ASTM D7304 - 14

    Standard Test Method for Determination of Denatonium Ion in Engine Coolant by HPLC

    Active Standard ASTM D7304 | Developed by Subcommittee: D15.04

    Book of Standards Volume: 15.05


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    Significance and Use

    4.1 This test method provides for the qualitative and quantitative determination of denatonium benzoate in engine coolant in milligrams per litre to low percent range and requires approximately 1 mL per test, with results available in less than 10 min. Denatonium benzoate is a compound composed of a quaternary ammonium cation, denatonium and an inert anion, benzoate. In solution the denatonium benzoate exists in equilibrium between the denatonium benzoate compound, the denatonium cation and benzoate anion. By slightly adjusting the pH of the solution to be more acidic ( pH 4.6) the equilibrium will be shifted to the direction of forming more denatonium and benzoate ions in the solution.

    1. Scope

    1.1 This test method covers the chemical analysis of engine coolant for denatonium benzoate (DNB) by high-performance liquid chromatography (HPLC). DNB is added to potentially render engine coolant unpalatable to animals and humans. This analytical method was designed for the analysis of DNB and is not valid for any other bittering agents such as denatonium saccharide.

    1.2 This test method is applicable to both new and used coolants.

    1.3 Coelution of other ions may cause interferences in the detection of the denatonium cation. In the case of unfamiliar formulations, identification verification should be performed by either or both fortification and dilution of the sample matrix with denatonium ion.

    1.4 The values stated in SI units are to be regarded as the standard. The values given in parentheses are for information only.

    1.5 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

    D1176 Practice for Sampling and Preparing Aqueous Solutions of Engine Coolants or Antirusts for Testing Purposes

    D1193 Specification for Reagent Water


    ICS Code

    ICS Number Code 71.100.45 (Refrigerants and antifreezes)

    UNSPSC Code

    UNSPSC Code 25174004(Engine coolant)


    Referencing This Standard

    DOI: 10.1520/D7304

    ASTM International is a member of CrossRef.

    Citation Format

    ASTM D7304-14, Standard Test Method for Determination of Denatonium Ion in Engine Coolant by HPLC, ASTM International, West Conshohocken, PA, 2014, www.astm.org

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