Polysaccharides
SUGAR KS-800 series are suited to the analysis of polysaccharides. Calibration curves for KS-800 series are shown here. Besides KS-800 series, OHpak SB-800 HQ series and Asahipak GS-HQ series also can be used for the analysis of polysaccharides.
Sample : Pullulan
Columns : Shodex SUGAR KS-800 (8.0mmID*300mm each) Eluent : H2O Detector : Shodex RI Column temp. : 80deg-C
SUGAR KS-800 series are suited to the analysis of polysaccharides. Calibration curves for the combination of KS-800 series columns of different exclusion limit are shown here.
Sample : Pullulan
Columns : Shodex SUGAR KS-802, 803, 804 (8.0mmID*300mm each) Eluent : H2O Detector : Shodex RI Column temp. : 80deg-C
Carboxymethylcellulose was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Carboxymethylcellulose Medium viscosity (CMC),
200micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaCl aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Carboxymethylcellulose was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Carboxymethylcellulose 145cp (CMC), 200micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaCl aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Hyaluonic acid was analyzed using OHpak SB-805 HQ (a column for GFC separation).
Sample : Hyaluronic acid
Columns : Shodex OHpak SB-805 HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
Hyaluronic acid was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Hyaluronic acid, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
The analytical results of hyaluronic acid are shown. In the analysis with OHpak SB-806 HQ (exclusion limit : 2.0×107g/mol), although HA 30 is larger than HA 20 by 106 in MW, both of them elute around exclusion limit. On the other hand, SUGAR KS-807 (exclusion limit : 2.0×108g/mol) anables us to distinguish them clearly.
Sample : Hyaluronic acid, 100micro-L
0.028% HA 30 (MW:3.0x 106)
0.026% HA 20 (MW:2.0×106)
0.027% HA 12 (MW:1.2×106)
(offered by Shiseido Company, Limited)
Sample : Hyaluronic acid, 100micro-L
0.028% HA 30 (MW:3.0×106)
0.026% HA 20 (MW:2.0×106)
0.027% HA 12 (MW:1.2×106)
(offered by Shiseido Company, Limited)
(above) Column : Shodex OHpak SB-806 HQ (8.0mmID*300mm) Eluent : 0.2M NaCl aq. Flow rate : 0.5mL/min Detector : Shodex RI & MALS(Multi angle laser light scattering detector) Column temp. : Room temp. (below) Column : Shodex SUGAR KS-807 (8.0mmID*300mm) Eluent : 0.2M NaCl aq. Flow rate : 0.5mL/min Detector : Shodex RI & MALS(Multi angle laser light scattering detector) Column temp. : Room temp.
Three kinds of hyaluronic acid with MW of one million or more was analyzed with OHpak SB-807 HQ. Since estimated exclusion limit of SB-807 HQ is 5.0 x 108g/mol, Hyaluronic acids elute with size exclusion mode.
Sample : Hyaluronic acid, 100micro-L
Column : Shodex OHpak SB-807 HQ (8.0mmID*300mm) Eluent : 0.2M NaCl aq. Flow rate : 0.5mL/min Detector : Shodex RI Column temp. : 30deg-C
For the analysis of polysaccharides such as starch, which does not dissolve easily in water, samples can be analyzed using OHpak SB-806 HQ by GFC mode with an aqueous eluent after samples had been dissolved in DMSO. However, this method was unsatisfactory as undissolved components were left undetected.
Please refer to Corn Starch (2) (DMSO/DMF Eluent).
Sample : Starch in DMSO
1. Corn starch
2. Cationized corn starch N=0.13
3. Cationized corn starch N=0.24
4. Cationized corn starch N=0.35
Columns : Shodex OHpak SB-G (6.0mmID*50mm) + SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M LiBr aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 30deg-C
For the analysis of polysaccharides such as starch, which does not dissolve easily in water, samples used to be dissolved in DMSO before doing the analysis by GFC mode with an aqueous an eluent. However, this method was unsatisfactory as undissolved components were left undetected. In contrast with the above, if a mixed eluent of DMSO/DMF containing LiBr is used as an eluent, the solubility of starch in the eluent is increased allowing a more accurate GFC analysis. There are some cases in which starch has ionic functional groups such as carboxyl groups. An addition of LiBr to the eluent allows GFC analysis without interference from the ionic groups. The sample used in this analysis example was prepared by adding DMSO to starch and then heating the mixture to 120 deg.C to dissolve the starch using OHpak SB-806 HQ (a column for GFC separation).
Please refer to Corn Starch (1) (Aqueous Eluent).
Sample : Starch
Corn starch in DMSO
Column : Shodex OHpak SB-806 HQ (8.0mmID*300mm)
Eluent : DMSO/DMF=75/25
5mM LiBr in DMSO/DMF=75/25
Flow rate : 0.25mL/min
Detector : Shodex RI
Column temp. : 40deg-C
Chondroitin sulfate A was analyzed using SUGAR KS-804 (a column for saccharides analysis).
Sample : Chondroitin sulfate A
Columns : Shodex SUGAR KS-G (6.0mmID*50mm) + KS-804 (8.0mmID*300mm) x 2 Eluent : 0.05M NaCl aq. Flow rate : 0.7mL/min Detector : Shodex RI Column temp. : 80deg-C
Chondroitin sulfate Na was analyzed using SUGAR KS-801 and KS-804 (columns for saccharides analysis).
Sample : 20mg/5mL Chondroitin sulfate in H2O , 25micro-L
Columns : Shodex SUGAR KS-G (6.0mmID*50mm) + KS-804 (8.0mmID*300mm) + KS-801 (8.0mmID*300mm) Eluent : 10mM NaCl aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 80deg-C
Chondroitin sulfate A was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Chondroitin sulfate, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40 deg-C
Chitosan was analyzed using OHpak SB-805 HQ and SB-806 HQ (columns for GFC separation).
Sample : Chitosan
Columns : Shodex OHpak SB-G (6.0mmID*50mm) + SB-806 HQ + SB-805 HQ (8.0mmID*300mm each) Eluent : 0.5M Acetic acid + 0.5M Sodium acetate aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Hydroxyethyl cellulose was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Hydroxyethyl cellulose, 200micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaCl aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Pullulan and Oligosaccharides were analyzed using SUGAR KS-804 (a column for saccharides analysis).
Sample : Pullulan and oligosaccharides
1. P-500 (MW: 380,000)
2. P-200 (MW: 180,000)
3. P-100 (MW: 100,000)
4. P-50 (MW: 48,000)
5. P-20 (MW: 23,700)
6. P-10 (MW: 12,200)
7. P-5 (MW: 5,800)
8. Maltoheptaose
9. Maltopentaose
10. Maltotriose
11. Maltose
12. Glucose
13. Ethylene glycol
Column : Shodex SUGAR KS-804 (8.0mmID*300m) Eluent : H2O Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 80deg-C
Pllulan, trisaccharide, disaccharide, monosaccharide and ethanol are separated using SUGAR KS-802 by GFC mode.
Sample :
1. Pullulan (P-50 MW: 46,700)
2. Pullulan (P-5 MW: 5,300)
3. Raffinose
4. Sucrose
5. Fructose
6. Ethanol
Column : Shodex SUGAR KS-802 (8.0mmID*300mm) Eluent : H2O Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 80deg-C
Heparin was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Heparin, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Glycogen was analyzed using OHpak SB-806M HQ (a column for GFC separation) from oyster.
Sample : 0.1% Glycogen from oyster, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
CS-glucan in coix seed was analyzed using Asahipak GS-510 7G and GS-320 7G.
GS-510 7G is an old type of GF-510 HQ and GS-320 7G is an old type of GS-320 HQ.
Sample : CS-Glucan
Columns : Shodex Asahipak GS-510 7G + GS-320 7G (7.5mmID*500mm each) Eluent : H2O Detector : Shodex RI
Courtesy of Dr. Yamada, The Kitasato Institute
Inulin was analyzed using SUGAR KS-806 (a column for saccharides analysis) with the two kinds of the eluent. When inulin was analyzed with H2O as the eluent, Inulin was eluted later in order to that inulin was absorbed to the gel material and further some peaks are appeared. On the other hand, when 0.05M NaNO3 aqueous solution is used as the eluent, we could obtain the good peak shape of inulin. So we recommend using a saline aqueous solution as the eluent for the analysis of inulin when KS-800 series columns are used.
Sample : Inulin
Column : Shodex SUGAR KS-806 (8.0mmID*300mm)
Eluent : (Left); 0.05M NaNO3 aq.
(Right); H2O
Flow rate : 1.0mL/min
Detector : Shodex RI
Column temp. : 80deg-C
Inulin was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Inulin, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Mucin was analyzed using OHpak SB-806M HQ (a column for GFC separation) from pig stomach.
Sample : 0.1% Mucin from pig stomach, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40 deg-C
Pectin was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Pectin from citrus, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Sodium Alginate was analyzed using OHpak SB-806M HQ (a column for GFC separation).
Sample : 0.1% Sodium alginate, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Carrageenan was analyzed using SB-806M HQ (a column for GFC separation).
Sample : 0.1% Carrageenan, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Keratan sulfate was analyzed using OHpak SB-806M HQ (a column for GFC separation) from bovine.
Sample : 0.1% Keratan sulfate from bovine, 100micro-L
Columns : Shodex OHpak SB-806M HQ (8.0mmID*300mm) x 2 Eluent : 0.1M NaNO3 aq. Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 40deg-C
Corn starch was analyzed using organic SEC column GPC KD-806M with DMSO/DMF eluent containing LiBr. Adding LiBr in eluent can suppress the influence of ionic group like carboxyl group, which starch includes. Corn starch was dissolved at 80deg-C.
Sample : 100micro-L
Corn starch (Starch) in Eluent 0.1%
Column : Shodex GPC KD-806M (8.0mmID*300mm) Eluent : 10mM LiBr in DMSO/DMF=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 50deg-C
Potato starch was analyzed using organic SEC column GPC KD-806M with DMSO/DMF eluent containing LiBr. Adding LiBr in eluent can suppress the influence of ionic group like carboxyl group, which starch includes. Potato starch was dissolved at 80deg-C.
Sample : 100micro-L
Potato starch (Starch) in DMSO 0.1%
Column : Shodex GPC KD-806M (8.0mmID*300mm) Eluent : 10mM LiBr in DMSO/DMF=75/25 Flow rate : 1.0mL/min Detector : Shodex RI Column temp. : 50deg-C
Organic solvent SEC (GPC) column, GPC KD-806M was used to analyze cellulose. Sample preparation (to dissolve cellulose into solvent) is highly dependent on the type of solvent and characteristics of the sample (crystallization, MW). Some may take up to 60 days for the preparation.
Sample : Cellulose ca. 0.05% each, 100micro-L
Column : Shodex GPC KD-806M (8.0mmID*300mm) Eluent : 1% LiCl in *DMI Flow rate : 0.5mL/min Detector : MALS(Multi Angle Laser Light Scattering), RI Column temp. : 60deg-C * 1,3-dimethyl-2-imidazolidinone
Data provided by Dr. Masahiko Yanagisawa, Isogai group, Graduate School of Agricultural and Life Sciences, The University of Tokyo