Phenol, paraalkylation products with C10-15 branched olefins ( C12 rich) derived from propene oligomerization, calcium salts, sulfurized, including distillates (petroleum), hydrotreated, solvent-refined, solvent-dewaxed, or catalyc dewaxed, light or heavy paraffinic C15-C50

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study Initiation Date:October 2, 1996 Study Completion Date: January 15, 1997. In life dates November 15-17 1996.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study following GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: This definitive test used a dilution water control and the WAF of five concentrations of the test material: 130, 220, 360, 600, and 1,000 mg/L.
- Sampling method: Samples were collected in duplicate from the control and the highest and lowest test concentrations prior to distribution to test vessels and from each of the two control replicates and the two replicates of the highest and lowest test concentrations at the end of the test.
- Sample storage conditions before analysis: Prior to use the test substance was stored at room temperature.

Vehicle:

not specified

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
Each of the five WAFs was prepared by combining the appropriate amount of test substance and dilution water in a mixing vessel equipped with a small magnetic stirrer (the vortex extended from the surface approximately 25% of the way to the bottom of the mixing vessel), stirring these mixtures for approximately 20 hours, settling the mixtures for approximately four hours, and siphoning the water phase containing the WAF.

Test organisms (species):

Daphnia magna

Details on test organisms:

Test species: Juvenile daphnids less than 24-hours old were produced from laboratory in-house culture.
TEST ORGANISM
- Common name: Daphnia magna
- Source: in-house culture, the original parental culture was procured from Aquatic Research Organisms, Hampton, New Hampshire on October 11, 1996
- Age at study initiation (mean and range, SD): less than 24 hours old
- Feeding during test: Daphnids were not fed during the test.

ACCLIMATION
- Acclimation period: During the 14 day period before the start of the test the culture temperature was 19.0 to 20.9°C and the dissolved oxygen concentration was always at least 7.3 mg/L. A subsample of daphnids from this culture produced at least 3 young/female/day during the seven day period preceding the definitive toxicity test.
- Type and amount of food: Daphnid cultures were supplied with the freshwater alga, Selenastrum capricornutum, and a yeast/trout chow mixture daily throughout the acclimation period.
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no mortality was observed during the 48 hours preceding the start of the toxicity test. During acclimation daphnids were not treated for disease and they were free of apparent sickness, injuries, and abnormalities at the beginning of the test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

Water used for the test had a hardness of 160-164 mg/L as CaCO3 and an alkalinity of 122 mg/L as CaCO3, and it contained <0.01 mg/L residual chlorine.

Test temperature:

20.1 - 21.0 deg C

pH:

7.4 - 8.4

Dissolved oxygen:

7.4 - 8.5 mg/L

Nominal and measured concentrations:

Control, 130, 220, 360, 600, & 1,000 mg/L WAF loading rates

Details on test conditions:

TEST SYSTEM
- Test vessel: 300 ml glass beakers
- Type (delete if not applicable): loosely covered
- Material, size, headspace, fill volume: containing 250 ml of test solution
- Aeration: Aeration was not required to maintain acceptable dissolved oxygen concentrations.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Water used for acclimation of test organisms and for all toxicity testing was deionized tap water collected at Marblehead, Massachusetts. Water was adjusted to a hardness of 160 to 180 mg/L as CaC03 and a pH of <8 with hydrochloric acid. Water was stored in 500-gallon polyethylene tanks where it was aerated and continuously passed through a particle filter, ultraviolet sterilizer, and activated carbon. Water used for the definitive test had a hardness of 160 to 164 mg/L and an alkalinity of 122 mg/L as CaCO3, and it contained less than 0.01 mg/L residual chlorine.
- Conductivity: 600 umhos/cm.

OTHER TEST CONDITIONS
- Adjustment of pH: The pH of the dilution water was adjusted to <8 with hydrochloric acid to assure that the pH was acceptable for daphnid survival.
- Photoperiod: A 16 hour light and 8 hour dark photoperiod was automatically maintained with cool-white fluorescent lights
- Light intensity: ~6 µEin/m2sec

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
The number of surviving organisms and the presence of sublethal effects was determined visually and recorded initially and after 3, 24, and 48 hours.

TEST CONCENTRATIONS
- Range finding study:
A range-finding test was conducted from October 29 to 31, 1996 with a dilution water control and the WAF of three concentrations of the test material: 10, 100, and 1,000 mg/L. The range-finding test was conducted with a single replicate of each concentration, each containing ten organisms. No insoluble material was observed during the test. At the end of the test there was 100% survival in the control and at 10 and 100 mg/L, and 0% survival at 1,000 mg/L.
- Test concentrations: 10, 100, and 1,000 mg/L

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Observations on body length and weight: At the conclusion of the test the control daphnids had an average wet weight of 0.19 mg (loading rate = 7.6 mg/L).

Reported statistics and error estimates:

Results of the toxicity tests could not be interpreted by standard statistical techniques (Stephan, 1983) because at least 90% survival occurred in all tested concentrations. The no observed effect concentration is the highest concentration of test substance that allowed at least 90% survival of exposed organisms and did not cause sublethal effects.

Nominal concentrations: the 24 and 48-h EL50s (stated “EC50s” in report) >1,000 mg/L loading rate. The 24 and 48-h NOELR (stated as “NOECs” in report) = 1000 mg/L based on WAF loading rates.

Test Findings: 100% survival in the controls through 48 hours. At 24 hours, one dead or immobilized organism was noted for both 130 mg/L treatments and one of the 600 mg/L treatments. No additional immobilization or mortality was noted at 48 hours. Therefore, survival in all treatments was >90%.

Test results reported in original study as “effect concentrations” (EC) are reported in this summary as “effective loading” (EL), because test results are based on WAF loading rates and not measured concentrations. Similarly, the “no observed effect concentration (NOEC) in the report is presented as “no observed effect loading rate” (NOELR) in this summary.

Significant immobilization observed in preliminary tests at 1000 mg/L was likely due to insoluble material in the test vessels due to improper WAF preparation. WAF preparation for the final test did not exhibit insoluble material, and immobilization was not observed.

Validity criteria fulfilled:

yes

Conclusions:

The 24 and 48-h EL50s (stated “EC50s” in report) >1,000 mg/L loading rate. The 24 and 48-h NOELR (stated as “NOECs” in report) = 1000 mg/L based on WAF loading rates.

Executive summary:

The acute toxicity of the water accommodated fraction (WAF) of five mixtures of the test material and water to the daphnid, Daphnia magna, was investigated during a study conducted at T.R. Wilbury Laboratories, Inc. in accordance with OED guideline 202 under conditions of GLP. The test, which was designed to determine the toxicity of the test substance, was performed from November 13 to 15, 1996.

The test was performed at 20 ± 1°C under static conditions with a control and five nominal concentrations of test substance (130, 220, 360, 600, and 1,000 rng/L). The dilution water was carbon filtered deionized water collected at Marblehead, Massachusetts and adjusted to a hardness of 160 to 164 mg/L as CaCO₃ and a pH of <8.0. Water quality was within acceptable limits throughout the test and no insoluble material was noted during the test. Juvenile daphnids (less than 24 hours old) produced by an in-house culture that was maintained at test conditions were used to initiate the test.

Exposure of daphnids, Daphnia magna, to the WAFs resulted in a 48 hour median effective concentration (EC50) greater than 1,000 mg/L (the highest tested concentration), based on nominal concentrations. The 48 hour no observed effect concentration (NOEC) is 1,000 mg/L, based on nominal concentrations.

The 24 and 48-h EL50s (stated “EC50s” in report) >1,000 mg/L loading rate. The 24 and 48-h NOELR (stated as “NOECs” in report) = 1000 mg/L based on WAF loading rates.

Test results reported in original study as “effect concentrations” (EC) are reported in this summary as “effective loading” (EL), because test results are based on WAF loading rates and not measured concentrations. Similarly, the “no observed effect concentration (NOEC) in the report is presented as “no observed effect loading rate” (NOELR) in this summary.

Description of key information

In the key study (Ward, 1997), the acute toxicity of the water accommodated fraction (WAF) of the test material to the daphnid, Daphnia magna, was conducted according to the OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test). The results were 48h EL50 > 1,000 mg/L and a NOELR = 1,000 mg/L. The reliability rating for this study is 1, according to the criteria of Klimisch, 1997 and was chosen as the key study as it was the most recent study and is the most consistent with the results from the alkyl phenate sulfide category. Specifically, the supporting study (Ward, 1993) also had an LL50 > 1000 mg/L but a NOELR of 100 mg/L due to immobilization observed at doses of 300 and 1,000 mg/L. However, multiple category members, including EC 272-388-1 and EC 701-251-5, which bracket the registered substance, both had NOELRs of 1,000 mg/L. Therefore, the immobilization in the Ward (1993) study may have been due to insoluble test material. Regardless, all studies demonstrate that EC 701-249-4 does not require hazard classification for aquatic toxicity.
  
    
    
    

Regarding the study conducted under marine conditions (Douglas M.T. and Sewell I.G., 1988 study (HRC report number: CMA 2(1)/88925), it was considered to be unreliable. During the conduction of the study, the test solution is considered to have been improperly prepared in that test material was added to solution in amount that was far in excess of water solubility. Furthermore, the test substance formed a very poor dispersion in the water column and adhered to the mesh screens around the propeller shield and a small number of globules of the test substance were observed circulating in the water. As such the study has been assigned a reliability rating of 3, according to the criteria of Klimisch, 1997.











 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1 000 mg/L