Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study without detailed documentation

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report date:
2004

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N'-[dithiobis[2-(2-hydroxyethyl)-1-methylvinylene]]bis[N-[(4-amino-2-methyl-5-pyrimidinyl)methyl]formamide
EC Number:
200-644-4
EC Name:
N,N'-[dithiobis[2-(2-hydroxyethyl)-1-methylvinylene]]bis[N-[(4-amino-2-methyl-5-pyrimidinyl)methyl]formamide
Cas Number:
67-16-3
Molecular formula:
C24H34N8O4S2
IUPAC Name:
N-[(4-amino-2-methylpyrimidin-5-yl)methyl]-N-[(2E)-3-{[(2E)-2-{N-[(4-amino-2-methylpyrimidin-5-yl)methyl]formamido}-5-hydroxypent-2-en-3-yl]disulfanyl}-5-hydroxypent-2-en-2-yl]formamide
Test material form:
solid: particulate/powder

Sampling and analysis

Analytical monitoring:
not specified
Details on sampling:
Not specified

Test solutions

Vehicle:
not specified
Details on test solutions:
A solution containing a maximum of 100 mg/L of the test item plus two other range-finding concentrations was prepared. Test vessels (aquariums) was filled directly from the test solution containers immediately after preparation. The same vessels was used throughout the test. The pH range considered as appropriate for a normal life of test organisms is circa 6 to 9. Consequently, the tesst solution « with a pH < 5 R or > 0.2 was tested with pH adjustment.

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Justification of this choice: the rainbow trout is a fish species generally accepted by regulatory authorities for this type of study and is recommended in OECD and EEC guidelines.
Origin: CIT, BP 563, 27005 Evreux, France; the eggs are obtained from Viviers de France, Pisciculture des Fontaines, 64490 Sarrance, France.
Allocation to study: animals was selected using a hand-net, without preference, and randomly assigned to test vessels.
Number of animals: four groups of seven fish.

Study design

Test type:
static
Water media type:
freshwater
Total exposure duration:
96 h

Test conditions

Hardness:
151 to 157 mg/L of CaCO3 in the control
Test temperature:
14.1 to 14.6°C
pH:
7.51 (mini) to 8.04 (maxi)
Dissolved oxygen:
6.9 mg/L (min) to 8.5 (max)
Salinity:
Not specified
Conductivity:
Not specified
Nominal and measured concentrations:
Nominal concentrations : 1, 10 & 100 mg/L
Details on test conditions:
Test solutions was not changed over the test period.
Light/dark cycle: 16/8 hours.
Duration of test: 96 hours.
Loading: up to 1g of fish per liter of test solution. The animals were between 4 and 6 cm in length.
Oxygen concentration: all attempts was made to maintain dissolved oxygen at or above 60% of the air saturation value at that temperature.

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Key result
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Results with reference substance (positive control):
Not specified

Any other information on results incl. tables

Sublethal observations / clinical signs:

 Table 1: Physicochemical paramters

Time (hours) Temperature (°C)   pH  Oxygen content (mg/L)  Water hardness (mg/L of CaCO3)
 Control          
 0 14.2  7.51  8.5  151 
 24 14.3  7.71  8.4   
 48 14.2  7.62  7.4   
 72 14.4  8.03  7.8   
 96 14.6  7.88  6.9  157 
 Highest nominal rate charge : 100mg/L            
 0 15.6  8.01  8.3   
 24 14.1  7.98  8.4   
 48 14.4  8.04  7.7   
 72 14.4  8.04  7.7   
 96 14.5  7.91  7.2   

Table 2: Mortality summary

    Observation time (hours)

          Mortality at each nominal concentrations (mg/L)

n fishes on 7 fishes

 control 10  100 
 0
 24
 48
 72
 96

Applicant's summary and conclusion

Conclusions:
The highest loading rate without clinical effect and mortality at T96h was 100 mg/L. The lowest loading rate with 100% mortality at T96h was> 100 mg / L.
LC50s at 24h, 48h, 72h and 96h were> 100 mg / L (loading rate).
Executive summary:

Acute short-term toxicity test on fish (rainbow trout) under static conditions was conducted accorting to guidline OCDE.

The highest loading rate without clinical effect and mortality at T96h was 100 mg/L. The lowest loading rate with 100% mortality at T96h was> 100 mg / L.

LC50s at 24h, 48h, 72h and 96h were> 100 mg / L (loading rate).