ASTM F2605 - 16

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

    Active Standard ASTM F2605 | Developed by Subcommittee: F04.42

    Book of Standards Volume: 13.02

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    Significance and Use

    4.1 The composition and sequential structure of alginate, as well as the molar mass and molar mass distribution, determines the functionality of alginate in an application. For instance, the gelling properties of an alginate are highly dependent upon the composition and molar mass of the polymer.

    4.2 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 to obtain 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.

    4.3 Multi-angle laser light scattering (MALS) is a technique where measurements are made simultaneously over a range of different angles and used to determine the scattering at 0°, which directly relates to molecular weight. MALS detection can be used to obtain information on molecular size, since this parameter is determined by the angular variation of the scattered light. This can be related to branching, aggregation, and molecular conformation. Molar mass can also be determined by detecting scattered light at a single low angle (LALS) and assuming that this is not significantly different from the scattering at 0°.

    4.4 Size exclusion chromatography uses columns, which are typically packed with polymer particles containing a network of uniform pores into which solute and solvent molecules can diffuse. While in the pores, molecules are effectively trapped and removed from the flow of the mobile phase. The average residence time in the pores depends upon the size of the solute molecules. Molecules that are larger than the average pore size of the packing are excluded and experience virtually no retention; these are eluted first, in the void volume of the column. Molecules which penetrate the pores will have a larger volume available for diffusion; their retention will depend on their molecular size, with the smaller molecules eluting last.

    4.5 For polyelectrolytes, dialysis against the elution buffer has been suggested, in order to eliminate Donnan-type artifacts in the molar mass determination by light scattering (1, 2).6 However, in the present method, the size exclusion chromatography step preceding the light scatter detection is an efficient substitute for a dialysis step. The sample is separated on SEC columns with large excess of elution buffer for 30 to 40 min, and it is therefore in full equilibrium with the elution buffer when it reaches the MALS detector.

    1. Scope

    1.1 This test method covers the determination of the molar mass (typically expressed as grams/mole) of sodium alginate 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 alginate has been published as Guide F2064.

    1.2 Alginate 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 alginate for their particular application. Alginate 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 and health practices and determine the applicability of regulatory limitations prior to use.

    2. Referenced Documents (purchase separately) The documents listed below are referenced within the subject standard but are not provided as part of the standard.

    ASTM Standards

    F2064 Guide for Characterization and Testing of Alginates as Starting Materials Intended for Use in Biomedical and Tissue Engineered Medical Product Applications

    F2315 Guide for Immobilization or Encapsulation of Living Cells or Tissue in Alginate Gels

    United States Pharmacopeia/National Formulary


    National Institute of Standards and Technology

    NIST SP811 Special Publication: Guide for the Use of the International System of Units

    ISO Standards

    ISO 31-8

    ICS Code

    ICS Number Code 11.120.10 (Medicaments)

    UNSPSC Code

    UNSPSC Code 42312000(Medical tissue closure and related products)

    Referencing This Standard
    Link Here
    Link to Active (This link will always route to the current Active version of the standard.)

    DOI: 10.1520/F2605-16

    Citation Format

    ASTM F2605-16, Standard Test Method for Determining the Molar Mass of Sodium Alginate by Size Exclusion Chromatography with Multi-angle Light Scattering Detection (SEC-MALS), ASTM International, West Conshohocken, PA, 2016,

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