GB/T 40173-2021 PDF in English
GB/T 40173-2021 (GB/T40173-2021, GBT 40173-2021, GBT40173-2021)
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Determination of reducing-end sugar in water-soluble chitosan - Spectrophotometric method
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Standards related to (historical): GB/T 40173-2021
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GB/T 40173-2021: PDF in English (GBT 40173-2021) GB/T 40173-2021
GB
NATIONAL STANDARD OF THE
PEOPLE’S REPUBLIC OF CHINA
ICS 07.080
CCS A 40
Determination of reducing-end sugar in water-soluble
chitosan - Spectrophotometric method
ISSUED ON: MAY 21, 2021
IMPLEMENTED ON: DECEMBER 01, 2021
Issued by: State Administration for Market Regulation;
Standardization Administration of the People’s Republic of
China.
Table of Contents
Foreword ... 3
1 Scope ... 4
2 Normative references ... 4
3 Terms and definitions ... 4
4 Principle ... 5
5 Reagents and materials ... 5
6 Instruments and equipment... 6
7 Preparation of sample solution ... 6
8 Determination steps ... 6
9 Result calculation and expression ... 7
10 Precision ... 8
Determination of reducing-end sugar in water-soluble
chitosan - Spectrophotometric method
1 Scope
This document specifies the spectrophotometric method for the determination
of reducing-end sugar in water-soluble chitosan.
This document applies to the detection of reducing-end sugar in water-soluble
chitosan (content ≥90%, degree of deacetylation ≥90%).
The detection limit of this method is 0.006 mmol/g; the quantitation limit is 0.023
mmol/g.
2 Normative references
The contents of the following documents constitute the indispensable clauses
of this document through normative references in the text. For dated references,
only the version corresponding to that date is applicable to this document; for
undated references, the latest version (including all amendments) is applicable
to this document.
GB/T 6682, Water for analytical laboratory use - Specification and test
methods
3 Terms and definitions
3.1 Chitosan
The derivative, which is obtained by deacetylation of chitin, that is insoluble in
water and soluble in most dilute acids. The chemical name is β-1,4-2-amino-2-
deoxy-D-glucan.
3.2 Degree of deacetylation
The percentage of the number of sugar residues that are removed from the
acetyl group in the chitosan molecule TO the total number of sugar residues in
the chitosan molecule.
3.3 Water-soluble chitosan
Chitosan of low degree of polymerization, which is soluble in acidic, neutral and
alkaline solutions.
4 Principle
The reducing-end sugar at the end of the chitosan molecule reacts with 3-
methyl-2-MBTH under alkaline conditions to form azine. Under acidic conditions,
excess MBTH is oxidized by Fe3+ to cations; then, it reacts with azine to produce
a cyan compound. The concentration of the reducing-end sugar is in a linear
relationship with its absorbance at 590 nm within a certain range. Quantify by
external standard method by measuring the absorbance value at 590 nm.
5 Reagents and materials
Unless otherwise specified, the reagents which are used in this method are
analytical reagents. The water is grade-3 water that meets the requirements of
GB/T 6682.
5.1 3-Methyl-2-benzothiazolinone hydrazone hydrochloride monohydrate
(MBTH, C8H9N3S·HCl·H2O): CAS number: 38894-11-0, purity ≥98%.
5.2 DL-Dithiothreitol [DTT, HSCH2CH(OH)CH(OH)CH2SH]: CAS number:
3483-12-3, purity ≥98%.
5.3 D-Glucosamine hydrochloride (C6H13NO5·HCl): CAS number: 66-84-2,
purity ≥99%, or a standard substance that is certified by the country and
awarded with a certificate of standard substance.
5.4 Concentrated hydrochloric acid.
5.5 Sodium hydroxide solution (0.5 mol/L): Weigh 2 g of sodium hydroxide; add
water to dissolve; use water to dilute and fix volume to 100 mL; mix well.
5.6 MBTH solution: Weigh 0.3 g of MBTH; add water to dissolve and dilute to
100 mL; mix well; store in the dark at 4 °C; it is valid for 10 days.
5.7 DTT solution: Weigh 0.080 g of DTT; add water to dissolve and dilute to 100
mL; mix well; store in the dark at 4 °C; it is valid for 10 days.
5.8 MBTH reagent: Mix MBTH solution (5.6) and DTT solution (5.7) in equal
volumes to obtain MBTH reagent. Prepare when necessary.
5.9 Ferric ammonium sulfate reagent: Weigh 5 g of ferric ammonium sulfate
[NH4Fe(SO4)2·12H2O] and 5 g of sulfamic acid; add 41.8 mL of concentrated
......
Source: Above contents are excerpted from the PDF -- translated/reviewed by: www.chinesestandard.net / Wayne Zheng et al.
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