N,N''-(methylenedi-4,1-phenylene)bis[N'-octyl]urea

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:

21 October 2002 - 25 October 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The test was conducted according to OECD Guideline 203 and in compliance with GLP. The stock solutions were prepared by agitation for 20-23 hours and then filtered twice (porosity of 4-7 µm and then of 0.45 µm). The filtered solution was then further used to prepare the test. The measured concentrations of test substance in these solutions were below the limit of quantification (0.5 mg/L) at all measurement times. Based on the fact that the substance is highly insoluble in water (water solubility 4 µg/L) and has a high adsorptive potential (Koc of one constituent 4.8 and of the two other constituents> 5.6), it can only be concluded that the LC50 > maximum solubility of the test substance in medium.

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)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 1000 mg/L
- Sampling method: samples taken at the begining of the test, before and after each solution change and at the end of the test.
- Sample storage conditions before analysis: at -20 °C

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:stirring the test item in water for 20 to 23 h
- Evidence of undissolved material (e.g. precipitate, surface film, etc): the solution was filtered through a filter with 4-7 µm porosity, followed by filtration through 0.45 µm

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Strain: Salmonidae
- Source: Viviers de France, Piscicultures des Fonatines, Sarrance, France
- Age at study initiation (mean and range, SD): not reported
- Length at study initiation (length definition, mean, range and SD): average 46 mm
- Weight at study initiation (mean and range, SD): avearge 1.08 g
- Method of breeding: during the culture the fish were mantained under flow-through system in water of the same quality and temperature as that used during the test. During culture the animals were fed twice daily, ceasing 24 h before the start of the test.
- Feeding during test none

ACCLIMATION
- Acclimation period: not reported
- Acclimation conditions (same as test or not): yes
- Type and amount of food: trout chow
- Feeding frequency: twice daily, up to 24 h before the start of test
- Health during acclimation (any mortality observed): not reported

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

161-167 mg CaCO3/L

Test temperature:

12.9-13.9 °C

pH:

7.86-8.14

Dissolved oxygen:

6.4 - 8.3 mg/L

Nominal and measured concentrations:

Limit test nominal: 0 and 1000 mg/L
Limit test measured: < 0.5 mg/L (< LOQ)
Range finding test nominal: 1; 10 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: not reported
- Type: not reported
- Material, size, headspace, fill volume: not reported
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): every 24 h
- No. of organisms per vessel: for limit test 7 and for range finding 5
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates):1
- Biomass loading rate: 0.95 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water
- Total organic carbon: not reported
- Particulate matter: not reported
- Metals: not reported
- Pesticides: not reported
- Chlorine: not reported
- Alkalinity: not reported
- Ca/mg ratio: 41.8/10.5 mg/L
- Conductivity: not reported
- Culture medium different from test medium: no
- Intervals of water quality measurement:temperature, hardness, oxygen and pH daily

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light and 8 h dark
- Light intensity: not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : LC50 (mortality)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10 (range finding)
- Range finding study yes
- Test concentrations (range finding): 1, 10 and 100 mg/L
-Limit test: 0 and 1000 mg/L

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: The LC50 was > maximum soluble concentration of test substance in medium.

Details on results:

- Behavioural abnormalities: no
- Observations on body length and weight: not reported
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: not reported
- Effect concentrations exceeding solubility of substance in test medium: no
- analytics : < 0.5 mg/L ( < LOQ)

The authors concluded that the 96h LC50 was > 1000 mg/L (initial loading rate). The French Authorities reported the 96h LC50 as > 4.38 µg/L (water solubility limit).

Validity criteria fulfilled:

yes

Conclusions:

No effects were observed in the limit test, conducted with the filtrate of a 1000 mg/L loading rate. The measured concentrations of test substance in these solutions were below the limit of quantification (0.5 mg/L) at all measurement times. Based on the fact that the substance is highly insoluble in water (4 µg/L) and has a high adsorptive potential (Koc of one constituent 4.8 and of the two other constituents> 5.6), it can only be concluded that the LC50 > maximum solubility of the test substance in medium.
The Competent Authority of France concluded in 2003 that the LC50 for freshwater fish should be >4.38 µg/L (the water solubility limit).

Executive summary:

The acute toxicity of KY-EU to rainbow trout was examined in a semi-static system according to OECD guideline 203 and GLP.

Fish were exposed to a WAF that was prepared at a loading rate of 1000 mg/L by stirring for 20-23h, followed by filtration through a filter with 4-7 µm porosity, and a filter with 0.45 µm porosity, subsequently. The media were refreshed every 24h. Samples were taken at the begining of the test, before and after each solution change and at the end of the test and stored at -20°C until analysis. Samples were analysed with HPLC-UV. The analysis was based on only one component out of 3 of KY-EU.

No effects of the test item to fish were observed.

The measured concentrations of test substance in these solutions were below the limit of quantification (0.5 mg/L) at all measurement times. Based on the fact that the substance is highly insoluble in water (4 µg/L) and has a high adsorptive potential (Koc of one constituent 4.8 and of the two other constituents> 5.6), it can only be concluded that the LC50 > maximum solubility of the test substance in medium.

Description of key information

No effects were observed in the limit test, conducted with the filtrate of a 1000 mg/L loading rate. The measured concentrations of test substance in these solutions were below the limit of quantification (0.5 mg/L) at all measurement times. Based on the fact that the substance is highly insoluble in water (4 µg/L) and has a high adsorptive potential (Koc of one constituent 4.8 and of the two other constituents> 5.6), it can only be concluded that the LC50 > maximum solubility of the test substance in medium.

Additional information

The acute toxicity of KY-EU to rainbow trout was examined in a semi-static system according to OECD guideline 203 and GLP.

Fish were exposed to a WAF that was prepared at a loading rate of 1000 mg/L by stirring for 20-23h, followed by filtration through a filter with 4-7 µm porosity, and a filter with 0.45 µm porosity, subsequently. The media were refreshed every 24h. Samples were taken at the begining of the test, before and after each solution change and at the end of the test and stored at -20°C until analysis. Samples were analysed with HPLC-UV. The analysis was based on only one component out of 3 of KY-EU.

No effects of the test item to fish were observed.

The measured concentrations of test substance in these solutions were below the limit of quantification (0.5 mg/L) at all measurement times. Based on the fact that the substance is highly insoluble in water (4 µg/L) and has a high adsorptive potential (Koc of one constituent 4.8 and of the two other constituents> 5.6), it can only be concluded that the LC50 > maximum solubility of the test substance in medium