Registration Dossier

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

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:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
published in O.J. L 142 (2008)
Deviations:
yes
Remarks:
See Any othfer information on materials and methods
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Test substance name: Acid Brown 235
- Other name: Korostan Brown DGR
- Source and lot/batch No. of test material: 7012/2007
- Expiration date of the lot/batch: May 2018
- Appearance: dark brown powder
Analytical monitoring:
yes
Details on sampling:
The analytical determination of the test substance concentrations were performed at the beginning of the test and at the end of the test. The samples for analysis were taken from the highest (100 mg/L) and the lowest test concentration (10 mg/L). The samples for analysis (0 hours) were prepared at the beginning of the test and immediately delivered in transport box to analytical laboratory. The samples were analyzed on the day of delivery. The samples for analysis at the end of the test (96 hours) were delivered to analytical laboratory immediately after the end of testing. The samples were analyzed on the day of delivery. All samples were stored at laboratory temperature.
Vehicle:
no
Details on test solutions:
The stock solution of the test substance was prepared in the dilution water. 400 mg of the test substance was weighed into 2000 mL of the dilution water for the preliminary test. The concentrations of solutions used in the preliminary test were obtained by dilution of the stock solution with dilution water.
Test organisms (species):
Poecilia reticulata
Details on test organisms:
TEST ORGANISM
- Common name: Poecilia reticulata
- Source: certified supplier EMPLA s.r.o., Hradec Králové (Czech Republic)
- Age at study initiation: 3 - 4 months
- Length at study initiation: 2 - 3 cm:

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: TetraMin – complete feed for aquarium fish (manufacturer: Tetra GmbH, Germany)
- Feeding frequency during acclimation: once a day
- Health during acclimation: mortality 0.33 % (1 fish from 300 individuals)
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Test temperature:
22 ± 2°C, with a variation of ± 1°C at each individual test
pH:
7.5 - 8.1
Dissolved oxygen:
6.8 - 8.6 mg/L
Nominal and measured concentrations:
nominal concentration 10 mg/L:
9.60 mg/L (0 h)
9.54 mg/L (96 h)
nominal concentration 100 mg/L:
89.8 mg/L (0 h)
99.3 mg/L (96 h)
Details on test conditions:
TEST SYSTEM
- Aeration: without aeration
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
Standard dilution water was prepared just before the test by admixing 2.5 mL of stored solutions A, B, C and D in 1000 mL of deionized water.
A: 117.6 g CaCl2·2H20 in 1L deionized water
B: 49.3 g MgSO4·7H2O in 1L deionized water
C: 25.9 g NaHCO3 in 1L deionized water
D: 2.3 g KCl in 1L deionized water
The dilution water should have a pH 7.8 ± 0.2, value measured in this study was 8.0 in the preliminary test. The content of sum Ca + Mg ions should have the value of 2.50 ± 0.10 mmol/L; value measured in this study was 2.55 mmol/L in the preliminary test.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: daily light

TEST CONCENTRATIONS
- Test concentrations: 100, 80, 40, 20 and 10 mg/L
Reference substance (positive control):
yes
Remarks:
potassium dichromate (CAS: 7778-50-9)
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
72 h
Dose descriptor:
LC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
no animals died
see Table 1
Results with reference substance (positive control):
- Positive control substance: potassium dichromate
- LC50 = 110 mg/L (Poecilia reticulata, 96 h)
- Interlaboratory test data: LC50 = 86 - 200 mg/L (Poecilia reticulata, 96 h)

Table 1:Fish mortality

Nominal concentration

mg/L

100

80

40

20

10

Control

Number of fish alive

2 hours

7

7

7

7

7

7

Mortality %

0

0

0

0

0

0

Number of fish alive

24 hours

7

7

7

7

7

7

Mortality %

0

0

0

0

0

0

Number of fish alive

48 hours

7

7

7

7

7

7

Mortality %

0

0

0

0

0

0

Number of fish alive

72 hours

7

7

7

7

7

7

Mortality %

0

0

0

0

0

0

Number of fish alive

96 hours

7

7

7

7

7

7

Mortality %

0

0

0

0

0

0

Validity criteria fulfilled:
yes
Conclusions:
The experiment with the concentration of 100 mg/L included in the preliminary test has been performed using 7 fish, therethrough the conditions of the limit test have been fulfilled. It was demonstrated that the LC50 is higher than this concentration.
No mortality was observed in the preliminary test. The exact values of LC50 could not be calculated and the values of LC are given in the form of a range. The LC100 and LC0 values were identified by direct observation.
There is stated in the guideline that, if evidence is available to demonstrate that the concentration of the test substance has been satisfactorily maintained within ± 20 percent of the nominal or measured initial concentration throughout the test, the results can be based on nominal or measured initial values. We choose the nominal concentrations for the evaluation.
LC50 > 100 mg/L (Poecilia reticulata, 96 h) (nominal concentration)
LC50 > 100 mg/L (Poecilia reticulata, 72 h) (nominal concentration)
LC50 > 100 mg/L (Poecilia reticulata, 48 h) (nominal concentration)
LC50 > 100 mg/L (Poecilia reticulata, 24 h) (nominal concentration)

LC0 = 100 mg/L (Poecilia reticulata, 96 h) (nominal concentration)
LC0 = 100 mg/L (Poecilia reticulata, 72 h) (nominal concentration)
LC0 = 100 mg/L (Poecilia reticulata, 48 h) (nominal concentration)
LC0 = 100 mg/L (Poecilia reticulata, 24 h) (nominal concentration)

LC100 > 100 mg/L (Poecilia reticulata, 96 h) (nominal concentration)
LC100 > 100 mg/L (Poecilia reticulata, 72 h) (nominal concentration)
LC100 > 100 mg/L (Poecilia reticulata, 48 h) (nominal concentration)
LC100 > 100 mg/L (Poecilia reticulata, 24 h) (nominal concentration)
Executive summary:

The test substance, Acid Brown 235, was tested for acute toxicity on fish Poecilia reticulata. The test was performed according to method C.1 – Acute toxicity for fish, Council Regulation (EC) No. 440/2008, published in O.J.L 142, 2008.

The preliminary test was performed in a range of the test substance nominal concentrations 10 – 100 mg/L. Based on no toxicity of the test substance found in the preliminary test, the study was terminated. The experiment with the concentration of 100 mg/L included in the preliminary test has been performed using 7 fish, therethrough the conditions of the limit test have been fulfilled. It was demonstrated that the LC50is higher than this concentration.

The test was performed as static.

Samples for the analytical determination were taken at the beginning and at the end of the test. The analytical results from the preliminary test showed, that the test substance was stable in dilution water at conditions of the test.

There is stated in the guideline that, if evidence is available to demonstrate that the concentration of the test substance has been satisfactorily maintained within ± 20 percent of the nominal or measured initial concentration throughout the test, the results can be based on nominal or measured initial values.

For all evaluations and results the nominal concentrations of the test substance were used.

Test results:

LC50> 100 mg/L (Poecilia reticulata, 96 h) (nominal concentration)

LC50> 100 mg/L (Poecilia reticulata, 72 h) (nominal concentration)

LC50> 100 mg/L (Poecilia reticulata, 48 h) (nominal concentration)

LC50> 100 mg/L (Poecilia reticulata, 24 h) (nominal concentration)

 

LC0= 100 mg/L (Poecilia reticulata, 96 h) (nominal concentration)

LC0= 100 mg/L (Poecilia reticulata, 72 h) (nominal concentration)

LC0= 100 mg/L (Poecilia reticulata, 48 h) (nominal concentration)

LC0= 100 mg/L (Poecilia reticulata, 24 h) (nominal concentration)

 

LC100> 100 mg/L (Poecilia reticulata, 96 h) (nominal concentration)

LC100> 100 mg/L (Poecilia reticulata, 72 h) (nominal concentration)

LC100> 100 mg/L (Poecilia reticulata, 48 h) (nominal concentration)

LC100> 100 mg/L (Poecilia reticulata, 24 h) (nominal concentration)

Description of key information

Key value for chemical safety assessment

LC50 for freshwater fish:
100 mg/L

Additional information