ASTM D7644 - 10e2

    Standard Test Method for Determination of Bromadiolone, Brodifacoum, Diphacinone and Warfarin in Water by Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS)

    Active Standard ASTM D7644 | Developed by Subcommittee: D19.06

    Book of Standards Volume: 11.02


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    Significance and Use

    This test method has been developed by US EPA Region 5 Chicago Regional Laboratory (CRL).

    Bromadiolone, brodifacoum, diphacinone and warfarin are rodenticides for controlling mice, rats, and other rodents that pose a threat to public health, critical habitats, native plants and animals, crops, food and water supplies. These rodenticides also present human and environmental safety concerns. Warfarin and diphacinone are first-generation anticoagulants, while bromadiolone and brodifacoum are second-generation. The anticoagulants interfere with blood clotting, and death can result from excessive bleeding. The second-generation anticoagulants are especially hazardous for several reasons. They are highly toxic and persist a long time in body tissues. The second-generation anticoagulants are designed to be toxic in a single feeding, but time-to-death occurs in several days. This allows rodents to feed multiple times before death, leading to carcasses containing residues that may be many times the lethal dose.

    This method has been investigated for use with reagent, surface, and drinking water for the selected rodenticides.

    1. Scope

    1.1 This procedure covers the determination of bromadiolone, brodifacoum, diphacinone and warfarin (referred to collectively as rodenticides in this test method) in water by direct injection using liquid chromatography (LC) and detected with tandem mass spectrometry (MS/MS). These analytes are qualitatively and quantitatively determined by this method. This method adheres to multiple reaction monitoring (MRM) mass spectrometry.

    1.3 UnitsThe 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.

    TABLE 1 Detection Verification Level and Reporting Range

    AnalyteDVL
    (ng/L)
    Reporting Range
    (ng/L)
    Bromadiolone20125-2500
    Brodifacoum20125-2500
    Diphacinone20125-2500
    Warfarin20125-2500


    FIG. 1 Example Primary SRM Chromatograms Signal/Noise Ratios


    FIG. 2 Example Confirmatory SRM Chromatograms Signal/Noise Ratios


    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

    D1129 Terminology Relating to Water

    D1193 Specification for Reagent Water

    D2777 Practice for Determination of Precision and Bias of Applicable Test Methods of Committee D19 on Water

    D3694 Practices for Preparation of Sample Containers and for Preservation of Organic Constituents

    D3856 Guide for Management Systems in Laboratories Engaged in Analysis of Water

    D4841 Practice for Estimation of Holding Time for Water Samples Containing Organic and Inorganic Constituents

    D5847 Practice for Writing Quality Control Specifications for Standard Test Methods for Water Analysis

    E2554 Practice for Estimating and Monitoring the Uncertainty of Test Results of a Test Method in a Single Laboratory Using a Control Sample Program

    Other Documents

    EPApublicationSW-846 Test Methods for Evaluating Solid Waste, Physical/Chemical Methods Available from National Technical Information Service (NTIS), U.S. Department of Commerce, 5285 Port Royal Road, Springfield, VA, 22161 or at http://www.epa.gov/epawaste/hazard/testmethods/index.htm.


    ICS Code

    ICS Number Code 13.060.50 (Examination of water for chemical substances)

    UNSPSC Code

    UNSPSC Code


    DOI: 10.1520/D7644-10E02

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