GB/T 47142-2026 Determination of molecular weight of polysaccharides—Size exclusion chromatography with multi-angler laser light scattering method English, Anglais, Englisch, Inglés, えいご
This is a draft translation for reference among interesting stakeholders. The finalized translation (passing through draft translation, self-check, revision and verification) will be delivered upon being ordered.
ICS 13.220.10
CCS H 57
National Standard of the People's Republic of China
GB/T 47142-2026
Determination of molecular weight of polysaccharides - Size exclusion chromatography with multi-angler laser light scattering method
多糖分子量的测定 分子排阻色谱-多角度激光光散射法
Issue date: 2026-01-28 Implementation date: 2027-02-01
Issued by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China
the Standardization Administration of the People's Republic of China
Contents
Foreword
Introduction
1 Scope
2 Normative References
3 Terms and Definitions
4 Principle
5 Reagents, Materials, Instruments and Equipment
6 Experimental Procedure
Determination of Polysaccharide Molecular Weight Method of Size Exclusion Chromatography - Multi-Angle Laser Light Scattering
1 Scope
This document describes the method and test conditions for determining the molecular weight of pectin, glucan, and inulin using size exclusion chromatography - multi-angle laser light scattering.
This document applies to the determination of molecular weight for pectin, glucan, and inulin with purity ≥ 95% and a molecular weight determination range of 1 kDa to 1000 kDa.
2 Normative References
The following documents contain provisions which, through normative reference in this text, constitute essential provisions of this document. For dated references, only the edition cited applies. For undated references, the latest edition (including any amendments) applies.
GB/T 6682 Water for analytical laboratory use - Specification and test methods
3 Terms and Definitions
For the purposes of this document, the following terms and definitions apply.
3.1 polysaccharide
Natural high molecular polymers formed by the linear or branched polymerization of more than 10 monosaccharide molecules through glycosidic bonds.
4 Principle
After the sample is prepared into a solution, size exclusion chromatography is used to separate polysaccharide molecules of different molecular weights, which then sequentially enter a multi-angle laser light scattering detector and a differential refractive index detector. The multi-angle laser light scattering detector directly determines the absolute molecular weight of each component by measuring the light scattering signal. The differential refractive index detector provides the concentration of the corresponding component. The instrument's accompanying software combines the results from both detectors to calculate the weight-average molecular weight.
5 Reagents, Materials, Instruments and Equipment
5.1 Reagents and Materials
Unless otherwise specified, all reagents are of analytical grade, and the water is Grade 1 water as specified in GB/T 6682.
5.1.1 Dextran reference standard: Molecular weight 30 kDa ~ 40 kDa.
5.1.2 0.2 mol/L Sodium chloride solution: Weigh 11.69 g of sodium chloride, dissolve with ultrapure water, and dilute to 1 L.
5.1.3 Preservative: ProClin300.
5.1.4 Microporous membrane filter: with pore size 0.45 μm.
5.1.5 Microporous membrane filter: with pore size 0.22 μm.
5.1.6 Membrane filter for vacuum filtration: with pore size 0.22 μm.
5.1.7 Volumetric flask: 25 mL.
5.1.8 Flat-tip syringe: 50 mm.
5.2 Instruments and Equipment
5.2.1 Electronic balance: with a scale division of 0.1 mg.
5.2.2 High-performance liquid chromatograph: equipped with a multi-angle laser light scattering detector and a differential refractive index detector, as well as associated data acquisition and processing software.
5.2.3 Gel chromatography column: resistant to aqueous mobile phases, suitable for size exclusion separation.
6 Experimental Procedure
6.1 Preparation of Sample Solution
Accurately weigh 12.5 mg ~ 250 mg of the sample (accurate to 0.1 mg) into a 25 mL centrifuge tube. Add 15 mL of the mobile phase, shake until dissolved, and let the centrifuge tube stand overnight to ensure the sample is fully dissolved. Transfer the sample solution to a 25 mL volumetric flask. Rinse the inner wall of the centrifuge tube multiple times with the mobile phase, combine the rinses, dilute to the mark with the mobile phase, and mix well. Filter the solution through a microporous membrane filter before analysis. The solution concentration can be adjusted according to the solubility of the sample.
NOTE: Polysaccharide samples with different molecular weights require different preparation concentrations. Recommendation: When the polysaccharide molecular weight is ≤ 5 kDa, the sample solution concentration should be 5 mg/mL ~ 10 mg/mL; when 5 kDa < polysaccharide molecular weight ≤ 100 kDa, the sample solution concentration should be 1 mg/mL ~ 5 mg/mL; when 100 kDa < polysaccharide molecular weight ≤ 1000 kDa, the sample solution concentration should be 0.5 mg/mL ~ 1 mg/mL.
6.2 Reference Conditions for Liquid Chromatography
The reference conditions for liquid chromatography are as follows:
a) Chromatographic column: Hydrophilic silicified polymer gel chromatography column (300 mm × 7.8 mm, 10 μm), or a styrene-divinylbenzene copolymer gel chromatography column suitable for the polysaccharide molecular weight range;
6.3 Normalization Procedure
Place 20.0 mg of the dextran reference standard into a 10 mL volumetric flask. Dissolve and dilute to volume with the mobile phase to prepare a 2 mg/mL dextran solution. Inject this solution into the liquid chromatograph to perform the normalization procedure. Save this normalized data as an analysis template for the subsequent determination of molecular weight and molecular weight distribution of samples.
6.4 Sample Analysis
8 Precision
Under repeatability conditions, the absolute difference between two independent determination results for Mw shall not exceed 10 % of the arithmetic mean.
Standard
GB/T 47142-2026 Determination of molecular weight of polysaccharides—Size exclusion chromatography with multi-angler laser light scattering method (English Version)
Standard No.
GB/T 47142-2026
Status
to be valid
Language
English
File Format
PDF
Word Count
7000 words
Price(USD)
210.0
Implemented on
2026-5-1
Delivery
via email in 1~3 business day
Detail of GB/T 47142-2026
Standard No.
GB/T 47142-2026
English Name
Determination of molecular weight of polysaccharides—Size exclusion chromatography with multi-angler laser light scattering method
GB/T 47142-2026 Determination of molecular weight of polysaccharides—Size exclusion chromatography with multi-angler laser light scattering method English, Anglais, Englisch, Inglés, えいご
This is a draft translation for reference among interesting stakeholders. The finalized translation (passing through draft translation, self-check, revision and verification) will be delivered upon being ordered.
ICS 13.220.10
CCS H 57
National Standard of the People's Republic of China
GB/T 47142-2026
Determination of molecular weight of polysaccharides - Size exclusion chromatography with multi-angler laser light scattering method
多糖分子量的测定 分子排阻色谱-多角度激光光散射法
Issue date: 2026-01-28 Implementation date: 2027-02-01
Issued by the General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China
the Standardization Administration of the People's Republic of China
Contents
Foreword
Introduction
1 Scope
2 Normative References
3 Terms and Definitions
4 Principle
5 Reagents, Materials, Instruments and Equipment
6 Experimental Procedure
Determination of Polysaccharide Molecular Weight Method of Size Exclusion Chromatography - Multi-Angle Laser Light Scattering
1 Scope
This document describes the method and test conditions for determining the molecular weight of pectin, glucan, and inulin using size exclusion chromatography - multi-angle laser light scattering.
This document applies to the determination of molecular weight for pectin, glucan, and inulin with purity ≥ 95% and a molecular weight determination range of 1 kDa to 1000 kDa.
2 Normative References
The following documents contain provisions which, through normative reference in this text, constitute essential provisions of this document. For dated references, only the edition cited applies. For undated references, the latest edition (including any amendments) applies.
GB/T 6682 Water for analytical laboratory use - Specification and test methods
3 Terms and Definitions
For the purposes of this document, the following terms and definitions apply.
3.1 polysaccharide
Natural high molecular polymers formed by the linear or branched polymerization of more than 10 monosaccharide molecules through glycosidic bonds.
4 Principle
After the sample is prepared into a solution, size exclusion chromatography is used to separate polysaccharide molecules of different molecular weights, which then sequentially enter a multi-angle laser light scattering detector and a differential refractive index detector. The multi-angle laser light scattering detector directly determines the absolute molecular weight of each component by measuring the light scattering signal. The differential refractive index detector provides the concentration of the corresponding component. The instrument's accompanying software combines the results from both detectors to calculate the weight-average molecular weight.
5 Reagents, Materials, Instruments and Equipment
5.1 Reagents and Materials
Unless otherwise specified, all reagents are of analytical grade, and the water is Grade 1 water as specified in GB/T 6682.
5.1.1 Dextran reference standard: Molecular weight 30 kDa ~ 40 kDa.
5.1.2 0.2 mol/L Sodium chloride solution: Weigh 11.69 g of sodium chloride, dissolve with ultrapure water, and dilute to 1 L.
5.1.3 Preservative: ProClin300.
5.1.4 Microporous membrane filter: with pore size 0.45 μm.
5.1.5 Microporous membrane filter: with pore size 0.22 μm.
5.1.6 Membrane filter for vacuum filtration: with pore size 0.22 μm.
5.1.7 Volumetric flask: 25 mL.
5.1.8 Flat-tip syringe: 50 mm.
5.2 Instruments and Equipment
5.2.1 Electronic balance: with a scale division of 0.1 mg.
5.2.2 High-performance liquid chromatograph: equipped with a multi-angle laser light scattering detector and a differential refractive index detector, as well as associated data acquisition and processing software.
5.2.3 Gel chromatography column: resistant to aqueous mobile phases, suitable for size exclusion separation.
6 Experimental Procedure
6.1 Preparation of Sample Solution
Accurately weigh 12.5 mg ~ 250 mg of the sample (accurate to 0.1 mg) into a 25 mL centrifuge tube. Add 15 mL of the mobile phase, shake until dissolved, and let the centrifuge tube stand overnight to ensure the sample is fully dissolved. Transfer the sample solution to a 25 mL volumetric flask. Rinse the inner wall of the centrifuge tube multiple times with the mobile phase, combine the rinses, dilute to the mark with the mobile phase, and mix well. Filter the solution through a microporous membrane filter before analysis. The solution concentration can be adjusted according to the solubility of the sample.
NOTE: Polysaccharide samples with different molecular weights require different preparation concentrations. Recommendation: When the polysaccharide molecular weight is ≤ 5 kDa, the sample solution concentration should be 5 mg/mL ~ 10 mg/mL; when 5 kDa < polysaccharide molecular weight ≤ 100 kDa, the sample solution concentration should be 1 mg/mL ~ 5 mg/mL; when 100 kDa < polysaccharide molecular weight ≤ 1000 kDa, the sample solution concentration should be 0.5 mg/mL ~ 1 mg/mL.
6.2 Reference Conditions for Liquid Chromatography
The reference conditions for liquid chromatography are as follows:
a) Chromatographic column: Hydrophilic silicified polymer gel chromatography column (300 mm × 7.8 mm, 10 μm), or a styrene-divinylbenzene copolymer gel chromatography column suitable for the polysaccharide molecular weight range;
6.3 Normalization Procedure
Place 20.0 mg of the dextran reference standard into a 10 mL volumetric flask. Dissolve and dilute to volume with the mobile phase to prepare a 2 mg/mL dextran solution. Inject this solution into the liquid chromatograph to perform the normalization procedure. Save this normalized data as an analysis template for the subsequent determination of molecular weight and molecular weight distribution of samples.
6.4 Sample Analysis
8 Precision
Under repeatability conditions, the absolute difference between two independent determination results for Mw shall not exceed 10 % of the arithmetic mean.
