Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Version / remarks:
OECD Guideline for Testing of Chemicals No. 202, adopted 13. Apr. 2004:
”Daphnia sp., Acute Immobilisation Test
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Version / remarks:
Council Regulation (EC) No. 440/2008, Method C.2. “Daphnia sp. Acute Immobilisa-tion Test”, adopted 30. May 2008
OECD guidance document no. 23, GUIDANCE DOCUMENT ON AQUATIC TOXICI-TY TESTING OF DIFFICULT SUBSTANCES AND MIXTURES, adopted 14. Dec. 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
all tests were made in accordance with the „Revised OECD Principles of Good Laboratory Practice“ (Paris, 1997) Chemicals Act of the Federal Republic of Germany (ChemG) §19a and §19b and an-nexes 1 and 2 from 28. Aug. 2013,
Analytical monitoring:
yes
Details on sampling:
- Concentrations:
Dilution water (Daphnia medium) with the following specification will be used (stated in SOP 11800201):
Table 6.4 a Dilution water specification
Parameter Concentration in mg/L
CaCl2*2H2O 293.80
MgSO4*7H2O 123.30
NaHCO3 64.80
KCl 5.80
Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: For this limit test, a stock solution containing 101 mg/L test item in dilution water was pre-pared and homogenised through vigorous shaking
- Eluate: water
Test organisms (species):
Daphnia magna
Details on test organisms:
6.3.1 Specification
Species Daphnia magna
Authority STRAUS
Strain Berlin
Sex female
Age between 0 and 24 hours
Origin Umweltbundesamt Berlin
In-house breeding since 27. September 2007
Selection of the test system was made following the proposal of the guidelines

6.3.2 Animal Husbandry
Daphnia magna is bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. They multiply by parthenogenesis, thus being genetically identi-cal. The husbandry is performed similar to the method described in the OECD guideline, following SOP 115 002 01 („Zucht und Hälterung von Daphnia magna STRAUS“), version 12 from 02. Feb. 2015.
Vessels preserving glasses, nominal volume 2 L
Medium M4-Medium (recipe of ELENDT)
Food unicellular green algae (Desmodesmus subspicatus)
Medium renewal twice a week
Photo period 16/8 hours, using neon tubes
Temperature 20  2 °C

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
19 hours 40 minutes before the start of the test, the adult animals were separated from the young. 40 minutes before test start, the adults were caught with the help of a glass tube, and the newborn daphnia, aged between 0 and 19 hours, were sieved from the medium and immediately placed into a beaker containing dilution water. After the settling-in peri-od, animals which showed no apparent damage were used for the test
Hardness:
Resulting hardness in mmol/L: 2.502
Resulting hardness in mg CaCO3/L: 250
Test temperature:
20°C - 22°C
pH:
7.8
Dissolved oxygen:
Nominal Concentration in mg/L pH O2-Concentration in mg/L
0 h 48 h 0 h 48 h
Blank control 7.8 7.9 8.8 8.2
100 7.8 7.8 8.6 8.2
Conductivity:
conductivity < 0.4 µS/cm
Nominal and measured concentrations:
nominal oncentration test item: 100 mg/l
measured concentration at t=0h: 101.48 / (102.95)*
measured concentration at t=48h 111.00


*Value in italic and in brackets is the measured concentration at the beginning of the test where the sampling vials were too full and too tightly closed and therefore not further considered for evaluation
Details on test conditions:
TEST SYSTEM
- Test vessel: glass beakers, nominal volume 50 mL, tall shape
Type: open glass beakers
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 vessels, each containing 20 ± 5 mL test solution and 5 Daphnia
- No. of vessels per control (replicates): 4 vessels, each containing 20 ± 5 mL dilution water and 5 Daphnia
- No. of vessels per vehicle control (replicates):
- Biomass loading rate:

OTHER TEST CONDITIONS
- Adjustment of pH: pH is adjusted to 7.8  0.2 with NaOH or HCl.
- Photoperiod: 16/8 hours, using neon tubes


VEHICLE CONTROL PERFORMED: yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)
Key result
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
> 48 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
The study was performed as a limit test using 1 concentration at 100 mg/L. For the test concentration, 20 daphnia were exposed to the test item for 48 hours in a static test sys-tem. After 24 and 48 hours, the immobilised daphnia were counted.
Only one daphnia was immobilised in the tested concentration which can be stated as not significant. None of the animals was immobilised in the blank control
Results with reference substance (positive control):
- Results with reference substance valid?
- Relevant effect levels: NOEC LOEC EC50 for 24h & 48h
- Limit test: using 1 concentration at 100 mg/
-
Reported statistics and error estimates:
No relevant toxicity occurred in any treatment or in the blank control. Therefore, no statis-tical evaluation was conducted

1.1     Immobility

Only one daphnia was immobilised in the tested concentration which can be stated as not significant (below validity criterion of maximum 10% immobility in blank control). In the blank control, none of the daphnia were immobilised (see table below).

Table8.1‑a      Immobility

Nominal Concentration in mg/L

Immobility 24 hours

Immobility 48 hours

absolute

in %

absolute

in %

Blank control

0

0

0

0

0

0

0

0

0

0

100

0

1

0

0

5

0

1

0

0

5

 

1.2     pH and O2

The pH values in the test media and the blank control and the concentration of dissolved oxygen are given in the following tables:

Table8.2‑a      pH and O2-values

Nominal Concentration in mg/L

pH

O2-Concentration in mg/L

0 h

48 h

0 h

48 h

Blank control

7.8

7.9

8.8

8.2

100

7.8

7.8

8.6

8.2

The measured concentrations for treatment and blank control are given in the following table:

Table8.3‑a      Measured Concentrations

Nominal

concentration

test item 

Measured

concentration

Measured

concentration

% of nominal concentration

% of nominal concentration

 t = 0 h

 t = 48 h

 t = 0 h

 t = 48 h

mg/L

mg/L

mg/L

%

%

Blank control

0

0

--

--

100

101.48 /(102.95)*

111.00

101.48

111.00

 

Validity criteria fulfilled:
yes
Remarks:
Immobilisation in the controls may not exceed 10 %. Immobilisation in the controls was 0 %.
Conclusions:
The study was performed as a limit test using 1 concentration at 100 mg/L. For the test concentration, 20 daphnia were exposed to the test item for 48 hours in a static test sys-tem. After 24 and 48 hours, the immobilised daphnia were counted.
Only one daphnia was immobilised in the tested concentration which can be stated as not significant. None of the animals was immobilised in the blank control
The EC50 is > to 100mg/L, accordingly test substance Blendazol Red Blendwell is not toxic to aquatic invertebrates Daphnia magn
Executive summary:

In a GLP compliant 48 h acute toxicity test, the test substance was assessed according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test); and EU Method C.2 (Acute Toxicity for Daphnia).Daphnia magna were exposed to the test substance at concentrations of 100 mg/l and observed at 24 and 48 h for mobility. The EC50 of the test item in Daphnia magna after 48 hours is determined to be superior to 100 mg/L.

The EC50 of the test substance to Daphnia magna is >100 mg/l. Hence, the Blendazo Red is not toxic to aquatic invertebrates Daphnia magna.

Description of key information

The biological results are presented in the following table:

Table9.1‑a      Biological Results Test Item

Parameter

Value

48h NOEC

≥100 mg/L

48h LOEC

> 100 mg/L

24h-EC50

> 100 mg/L

48h-EC50

> 100 mg/L

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
100 mg/L
EC50/LC50 for marine water invertebrates:
100 mg/L

Additional information

The prupose of the study was to to evaluate the toxic potential of Blendazol Red Blendwell towards freshwater flea, using the species Daphnia magna Straus, which was chosen in the guideline as a typical part of zoo-plankton.

Young daphnia, aged less than 24 hours at the start of the test, were exposed to the test item at a range of concentrations for a period of 48 hours.Immobilisation was recorded after 24 hours and 48 hours and compared with control values. The results were analysed in order to calculate the EC50at 24h and 48h.

The EC50 24h and 48h was > 100 mg/L, accordingly the test item is not toxic to aquatic invertebrates


Route: .live1