1,3,3-trimethyl-N-(2-methylpropylidene)-5-[(2-methylpropylidene)amino]cyclohexanemethylamine

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:

2011-07-11 to 2011-07-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 100 mg/L
- Sampling method: Samples taken from test container (aquaria)
- Sample storage conditions before analysis: Samples were taken, immediately transferred to the analytical laboratory and analysed on the same day.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test substance was directly weighed into the test vessels after adding the test water.
- Two barrels (30 L) were prepared: one was filled with 10.5 L of dilution wtaer for the control. The 2nd barrel was also filled with 10.5 L dilution water, stirred with a magnetic stirrer (300 rpm) and 1050 mg test substance were added (= 100 mg/L). The test medium was stirred for 24 h. Afterwards, undissolved particles were removed by filtration
- Controls: control preparation without test substance was treated the same way.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): precipitate removed by filtration.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Strain: Hamilton-Buchanan
- Source: Raiffeisenmarkt, 47906 Kempen, Germany
- Length at study initiation (length definition, mean, range and SD): 2 +/- 1 cm
- Feeding during test: There was no feeding throughout the exposure period in the test.

ACCLIMATION
- Acclimation period: at least 12 days
- Acclimation conditions: in 300 L glass tanks, illumination 12/12 hours light/dark cycle, aeration with compressed oil free air (at least 80% air saturation)
- Type and amount of food: The fish were fed with “Tetra Min” dry food from Tetra, Melle, Germany.
- Feeding frequency: daily
- Health during acclimation (any mortality observed): Their state of health was controlled routinely during holding.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

1.818 mmol/L (total hardness)

Test temperature:

21.7 - 22.2 °C

pH:

8.01 - 8.68

Dissolved oxygen:

71.0 - 76.8 %

Nominal and measured concentrations:

nominal (measured DOC at 0 h in brackets): 100 mg/L ( 53.7 mg/L)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: all glas aquaria, 15 L volume, filled with 10.5 L of test solution
- Aeration: dilution water was aerated before use in the experiment
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 1 g/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The holding and dilution water for the fish was prepared separately in a silicone sealed PVC tank (250 L) by mixing demineralised water with de-chlorinated tap water from municipal source (SWK Aqua GmbH, Krefeld, Germany) in a ratio of 3:2 and 9 mL of sulphuric acid.
- Total organic carbon: 0 mg/L
- Alkalinity: 0.75 mmol/L
- Conductivity: 358 µS/cm
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 12 h / 12 h light / dark cycle

EFFECT PARAMETERS MEASURED : mortality, symptoms of toxicity at 0, 24, 48, 72 and 96 h

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 53.7 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

dissolved

Remarks:

(DOC)

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 53.7 mg/L

Nominal / measured:

nominal

Conc. based on:

dissolved

Remarks:

(DOC)

Basis for effect:

mortality (fish)

Details on results:

No mortality was observed during the study. Fish did not show any symptoms of toxicity.

Reported statistics and error estimates:

Statistical analysis was done with ToxRat Professional (ToxRat Solutions GmbH, Alsfeld, Germany)

Sublethal observations / clinical signs:

Summary of mortality data

 

Nominal Concentration of test substance	Number of dead fish Exposure period						Cumulative frequency (%) Exposure period
[mg/L]	3 h	6 h	24 h	48 h	72 h	96 h		3 h	6 h	24 h	48 h	72 h	96 h
(0) Control	0	0	0	0	0	0		0	0	0	0	0	0
100	0	0	0	0	0	0		0	0	0	0	0	0

 

Analytical measurements

Time		Control (dilution water)				100 mg/L
			TC (mg/L)	TIC (mg/L)	DOC^1 (mg/L)		TC (mg/L)	TIC (mg/L)	DOC (mg/L)^2	Recovery in %
	0h		11.4	11.3	0.1		69.2	15.4	53.7
	24 h		11.4	11.4	≤0		67.3	17.1	50.2	93
	48 h		11.4	11.5	≤0		66.3	17.2	49.1	91
	72 h		11.3	11.9	≤0		72.0	17.5	54.5	101
	96 h		11.4	11.8	≤0		70.8	17.7	53.1	99

 ¹DOC = TC – TIC

²DOC = DOC minus DOC Control

Recovery rate for DOC in % after 24h, 48h, 72h and 96h relating to 0h values.

Percent values were rounded.

Validity criteria fulfilled:

yes

Conclusions:

The LC50 (96 h)of based on nominal concentration is > 100 mg/L and based on the measured DOC >53.7 mg/L and indicating that the test substance does not pose a hazard to fish.

Executive summary:

The test substance was tested in the Fish Acute Toxicity Test according to the OECD Guideline for the Testing of Chemicals No. 203 (July 1992) using the zebra fish, Danio rerio (Hamilton-Buchanan) as test organism. The experiment was carried out as a static limit test, i.e. the test substance preparations were not exchanged for the whole exposure period of 96 hours. The results of this study show that the test substance does not have any impact on Danio rerio at the tested concentration of 100 mg/L (nominal). Hence the LC₅₀value after 96 h of exposure was determined a >100 mg/L, based on nominal concentration. Based on the measured DOC value the LC50 value after 96 h was > 53.7 mg/L.

Description of key information

The test substance was tested in the Fish, Acute Toxicity Test according to the OECD Guideline for the Testing of Chemicals No. 203 (July 1992) using the zebra fish, Danio rerio (Hamilton-Buchanan) as test organism. The experiment was carried out as a static limit test, i.e. the test substance preparations were not exchanged for the whole exposure period of 96 hours. The results of this study show that the test substance does not have any impact on Danio rerio at the tested concentration of 100 mg/L (nominal). Hence the LC50 value after 96 h of exposure was determined a >100 mg/L, based on nominal concentration. Based on the  measured DOC value the LC50 value after 96 h was > 53.7 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

EC50

Effect concentration:

53.7 mg/L