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 fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997-05-30 - 1998-01-02

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study (OECD TG 203) without detailed description

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity 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 48 hours (old and new
solutions), and test termination. Water samples (100 mL per composited test solution) were taken using volumetric pipets.

Vehicle:

not specified

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 stock for preparing the three test concentrations at targeted concentrations of 0.020, 0.067 and 0.200 mg/L. 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 over the weekend to permit the test substance to coat the glassware. These solutions were discarded when the test was initiated and fresh solutions were added after all test chambers were rinsed with deionized water).

LOADING
- Loading was calculated to be 0.47 grams of fish tissue per liter of test solution.

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name/age: Juvenile fathead minnow
- Source: Toxikon Environmental Sciences in-house culture
- Length at study initiation: 31 +/- 2 mm (mean)
- Weight at study initiation: 0.42 +/- 0.11 g (mean)
- Feeding during test: None
- Adaptation: 7 days / no food 24 hours prior to exposure

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96

Remarks on exposure duration:

h

Hardness:

68 mg CaCO3/L

Test temperature:

21.9 - 23.5°C

pH:

Start: Test solution: 7.1 - 7.2; 6.6 - 7.2 during the remainder of the test

Dissolved oxygen:

Start: Test solution: 7.8 and 8.7 mg/L (91 to 101% of saturation); Control: 8.7 mg/L (101% of saturation)
End: Test solution: 4.9 mg/L to 8.5 mg/L (57 to 99% of saturation); 5.8 to 8.4 mg/L (67 to 96% of saturation)

Nominal and measured concentrations:

- Loading dose: 100 mg/L
- Nominal: 0, 0.020, 0.067 and 0.200 mg test item/L
- Actual:
0.027, 0.145, 0.216, 0.219 mg/L (0 hrs)
0.159(b), 0.111(b),0.258 (b), 0.409 (b) (96 hrs)
(a) One-half of the MVL (minimum validation level) of 0.025 mg/Lwas used for calculating the mean value.
(b) Sample bottles broken during sample preservation, label dye contaminated the samples and caused interference which gave the high numbers. Numbers were not used in the mean calculations.

Details on test conditions:

TEST SYSTEM
- Test vessel: 3.5-gallon glass jars, covered
- Fill volume: 9 L
- Aeration: initiated on day 1 in all test solutions due to dissolved oxygen levels falling below 60% saturation
- No. of organisms per vessel: 10
- No. of vessels per concentration: 1
- No. of vessels per vehicle control: 1
- The test solutions were renewed on day 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. The dilution water, at test initiation, possessed a hardness of 68 mg/L as calcium carbonate (CaCO2) , alkalinity of 9 mg/L as CaCO2, and specific conductivity of 399 microsiemens per centimeter (µS).
OTHER TEST CONDITIONS
- Photoperiod: Fluorescent light, 16 hours daily

EFFECT PARAMETERS MEASURED:
- Survival of fathead minnow was monitored daily and any dead fish observed were removed.
- Any abnormalities in the behavior or physical appearance of the fathead minnow were also noted.
- pH, oxygen concentration and temperature: at 0, 24, 48, 72 and 96 hours
- Specific conductivity, total alkalinity, and total hardness of the dilution water: at 0 and 96 hours

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.268 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.268 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

behaviour

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: WAF

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: WAF

Details on results:

No abnormal responses of the fish were observed.

Mortality of fathead minnow exposed for 96 hours to the test substance was zero percent in all the test concentrations and control.

Test concentrations:
- Nominal: 0, 0.020, 0.067 and 0.200 mg test substance/L
- Actual: >0.020, 0.085, 0.21, and 0.211 mg test substance /L (48 h old test solutions)

Reported statistics and error estimates:

Based on the results of the test, the 96-hour LC50 value, its 95 percent confidence limits, and the slope, of the concentration/mortality response curve could not be calculated.

Executive summary:

In a 96-h acute toxicity study, fathead Minnow were exposed to the test item (as described in section 1.2) at nominal concentrations of 0 (control) and 0.020, 0.067 and 0.200 mg test item/L under semi-static conditions. All test solutions were clear and colorless throughout the test. Mean measured concentrations of the test item ranged from 0.080 to 0.268 mg/L. Mortality of fathead minnow exposed for 96 hours to the test item was zero percent in all the test concentrations and control. The 96-hour LC50 was >0.268 mg/L and the no-observed-effect-concentration (NOEC) was 0.268 mg/L based upon the lack of mortality and sublethal effects observed at this and lower test concentrations. This toxicity study is classified as acceptable and satisfies the guideline requirement for an acute toxicity test on fish in general.

Description of key information

The 96-h LC50 was > 0.268 mg /L measured concentration; with regard to the loading dose the LC50 was > 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

>= 100 mg/L

Additional information

The acute toxicity to fish was assessed according to OECD guideline 203. Fathead Minnow were exposed for 96 hours to the test substance at nominal concentrations of 0 (control) and 0.020, 0.067 and 0.200 mg test substance/L under semi-static conditions. The test substance solutions were prepared from the water accomodated fraction obtained from a loading dose of 100 mg test substance/L. All test solutions were clear and colorless throughout the test. Mean measured concentrations of the test substance ranged from 0.080 to 0.268 mg/L. Mortality of fathead minnow exposed for 96 hours to the test substance was zero percent in all the test concentrations and control. The 96-hour LC50 was >0.268 mg/L and the no-observed-effect-concentration (NOEC) was 0.268 mg/L.