Recommended Procedures Regarding the CPSC's Policy on Animal Testing

November 26, 2012

 

The Federal Hazardous Substances Act (FHSA) is one of the laws administered by CPSC. One of the primary functions of the law is to require cautionary labeling on certain household products that contain a hazardous substance. Hazardous substances are those that are toxic, corrosive, an irritant, flammable, combustible or a strong sensitizer and that may cause substantial personal injury or illness as a result of reasonably foreseeable use. In order to determine the appropriate cautionary labeling it is necessary to have objective criteria by which the existence of a hazard can be determined. Neither the FHSA nor the regulations issued thereunder require animal testing to determine whether a hazard exists. Under the FHSA animal testing is one possible option that can be used to determine the biological response and appropriate cautionary labeling. The Commission's policy is to find alternatives to traditional animal testing that replace animals, reduce the number of animals tested, and decrease the pain and suffering in animals associated with testing household products. As such, the Commission and CPSC staff strongly encourage the use of scientifically validated alternatives to animal testing and the use of existing information, including expert opinion, prior human experience, and prior animal testing results, in the determination of hazard. The CPSC is a member of the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), and as suand supports that Committee’s development and use of validated alternative test methods. ICCVAM test methods that have been approved by the CPSC for hazard determination under the FHSA are listed on this Web page.

 

The CPSC has codified its policy on animal testing at 16 CFR § 1500.232 (77 FR 73286). In addition, the CPSC has amended and updated its animal testing regulations to allow alternatives to animal testing, whenever possible, under 16 CFR part 1500 (77 FR 73289). The information on this page is intended to supplement the CPSC's statement of policy on animal testing to help industry determine the alternative tests that are acceptable and available currently to determine the hazards addressed by the FHSA.

 

This page also references other acceptable alternative methods of testing to identify specific hazards under the FHSA. If a manufacturer or other entity performs a hazard test for FHSA labeling purposes that has not been previously approved by the CPSC, that manufacturer may submit its data to the agency and CPSC staff will consider these data on a case-by-case basis. In particular, data generated using Organization for Economic Cooperation and Development (OECD) Guidelines may be considered by the CPSC in making a hazard determination as a result of the Mutual Acceptance of Data (MAD) agreement. The MAD agreement allows OECD member countries to mutually accept data that comply with OECD Test Guidelines and Principles of Good Laboratory Practice. Data from methods other than OECD that are scientifically supportable may be considered as well. However, neither the MAD agreement, nor CPSC staff’s decision to consider any test data, guarantees that CPSC staff will accept data from alternative tests for purposes of FHSA classification and/or labeling. The use of an alternative test will be considered to be acceptable by the CPSC only if it is posted on this Web page after review.


Acute Toxicity Testing

 

For each of three exposure routes (oral, dermal, and inhalation), the FHSA distinguishes two levels of acute toxicity, highly toxic and toxic, from substances that are not toxic, and therefore, do not require labeling. These terms are defined in 16 CFR § 1500.3, along with a description of the traditional method for determining the acute toxicity endpoint, the LD50 or the LC50.

Oral acute toxicity testing

 

16 CFR § 1500.3(c)(1)(ii)(A) defines highly toxic by the oral exposure route and § 1500.3(c)(2)(i)(A) gives the FHSA definition of toxic by the oral exposure route. These sections also contain guidelines for a traditional acute toxicity test. CPSC staff recommends the revised oral Up-and-Down Procedure (UDP) for determining acute oral toxicity for the purpose of classification and labeling under the FHSA. The UDP was issued by U.S. Environmental Protection Agency's Office of Prevention, Pesticides and Toxic Substances (OPPTS) as OPPTS Harmonized Test Guideline 870.1100. The scientifically validated test guideline is referenced in CPSC staff's 2011 Response (pdf) to ICCVAM on Acute Toxicity Testing.


In addition, CPSC staff has determined that in vitro basal cytotoxicity tests are appropriate for determining a starting dose for the oral LD50 test. The test guidelines are referenced in CPSC staff's 2008 Response (pdf) to ICCVAM on the Use of In Vitro Basal Cytotoxicity Test Methods for Estimating Starting Doses for Acute Oral Systemic Toxicity Testing.

Inhalation acute toxicity testing

 

16 CFR § 1500.3(c)(1)(ii)(B) defines highly toxic by the inhalation exposure route, and § 1500.3(c)(2)(i)(B) gives the FHSA definition of toxic by the inhalation exposure route. Currently, there are no Commission-approved non-animal methods for inhalation toxicity testing.


Dermal acute toxicity testing

16 CFR § 1500.3(c)(1)(ii)(C) defines highly toxic by the dermal exposure route, and § 1500.3(c)(2)(i)(C) gives the FHSA definition of toxic by the dermal exposure route. Section 1500.40 of 16 CFR details an in vivo method for testing the acute dermal toxicity of substances. Currently, there are no Commission-approved non-animal methods for dermal toxicity testing.
 

Ocular Irritation Testing

  • In Vitro Ocular Irritation Testing Methods
    The Commission has voted to accept the recommendations of ICCVAM on several in vitro alternatives to the Draize rabbit eye test. The strengths and limitations of each method with respect to FHSA labeling requirements are described briefly here.
    • The isolated chicken eye (ICE) test can be used as a screening tool for severe irritants/corrosive substances. When the ICE test indicates a substance is a severe ocular irritant/corrosive, that substance can be labeled "irritant" under FHSA with no additional testing. However, this assay cannot reliably distinguish all substances that require the FHSA irritant label (which include mild and moderate irritants) from substances not requiring the label. Therefore, if this screening test indicates the substance is not a strong irritant/corrosive, additional testing (i.e., a Draize test) would be necessary for validation and subsequent determination of labeling. The ICE test cannot be used to distinguish substances not requiring the FHSA irritant label from those requiring it. The ICE test cannot be used with alcohols, surfactants, or solids.
    • The Bovine Corneal Opacity and Permeability (BCOP) test can be used as a screening tool for severe irritants/corrosive substances. When the BCOP test indicates a substance is a strong irritant/ocular corrosive, that substance can be labeled irritant under the FHSA with no additional testing. However, the BCOP test cannot reliably distinguish all substances that require the FHSA irritant label (which include mild and moderate irritants) from substances not requiring the label. If screening indicates the substance is anything but a strong irritant/corrosive, additional testing (i.e., a Draize test) would be necessary for validation and subsequent determination of labeling. The BCOP test cannot be used with alcohols, ketones, or solids.
    • Cytosensor Microphysiometer (CM) test method can be used with water-soluble surfactant chemicals and certain types of surfactant-containing formulations (e.g., cosmetics and personal care product formulations, but not pesticide formulations) to distinguish substances not requiring the FHSA irritant label from those requiring the irritant label under FHSA. However, due to a substantial rate of false positive outcomes when the CM test method is being used for this distinction, a substance identified as an irritant would require additional testing in a valid test system (i.e., the Draize test) that can accurately characterize whether it requires hazard labeling.


  • In Vivo Ocular Irritation Testing Methods
    The Commission has approved a modified version of the traditional Draize rabbit eye test for ocular irritants. The traditional test guidelines are outlined in § 1500.42 of 16 CFR. Modifications to this method comprise a balanced three-part preemptive pain management strategy using topical anesthetics, systemic analgesics, and humane endpoints to avoid or minimize pain and distress associated with the traditional Draize method.
    • Topical Anesthetics: Part one of the modified Draize eye test involves pretreatment of animals with a topical anesthetic and systemic analgesic before applying test substances. The ICCVAM-recommended protocol calls for administration of 0.01 mg/kg buprenorphine by subcutaneous injection 60 minutes before application of the test substance, plus 1 to 2 drops per eye of 0.5 percent proparacaine hydrochloride or 0.5 percent tetracaine hydrochloride five minutes before applying the test substance. This topical application can be repeated in 5-minute intervals, as needed, before test substance administration. However, ICCVAM qualified this recommendation with a warning that multiple applications could increase the severity or longevity of ocular lesions.
    • Systemic Analgesics: The second part of the pain management strategy is adherence to a routine schedule of systemic analgesia after test substance administration. ICCVAM recommended a "rescue" dose of 0.03 mg/kg buprenorphine, given every eight hours, and 0.5 mg/kg meloxicam every 12 hours following test substance administration for a distressed animal. If the test animal is not in distress, ICCVAM's protocol called for 0.01 mg/kg buprenorphine, plus 0.5 mg/kg meloxicam, to be delivered subcutaneously 8 hours after test article administration. If ocular lesions and/or clinical signs of pain and distress persist after this combination dose, another dose of buprenorphine could be given at 12-hour intervals and another dose of meloxicam at 24-hour intervals.
  • Humane Endpoints: The third part of ICCVAM's recommended pain management strategy was scheduled observations, monitoring, and recording of the nature, severity, and progression of all eye injuries. (One recommended eye irritation scoring system is the U.S. EPA's Test Guideline, OPPTS 870.2400: Acute Eye Irritation) (pdf).


    ICCVAM recommended maintaining a written record of all observations to facilitate decisions on the progression or resolution of ocular lesions. Specific methods emphasized by ICCVAM to help detect and measure ocular endpoints included fluorescein staining, slit-lamp biomicroscopy, and digital photography. ICCVAM also identified specific ocular lesions indicative of a severity and irreversibility of damage that should warrant termination of studies before the end of the scheduled 21-day observation period, including:

    • Draize corneal opacity score of 4 that persists for 48 hours;
    • Corneal perforation or significant corneal ulceration, including staphyloma;
    • Blood in the anterior chamber of the eye;
    • Absence of light reflex that persists for 72 hours;
    • Ulceration of the conjunctival membrane;
    • Necrosis of the conjunctiva or nictitating membrane;
    • Sloughing;
    • Destruction of more than 75 percent of the limbus;
    • Depth of injury to the cornea (routinely using slit-lamp and fluorescein staining)), in which corneal ulceration extends beyond superficial layers of the stroma;
    • Vascularization of the corneal surface (i.e., pannus);
    • No diminishment in area of fluorescein staining and/or increase in depth of injury over time; and
    • Lack of re-epithelialization 5 days after application of the test substance.


    Dermal Irritation Testing

  • The traditional in vivo method for testing primary irritants is described in 16 CFR § 1500.41.
  • In vitro methods assessing skin irritation and corrosivity have not been assessed by CPSC staff. The Organization for Economic Cooperation and Development (OECD) has written guidelines outlining in vitro methods for determining skin irritation and corrosivity, which are described in OECD Test Guidelines 430 and 431. Data generated using OECD Guidelines can be considered by CPSC in making a hazard determination as a result of the Mutual Acceptance of Data (MAD) agreement. The MAD agreement allows OECD member countries to mutually accept data that comply with OECD Test Guidelines and Principles of Good Laboratory Practice.

 

Sensitization Testing

16 CFR § 1500.3(b)(9) and 16 CFR § 1500.3(c)(5)(i) define the term strong sensitizer. The supplemental definition at 16 CFR § 1500.3(c)(5)(ii) references testing on humans and animals, as well as indicates using other types of data to support the determination of a substance as a strong sensitizer.


Alternative methods for testing sensitization that have been approved by the Commission include:



Commission Votes and other CPSC Approvals Pertinent to Alternatives to Animal Testing

The following are Commission votes or approvals made on behalf of the Commission, with respect to animal testing policy:

 

List of Commission Votes or CPSC Approvals Pertinent to the Alternatives to Animal Testing Policy
DateVote or Approval ItemOutcome
08/29/2003 Recommended response to ICCVAM on acute toxicity testing: (1) the revised Up-and-Down Procedure (UDP) for determining acute oral toxicity for the purpose of classification and labeling, and (2) in vitro methods for determining the starting dose for acute systemic toxicity testing. Accepted
01/29/2008 Staff recommendation on response to ICCVAM on four in vitro ocular toxicity test methods: (1) the isolated rabbit eye (IRE) test; (2) the isolated chicken eye (ICE) test; (3) the bovine corneal opacity and permeability (BCOP) test; and (4) the hen's egg test - chorioallantoic (HET-CAM). Accepted
08/28/2008 Staff recommendation on response to ICCVAM on the use of in vitro basal cytotoxicity test methods for estimating starting doses for acute oral systemic toxicity testing. Accepted
01/05/2009 Staff response to ICCVAM recommendations for five in vitro methods for assessing pyrogenicity of pharmaceuticals and other products. Determined irrelevant to consumer products/ CPSC mission
03/09/2010 Staff recommendation on response to ICCVAM revisions to the murine local lymph node assay: (1) updates to the test method protocol; (2) establishment of performance standards; and (3) a modified form of the assay, the reduced Local Lymph Node Assay (rLLNA). Accepted
01/26/2011 Staff response to ICCVAM recommendations on revisions to the murine local lymph node assay: (1) the Bromodeoxyuridine Enzyme-linked Immunosorbent Assay (BrdU-ELISA); (2) the Daicel Chemical Industries version (LLNA:DA); and (3) an update on the LLNA's applicability domain, particularly its effectiveness in testing pesticide formulations, metals, and substances in aqueous solutions. Accepted
03/02/2011 Staff response to ICCVAM recommendations on four test method evaluation reports regarding ocular toxicity testing: (1) the routine use of topical anesthetics, systemic analgesics, and humane endpoints to avoid or minimize pain and distress in ocular safety testing; (2) the current validation status of five in vitro test methods proposed for identifying eye injury hazard potential of chemicals and products; (3) the discontinuation of the use of the low volume eye test for ocular safety testing; and (4) the current validation status of a proposed in vitro testing strategy for the U.S. Environmental Protection Agency ocular hazard classification and labeling of antimicrobial cleaning products. 3 of 4 Accepted; one not considered because determined irrelevant to CPSC mission
07/23/2012 CPSC Staff Response to the ICCVAM Test Method Recommendations on the Usefulness and Limitations of the LUMI-CELL® Estrogen Receptor (BG1LucERTA) Screening Assay: A Test Method for the Identification of Chemical Substances with In Vitro Agonist (Activation) or Antagonist (Inactivation) Activity to the Human Estrogen Receptor. Accepted