ASTM D6499 - 12

    Standard Test Method for The Immunological Measurement of Antigenic Protein in Natural Rubber and its Products

    Active Standard ASTM D6499 | Developed by Subcommittee: D11.40

    Book of Standards Volume: 09.02


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    Significance and Use

    Type 1 latex allergy most commonly manifests as localized urticaria after contact of skin with natural rubber but can also include symptoms of allergic rhinoconjunctivitis, asthma and rarely anaphylaxis. This immediate (Type I) allergy is caused by natural proteins inherent to the rubber tree, which remain on the finished natural rubber products. The quantification of protein levels in NRL products using the standard colorimetric protein assays may give spurious results due to chemical additives in the latex formulations that interfere with the assay (2,3). Furthermore, the amount of protein found in NRL products are often below the detection limits of the standard colorimetric protein assay (4,5).

    This test method describes an immunological method for quantitation of natural rubber latex proteins using rabbit anti-NRL serum. Rabbits immunized with NRL proteins react to the majority of the proteins present, and their sera have the capability to detect most if not all of the proteins in NRL. Therefore, although rabbit antibody reacts with antigenic material, this should not be considered as quantitative measure of total protein levels.

    1. Scope

    1.1 This test method covers an immunological method to determine the amount of antigenic protein in natural rubber and its products using rabbit antisera specific for natural rubber latex (NRL) proteins. This immunoassay procedure quantitatively measures the level of antigenic latex proteins in solution using an inhibition format. The samples may include glove or other rubber product extracts which have been collected in order to measure the latex protein levels. Although this method detects antigenic proteins, it should not be considered as a measure of allergenic proteins. Correlation of protein/antigen levels with the level of allergenic proteins has not been fully established.

    1.2 For the purpose of this test method, the range of protein will be measured in terms of microgram to milligram quantities.

    1.3 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

    D4483 Practice for Evaluating Precision for Test Method Standards in the Rubber and Carbon Black Manufacturing Industries

    D5712 Test Method for Analysis of Aqueous Extractable Protein in Natural Rubber and Its Products Using the Modified Lowry Method

    E691 Practice for Conducting an Interlaboratory Study to Determine the Precision of a Test Method


    ICS Code

    ICS Number Code 11.100 (Laboratory medicine)

    UNSPSC Code

    UNSPSC Code 13101500(Natural rubber)


    Referencing This Standard

    DOI: 10.1520/D6499-12

    ASTM International is a member of CrossRef.

    Citation Format

    ASTM D6499-12, Standard Test Method for The Immunological Measurement of Antigenic Protein in Natural Rubber and its Products, ASTM International, West Conshohocken, PA, 2012, www.astm.org

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