§79.60. Good laboratory practices (GLP) standards for inhalation exposure health effects testing. — Inbound Citations

(2) Laboratory facilities shall perform testing in compliance with Good Laboratory Practice (GLP) requirements as those requirements apply to inhalation toxicology studies. All studies shall be monitored by the facilities' Quality Assurance units (as specified in § 79.60).

(a) Subject to the provisions of § 79.53(b) through (d), the combustion emissions of each fuel or fuel additive subject to testing under this subpart must be tested in accordance with each of the testing guidelines in §§ 79.60 through 79.68, except that fuels and additives in the methane and propane fuel families (pursuant to § 79.56(e)(1)(v) and (vi)) need not undergo the Salmonella mutagenicity assay in § 79.68). Similarly, subject to the provisions of § 79.53(b) through (d), the evaporative emissions of each designated evaporative fuel and each designated evaporative fuel additive subject to testing under this subpart must be tested according to each of the testing guidelines in §§ 79.60 through 79.67 (excluding § 79.68, Salmonella typhimurium Reverse Mutation Assay).

(C) The testing facility shall allow an audit of its premises, the qualifications, e.g., curriculum vitae, of its staff assigned to testing, and the specimens and records of the testing for registration purposes (as specified in § 79.60).

(ii) Summary of procedures (including quality assurance, quality control and compliance with Good Laboratory Practice Standards as specified in § 79.60), findings, and conclusions, including references if used to support such results and conclusions,

(i) All data developed within this study shall be in accordance with good laboratory practice provisions under § 79.60.

(d) If a test at one dose level of the highest concentration that can be achieved while maintaining a particle size distribution with a mass median aerodynamic diameter (MMAD) of 4 micrometers (µm) or less, using the procedures described in section 79.60 of this part produces no observable toxic effects and if toxicity would not be expected based upon data of structurally related compounds, then a full study using three dose levels might not be necessary. Expected human exposure though may indicate the need for a higher dose level.

(i) All data developed within this study shall be in accordance with good laboratory practice provisions under § 79.60.

(i) All data developed within this study shall be in accordance with good laboratory practice provisions under § 79.60.

(i) All data developed within this study shall be in accordance with good laboratory practice provisions under § 79.60.

(i) All data developed within this study shall be in accordance with good laboratory practice provisions under § 79.60.

(e) All data developed within this study shall be in accordance with good laboratory practice provisions under § 79.60.

(1) When testing with metabolic activation, test solution, bacteria, and 0.5 ml of activation mixture containing an adequate amount of postmitochondrial fraction shall be added to the liquid overlay agar and mixed. This mixture is poured over the surface of a selective agar plate. Overlay agar shall be allowed to solidify before incubation. At the end of the incubation period, revertant colonies per plate shall be counted. When testing without metabolic activation, the test sample and 0.1 ml of a fresh bacterial culture shall be added to 2.0 ml of overlay agar.

(2) When testing with metabolic activation, 0.5 ml of activation mixture containing 150 µl of postmitochondrial fraction and 0.1 ml of bacterial culture shall be added to a test tube kept on ice. 0.1 ml of test solution shall be added, and the tubes shall be incubated with shaking at 30 °C for 30 minutes. At the end of the incubation period, 2.0 ml of agar shall be added to each tube, the contents mixed and poured over the surface of a selective agar plate. Overlay agar shall be allowed to solidify before incubation. At the end of the incubation period, revertant colonies per plate shall be counted. For tests without metabolic activation, 0.5 ml of buffer shall be used in place of the 0.5 ml of activation mixture. All other procedures shall be the same as those used for the test with metabolic activation.

(3) Other methods/modifications may also be appropriate.

(4) An appropriate selective medium with an adequate overlay agar shall be used.

(5) All plates within a given experiment shall be incubated for the same time period. This incubation period shall be for 48-72 hours at 37 °C.

(6) All plating shall be done at least in triplicate.

(B) Data or information submitted to EPA under part 79, including the statement required by § 79.60(a)(4), include information or data that are false or misleading, contain significant omissions, or otherwise do not fulfill the requirements of this part; or

(C) Entry in accordance with § 79.60(a)(5) for the purpose of auditing test data is denied.

(v) Summaries of training and experience and job descriptions required to be maintained by § 79.60(b)(1)(ii) may be retained along with all other testing facility employment records for the length of time specified in paragraph (h)(3)(ii) of this section.

(i) Designate a study director as described in § 79.60(b)(3) before the study is initiated.

(iii) Assure that there is a quality assurance unit as described in § 79.60(b)(4).

(N) The statement, prepared and signed by the quality assurance unit, as described in § 79.60(b)(4)(i)(G).

(iv) The master schedule sheet, copies of protocols, and records of quality assurance inspections, as required by § 79.60(b)(4)(iii) shall be maintained by the quality assurance unit as an easily accessible system of records for the period of time specified in paragraph (h)(3)(ii) of this section.

(vi) Records and reports of the maintenance and calibration and inspection of equipment, as required by § 79.60(d)(2) (ii) and (iii), shall be retained for the length of time specified in paragraph (h)(3)(ii) of this section.

(ii) The written standard operating procedures required under § 79.60(e)(1)(ii)(K) shall set forth in sufficient detail the methods, materials, and schedules to be used in the routine inspection, cleaning, maintenance, testing, calibration, and/or standardization of equipment, and shall specify, when appropriate, remedial action to be taken in the event of failure or malfunction of equipment. The written standard operating procedures shall designate the person responsible for the performance of each operation.

(2) As used in this section, the following terms shall have the meanings specified:

Batch means a specific quantity or lot of a test fuel, additive/base fuel mixture, or reference substance that has been characterized according to § 79.60(f)(1)(i).

CAA means the Clean Air Act.

Carrier means any material which is combined with engine/motor vehicle emissions or a reference substance for administration to a test system. “Carrier” includes, but is not limited to, clean, filtered air, water, feed, and nutrient media.

Control atmosphere means clean, filtered air which is administered to the test system in the course of a study for the purpose of establishing a basis for comparison with the test atmosphere for chemical or biological measurements.

Experimental start date means the first date the test atmosphere is applied to the test system.

Experimental termination date means the last date on which data are collected directly from the study.

Person includes an individual, partnership, corporation, association, scientific or academic establishment, government agency, or organizational unit thereof, and any other legal entity.

Quality assurance unit means any person or organizational element, except the study director, designated by testing facility management to perform the duties relating to quality assurance of the studies.

Raw data means any laboratory worksheets, records, memoranda, notes, or exact copies thereof, that are the result of original observations and activities of a study and are necessary for the reconstruction and evaluation of the report of that study. In the event that exact transcripts of raw data have been prepared (e.g., tapes which have been transcribed verbatim, dated, and verified accurate by signature), the exact copy or exact transcript may be substituted for the original source as raw data. “Raw data” may include photographs, videotape, microfilm or microfiche copies, computer printouts, magnetic media, including dictated observations, and recorded data from automated instruments.

Reference substance means any chemical substance or mixture, analytical standard, or material other than engine/motor vehicle emissions and/or its carrier, that is administered to or used in analyzing the test system in the course of a study. A “reference substance” is used to establish a basis for comparison with the test atmosphere for known chemical or biological measurements, i.e., positive or negative control substance.

Specimen means any material derived from a test system for examination or analysis.

Sponsor means person who initiates and supports, by provision of financial or other resources, a study or a person who submits a study to EPA in response to the CAA Section 211(b) or 211(e) Fuels and Fuel Additives Registration Rule or a testing facility, if it both initiates and actually conducts the study.

Study means any experiment, at one or more test sites, in which a test system is exposed to a test atmosphere under laboratory conditions to determine or help predict the health effects of that exposure in humans, other living organisms, or media.

Study completion date means the date the final report is signed by the study director.

Study director means the individual responsible for the overall conduct of a study.

Study initiation date means the date the protocol is signed by the study director.

Test substance means a vapor and/or aerosol mixture composed of engine/motor vehicle emissions and clean, filtered air which is administered directly, or indirectly, by the inhalation route to a test system in a study which develops data to meet the registration requirements of CAA section 211(b) or (e).

Test system means any animal, microorganism, chemical or physical matrix, to which the test, control, or reference substance is administered or added for study. This definition also includes appropriate groups or components of the system not treated with the test, control, or reference substance.

Testing facility means a person who actually conducts a study, i.e., actually uses the test substance in a test system. “Testing facility” encompasses only those operational units that are being or have been used to conduct studies.

TSCA means the Toxic Substances Control Act (15 U.S.C. 2601 et seq.).

(i) The protocol, including any changes, is approved as provided by § 79.60(g)(1)(i) and is followed;