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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report date:
2006

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
yes
Remarks:
(the temperature of the waterbath was not maintained between 19 and 21 degrees C)
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
1,1,3,3-tetramethylbutylamine
EC Number:
203-491-1
EC Name:
1,1,3,3-tetramethylbutylamine
Cas Number:
107-45-9
Molecular formula:
C8H19N
IUPAC Name:
2,4,4-trimethylpentan-2-amine
Details on test material:
- Name of test material (as cited in study report): Primene(TM) TOA
- Physical state: liquid
- Analytical purity: 97.45%
- Lot/batch No.: 2377627

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: all test concentrations
- Sampling method: For each treatment sample, a 15-mL sample was collected and centrifuged for 15 minutes. Following centrifugation, a 9-mL aliquot was removed and placed in a separate 15-mL culture tube. One drop of 50% NaOH was added to the culture tube to adjust the pH to basic. Three milliliters of methylene chloride was added to the culture tube and vortexed for approximately one minute. The sample was centrifuged for two minutes. The methylene chloride layer was removed following centrifugation and used for analysis.
- Sample storage conditions before analysis: no data

Test solutions

Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A 0.10-mg whole productImL primary standard was prepared by weighmg 0.2000 g of the Primene TOA into a 2,000 mL glass volumetric flask and bringing to volume with dilution water. Aliquots of the primary standard were then appropriately diluted with 1.0 L of dilution water to obtain nominal concentrations of 3.3, 6.5, 13, 25, 50, and 100 mg whole product/L.
- Controls: dilution water only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dilution water
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Strain: magna
- Source: in-house culture
- Age at study initiation (mean and range, SD): <24 hours old

- Method of breeding: All daphnids were cultured in a temperature-controlled area at approximately 20 deg C. One day prior to neonate selection, the adult daphnids were isolated, by transferring the adults to a ftesh culture with a waterlfood suspension. The adults were considered acceptable with no signs of stress, disease or physical damage. No adult mortality occurred during the 48-hour period immediately prior to production of neonates used in test initiation.
- Feeding during test: no
- Food type: suspension of the algal species Pseudokirchneriella subcapitata supplemented by an artificial diet
- Frequency: once daily

ACCLIMATION
- Acclimation period: Since the culturing and testing environmental parameters were equivalent (i.e. temperature, dilution water, and lighting), no acclimation period was necessary.
- Acclimation conditions (same as test or not): yes

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
144 mg CaCO3/L
Test temperature:
19.4 to 19.6 degC
pH:
8.3 to 9.4
Dissolved oxygen:
8.4 to 9.0 mg/L
Nominal and measured concentrations:
3.3,6.5, 13,25, 50, and 100 mg whole product/L nominal
Details on test conditions:
TEST SYSTEM
- Test vessel: 250-mL glass beakers
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 250-mL glass beakers
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: aged laboratory freshwater prepared by blending naturally hard well water with well water that was demineralized by reverse osmosis. The water was passed through a sediment filter and UV sterilizer prior to use.
- Alkalinity: 152 mg CaCO3
- Conductivity: 324 uS
- Culture medium different from test medium: no
- Intervals of water quality measurement: at test initiation; If 100-percent mortality was observed in a test concentration, water quality data was recorded at that time; temperature was measured daily

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16-hour daylight photoperiod with 30-minute simulated dawn and dusk periods
- Light intensity: 563 lux.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : observations for immobility and sublethal responses were made at approximately 24 and 48 hours (+/- 1 hour of test initiation).

TEST CONCENTRATIONS
- Range finding study 48-hour static range-finding test
- Test concentrations: 0 (control), 0.010, 0.10, 1.0, 10, and 100 mg whole product/L
- Results used to determine the conditions for the definitive study: The control and all test solutions were clear and colorless with no visible precipitate, particulate matter, or surface film. After 48 hours, immobility was 0, 0, 0, 0, 10, and 100% in the 0 (control), 0.010, 0.10, 1.0, 10, and 100 mg whole product/L treatments, respectively.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
9.49 mg/L
Nominal / measured:
estimated
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95 CL 7.05 to 12.5 mg/L
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
3.26 mg/L
Nominal / measured:
meas. (not specified)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Behavioural abnormalities: Of the 13 surviving daphnids in the 11.2 mg whole product/L, treatment, five daphnids were observed as quiescent. Of the four surviving daphnids in the 22.9 mg whole product/L, treatment, three daphnids were observed to be quiescent.
- Observations on body length and weight: no data
- Other biological observations: immobility (mortality) was 15, 45, 35, 80, 100, and 100% in 3.26, 6.08, 11.2, 22.9, 43.2, and 91.2 mg whole product/L treatments, respectively
- Mortality of control: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
All statistical analyses were performed with SAS software. Estimates of ECso values and their 95% confidence limits were calculated using the probit method and Trimmed Spearman-Karber method. When the P value for Goodness of Fit was >0.05 and there was no other evidence of questionable convergence, the probit method was selected for reporting. When this criterion was not achieved, the Trimmed Spearman-Karber method was selected for reporting. The no observed-effect concentration (NOEC) was determined by using a Fisher's exact test. A Hochberg adjustment was used to control the experiment wise error rate for the Fisher's test at the same alpha level.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The 24-hour EC50 value for Daphnia magna exposed to the test substance is estimated to be 38.2 mg whole product/L. The 48 hour EC50 for Daphnia magna exposed to the test substance is estimated to be 9.49 mg whole product/L. The slope of the 48-hour concentration-response line was 2.4. The 48-hour no-observed-effect concentration (NOEC) was 3.26 mg whole product/L, based on the lack of statistically significant immobility and sublethal effects at this concentration.
Executive summary:

A test was conducted to estimate the potential acute toxicity of PrimeneTMTOA (CAS 107-45-9) to the water flea, Daphnia magna. Daphnids were exposed for 48 hours under static conditions to nominal concentrations of 0 (control), 3.3, 6.5, 13,25, 50, and 100 mg whole product/L of the test substance.

Test treatments were prepared on a weight/volume basis based on whole product. Mean measured concentrations were <MQL (control), 3.26, 6.08, 11.2, 22.9, 43.2, and 91.2 mg whole product/L, which represent 99, 94, 86, 92 , 86, and 91% of the nominal concentrations, respectively. Water quality characteristics of dissolved oxygen concentration, temperature, and pH were within acceptable limits throughout the exposure. The controls and all treatment solutions were clear and colorless with no visible particulates, surface film, undissolved test substance, or precipitate throughout the exposure.

After 48 hours, immobility (mortality) was 0, 15, 45, 35, 80,100, and 100% in <MQL (control), 3.26, 6.08, 11.2, 22.9, 43.2, and 91.2 mg whole product/L treatments, respectively. Of the 13 surviving daphnids in the 11.2 mg whole product/L treatment, five daphnids were observed as quiescent. Of the four surviving daphnids in the 22.9 mg whole product/L, treatment, three daphnids were observed to be quiescent. The quiescent animals could weakly swim after gentle perturbation and thus were not considered to be immobile. The 24-hour EC50 value for Daphnia magna exposed to the test substance is estimated to be 38.2 mg whole product/L

(95% confidence limits: 30.3 to 48.2 mg whole product/L). The 48-hour EC50 value for Daphnia magna exposed to the test substance is estimated to be 9.49 mg whole product/L, (95% confidence limits: 7.05 to 12.5 mg whole product/L). The slope of the 48-hour concentration-response line was 2.4. The 48-hour no-observed-effect concentration (NOEC) was 3.26 mg whole product/L, based on the lack of statistically significant immobility and sublethal effects at this concentration.