Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1982-08-30 - 1982-09-03
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study. For justification of read-across within the category please see chapter 1 of the Chemical Safety Report.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
GLP compliance:
yes
Remarks:
Principles of Good Laboratory Practice (GLP) § 19a, secttion 1 German Chemical Act (ChemG)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
Name: Hoechst-Wachs E
Category: Lubricant for plastics
Content: technical pure, approx. 100%
Appearance: slight yellow powder
Date of production: June 21, 1996
Date of receiving: November 25, 1996
Storage conditions: darkness at approximately 20 °C in a fume cupboard
Solubility in water: < 1 g/L (information by sponsor), < 0.02 mg/L (informal value, Analytical Toxicology)

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
no
Details on test solutions:
Preparation of the test concentrations:
The test substance was weighed into a beaker. The substance was transfered into the test chamber. By heating a thin layer of the test substance was formed to increase the surface volume ratio. Water for dilution was added and then the chamber contents were stirred for approximately 24 hours with a KPG-stirrer and a glass rod.
At first the tested concentration showed a strong turbidity which turned during the exposure time to a light turbidity due to settling down of dispersed material. Particulate matters an the water surface as well as on the bottom were observed in the test vessel.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
Species : zebrafish, Brachydanio rerio (HAMILTON-BUCHANAN)
Origin : Hoechst AG - Hoechst Marion Roussel Preclinical Development Drug Safety
Date of hatching: January 13, 1997
Delivery date: February 19, 1997
The fish were kept for 12 days before the start of the study in water for dilution under the following conditions:
Temperature : 22 ± 1 °C
Oxygen content : >= 80 % of the saturation value
Duration of light period : 12 hours daily
Population density: <= 1 g fish/L water
Feeding : twice daily ad libitum
Food : Tetra Min, Tetra Werke, Melle (Germany)
The body length of 7 representative fish from each batch was measured:
Batch No.: 3/97/L1 (1997-04-18); n=7; Body length (Variation range): 2.6 ¿ 2.9; (cm) mean: 2.7; s: ± 0.12

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
Not applicable

Test conditions

Hardness:
The total hardness of the dilution water was determined weekly and was in the range of 2.1 - 2.3 mmol CaE(2+) + MgE(2+)/L during the study.
Test temperature:
Temperature (°C)
Concentration group: 22.7 - 22.8
Control group: 22.2 - 22.9
pH:
Concentration group: 7.5 - 7.9
Control group: 7.7 - 8.0
Dissolved oxygen:
Oxygen content (mg/L):
Concentration group: 6.1* to 9.7
Control group: 7.6 to 9.8
* The chamber was aerated from 24 hours after study start on.
Salinity:
Water for dilution:
Reconstituted water, composition according to ISO/DIS 7346/1 was used as water for dilution.
Preparation was carried out in a unit consisting of two Hostalen - lined steel vessels with a capacity of 1000 litres each.
The dilution water was prepared as described and was aerated until oxygen saturation. The pH was measured before the use of each study batch. It was 8.0.
Nominal and measured concentrations:
One test group and one control group with 7 fishes, each, had been evaluated:
0 mg/L control group
10 g/L (nominal) test group
Details on test conditions:
The study was conducted in a static system. The test chambers, which were calibrated to 4 liters, were made of glass (length 20 cm, width 15 cm, height 20 cm) and stood in a water bath made of Hostalit Z® with a Plexiglas viewing panel. The temperature of the water bath was regulated by a thermostat to 22 ± 1 °C. The chambers were illuminated from above from 06.00 a.m. to 06.00 p.m. The light intensity directly over the chambers was approximately 700 lux.
The test chambers were not aerated during the course of the study except in the case of the 10000 mg/L group, which was aerated 24 hours after study start. From then on, the maintenance of an oxygen content of at least 60% of the saturation value was not ensured. Therefore aeration was carried out using a glass capillary with a bubble-frequency of 1 - 3 per second..

Test procedure
After the test concentrations had been prepared and water parameters recorded, 7 fish were assigned to each test and control chamber. The fish received no feed for the entire study period. Inspection of the fish took place after 3, 6, 24, 48, 72 and 96 hours and involved recording the lethality and visible changes in appearance and behavior. Dead fish were removed from the chambers. Fish were considered dead when there was a lack of opercular movement and no response to slight mechanical stimulus. The water parameters were measured and recorded before study start and after 0, 24, 48, 72 and 96 hours.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
LC0
Effect conc.:
10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC0
Effect conc.:
10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC0
Effect conc.:
10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
> 10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 10 g/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
Observations
The observation of symptoms was disturbed because of the turbidity.
Some fish of the concentration group showed slight changes in appearance and behavior (swimming posture tail-heavy, projecting opercula) only transiently at the second day of exposure.

Lethality
In this 96-hour acute toxicity study of the test item in zebra fish (Brachydanio rerio) no lethality occurred in the 10000 mg/L-group (nominal concentration) and in the control group. The tested nominal concentration was orders of magnitude above the solubility limit of the test substance in the test water.
Results with reference substance (positive control):
not applicable
Reported statistics and error estimates:
not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
A 96 hour acute toxicity test of Hoechst-Wachs E on Brachydanio rerio according to relevant guidelines and compliant to GLP (reliability category 1) had been performed. Under the conditions of this test (limit test due to the very low solubility of the test item in water) the LC50 of the test item after 24, 48, 72 and 96 hours was > 10 g/L (based on nominal concentration), the LC0 was equal to 10 g/L.
Executive summary:

Hoechst-Wachs E

was tested in zebra fish (Brachydanio rerio) over 96 hours in astatic system.

The concentrations tested was 10000 mg/L and a negative control (0 mg/L).

The test substance was weighed into a beaker. The substance was transfered into the test chamber. By heating a thin layer of the test substance was formed to increase the surface volume ratio. Water for dilution was added and then the chamber contents were stirred for approximately 24 hours with a KPG-stirrer and a glass rod.

At first the tested concentration showed a strong turbidity which turned during the exposure time to a light turbidity due to settling down of dispersed material. Particulate matters on the water surface as well as on the bottom were observed in the test vessel.

An analytical determination of Hoechst-Wachs E in the test water was not possible, because the solubility limit of the test substance is below the analytical detection limit (0.02 mg/L test water).

 

after 24
hours

after 48
hours

after 72
hours

after 96
hours

LC0 (mg/L)

10000

10000

10000

10000

LC50 (mg/L)

> 10000

> 10000

> 10000

> 10000

LC100 (mg/L)

not determined

not determined

not determined

not determined

The 10000 mg/L group exhibited reversible symptoms but no lethality.