2-Butenedioic acid (Z)-, ester with 1,2-propanediol, compd. with 2-(dibutylamino)ethanol

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: According to international guideline(s), GLP-compliant, performed in recognized contract research organization, no restrictions, fully adequate for assessment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: nominal 100 , 50, 25, 12.5, 6.25 mg/L and control
- Sampling intervals: start and end of the first and the last renewal period
- Sampling method: samples were taken from flasks containing no animals
- Sample storage conditions before analysis: no storage prior to analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The highest test concentration of 100 mg/L and dilutions thereof were prepared in test medium.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: “Black Molly” Fish Farm, Veszprém, Hungary
- Length at study initiation: 2.6 – 3.1 cm
- Weight at study initiation (mean and range, SD): 0.36 g ± 0.01 g (0.35-0.38 g)
- Feeding during test: none

ACCLIMATION
- Acclimation period: > 12 days
- Acclimation conditions (same as test or not): same water quality (circulated and filtered tap water)
- Type and amount of food: not reported
- Feeding frequency: daily, until 24 hours before the start of the test
- Health during acclimation (any mortality observed): The fish were healthy. There was no mortality of the population for seven days before the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

203 mg/L as CaCO3 (measured)

Test temperature:

23.1-23.6 °C

pH:

7.9-8.1

Dissolved oxygen:

95-96% air saturation value

Nominal and measured concentrations:

nominal concentrations: 100, 50, 25, 12.5, and 6.25 mg/L
freshly prepared test media:87-126% of nominal
aged test media (24 h): 100-137% of nominal (94-128% of initially measured)
The analytically measured concentrations deviated more than 20% from the nominal values in some cases, thus the biological results are based on the geometric mean measured concentrations: 108.9, 56.9, 30.1, 15.0, and 6.4 mg/L (102-120% of nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: aquaria, containing 5 litres of medium
- Type: not reported (the test material is not volatile)
- Aeration: not reported (oxygen concentration was sufficient during exposure)
- Renewal rate of test solution: daily
- No. of organisms per vessel: 7 (except the 50 mg/L test group which contained 8 fish)
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.49 - 0.56 g body weight/L (mean 0.52 ± 0.02 g bw/L)

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: circulated and filtered tap water
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light, 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and sub-lethal effects; recorded at approx. 3, 6, 24, 48, 72 and 96 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Range finding study
- Test concentrations: nominal 100, 10, 1, and 0.1 mg/L
- Results used to determine the conditions for the definitive study: 100% mortality after 96 hours at 100 mg/L; no mortality at the lower test concentrations

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

36 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% confidence limits: 29.3-60.3 mg/L

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

15 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

30 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: one dead fish after 48 h

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

15 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

57 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: swimming problems and decreased activity at the highest test concentration (at the 3-hour observation); no abnormalities observed at any other concentration and/or time
- Mortality of control: zero
- Other adverse effects control: none

For cumulative mortality data Table 1.

Reported statistics and error estimates:

The 24, 48, 72 and 96 hours LC50 values were calculated by Probit analysis with 95% confidence limits using SPSS PC+ software.
The NOEC, LOEC and LC100 were determined directly from the raw data.

Sublethal observations / clinical signs:

Table 1: Cumulative mortality data:

 

Concentration (mg/L)*	Cumulative mortality [number dead of fish] **
	3 h	6 h	24 h	48 h	72 h	96 h
Control	0	0	0	0	0	0
6.4 (6.25)	0	0	0	0	0	0
15.0 (12.5)	0	0	0	0	0	0
30.1 (25)	0	0	0	1	1	1
56.9 (50)	0	0	0	8	-	-
108.9 (100)	0***	0	3	7	-	-

 

*:   geometric mean measured (nominal)

**:  initial population = 7 fish, except 8 fish at nominal 50 mg/L

***: swimming problems and decreased activity

Validity criteria fulfilled:

yes

Remarks:

no mortality of control animals; concentration of dissolved oxygen in all test vessels >60% saturation

Conclusions:

The test material is toxic to freshwater fish with a 96-hour LC50 between 10 and 100 mg/L (calculated value: 36 mg/L).

Description of key information

96-h LC50 (based on geometric mean concentrations) 36 mg/L for Brachydanio rerio (new name Danio rerio) (OECD 203, EU C.1)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

36 mg/L

Additional information

The results indicate that the test material is toxic to freshwater fish with a 96-hour LC50 between 10 - 100 mg/L (calculated value: 36 mg/L).