Registration Dossier

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

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:
1996-12-16 to 1996-12-20 (experimental phase)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP guideline study with acceptable restrictions
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
EEC Directive 84/449
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: H 50541 H x B
- Analytical purity: 98.4% (analytical report ZAX 96L00027)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: no data
- Stability under test conditions: The stability of the test substance over the study period was proven by reanalysis.

OTHER SPECIFICS:
- Name of test material (as cited in study report): 2,2'-Dimorpholinodiethylether
- Substance type: pure active substance
- Physical state: liquid, slightly yellow, clear
Analytical monitoring:
no
Details on sampling:
- Concentrations: 0, 100, 215, 464. 1000, and 2150 mg/L
- Sample storage conditions before analysis: analysis performed on day of sampling
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance was added directly to the test water of each concentration and mixed using an ultra-turrax stirrer. 25 min were needed from the preparation of the test solutions to placing the fish in the aquaria.
- Controls: negative control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish
- Source: Charles River Aquatics, Someren , The Netherlands
- Length at study initiation (length definition, mean, range and SD): 3.5 cm (3.1 to 4.2 cm)
- Weight at study initiation (mean and range, SD): 0.33 g (0.15 to 0.82 g)
- Feeding during test: no

ACCLIMATION
- Acclimation period: 3 months
- Acclimation conditions (same as test or not): water conditions similar (tap water, not chlorinated, passed through an active carbon filter, aerated with oil-free air) but flow through conditions were used
- Type and amount of food: Tetra Min" standard feed for aquarium fish, Tetra-Werke, Melle, Germany; ad libitum; on workdays live and frozen Artemia
- Feeding frequency: daily
- Health during acclimation (any mortality observed): 0% during the last 2 weeks
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
total hardness: about 2.5 mmol/L
Test temperature:
22 °C (temperature-controlled room)
pH:
7.9 to 8.3
Dissolved oxygen:
4.1 to 7.6 mg/L
Nominal and measured concentrations:
nominal: 0 (control), 100, 215, 464, 1000, and 2150 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: glass aquaria
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: all-glass aquaria, 30 x 22 x 24 cm, 10 L fill volume (water depth about 17 cm)
- Aeration: slight
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.3 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: municipal water of the city of Frankenthal (Germany), not chlorinated and passed through a charcoal filter
- Ca/Mg ratio: 9:1 (mass ratio)
- Acid capacity: 5.5 mmol/L
- Total hardness: 2.5 mmol/L
- pH: about 8.0 to 8.6


OTHER TEST CONDITIONS
- Photoperiod: 16 hours light : 8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and symptoms after 1, 4, 24, 48, 72, and 96 h
- mortality and symptoms: after 1, 4, 24, 48, 72, and 96 h
- pH, oxygen, and temperature: after 1, 4, 24, 48, 72, and 96 h
- chemical analysis: after 1, 24, 48 and 96 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: approximately 2.2
- Range finding study
- Test concentrations: based on results of four range-finding studies
- Results used to determine the conditions for the definitive study: 96-h LC50: 1000-10000 mg/L
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 2 150 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 1% significance level
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
215 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
2 150 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 2 337.5 mg/L
Nominal / measured:
meas. (arithm. mean)
Conc. based on:
act. ingr.
Basis for effect:
mortality
Remarks on result:
other: 1% significance level
Details on results:
- Behavioural abnormalities: see "Any other information on results incl. tables"
- Mortality of control: 0%
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no

Mortality

No fish died during exposure.

Symptoms

Test item conc.

Symptoms after

 

 

 

mg/L; nominal

1 h

24 h

48 h

72 h

96 h

0

 

 

 

 

 

100

 

 

 

 

 

215

 

 

 

 

 

464

 

A, D7

A

a

a

1000

 

A, D

A, D

A, D9

A, N

2150

 

A, D

A, D

A, N

A, N, D5

A: apathy

a: slight to very slight apathy

N: narcotic-like state

D: swimming near bottom

number behind symbol indicates number of affected fish

pH values

Test item conc.

pH values after

mg/L; nominal

1 h

24 h

48 h

72 h

96 h

0

7.9

8.3

8.2

8.2

8.2

100

7.9

8.3

8.2

8.1

8.2

215

7.9

8.3

8.2

8.1

8.2

464

7.9

8.3

8.2

8.1

8.1

1000

7.8

8.3

8.2

8.2

8.2

2150

7.8

8.3

8.2

8.2

8.2

Oxygen content

Test item conc.

Oxygen content (mg/L) after

mg/L; nominal

1 h

24 h

48 h

72 h

96 h

0

7.2

6.5

5.6

4.8

4.2

100

7.1

6.5

5.7

4.2

4.1

215

7.4

6.7

6.1

4.8

4.2

464

7.8

6.8

6.2

5.1

4.3

1000

7.6

6.5

6.4

5.3

4.4

2150

7.6

6.5

6.1

4.6

4.2

Chemical analysis

Test item conc.

Analytically detected concentrations after

mg/L; nominal

1 h

24 h

48 h

96 h

 

mg/L

%

mg/L

%

mg/L

%

mg/L

%

0

< 10

---

< 10

---

< 10

---

< 10

---

100

100

100.0

95

95.0

101

101.0

101

101.0

215

216

100.5

218

101.4

219

101.9

220

102.3

464

486

104.7

490

105.6

497

107.1

500

107.8

1000

1010

101.0

1030

103.0

1040

104.0

1050

105.0

2150

2300

107.0

2330

108.4

2360

109.8

2360

109.8

Conclusions:
In this study, the acute toxicity of DMDEE to Danio rerio was investigated in a 96-h static study (nominal concentrations of 100, 215, 464, 1000, and 2150 mg/L). No mortalities were observed throughout the study, resulting in a 96-h LC50 of 2150 mg/L.

Description of key information

The key value was derived from a study performed under GLP conditions and according to OECD 203 and EU Method C.1. An LC50 was found for zebrafish (Danio rerio) of > 2150 mg/l after an exposure of 96 hours in a static test. 

Key value for chemical safety assessment

LC50 for freshwater fish:
2 150 mg/L

Additional information

Two studies investigating the acute toxicity on fish were available. In the BASF study (1998) zebra fish (Danio rerio) were exposed to the test substance during 96 hours in a static test, whereas the study of Kurume Research Laboratories (Yakata, 1994) was a limit test and exposed orange-red killifish (Oryzias latipes) to the test substance during 48 hours in a semi-static design. Although both studies were conducted under GLP conditions and found reliable with acceptable restrictions (K2), the BASF study was selected as key study. The main reasons for this decision are the shorter exposure period (48 hours instead of 96 hours) in the study of Kurume Research Laboratories (Yakata, 1994), and the fact that the result of this limit test was a LC50(48h) of > 1000 mg/l, whereas the LC50 (96h) of the BASF study appeared to be > 2150 mg/l.