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:
04 - 06 Aug 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. certificate)
Remarks:
Landesamt für Umwelt, Wasserwirtschaft und Gewerbeaufsicht, Mainz, Germany

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): PM-5927
- Substance type: Whitish/Crystalline powder
- Physical state: solid
- Lot/batch No.: 14-121
- Expiration date of the lot/batch: 01 June 2016
- Storage condition of test material: Room Temperature (20 ± 5 °C); Keep away from light

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
At the beginning and at the end of the test, the content of the test item in the test solutions was determined using HPLC-determination by measuring Sulfonium, tris(4-chlorophenyl)-(cation) and 5-benzoyl-4-hydroxy-2-methoxybenzene-sulfonic acid (anion).

Test solutions

Vehicle:
no

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: originally from Umweltbundesamt Berlin, Germany; in-house breeding since 27 Sep 2007
- Age at study initiation (mean and range, SD): < 24 hours
- Method of breeding: 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 husbandry is performed similar to the method described in EN ISO 6341. Vessels: preserving glasses, nominal volume 2 L; Medium: M4-Medium (recipe of ELENDT); Medium renewal twice a week; Photo period: 16/8 hours, using neon tubes; Temperature 20 ± 2 °C
- Feeding during test: none
- Food type: unicellular green algae (Desmodesmus subspicatus)

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Test temperature:
19.8 - 21.0 °C
pH:
7.5 - 7.8
Dissolved oxygen:
8.5 - 8.8 mg O2/L
Nominal and measured concentrations:
control, 10, 18, 32, 54, and 100 mg/L (nominal)
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: Beakers, glass, nominal volume 50 mL
- Aeration: After preparation, the dilution water was aerated
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Deionised water with an enrichment of certain minerals (according to guidelines)

OTHER TEST CONDITIONS
- Adjustment of pH: yes, to 7.9
- Light intensity: diffuse lighting

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): 24 and 48 hours, the immobilised Daphnia were counted

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 1.8
- Range finding study:
Test concentrations: 1, 10, and 100 mg/L
- Results used to determine the conditions for the definitive study: At the concentration 100 mg/L, 100 % immobilization was observed. In the lower concentrated treatments none of the Daphnia were immobilized.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
19 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
18 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Results with reference substance (positive control):
Potassium dichromate K2Cr2O7 was used as positive control in a current reference study. The 24h-EC50 value was determined as 1.6 mg/L, lying within the demanded range of 0.6 – 1.7 mg/L.
Reported statistics and error estimates:
Calculation of results was performed with the help of validated software (Microsoft Excel®). The estimation of the biological data was accomplished using the software ToxRat® Professional, version 3.1.

Any other information on results incl. tables

Analytical Determinations:

The determined concentrations of the cation at the start and the end of the test were in the range of 97 % - 110 % of the nominal concentration. The determined concentrations of the anion at the start and the end of the test were in the range of 98 % - 109 % of the nominal concentration. Therefore, the determination of the biological results was based on the nominal concentration.

Table 1: Measured concentrations (cation)

Nominal Concentration test item

Measured Concentration in mg/L (t = 0 h)

Measured Concentration in mg/L (t = 48 h)

% of Nominal Concentration (t = 0h)

% of Nominal Concentration (t = 48h)

mg/L

mg/L

mg/L

 mg/L

mg/L

Blank control

n.d.

n.d.

n.d.

n.d.

10

9.99

10.74

100%

107%

18

17.52

18.95

97%

105%

32

32.61

35.26

102%

110%

54

54.31

58.59

101%

109%

100

100.87

108.66

101%

109%

Table 2: Measured concentrations (anion)

Nominal Concentration test item

Measured Concentration in mg/L (t = 0 h)

Measured Concentration in mg/L (t = 48 h)

% of Nominal Concentration (t = 0h)

% of Nominal Concentration (t = 48h)

mg/L

mg/L

mg/L

 mg/L

mg/L

Blank control

n.d.

n.d.

n.d.

n.d.

10

10.01

10.70

100%

107%

18

17.57

18.84

98%

105%

32

32.39

35.03

101%

109%

54

53.69

57.93

99%

107%

100

100.03

107.72

100%

108%

Biological Findings:

Table 3: 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

10

0

0

0

0

0

0

0

0

0

0

18

0

0

0

0

0

1

2

1

2

30

32

3

1

1

4

45

5

5

5

5

100

54

5

5

4

5

95

5

5

5

5

100

100

5

5

5

5

100

5

5

5

5

100

Validity:

- The 24h-EC50 of K2Cr2O7 should lie between 0.6 and 1.7 mg/L. The 24h-EC50 of K2Cr2O7 was determined as 1.6 mg/L in a separate GLP study.

- Immobilisation in the controls may not exceed 10 %. Immobilisation in the controls was 0 %.

- The concentration of dissolved oxygen at the end of the test must be at least 3 mg/L. The lowest concentration of dissolved oxygen at the end of the test was 8.5 mg/L.

- The pH-value in the test solutions should not vary by more than 1.5 units during the test. The highest variation was 0.3 units.

Applicant's summary and conclusion