Reaction products of N,N'-ethane-1,2-diylbis(1,3-propanediamine), cyclohexane, peroxidized 4-butylamino-2,2,6,6-tetramethylpiperidine and 2,4,6-trichloro-1,3,5-triazine

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:

1997-05-30 - 1998-01-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study (OECD test guideline 202)

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Test item concentrations were determined in the control and all test treatments at test initiation, at 24 hours (old and new solutions), and test termination. Water samples (100 mL percomposited test solution) were taken using volumetric pipets.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- The stock solution was prepared by sonicating a 100 mg/L solution for 15 minutes and then filtering the solution through a 0.2-µm filter to remove any undissolved test substance. These procedures produced a super-saturated solution approximately ten times greater than the natural solubility limit (under conditions of OECD 105) that appeared to be free of undissolved material. This solution was utilized as the maximum concentration tested and for preparing the four lower test concentrations (a 50% dilution series).A dilution water control was maintained concurrently with the test solutions.
- All test solutions were prepared by mixing the appropriate amount of prepared test substance stock solution with the appropriate amount of dilution water to obtain test solutions.
(- Evidence of undissolved material: All test chambers were filled with appropriate test solutions (without filtration) and allowed to sit overnight to permit the test substance to coat the glassware. These solutions were discarded when the test was initiated and fresh solutions were added).

RANGE-FINDING TEST
A static range-finding test was conducted at nominal concentrations of 0.10, 1.0, 10.0 and 100 mg/L prior to performing the main test.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: Toxikon Environmental Sciences in-house
- Subculture of adults was isolated from these cultures and maintained prior to testing
- Age at study initiation: Neonates, age of less than 24 hours
- Method of breeding: Test organisms fed with a suspension of green algae
- Feeding during test: None
- 24 hours prior to test initiation adults were re-isolated in food-free dilution water
- Daphnia magna were cultured and isolated in moderately hard freshwater at 19.9 to 20.5°C during the seven days preceding test initiation. No ephippia were produced during culture.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

80 mg/L CaCO3

Test temperature:

19.0 to 20.5°C

pH:

6.6 - 7.2

Dissolved oxygen:

Test solutions: 6.1 mg/L - 8.8 mg/L (dissolved oxygen concentrations remained above 60% of saturation in all test chambers throughout the test)
Control: 8.3 to 9.0 mg/L (88 to 98 % of saturation throughout the test)

Nominal and measured concentrations:

Nominal concentrations: 0, 6.25, 12.5, 25.0, 50.0, and 100 mg/L.
Mean measured concentrations: 0.0, 30.7, 57.9, 108, 226, and 312 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 400 mL glass beakers, covered
- Fill volume: 400 mL solution per beaker
- Aeration: none
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per vehicle control: 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: moderately hard freshwater, originated from the Town of Jupiter. It was treated by vigorous aeration, filtered to 20 micrometers, passed through activated carbon and re-aerated prior to use.

OTHER TEST CONDITIONS
- Photoperiod: Fluorescent light, 16 hours daily, in an environmental chamber (20 ± 1°C)

EFFECT PARAMETERS MEASURED:
- Test organisms: Immobilizations were recorded after 24 and 48 hours exposure
- pH, oxygen concentration and temperature: at 0, 24, and 48 hours
- Specific conductivity, water hardness, and alkalinity

TEST CONCENTRATIONS
Nominal concentrations: 0, 6.25, 12.5, 25.0, 50.0, and 100 mg/L
Mean measured concentrations: 0.0, 30.7, 57.9, 108, 226, and 312 µg/L

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 312 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

312 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

and sublethal effects

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: WAF

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks:

and sublethal effects

Remarks on result:

other: WAF

Details on results:

All test solutions were clear and colorless throughout the test. Test concentrations tended to decrease over each 24-hour exposure period possibly related to precipitation of very fine particles or depositions from the super-saturated solution.
Mortality of the water flea exposed for 48 hours to test item was zero or 5 percent at all concentrations tested. Control mortality was zero percent.

Executive summary:

To determine the toxicity of the test item (as described in section 1.2) to freshwater invertebrates, a GLP-conform static study using Daphnia magna was performed. The water accomodated fraction (derived from a suspsension containing the test item loading concentration of 100 mg/L) was used to prepare solutions with the nominal test concentrations of 0, 6.25, 12.5, 25.0, 50.0, and 100 mg test item/L. All test solutions were clear and colorless throughout the test. Mortality was zero or 5 percent at all concentrations tested. Control mortality was zero percent. The EC50 was > 312 µg mg test item/L for 48 hours and the NOEC 312 µg test item/L, related to the analytically verified concentrations (0.0, 30.7, 57.9, 108, 226, and 312 µg/L). This toxicity study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates (Daphnia sp., Acute Immobilisation Test OECD TG 202).

Description of key information

The 48-h EC50 was > 312 µg test substance/L. With regard to the nominal concenetration the 48-h EC50 was > 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

>= 100 mg/L

Additional information

The acute toxicity to daphnia was assessed according to OECD guideline 202. Daphnids were exposed to the test system under static conditions for 48 hours. The water accomodated fraction (derived from a suspsension containing the test substance loading concentration of 100 mg/L) was used to prepare solutions with the nominal test concentrations of 0, 6.25, 12.5, 25.0, 50.0, and 100 mg test substance/L. All test solutions were clear and colorless throughout the test. Mortality was zero or 5 percent at all concentrations tested. Control mortality was zero percent. The 48 hour EC50 was > 312 µg/L and the NOEC was 312 µg/L, related to the analytically verified concentrations (0.0, 30.7, 57.9, 108, 226, and 312 µg/L).