ASTM F2602-18

Standard Test Method for Determining the Molar Mass of Chitosan and Chitosan Salts by Size Exclusion Chromatography with Multi-angle Light Scattering Detection (SEC-MALS)

SIGNIFICANCE AND USE
4.1 The degree of deacetylation of chitosan, as well at the molar mass and molar mass distribution, determines the functionality of chitosan in an application. For instance, functional and biological effects are highly dependent upon the composition and molar mass of the polymer.  
4.2 This test method describes procedures for measurement of molar mass of chitosan chlorides and glutamates, and chitosan base, although it in principle applies to any chitosan salt. The measured molar mass is that for chitosan acetate, since the mobile phase contains acetate as counter ion. This value can further be converted into the corresponding molar mass for the chitosan as a base, or the parent salt form (chloride or glutamate).  
4.3 Light scattering is one of very few methods available for the determination of absolute molar mass and structure, and it is applicable over the broadest range of molar masses of any method. Combining light scattering detection with size exclusion chromatography (SEC), which sorts molecules according to size, gives the ability to analyze polydisperse samples, as well as obtaining information on branching and molecular conformation. This means that both the number-average and mass-average values for molar mass and size may be obtained for most samples. Furthermore, one has the ability to calculate the distributions of the molar masses and sizes.  
4.4 Multi-angle laser light scattering (MALS) is a technique where measurements of scattered light are made simultaneously over a range of different angles. MALS detection can be used to obtain information on molecular size, since this parameter is determined by the angular variation of the scattered light. Molar mass may in principle be determined by detecting scattered light at a single low angle (LALLS). However, advantages with MALS as compared to LALLS are: (1) less noise at larger angles, (2) precision of measurements is improved by detecting at several angles, and (3) the ability to detect angular v...
SCOPE
1.1 This test method covers the determination of the molar mass of chitosan and chitosan salts intended for use in biomedical and pharmaceutical applications as well as in tissue engineered medical products (TEMPs) by size exclusion chromatography with multi-angle laser light scattering detection (SEC-MALS). A guide for the characterization of chitosan salts has been published as Guide F2103.  
1.2 Chitosan and chitosan salts used in TEMPs should be well characterized, including the molar mass and polydispersity (molar mass distribution) in order to ensure uniformity and correct functionality in the final product. This test method will assist end users in choosing the correct chitosan for their particular application. Chitosan may have utility as a scaffold or matrix material for TEMPs, in cell and tissue encapsulation applications, and in drug delivery formulations.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

General Information

• Status: Published
• Publication Date: 31-May-2018
• ICS: 11.120.10 - Medicaments
• Technical Committee: F04 - Medical and Surgical Materials and Devices
• Drafting Committee: F04.42 - Biomaterials and Biomolecules for TEMPs

Relations

• Refers: ASTM F2103-18 - Standard Guide for Characterization and Testing of Chitosan Salts as Starting Materials Intended for Use in Biomedical and Tissue-Engineered Medical Product Applications
• Effective Date: 01-Jun-2018

Standards Content (Sample)

This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: F2602 − 18
Standard Test Method for
Determining the Molar Mass of Chitosan and Chitosan Salts
by Size Exclusion Chromatography with Multi-angle Light
1
Scattering Detection (SEC-MALS)
This standard is issued under the fixed designation F2602; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 2. Referenced Documents
2
1.1 This test method covers the determination of the molar 2.1 ASTM Standards:
mass of chitosan and chitosan salts intended for use in F2103 Guide for Characterization and Testing of Chitosan
biomedical and pharmaceutical applications as well as in tissue Salts as Starting Materials Intended for Use in Biomedical
engineered medical products (TEMPs) by size exclusion chro- and Tissue-Engineered Medical Product Applications
3
matography with multi-angle laser light scattering detection
2.2 National Institute of Standards and Technology:
(SEC-MALS).Aguideforthecharacterizationofchitosansalts
NIST SP811 Special Publication: Guide for the Use of the
has been published as Guide F2103.
International System of Units (SI)
4
2.3 ISO Document:
1.2 Chitosan and chitosan salts used in TEMPs should be
well characterized, including the molar mass and polydisper- ISO 80000-9:2009 Quantities and units – Part 9: Physical
chemistry and molecular physics
sity(molarmassdistribution)inordertoensureuniformityand
correct functionality in the final product. This test method will
3. Terminology
assist end users in choosing the correct chitosan for their
particular application. Chitosan may have utility as a scaffold
3.1 Definitions:
or matrix material for TEMPs, in cell and tissue encapsulation
3.1.1 chitosan, n—a linear polysaccharide consisting of
applications, and in drug delivery formulations.
β(1→4) linked 2-acetamido-2-deoxy-D-glucopyranose (Glc-
NAc) and 2-amino-2-deoxy-D-glucopyranose (GlcN). Chito-
1.3 The values stated in SI units are to be regarded as
san is a polysaccharide derived by N-deacetylation of chitin.
standard. No other units of measurement are included in this
standard.
3.1.1.1 chitin, n—a linear polysaccharide consisting of
β(1→4) linked 2-acetamido-2-deoxy-D-glucopyranose.
1.4 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
3.1.2 degree of deacetylation, n—the fraction or percentage
responsibility of the user of this standard to establish appro-
of glucosamine units (GlcN: deacetylated monomers) in a
priate safety, health, and environmental practices and deter-
chitosan polymer molecule.
mine the applicability of regulatory limitations prior to use.
3.1.3 molar mass average, n—the given molar mass (M) of
1.5 This international standard was developed in accor-
a chitosan will always represent an average of all of the
dance with internationally recognized principles on standard-
molecules in the population. The most common ways to
ization established in the Decision on Principles for the
Development of International Standards, Guides and Recom-
mendations issued by the World Trade Organization Technical 2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Barriers to Trade (TBT) Committee. contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
1 3
This test method is under the jurisdiction ofASTM Committee F04 on Medical Available from National Institute of Standards and Technology (NIST), 100
and Surgical Materials and Devices and is the direct responsibility of Subcommittee Bureau Dr., Stop 1070, Gaithersburg, MD 20899-1070, http://physics.nist.gov/cuu/
F04.42 on Biomaterials and Biomolecules for TEMPs. Units/bibliography.html.
4
Current edition approved June 1, 2018. Published August 2018. Originally Available from International Organization for Standardization (ISO), ISO
approved in 2008. Last previous edition approved in 2013 as F2602–13. DOI: Central Secretariat, BIBC II, Chemin de Blandonnet 8, CP 401, 1214 Vernier,
10.1520/F2602-18. Geneva, Switzerland, http://www.iso.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
F2602 − 18
¯
expressthemolarmassareasthe number average(M )andthe (1) less noise at larger angles, (2) precision of measurements is
n
¯
mass average (M ). The two aver
...

This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: F2602 − 13 F2602 − 18
Standard Test Method for
Determining the Molar Mass of Chitosan and Chitosan Salts
by Size Exclusion Chromatography with Multi-angle Light
1
Scattering Detection (SEC-MALS)
This standard is issued under the fixed designation F2602; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This test method covers the determination of the molar mass of chitosan and chitosan salts intended for use in biomedical
and pharmaceutical applications as well as in tissue engineered medical products (TEMPs) by size exclusion chromatography with
multi-angle laser light scattering detection (SEC-MALS). A guide for the characterization of chitosan salts has been published as
Guide F2103.
1.2 Chitosan and chitosan salts used in TEMPs should be well characterized, including the molar mass and polydispersity
(molar mass distribution) in order to ensure uniformity and correct functionality in the final product. This test method will assist
end users in choosing the correct chitosan for their particular application. Chitosan may have utility as a scaffold or matrix material
for TEMPs, in cell and tissue encapsulation applications, and in drug delivery formulations.
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility
of the user of this standard to establish appropriate safety safety, health, and healthenvironmental practices and determine the
applicability of regulatory limitations prior to use.
1.5 This international standard was developed in accordance with internationally recognized principles on standardization
established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued
by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
2. Referenced Documents
2
2.1 ASTM Standards:
F2103 Guide for Characterization and Testing of Chitosan Salts as Starting Materials Intended for Use in Biomedical and
Tissue-Engineered Medical Product Applications
3
2.2 United States Pharmacopeia/National Formulary:
<621> Chromatography
3
2.2 National Institute of Standards and Technology:
NIST SP811 Special Publication: Guide for the Use of the International System of Units (SI)
4
2.3 ISO Document:
ISO 80000-9:2009 Quantities and units – Part 9: Physical chemistry and molecular physics
3. Terminology
3.1 Definitions:
1
This test method is under the jurisdiction of ASTM Committee F04 on Medical and Surgical Materials and Devices and is the direct responsibility of Subcommittee
F04.42 on Biomaterials and Biomolecules for TEMPs.
Current edition approved Aug. 1, 2013June 1, 2018. Published September 2013August 2018. Originally approved in 2008. Last previous edition approved in 20082013
ε1
as F2602–08–13. . DOI: 10.1520/F2602-13.10.1520/F2602-18.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
3
Available from United States Pharmacopeia and National Formulary, U.S. Pharmaceutical Convention, Inc. (USPC), Rockville, MD.
3
Available from National Institute of Standards and Technology (NIST), 100 Bureau Dr., Stop 1070, Gaithersburg, MD 20899-1070, http://physics.nist.gov/cuu/Units/
bibliography.html.
4
Available from International Organization for Standardization (ISO), ISO Central Secretariat, BIBC II, Chemin de Blandonnet 8, CP 401, 1214 Vernier, Geneva,
Switzerland, http://www.iso.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
F2602 − 18
3.1.1 chitosan, n—a linear polysaccharide consisting of β(1→4) linked 2-acetamido-2-deoxy-D-glucopyranose (GlcNAc) and
2-amino-2-deoxy-D-glucopyranose (GlcN). Chitosan is a polysaccharide derived by N-deacetylation of chitin.
3.1.1.1 chitin, n—a linear polysaccharide consisting of β(1→4) linked 2-acetamido-2-deoxy-D-glucopyranose.
3.1.2 degree of deacetylation, n—the fraction or percentage of
...