[Federal Register Volume 69, Number 201 (Tuesday, October 19, 2004)]
[Notices]
[Pages 61504-61505]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 04-23335]


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DEPARTMENT OF HEALTH AND HUMAN SERVICES

Public Health Service


National Institute of Environmental Health Sciences (NIEHS); 
National Toxicology Program (NTP); NTP Interagency Center for the 
Evaluation of Alternative Toxicological Methods (NICEATM): Availability 
of Updated Standardized In Vitro Cytotoxicity Test Method Protocols for 
Estimating Acute Oral Systemic Toxicity; Request for Existing In Vivo 
and In Vitro Acute Toxicity Data

    Summary: NICEATM announces the availability of two updated 
standardized in vitro cytotoxicity test method protocols to estimate 
acute oral systemic toxicity in rodents. These two test methods were 
previously recommended by the Interagency Coordinating Committee on the 
Validation of Alternative Methods (ICCVAM) for selecting starting doses 
for in vivo acute oral systemic toxicity tests (Federal Register Vol. 
66, No. 189, pages 49686-49687, September 28, 2001). This approach can 
reduce the number of animals required for acute oral toxicity testing. 
NICEATM also requests the submission of existing and future data on 
chemicals and products tested for both acute oral systemic toxicity and 
in vitro cytotoxicity using the standardized test method protocols 
mentioned in this notice. These data will be used to further evaluate 
the usefulness and limitations of cytotoxicity methods for estimating 
in vivo acute oral toxicity. The data will also be used to establish a 
database to support the investigation of other test methods necessary 
to improve the accuracy of in vitro assessments of acute systemic 
toxicity.

Availability of Standardized Test Method Protocols for Estimating 
Starting Doses for In Vivo Acute Oral Toxicity Tests

    Updated standardized protocols for two neutral red uptake assays 
using either BALB/c 3T3 cells or normal human keratinocytes are now 
available at: http://iccvam.niehs.nih.gov/methods/invitro.htm. These 
test method protocols have been improved to maximize intra- and inter-
laboratory reproducibility and are currently being used for the final 
phase of a joint NICEATM-European Center for the Validation of 
Alternative Methods (ECVAM) validation study. NICEATM recommends that 
these updated test method protocols be used in place of standard 
operating procedures previously recommended by ICCVAM for two 
cytotoxicity test methods to estimate starting doses for in vivo acute 
oral toxicity tests (ICCVAM, 2001b).

Submission of Chemical and Protocol Information/Test Data

    In vivo and in vitro acute toxicity testing data for chemicals or 
products should be sent by mail, fax or e-mail to NICEATM [Dr. William 
S. Stokes, Director, NICEATM, NIEHS, PO Box 12233, MD EC-17, Research 
Triangle Park, NC 27709, (phone) 919-541-2384, (fax) 919-541-0947, (e-
mail) [email protected]]. Data will be accepted at any time. Data 
submitted within the next 9 months will be considered during an 
evaluation of the validation status of the two cytotoxicity methods 
anticipated in late 2005. Chemical and protocol information/test data 
submitted in response to this notice may be incorporated in future 
NICEATM and ICCVAM reports and publications as appropriate.
    When submitting chemical and protocol information/test data, please 
reference this Federal Register notice and provide appropriate contact 
information (name, affiliation, mailing address, phone, fax, e-mail, 
and sponsoring organization, as applicable).
    NICEATM prefers data to be submitted as copies of pages from study 
notebooks and/or study reports, if available. Raw data and analyses 
available in electronic format may also be submitted. Each submission 
for a chemical should preferably include the following information, as 
appropriate:
     Common and trade name
     Chemical Abstracts Service Registry Number (CASRN)
     Chemical and/or product class
     Commercial source
     In vitro basal cytotoxicity test protocol used
     In vitro cytotoxicity test results
     In vivo acute oral toxicity test protocol used
     Individual animal responses at each observation time (if 
available)
     The extent to which the study complied with national or 
international Good Laboratory Practice (GLP) guidelines
     Date and testing organization
    Those persons submitting data on chemicals tested for in vitro 
basal cytotoxicity are referred to the standard test-reporting template 
recommended for the High Production Volume (HPV) program at http://www.epa.gov/chemrtk/toxprtow.htm or at http://iccvam.niehs.nih.gov/methods/invitro.htm. In vivo data for the same chemicals should be 
reported as recommended in the test reporting section of the current 
Environmental Protection Agency (EPA) guideline for acute oral toxicity 
(EPA, 2002).
    Submitted data will be used to further evaluate the usefulness and 
limitations of in vitro cytotoxicity data for estimating acute oral 
toxicity, and will be included in a database to support the 
investigation of other test methods necessary to improve the accuracy 
of in vitro assessments of acute systemic toxicity.

History

    In September 2001, the ICCVAM recommended that in vitro 
cytotoxicity test methods be considered as a tool for estimating 
starting doses for in vivo acute systemic toxicity testing studies 
(Federal Register Vol. 66, No. 189, pages 49686-49687, September 28, 
2001.) The recommendations were based on the Report of the 
International Workshop on In Vitro Methods for Assessing Acute Systemic 
Toxicity (ICCVAM, 2001a). The Guidance Document on Using In Vitro Data 
to Estimate In Vivo Starting Doses for Acute Toxicity (ICCVAM, 2001b) 
was

[[Page 61505]]

also made available at that time. The guidance document provided 
standard operating procedures for two cytotoxicity test methods and 
instructions for using these assays to estimate starting doses for in 
vivo testing.
    Federal agency responses to the ICCVAM test method recommendations 
were announced on March 10, 2004 (Federal Register Vol. 69, No. 47, 
pages 11448-11449). Federal agencies agreed to encourage, to the extent 
applicable, the use of in vitro tests for determining starting doses 
for acute systemic toxicity testing. Furthermore, EPA specifically 
encouraged those participating in the HPV Challenge Program to consider 
using the recommended in vitro tests as a supplemental component in 
conducting any new in vivo acute oral toxicity studies for the program 
(http://www.epa.gov/chemrtk/toxprtow.htm).
    A NICEATM-ECVAM validation study was initiated in 2002 to evaluate 
the usefulness of the two neutral red uptake cytotoxicity assays 
currently available for predicting starting doses for in vivo acute 
oral toxicity tests. During the pre-validation phases of the study, the 
test method protocols were further standardized and revised to improve 
their intra- and inter-laboratory reproducibility. NICEATM recommends 
using the revised test method protocols rather than the standard 
operating procedures outlined in the guidance document (ICCVAM, 2001b.) 
The guidance document should be consulted for the procedure for 
calculating starting doses using in vitro cytotoxicity data.

Background Information on ICCVAM and NICEATM

    ICCVAM is an interagency committee composed of representatives from 
fifteen Federal regulatory and research agencies that use, generate, or 
disseminate toxicological information. ICCVAM promotes the development, 
validation, regulatory acceptance, and national and international 
harmonization of toxicological test methods that more accurately assess 
the safety or hazards of chemicals and products, and test methods that 
refine, reduce and replace animal use. The ICCVAM Authorization Act of 
2000 (available at http://iccvam.niehs.nih.gov/about/PL106545.htm) 
established ICCVAM as a permanent interagency committee of the NIEHS 
under the NICEATM. NICEATM administers the ICCVAM and provides 
scientific support for ICCVAM and ICCVAM-related activities. NICEATM 
and ICCVAM work collaboratively to evaluate new and improved test 
methods applicable to the needs of Federal agencies. Additional 
information about ICCVAM and NICEATM can be found at the following Web 
site: http://iccvam.niehs.nih.gov/.

References

    EPA. 2002. Health Effects Test Guidelines, OPPTS 870.1100, Acute 
Oral Toxicity, EPA 712-C-02-190. Available at: http://www.epa.gov/opptsfrs/OPPTS_Harmonized/870_Health_Effects_Test_Guidelines/Series/870-1100.pdf.
    ICCVAM (Interagency Coordinating Committee on the Validation of 
Alternative Methods). 2001a. Report of the international workshop on 
in vitro methods for assessing acute systemic toxicity. NIH 
Publication 01-4499. Research Triangle Park, NC: National Institute 
for Environmental Health Sciences. Available at: http://iccvam.niehs.nih.gov/.

    ICCVAM. 2001b. Guidance document on using in vitro data to 
estimate in vivo starting doses for acute toxicity. NIH Publication 
01-4500. Research Triangle Park, NC: National Institute for 
Environmental Health Sciences. Available at: http://iccvam.niehs.nih.gov/.
    Dated: October 6, 2004.
Samuel H. Wilson,
Deputy Director, National Institute of Environmental Health Sciences.
[FR Doc. 04-23335 Filed 10-18-04; 8:45 am]
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