Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30/04/2020-16/07/2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2020
Report date:
2020

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
other: wet sticky solid
Specific details on test material used for the study:
Designation in Test Facility: 19101604G
Date of Receipt: 16. Oct. 2019
Condition at Receipt Ambient temperature, in proper conditions
6.1.1 Specification
The following information concerning identity and composition of the test item was provid-ed by the sponsor.
Name zinc chlorohydroxy sulphate and copper sulphide, precipitation
Batch no. 20190601
CAS no. unknown
Composition 2.9 % Cu; 18.0 % Zn; 0.5 % Fe; 0.1 % Pb; 0.7 % Cd; 0.4 % As; 0.3 % Sb; 0.1 % Sn; 0.0 % Bi; 0.0 % Br; 2.7 % Cl; 1.1 % F
Storage room temperature (20 ± 5 °C)
Expiry date 31. Dec. 2025
Stability stable under storage conditions
Appearance blackish-green, wet sticky solid
Purity 100 % (UVCB)
Homogeneity homogeneous
Production date not stated
EC no. unknown
Molecular formula not stated
Molecular weight not stated
Vapour pressure not stated
Solubility in solvents H2O: not stated; EtOH: not stated; acetone: not stated; CH3CN: not stated; DMSO: not stated
Stability in solvents H2O: not stated; EtOH: not stated; acetone: not stated; CH3CN: not stated; DMSO: not stated
A certificate of analysis is provided by the sponsor and attached (in copy) in chapter 15. This analytical report is only for informative purpose. The composition might differ in re-gards of percentage values from batch to batch.

6.1.2 Storage
The test item is stored in the test facility in a closed vessel at room temperature (20 ± 5 °C).
6.1.3 Preparation
Since the test item is a poorly soluble UVCB the water-accommodated fraction (WAF) was prepared for the test. This was done by weighing each nominal load (1 / 2.2 / 4.6 / 10 / 22 mg/L nominal concentration), adding the corresponding amount of dilution water and shak-ing vigorously for 24 hours. After a settling time of 1 hour the resulting solution was filtrat-ed through 0.45 µm nylon filters.

The following table show the exact loads for preparation of the water-accomodated frac-tions
Table 6.1 a Exact loads for preparation of the water-accommodated fractions
Nominal concentration in mg/L Real load in mg/L
1 1.0
2.2 2.3
4.6 4.8
10 10.0
22 22.0

Sampling and analysis

Analytical monitoring:
yes

Test organisms

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 identical. The keeping is performed similar to the method described in the OECD guideline, following SOP 115 002 01 („Zucht und Hälterung von Daphnia magna STRAUS“), version 13 from 11. Nov. 2019.
Vessels preserving glasses, nominal volume 2 L
Medium M4-Medium (recipe of ELENDT)
Food green algae (Desmodesmus subspicatus)
Medium renewal twice a week
Photo period 16/8 hours, using neon tubes
Temperature 20  2 °C

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Details on test conditions:
Date of performance 19. – 21. May 2020
Treatments 1 / 2.2 / 4.6 / 10 / 22 mg/L (nominal concentration)
The concentrations to be tested are based on the result of a non-GLP pre-test.
Temperature 19.7 – 20.7 °C
Photo period 16/8 hours, using neon tubes
Duration 48 hours
Observation times 24 and 48 hours
Medium renewal none
Test vessels glass beakers, nominal volume 50 mL, tall shape
Replicates (Treatments) 4 vessels, each containing 20 ± 5 mL test solution and 5 daphnia
Replicates (Blank control) 4 vessels, each containing 20 ± 5 mL dilution water and 5 daphnia
Reference substance (positive control):
yes
Remarks:
Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
3.32 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
3.19 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
2.2 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
The estimation of the biological results was accomplished using the software ToxRat® Professional, version 3.3.0. The details of calculation are stated in chapter 17.
Since the test item is a poorly soluble UVCB, it is not possible to calculate back from the Zn concentration to the test item concentration. Therefore, the nominal concentrations are used for statistical evaluation.

The biological results are presented in the following table:
Table 8.3 a Biological Results Test Item
Parameter Value 95%-confidence interval
24h EC50 3.32 mg/L 2.95 – 3.75 mg/L
48h EC50 3.19 mg/L 2.85 – 3.57 mg/L
48h NOEC 2.2 mg/L --
48h LOEC 4.6 mg/L --

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The estimation of the biological results was accomplished using the software ToxRat® Professional, version 3.3.0. The details of calculation are stated in chapter 17.
Since the test item is a poorly soluble UVCB, it is not possible to calculate back from the Zn concentration to the test item concentration. Therefore, the nominal concentrations are used for statistical evaluation.

The biological results are presented in the following table:
Table 8.3 a Biological Results Test Item
Parameter Value 95%-confidence interval
24h EC50 3.32 mg/L 2.95 – 3.75 mg/L
48h EC50 3.19 mg/L 2.85 – 3.57 mg/L
48h NOEC 2.2 mg/L --
48h LOEC 4.6 mg/L --
Executive summary:

One valid experiment was performed.

The study was performed using 5 concentrations ranging from 1 to 22 mg/L nominal concentration. For each test concentration and the blank control, 20 daphnia were exposed to the test item for 48 hours in a static test system. After 24 and 48 hours, the immobilised daphnia were counted.

Three concentrations showed toxicity of 100% immobilisation. None of the animals was immobilised in the blank control and the two lowest concentrations.

Potassium dichromate K2Cr2O7(CAS No. 7778-50-9) was used as positive controlin a current reference study to assure that the test conditions are reliable.

 

At the start and at the end of the test, the content of Zn as part of the test item in the test solutions was determined using ICP-analysis.

With the help of this determination method it could be proven that the test item was present in the test solutions. Since the test item is a poorly soluble UVCB, it is not possible to calculate back from the Zn concentration to the test item concentration. Therefore, the nominal concentrations are used for statistical evaluation.


The following results were determined for the test itemzinc chlorohydroxy sulphate and copper sulphide, precipitation
(species:Daphnia magna).

48h-NOEL =2.2 mg/L
48h-LOEL = 4.6 mg/L
24h-EL50= 3.32 mg/L
48h-EL50= 3.19 mg/L