Results from the test method suggest, within the confines of a controlled laboratory setting, the degree of aerobic aquatic biodegradation of a lubricant or components of a lubricant by measuring the evolved carbon dioxide upon exposure of the test material to an inoculum. The plateau level of CO₂ evolution in this test method will suggest the degree of biodegradability of the lubricant. Test substances that achieve a high degree of biodegradation in this test may be assumed to easily biodegrade in many aerobic aquatic environments.

Because of the stringency of this test, a low yield of CO₂ does not necessarily mean that the test substance is not biodegradable under environmental conditions, but indicates that further testing is necessary to establish biodegradability.

Information on toxicity to the inoculum of the test substance may be useful in the interpretation of low biodegradation results.

Activated sewage-sludge from a sewage-treatment plant that principally treats domestic waste is considered an acceptable active aerobic inoculum available over a wide geographical area in which to test a broad range of lubricants. An inoculum derived from soil or natural surface waters, or both, or any combination of the three sources, is also appropriate for this test method.

Note 1—Allowance for various and multiple inoculum sources provides access to a greater diversity of biochemical competency and potentially represents more accurately the capacity for biodegradation.

A reference or control substance known to biodegrade is necessary in order to verify the activity of the inoculum. The test must be regarded as invalid and should be repeated using a fresh inoculum if the reference does not demonstrate a biodegradation of > 60 % of the theoretical CO₂ evolution within 28 days.

A total CO₂ evolution in the blank at the end of the test exceeding 75 mg CO₂ per 3 L of medium shall be considered as invalidating the test.

The water solubility or dispersibility of the lubricant or component may influence the results obtained and hence the procedure may be limited to comparing lubricants or components with similar solubilities.

The ratio of carbon incorporated into cellular material to carbon released as CO₂ will vary depending on the organic substrate, on the particular microorganisms carrying out the conversion, and on the environmental conditions under which the conversion takes place. In principle, this variability complicates the interpretation of the results from this test method.

1.1 This test method covers the determination of the degree of aerobic aquatic biodegradation of fully formulated lubricants or their components on exposure to an inoculum under laboratory conditions.

1.2 This test method is intended to specifically address the difficulties associated with testing water insoluble materials and complex mixtures such as are found in many lubricants.

1.3 This test method is designed to be applicable to all lubricants that are not volatile and are not inhibitory at the test concentration to the organisms present in the inoculum.

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. Specific hazards are discussed in Section 10.