Registration Dossier

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1989-08-30 to 1991-07-05
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Cross-reference
Reason / purpose:
reference to same study
Reference
Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that the source and target substances have a similar toxicological profile due to their similar structure. Structural similarity is primarily based on common functional groups, given that the source substances are the precursors for the synthesis of the target substance as explained below. Furthermore, the prediction is supported by the fact that the target substances contains approximately 50% of one of the source substances in free form. Therefore, the toxicological profile of the target substance is anticipated to be largely determined by the toxicological profile of the source substance present in excess in the target substance both in free and bound form.
For more detailed information please refer to the'attached justification' section.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
The source substances m-aminobenzohydrazine (ABH) and 1,1'-(methylenedi-p-phenylene)bismaleimide (MDAB) are the starting materials in the manufacturing process of Compimide 183, in which they act as monomers.
ABH consists of benzol covalently bound to a hydrazide moeity at C1 and one further amine moeity bound to C3. MDAB consists of two aromatic rings each bound to a maleimide group, thus building together two N-phenylmaleimide moieties, and bridged by one methylene group.
The target substance Compimide 183 is the reaction product of MDAB and ABH. The underlying reaction mechanism comprises the Aza-Michael addition of either amine of ABH (i.e from the aminobenzyl or the benzohydrazide moiety) to either of the identical N-phenylmaleimide moieties of MDAB. Under the specific conditions of manufacture, the reaction leads to the formation of a mixture of oligomers, mainly from dimers to pentamers, where the former and the latter are the predominant oligomeric species.
Due to the excess of MDAB in the reaction mixture, and under the conditions of manufacture, the final reaction product contains ca. 50% of free MDAB, the remaining 42.4% being bound to at least one molecule of ABH.
For more detailed information please refer to the 'attached justification' section

3. ANALOGUE APPROACH JUSTIFICATION
The read-across hypothesis is based on the assumption that the potential toxicity of the target substance (Compimide 183), which is the reaction product of the both source substances MDAB and ABH, does not exceed the toxicity of its precursors. As indicated under Substance identity, the mol ratio of MDAB to ABH is 1:0.195. The reaction product thus consists of ca. 7.6% ABH bound to MDAB and ca. 92.4% MDAB (ca. 42.4% bound to ABH and ca. 50% free). Therefore, it is reasonable to assume that the toxicity profile of the target substance will most likely be similar to that of the source substance MDAB, given that N-phenylmaleimide is a common functional group present in the target substance both as free MDAB and at the end(s) of nearly all possible oligomeric reactions products.
The respective toxicological data (RL 1 or 2) are summarised in the table below; robust study summaries are included in the Technical Dossier in the respective sections.
For more detailed information please refer to the 'attached justification' section

Reason / purpose:
read-across source
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.91 mg/L
Validity criteria fulfilled:
no
Conclusions:
In the present study conducted according to OECD guideline 203 (adopted 1983) 10 fish each were exposed to 0.19, 0.34, 0.62, 1.11 and 2.0 mg/L of the test item for 96 h under semi-static conditions.The following LC50 (96 hours) was obtained: 0.91 mg/L (nominal). However, analytical data indicate that the substance is unstable in aqueous test medium.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1983
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
1984
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
For analytical purposes duplicate 50 mL samples were taken from the test tanks as follows:
On day 1 (0-20 minutes after treatment) of the tanks containing 0.19, 0.62 and 2 mg test article/L.
On day 2 (48 hours after treatment) of the tanks containing 0.19 and 0.62 mg/L.
On day 3 (0-20 minutes after treatment) and an day 4 (48 hours after treatment) of the tanks containing 0.19 and 0.62 mg/L.
In addition samples of the control with acetone were taken at the same times and a 10 and a 100 mL sample each from a tank without fish containing 2 mg/L were taken at 0 hrs and at 48 hrs. The samples were withdrawn from the center of the tanks without prior stirring. All samples were stored at -20 °C until analysis.

Test solutions

Vehicle:
yes
Remarks:
acetone
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 1000 mg of the test item were dissolved up to a final volume of 50 mL acetone by using a ultrasound source for 5 min. This solution was prepared on each respective treatment day. Of this solution, portions of 140 µL, 260 µL, 470 µL and 830 µL were each made up to 1500 µL with acetone and then added to 15 L test medium in the respective tanks.
- Controls: fish exposed to the test medium and fish exposed to the test medium + acetone. The concentration of acetone was 0.01% by volume in all instances.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): 0.01% v/v
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): Results of the analysis of concentrations revealed no detectable test item in the samples with the nominal concentration of 0.19 and 0.62 mg/L. the test item is considered to be unstable in aqueous solutions.

Test organisms

Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: Carp
- Strain: Cyprinus carpio
- Length at study initiation (length definition, mean, range and SD): 46 mm
- Weight at study initiation (mean and range, SD): 1.9 g

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation: Forellenfutter (KLIBA, Klingentalmühle AG, CH-4303 Kaiseraugst/Switzerland)
- Feeding frequency during acclimation: one daily until 1 day prior to treatment
- Health during acclimation (any mortality observed): 0% Mortality








Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
The total hardness of the water was 14 dH°
Test temperature:
19.5 to 21°C.
pH:
8.0 to 8.2.
Dissolved oxygen:
8.6 to 9.3 mg O2 per litre.
Nominal and measured concentrations:
nominal 0.19, 0.34, 0.62, 1.11 and 2 mg/L
Details on test conditions:
TEST SYSTEM
- Aeration: Acclimation: continuous-flow filtered tap water, Test Medium: continuous
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2
- Biomass loading rate: 0.45-0.85 fish biomass/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water with the following amounts of salt: 222.0 mg/L calcium chloride, 60.10 mg/L magnesium sulfate, 64.80 mg/L sodium hydrogencarbonate, 5.75 mg/L potassium chloride.
- Culture medium different from test medium: culture medium: Tap water, test medium: see above
- Intervals of water quality measurement: 6 month

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 12/12
- Light intensity: 500-1500 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality: daily, Clinical signs: 2, 24, 48, 72 and 96 h after start of the treatment, Fish size determination of size in 15 fish with a ruler, body weight: prior to treatment, pH: 2, 24, 48, 72 and 96 h after start of the treatment, dissolved oxygen content: 2, 24, 48, 72 and 96 h after start of the treatment, water temperature: 2, 24, 48, 72 and 96 h after start of the treatment

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8

Food was withheld from the carp one day before and throughout the exposure period of 96 hours.

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
< 0.19 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
<= 0.19 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
0.91 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Other biological observations: enhanced respiratory rate, irregular respiratory rate (observed for all concentrations), loss of equilibrium, convulsions (observed for the middle and high concentrations)
- Mortality of control: No
- Other adverse effects control: No clinical signs were observed in the control tanks.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Results of the analysis of concentrations revealed no detectable test item in the samples with the nominal concentration of 0.19 and 0.62 mg/L. the test item is considered to be unstable in aqueous solutions.
Reported statistics and error estimates:
Calculations:
All values are based on nominal concentrations. The Logit-Model was used to estimate the LC20, LC50 (including 95% confidence limits) and LC80. In addition, LC0, LC50 and LC100 were determined by linear regression of the values used for Logit estimation. NOEC and LOEC were also determined.

Any other information on results incl. tables

The stability test showes that MDAB is unstable in aqueous test medium.

Applicant's summary and conclusion

Validity criteria fulfilled:
no
Conclusions:
In the present study conducted according to OECD guideline 203 (adopted 1983) 10 fish each were exposed to 0.19, 0.34, 0.62, 1.11 and 2.0 mg/L of the test item for 96 h under semi-static conditions. The following LC50 (96 hours) was obtained: 0.91 mg/L (nominal). However, analytical data indicate that the substance is unstable in aqueous test medium.