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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 16 June 2003 (beginning of the study) to 20 June 2003 (end of the study)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
This test was performed according to international guidelines without GLP statement. All validity criteria were fulfilled except the percentage of the air saturation value throughout the test (comprised between 51.5% to 96% at the highest loading rates).
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report date:
2003

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
, 17th July 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
, 31st July 1992
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
2'-[2,4-dichloro-5-(1-methylethoxy)phenyl]-2,2-dimethylpropionohydrazide
EC Number:
257-027-8
EC Name:
2'-[2,4-dichloro-5-(1-methylethoxy)phenyl]-2,2-dimethylpropionohydrazide
Cas Number:
51167-18-1
Molecular formula:
C14H20Cl2N2O2
IUPAC Name:
N'-[2,4-dichloro-5-(propan-2-yloxy)phenyl]-2,2-dimethylpropanehydrazide
Test material form:
other: solid
Details on test material:
- Name of test material (as cited in study report): HTMA [1-TRIMETHYLACETYL-2-(2,4-DICHLORO-5-ISOPROPOXYPHENYL)-HYDRAZINE]
- Molecular weight: 319 mg/L
- Substance type: brown orange-colored pieces
PHYSICO-CHEMICAL PROPERTIES
- Water solubility: 40 ppm
- Density: 1.25

- Physical state: solid
- Stability under test conditions: no data
- Storage condition of test material: no data

Sampling and analysis

Analytical monitoring:
no
Details on sampling:
Not applicable

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test solutions were prepared by further dilution of the stock solution with water or directly from the test item. A solution containing a maximum of 100 mg/L of the test item plus two other range-finding concentrations were prepared. Test vessels were filled directly from the test solution containers immediately after preparation.
- Eluate: filtered and dechlorinated potable water
- Control: yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no data

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Strain: no data
- Source: CIT, BP 563, 27005 Evreux, France; the eggs are obtained from Viviers de France, Pisciculture des Fontaines, 64490 Sarrance, France.
- Age at study initiation (mean and range, SD): no data
- Length at study initiation (length definition, mean, range and SD): no data
- Weight at study initiation (mean and range, SD): no data
- Method of breeding: no data
- Feeding during test: no

ACCLIMATION
- Acclimation period: no data
- Acclimation conditions (same as test or not): no data
- Type and amount of food: no data
- Feeding frequency: during culture, animals were fed twice daily
- Health during acclimation (any mortality observed): no data

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Remarks on exposure duration:
none
Post exposure observation period:
None

Test conditions

Hardness:
145-149 mg CaCO3/L
Test temperature:
15.5-16.2°C
pH:
7.60-7.89
Dissolved oxygen:
5.2-9.7 mg/L
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal loading rates concentrations: 0, 1, 10 and 100 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: aquariums
- Type (delete if not applicable): no data
- Material, size, headspace, fill volume: no data
- Aeration: no data
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): not applicable
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): not applicable
- Biomass loading rate: up to 1 g of fish per liter of test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Filtered and dechlorinated potable water which is then softened by a Permo water softening system. Half of the water passes directly into a reservoir tank attached to the water delivery system for culture while the remaining half passes through a deioniser producing ultra-pure water which is then fed into the reservoir tank. This results in a 1:1 dilution of potable dechlorinated:deionised water giving a supply of the desirable hardness.

OTHER TEST CONDITIONS
- Adjustment of pH: not adjusted during the test, pH was between 7.6 and 7.89 during the study.
- Photoperiod: 16 h light/ 8h dark cycle
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : cumulative mortality and sub-lethal observations.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Remarks:
and behaviour
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 10 - < 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
See tables in "Any other information on results incl. tables" section.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
None

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 6.1.1/1: Cumulative mortality data

Observation time (hours)

Mortality at each range-finding loading rate

n fish out of 7 fish

Control

1

10

0

24

48

72

96

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Table 6.1.1/2: Sub-lethal observations

Nominal loading rate (mg/L)

Time (hours)

n fish out of 7 fish

Normal

Lost. equil

Fish at surface

Fish at bottom of aquarium

Fish hyper.

Swim vertical

Swim ups/dn

Spas swim

0

0 to 96

7/7

0

0

0

0

0

0

0

1

0 to 96

7/7

0

0

0

0

0

0

0

10

0 to 96

7/7

0

0

0

0

0

0

0

100

0

24

48 to 96

7/7

0

0

0

1

0

0

1

0

0

0

0

0

0

0

0

1

0

0

0

0

0

0

0

Lost equil. = mild loss of equilibrium while swimming

Hyper. = hyperactive swimming behavior

Swim vertical = severe loss of equilibrium - fish hung/swam vertically in the solution

Swim ups/dn = severe loss of equilibrium - fish swam on their backs

Spas. swim = spasmodic swimming

Applicant's summary and conclusion

Validity criteria fulfilled:
no
Remarks:
the dissolved oxygen concentration was not maintaine at >= 60% of the air saturation throughout the test (at the highest loading rates, the oxygen concentration was comprised between 5.2 and 9.7 mg/L, corresponding to 51.5% and 96% of the air saturation).
Conclusions:
The highest concentration without clinical effects and mortality (NOEC) at 96 hours was 10 mg/L. The 96h-LC50 was estimated to be between 10 and 100 mg/L.
Executive summary:

This test was performed according to international guidelines without GLP statement.

The objective of this screening test is to assess the acute toxicity of the test substance to the rainbow trout (Oncorhynchus mykiss), in a 96 hour static test.

3 nominal loading rates concentrations (1, 10 and 100 mg/L) were used plus one control. No analytical monitoring was performed and the cumulative mortality and the sub-lethal effects were recorded throughout the test.

At the nominal loading rates of 1 and 10 mg/L, 0% mortality and sub-lethal effects were observed. However, at the highest loading rate of 100 mg/L, 100% mortality was observed.

Based on these results, the highest concentration without clinical effects and mortality (NOEC) at 96 hours was 10 mg/L. The 96h-LC50 was estimated to be between 10 and 100 mg/L.

All validity criteria were fulfilled except the percentage of the air saturation value throughout the test (comprised between 51.5% to 96% at the highest loading rates). This study corresponds to a screening test and no defined LC50 value was determined.