns such as NH2P allow the separation of saccharides without causing the anomer separation as the column can be used under alkaline environment.
3) For other columns, refer to the conditions described in the chromatograms.
It has been reported that the anomer separation does not take place at high temperature. Therefore, when using SUGAR series (SP0810, SC1011, KS-801, SZ5532 or SC1211), set up the column temperature at 70-80 deg-C.
Please refer to Prevention of Anomer Separation.
Sample : 0.5% each, 10micro-L
Glucose, Mannose, Xylose, Galactose, Arabinose
Column : Shodex SUGAR SC1011 (8.0mmID*300mm) Eluent : H2O Flow rate : 0.7mL/min Detector : Shodex RI Column temp. : 29deg-C, 70deg-C
It has been reported that the anomer separation does not take place under strong alkaline conditions. Amino columns such as Asahipak NH2P-50 4E allow the separation of saccharides without causing the anomer separation as they can be used under alkaline conditions at room temperature.(Please refer to Prevention of Anomer Separation.)
Amide columns are also used for the separation of saccharides. However, acryl-amide goups are not alkaline, it is necessary to perform the separation at elevated temperature to prevent anomer separation.
Sample : 5mg/mL each, 10micro-L
1. Fructose
2. Glucose
3. Sucrose
4. Maltose
Column : Shodex Asahipak NH2P-50 4E, Amide column from other manufacturer (4.6mmID*250mm each) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
Comparison of columns with different counter ions, Pb2+ (SP0810), Ca2+ (SC1011) and Na+ (KS-801) is shown. For the separation of monosaccharides(glucose and fructose), the separation by KS-801 is not sufficient because the separation mode of KS-801 is mainly GFC mode and their molecular weights are too close to be separated by GFC mode. When SP0810 and SC1011 are used, monosaccharides can be separated well because ligand exchange mode works for these columns.
For the separation of oligosaccharides(maltotriose and maltose), KS-801 is the best suited. The difference is caused by the difference of peak width. In case of SC1011 and SP0810, ligand exchange mode works and anomers are separated by the mode. And, as the consequence, the peak width become larger. Since the ligand exchange mode of SP0810 is stronger than that of SC1011, the separation of oligosaccharides by SP0810 is the worst.
As the conclusion, we can say that monosaccharides can be separated well by ligand exchange mode and oligosaccharides can be separated well by GFC mode.
Sugar alcohol(sorbitol) is separated by the ligand exchange mode. In case of SP0810, the elution of sorbitol is too slow because ligand exchange mode is working too strongly.
Sample :
1. Pullulan
2. Maltotriose
3. Maltose
4. Glucose
5. Sorbitol
6. Fructose
7. Glycerol
8. Ethanol
Columns : Shodex SUGAR SP0810, SC1011, KS-801 (8.0mmID*300mm each) Eluent : H2O Flow rate : 0.6mL/min Detector : Shodex RI Column temp. : 80deg-C
When Asahipak NH2P-50 columns are used, the retention time of sugar alcohols as well as saccharides becomes longer as the acetonitrile concentration in the eluent becomes higher.
Sample :
Arabitol
Fructose
Galactose
Glucose
Glycerol
Lactose
Maltose
Mannitol
Mannose
Raffinose
Rhamnose
Ribitol
Sorbitol
Sucrose
Xylitol
Xylose
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
Same as packing materials for silica based amino columns, the packing material for Asahipak NH2P-50 columns has amino groups which are bonded to weak anion exchange resins.
The amino groups exist in the equilibrium of free type and acid type. (Please refer to the right figure.) The state of the equilibrium(free base ratio) depends on pH and ionic composition of the eluent.
To investigate the effect of free base ratio, NH2P-50 4E columns are equilibrated with ammonium acetate solvents of different pH to prepare columns of different free base ratio. Using the columns, analyses were performed under the same conditions. The result shows that the elution times became shorter and peak shapes became sharper as free base ratio increased.
.
| pH | 2.84 | 8.50 | 9.30 | Plate number vs free base ratio |
|---|---|---|---|---|
| Free base ratio |
32% | 82% | 93% | |
| Chromato- gram |
 |
 |
 |
 |
Sample : 1. Fructose, 2. Glucose, 3. Sucrose, 4. Maltose
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
The separation of saccharides using Asahipak NH2P-50 column can be controlled by changing mixing ratio of acetonitrile and water in the eluent. Since the NH2P-50 column is free from non-specific adsorption, sharp and symmetrical peaks can be obtained for most of saccharides.
When the NH2P-50 column is used, as same as when silica-based amino columns are used, the retention time of saccharides becomes longer as the acetonitrile concentration in the eluent becomes higher. The elution order of saccharides using the NH2P-50 columns is almost as same as that using silica-based amino columns though there are some exception such as the order between galactose and glucose and between lactose and maltose.
Sample :
1. Rhamnose
2. Xylose
3. Fructose
4. Glucose
5. Sucrose
6. Maltose
7. Lactose
8. Galactose
9. Raffinose
10. Mannose
Columns : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) , Silica based amino column(other manufacturer) Eluent : CH3CN/H2O Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
The base material of the packing material of SUGAR SZ5532 is the gel developed for adsorption & partition mode. It is highly cross-linked and its swelling and shrinkage is very small. The counter ion is Zn2+. Zn2+ forms a complex with saccharides but, when the counter ion is Zn2+, saccharides are not so strongly reteined by ligand exchange mode. And, when organic solvents are added in the eluent, saccharides and sugar alcohols are reteined by adsorption & partition mode and their elution volumes become larger by inceasing organic solvent concentration.
Sample :
Rhamnose
meso-Erythritol
Glucose
Xylose
Sucrose
Sorbitol
Isomaltose
myo-Inositol
Maltotriose
Raffinose
Column : Shodex SUGAR SZ5532 (6.0mmID*150mm) Eluent : CH3CN/H2O Flow rate : 0.6mL/min Detector : Shodex RI Column temp. : 60deg-C
When Asahipak NH2P-50 column is used, plate number of saccharides becomes higher as the acetonitrile concentration in the eluent becomes higher.
When silica-based amino columns are used, plate number and detection sensitivity of saccharides are different according to the type of saccharides, and, the plate number of some specific saccharide such as galactose is extremely low.
Please refer to Comparison of NH2P and Silica-based Amino Column (2).
Sample :
1. Rhamnose
2. Xylose
3. Fructose
4. Glucose
5. Sucrose
6. Maltose
7. Lactose
8. Galactose
9. Raffinose
10. Mannose
Columns : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm), Silica based amino column (other manufacturer) Eluent : CH3CN/H2O Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
Testing procedure
(1)Testing(63hrs) Eluent : 0.01M H2SO4(pH2) Flow rate : 0.1mL/min Column temp. : Room temp. |
(2)Replacement(1hr) Eluent : H2O Flow rate : 0.5mL/min Column temp. : Room temp. |
(3)Equilibrium(1hr) Eluent : 0.1M NaOH Flow rate : 0.5mL/min Column temp. : Room temp. |
Comparison between before and after testing
| Sample | Before testing | After testing | ||
|---|---|---|---|---|
| Elution volume (mL) |
Plate number | Elution volume (mL) |
Plate number | |
| 1.Fructose | 6.73 | 9600 | 6.70 | 8400 |
| 2.Glucose | 8.59 | 7900 | 8.58 | 8500 |
| 3.Sucrose | 11.88 | 9600 | 11.93 | 9300 |
| 4.Maltose | 14.24 | 8500 | 13.90 | 8600 |
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
Testing procedure
(1)Equilibrium(1hr) Conditions before testing Eluent : 100mM Ammonium acetate(pH9.3) Flow rate : 0.1mL/min Column temp. : Room temp. |
(2)Analysis |
(3)Testing(160hrs) Eluent : 5mM NaOH(pH11.4)/CH3CN=25/75 Flow rate : 0.1mL/min Column temp. : 30deg-C |
(4)Replacement(0.5hr) Eluent : H2O Flow rate : 0.5mL/min Column temp. : Room temp. |
(5)Replacement(0.5hr) Eluent : H2O Flow rate : 0.5mL/min Column temp. : Room temp. |
(6)Analysis |
Comparison between before and after testing
| Sample | Before testing | After testing | ||
|---|---|---|---|---|
| Elution volume (mL) |
Plate number | Elution volume (mL) |
Plate number | |
| 1.Fructose | 7.52 | 7100 | 7.62 | 8100 |
| 2.Glucose | 9.63 | 6100 | 9.68 | 7700 |
| 3.Sucrose | 13.41 | 8400 | 13.40 | 9700 |
| 4.Maltose | 16.16 | 7200 | 16.08 | 7400 |
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
Since the swelling and shrinking of the packing material of Asahipak NH2P-50 columns is considerably small even when the eluent composition is changed, any mixing ratio of acetonitrile and water can be used with the NH2P-50 columns.
Comparison between before and after testing
| Sample | Before testing | After testing | ||
|---|---|---|---|---|
| Elution volume (mL) |
Plate number | Elution volume (mL) |
Plate number | |
| 1. Fructose | 7.61 | 11600 | 7.56 | 9800 |
| 2. Glucose | 9.80 | 7500 | 9.72 | 6200 |
| 3. Sucrose | 13.86 | 11700 | 13.75 | 9300 |
| 4. Maltose | 16.78 | 8300 | 16.66 | 6700 |
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
When a sample containing polysaccharides is injected into amino columns, polysaccharides are adsorbed by the packing material and it causes poor separation and/or broad peaks.
The figures below show the result when polysaccharide (pullulan, MW: 1,200,000) is injected into the Asahipak NH2P-50 4E column:
1) Figure (A) shows that there is little difference in elution time.
2) Figure (B) shows that the plate number decreses as the amount of adsorption increases.
3) Figure (C) shows the tailing in chromatogram after the adsorption.
Since it is not so easy to eliminate polysaccharides which are adorbed by the packing material, it is neccesary to pre-treat the sample and eliminate polysaccharides before injection.
Sample : Maltose, Lactose, Sucrose, Glucose, Fructose
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/10mM *Tetrapropylammonium buffer(pH10.0)=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
*How to make Tetrapropylammonium buffer
Please add acetic acid to 10mM Tetrapropylammonium hydroxide and adjust pH10.
Effect of sample solvent composition when Asahipak NH2P-50 is used is shown. When making sample solution to be injected, sample should be dissolved in a solvent whose composition is as close as the eluent, preferably in the eluent. When 75% acetonitrile solvent is used for the eluent and a sample cannot be dissolved in it, first, the sample should be dissolved in a solvent which dissolves the sample, and then, adjust the solvent composition to be acetonitrile aqueous solution of 50% or more.
Sample : 1mg/mL each, 20micro-L
1. meso-Erythritol 2. Xylitol 3. Fructose 4. Glucose 5. Sucrose 6. Lactose 7. Maltose
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 25deg-C
Effect of sample injection volume when Asahipak NH2P-50 4E is used is shown. When making sample solution to be injected, sample should be dissolved in a solvent whose composition is as close as the eluent, preferably in the eluent. When 75% acetonitrile aqueous solution is used for the eluent and a sample can be dissolved only in aqueous solution (or 50v/v% ethanol, etc.), injection volume should be as small as possible.
Sample : 20micro-g each in purified water
1. Fructose 2. Glucose 3. Sucrose 4. Lactose 5. Maltose
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 25deg-C
Effect of flow rate when analyzing saccharides using SUGAR SC1011 (a column for saccharide analysis) is shown here.
It is confirmed that the theoretical plate number and resolution are improved by decreasing the flow rate.
Sample : 0.5%each, 5micro-L
1. Isomaltotriose
2. Sucrose
3. Glucose
4. Lactitol
5. Fructose
6. meso-Erythritol
7. Mannitol
8. Dulcitol
9. Xylitol
Column : Shodex SUGAR SC1011 (8.0mmID*300mm) Eluent : H2O Detector : Shodex RI Column temp. : 80deg-C
Effect of temperature when analyzing saccharides using SUGAR SC1011 (a column for saccharides analysis) is shown here. Saccharides having carbonyl or aldehyde group may have anomers. It is possible to prevent anomer separation by analyzing at elavated temperature. It is also possible to have higher theoretical plate number and resolution by it. Analyzing at elevated temperature around 80 deg-C is recommended when using columns such as SP0810, SC1011, SC1821, KS-801 and KS-802.
Sample : 0.5%each, 5micro-L
1. Isomaltotriose
2. Sucrose
3. Glucose
4. Lactitol
5. Fructose
6. meso-Erythritol
7. Mannitol
8. Dulcitol
9. Xylitol
Column : Shodex SUGAR SC1011 (8.0mmID*300mm) Eluent : H2O Flow rate : 0.6mL/min Detector : Shodex RI
Raffinose was separated using SUGAR SH1011. Since raffinose can be easily hydrolyzed by acids, it should be analyzed at ordinary temperature. The situation is the same for other saccharides such as sucrose.
Sample : Raffinose
Column : Shodex SUGAR SH1011 (8.0mmID*300mm) Eluent : 0.005M H2SO4 aq. Flow rate : 0.6mL/min Detector : Shodex RI Column temp. : (above); 60deg-C, (below); 25deg-C
Sample : 20micro-L
|
Correlation Coefficient
(0.2 to 200micro-g) |
|
| Glucose | 0.9998 |
| Sorbitol | 0.9999 |
| Fructose | 0.9998 |
| Sucrose | 0.9999 |
| Lactose | 0.9997 |
| Maltose | 0.9998 |
| meso-Erythritol | 0.9999 |
| Xylitol | 0.9999 |
| Mannitol | 0.9999 |
| Maltitol | 0.9997 |
Column : Shodex Asahipak NH2P-50 4E (4.6mmID*250mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 25deg-C
Sample : 10micro-L
|
Correlation Coefficient
(1 to 200micro-g) |
|
| Glucose | 1.0000 |
| Sorbitol | 0.9999 |
| Sucrose | 1.0000 |
| Maltose | 1.0000 |
| Isomaltose | 0.9999 |
| Maltotriose | 1.0000 |
| Raffinose | 1.0000 |
| meso-Erythritol | 1.0000 |
| myo-Inositol | 0.9999 |
| Stevioside | 1.0000 |
Column : Shodex SUGAR SZ5532 (6.0mmID*150mm) Eluent : CH3CN/H2O=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 60deg-C