ASTM D6081 Standard Practice for Aquatic Toxicity Testing of Lubricants
ASTM D6081 Standard Practice for Aquatic Toxicity Testing of Lubricants: Sample Preparation and Results Interpretation
1. Scope
1.1 This practice covers procedures to be used in the preparation of lubricants or their components for toxicity testing in aquatic systems and in the interpretation of the results of such tests.

1.2 This practice is suitable for use on fully-formulated lubricants or their components which are not completely soluble at the intended test treat rates. It is also suitable for use with additives, if the additive is tested after being blended into a carrier fluid at the approximate concentration as in the intended fully-formulated lubricant. The carrier fluid shall meet the above solubility criterion, be known to be minimally toxic in the toxicity test in which the material will be tested, and be known to have a chemical composition similar to the rest of the intended fully-formulated lubricant.

1.3 Samples prepared in accordance with this practice may be used in acute or chronic aquatic toxicity tests conducted in fresh water or salt water with fish, large invertebrates, or algae. This practice does not address preparation of samples for plant toxicity testing other than algae.

1.4 Standard acute and chronic aquatic toxicity procedures are more appropriate for lubricants with compositions which are completely soluble at the intended test treat rates (1,2,3,4,5).

1.5 This practice is intended for use with lubricants or lubricant components of any volatility.

1.6 This practice does not address any questions regarding the effects of any lubricant or lubricant component on human health.

1.7 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.

2. Referenced Documents
2.1 ASTM Standards:
D1129 Terminology Relating to Water
D1193 Specification for Reagent Water
E 729 Guide for Conducting Acute Tests on Test Materials with Fishes, Macroinvertebrates, and Amphibians
E 943 Terminology Relating to Biological Effects and Environmental Fate

3. Terminology
3.1 Definitions:
3.1.1 acute toxicity test, n - a comparative toxicity test in which a representative subpopulation of organisms is exposed to different treat rates of a test material and is observed for a short period usually not constituting a substantial portion of their life span.

3.1.2 chronic toxicity test, n - a comparative toxicity test in which a representative subpopulation of organisms are exposed to different treat rates of a test material and is observed for a period of time which constitutes a major portion of their life span.

3.1.3 ecotoxicity, n - propensity of a test material to produce adverse behavioral, biochemical, or physiological effects in non-human organisms or populations.

3.1.4 effect load XX (ELXX), n - a statistically or graphically estimated loading rate of test material that is expected to cause one or more specified effects in XX % of a representative subpopulation of organisms under specified conditions.
3.1.4.1 Discussion - This terminology should be used instead of the standard ECXX when the test material is not completely soluble at the test treat rates.

3.1.5 inhibition load XX (ILXX), n - a statistically or graphically estimated loading rate of test material that is expected to cause a XX % inhibition of a biological process (such as growth or reproduction) which has an analog as opposed to a digital measure.
3.1.5.1 Discussion - An example of a digital measure would be alive/dead. This terminology (ILXX) should be used instead of the standard ICXX when the test material is not completely soluble at the test treat rates.

3.1.6 lethal load XX (LLXX), n - a statistically or graphically estimated loading rate of test material that is expected to be lethal to XX % of a representative subpopulation of organisms under specified conditions.
3.1.6.1 Discussion - This terminology should be used instead of the standard LCXX when the material is not completely soluble at the test treat rates.

3.1.7 loading rate, n - the ratio of test material to aqueous medium used in the preparation of WAF, WSF, or mechanical dispersion and in the interpretation of the results of a toxicity study with a poorly water-soluble lubricant or lubricant component.

3.1.8 mechanical dispersion, n - a low energy aqueous medium produced by continuous stirring of the test solution and containing both dissolved and undissolved components of the test material.

3.1.9 toxicity, n - the propensity of a test material to produce adverse behavioral, biochemical, or physiological effects in a living organism.

3.1.10 water accommodated fraction (WAF), n - the predominantly aqueous portion of a mixture of water and a material poorly soluble in water which separates in a specified period of time after the mixture has undergone a specified degree of mixing and which includes water, dissolved components, and dispersed droplets of the poorly soluble material.
3.1.10.1 Discussion - The composition of the WAF depends on the ratio of poorly soluble material to water in the original mixture as well as on the details of the mixing procedure.

3.1.11 water soluble fraction (WSF), n - the filtrate or centrifugate of the water accommodated fraction which includes all parts of the WAF, except the dispersed droplets of the poorly soluble material.

3.2 Definitions of Terms Specific to This Standard:
3.2.1 dilution water, n - the water used in the aquatic test under consideration which may contain organic or inorganic components depending on the test requirements.

3.2.2 exposure matrix, n - the WAF, WSF, or mechanical dispersion to which test organisms are exposed for toxicity testing.

3.2.3 test systems, n - the test organisms plus the exposure matrix and the test chambers themselves.

3.2.4 water, n - a reagent which is defined by Type II of Specification D1193.

3.3 Definitions of additional terms used in this practice can be found in Terminologies D1129 and E 943.