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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:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Qualifier:
according to guideline
Guideline:
other: OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures"
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
1-(methylamino)anthraquinone
EC Number:
201-417-2
EC Name:
1-(methylamino)anthraquinone
Cas Number:
82-38-2
Molecular formula:
C15H11NO2
IUPAC Name:
1-(methylamino)anthraquinone
Test material form:
solid: bulk

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Duplicate samples from the freshly prepared test medium of the only test concentration and the control were taken at the start of the test.
For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentration during the test period, duplicate samples from the test medium of the only test concentration and the control were collected at the end of the test (after 48 hours).
All samples were diluted by a factor of 1.25 with acetonitrile.
Additional samples of the control and the dilution solvent were taken at test start and test end without any sample treatment.
Storage: The samples from test start were stored in a freezer (≤ - 20 °C), protected from light until analysis was performed. The samples from test end were analysed directly after sampling. Afterwards the samples were again stored deep frozen (< -20 °C) and will be kept stored up to the date of the final report.

Test solutions

Vehicle:
no
Details on test solutions:
Test Concentrations: A filtrate of nominal 10 mg test item/L and a control, corresponding to 7.08 µg /L initial mean measured concentration of the test item. Due to the limit of solubility of the test item in the test medium no higher concentrations could be tested.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
Species: Daphnia magna (Straus), clone 5
Age at Test Start: From 2.25 to 19.25 hours old
Sex: Female
Origin: The daphnids introduced in the test were taken from ibacon's in house laboratory culture.
Breeding Conditions: The daphnids were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in Elendt M4 medium. The test organisms were not first brood progeny. The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of ibacon.

Study design

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

Test conditions

Hardness:
Water Hardness: 2.5 mmol/L (= 250 mg/L) as Calcium Carbonate
Test temperature:
20 to 21°C at test start;
21°C at test end
pH:
8.1 to 8.4 at test start;
7.8 to 7.9 at test end; and thus the pH-value did not vary by more than 1.5 units
Dissolved oxygen:
6.8 to 9.2 mg/L at test start;
8.3 to 8.5 mg/L at test end

Nominal and measured concentrations:
filtrate of nominal 10 mg test item/L and a control, corresponding to 7.08 µg /L initial mean measured concentration of the test item.
Details on test conditions:
Test Environment: Controlled environment room
Measurement of pH, Dissolved
Oxygen and Water Temperature:
The water temperature, pH-values and dissolved oxygen concentrations were determined at test start and test end in each treatment group.
Water Temperature: 20 to 21°C at test start;
21°C at test end
pH-Values: 8.1 to 8.4 at test start;
7.8 to 7.9 at test end; and thus the pH-value did not vary by more than 1.5 units

Dissolved Oxygen Concentration: 6.8 to 9.2 mg/L at test start;
8.3 to 8.5 mg/L at test end
Light Regime: 16 h light : 8 h dark
Light Intensity: The light intensity was 260 to 420 lux (measured once during the test).
Reference substance (positive control):
yes
Remarks:
For the evaluation of the quality of the Daphnia clone and the consistency of the experimental conditions, the reference item potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.

Results and discussion

Effect concentrationsopen allclose all
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 7.08 µg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
>= 7.08 µg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
LOEC
Effect conc.:
> 7.08 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
After 48 hours of exposure no immobilisation of the test animals was observed at the control and 1 animal was immobilised at the only test concentration of 7.08 µg test item/L initial mean measured. However the immobilisation rate of 5 % was not considered a significant toxic effect caused by the test item, because according to the test guideline OECD 202 an immobilisation rate of 10 % at the control is tolerated for the validity of a study. The nontoxicity of the test item is also confirmed by the results of the non-GLP range finder.
Results with reference substance (positive control):
In the most recent test with the reference item potassium dichromate the EC50 after 24 hours was determined to be 1.58 mg test item/L, indicating that the sensitivity of the Daphnia was consistent with the level proposed by the OECD 202 guideline (EC50-24 h between 0.6 and 2.1 mg potassium dichromate/L).
Reported statistics and error estimates:
No statistical analysis was performed. The ECx values could not be quantified due to the absence of toxicity of the test item and were determined directly from the raw data. The NOEC and the LOEC were determined directly from the raw data.

Any other information on results incl. tables

Influence of test item on the Mobility of Daphnia magna

Initial Mean Measured Concentration No. ofDaphniatested No. of immobilisedDaphniaafter  % of immobilisedDaphniaafter
[µg test item/L] 24 h 48 h 24 h 48 h
           
Control 20 0 0 0 0
7.08 20 0 1 0 5

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
Control Immobilisation Rate was 0 % and furthermore no daphnid showed signs of disease or stress. Dissolved Oxygen Concentration was above or equal 8.6 mg O2/L in the control and test vessels at the end of the test.
Conclusions:
The toxic effect of the test item Solvaperm-Rot PFS to Daphnia magna was assessed in a static limit test. The highest test concentration was a filtrate of nominal 10 mg test item/L corresponding to an initial mean measured concentration of 7.08 µg test item/L. Due to the limit of solubility of the test item in the test medium no higher concentrations could be tested.
The 48-hour NOEC was determined to be ≥ 7.08 µg test item/L. The 48-hour LOEC was determined to be > 7.08 µg test item/L and the 48-hour EC50 value was determined to be > 7.08 µg test item/L.
The test item has no toxic effect at its solubilty limit.
The quantification of the test item Solvaperm-Rot PFS in the test samples was performed using liquid chromatography with UV detection.
The concentration of the test item was determined in the duplicate test media samples from the test concentration (filtrate of 10 mg/L) and the duplicate control samples from all sampling times. All reported results refer to initial mean measured values since the concentration of the test item was within ± 20% of the initial mean measured concentration during the test.
Executive summary:

 

Title:

Test item: Acute Toxicity toDaphnia magnain a Static 48-hour Immobilisation Limit Test

Purpose:

The purpose of this study was to determine the influence of the test item on the mobility ofDaphnia magna. Young daphnids (< 24 hours old) were exposed in astatic test for 48 hours to test water containing the test item at the concentration of a filtrate of nominal 10 mg test item/L. This limit test was performed in compliance with the test guidelines to demonstrate that the test item has no toxic effect on daphnids up to at least this concentration.

The test method of application and the test speciesDaphnia magnaare recommended by the test guidelines.

The purpose of the analytical part of this study was to verify the concentration of the test item in the test medium.

Guidelines:

-     Commission Regulation (EC) No 440/2008, Annex, Part C, C.2.: "Daphniasp. Acute Immobilisation Test", Official Journal of the European Union (EN), dated May 30, 2008

-     OECD Guideline for Testing of Chemicals No. 202: "Daphniasp., Acute Immobilisation Test" adopted April 13, 2004

-     OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000

-     SANCO/3029/99 rev.4 11/07/00: Residues: Guidance for generating and reporting methods of analysis in support of pre-registration data requirements for Annex II (part A; Section 4) and Annex III (part A; Section 5) of directive 91/414

 

 

Material and Methods

 

Test Item:

Solvent Red 111

Test Species:

FemaleDaphnia magna, clone 5;2.25to 19.25 hoursold

Source: The daphnids introduced in the test were taken from ibacon's in-house laboratory culture.

Test Design:

This study encompassed two treatment groups: one control and a filtrate of a supersaturated stock suspension of nominal 10 mg test item/L was tested (no concentrations above the solubility limit of the test item in the used test water were tested to avoid physical effects of undissolved test item onto the daphnids). Each treatment group contained 20 individuals. The mobility of the daphnids was determinedin a static 48-hour testby visual observation after 24 and 48 hours.

Endpoints:

Number of immobile organisms after 24 and 48 hours

Test Concentrations:

A filtrate of nominal 10 mg test item/L and a control, corresponding to 7.08 µg /L initial mean measured concentration of the test item.Due to the limit of solubility of the test item in the test medium no higher concentrations could be tested.

Test Conditions:

Water temperature:20to 21 °C; pH value:7.8to 8.4; dissolved oxygen concentration:6.8to 9.2mg/L; photoperiod: 16 h light - 8 h dark; light intensity:260 to 420 lux; and thus were within the ranges requested by guideline OECD 202

 

 

Results:

 

Biological test results:

After 48 hours of exposure no immobilisation of the test animals was observed at the control and 1 animal was immobilised at the only test concentration of 7.08 µg test item/L initial mean measured.However the immobilisation rate of 5 % was not considered a significant toxic effect caused by the test item, because according to the test guideline OECD 202 an immobilisation rate of 10 % at the control is tolerated for the validity of a study. The nontoxicity of the test item is also confirmed by the results of the non-GLP range finder (Appendix IV, Table 10).

 

 

 

Table1. Summary of Biological Results

 

Initial Mean Measured Concentration

% of immobilised daphnids after

 

[µg test item/L]

24 hours

48 hours

 

Control

0

0

 

7.08

0

5

 

EC50[µg/L]:

> 7.08

> 7.08 

 

95 % CI [µg/L]:

n.d. 

n.d. 

 

NOEC [µg/L]:

≥ 7.08 

≥ 7.08 

 

LOEC [µg/L]:

> 7.08 

> 7.08 

 

Values refer to initial mean measured test concentrations
CI: Confidence interval
n.d.: not determinable
NOEC and LOEC were determined directly from the raw data.

 

 

Conclusion:

The toxic effect of the test item toDaphnia magnawas assessed in a static limit test. The highest test concentration was a filtrate of nominal 10 mg test item/L corresponding to an initial mean measured concentration of 7.08 µg test item/L. Due to the limit of solubility of the test item in the test medium no higher concentrations could be tested.

The 48-hour NOEC was determined to be≥ 7.08 µg test item/L. The 48-hour LOEC was determined to be > 7.08 µg test item/Land the 48-hour EC50value was determined to be > 7.08 µg test item/L.

The test item has no toxic effect at its solubilty limit.

The quantification of the test item Solvaperm-Rot PFS in the test samples was performed using liquid chromatography with UV detection.

The concentration of the test item was determined in the duplicate test media samples from the test concentration (filtrate of 10 mg/L) and the duplicate control samples from all sampling times. All reported results refer to initial mean measured values since the concentration of the test item was within±20% of the initial mean measured concentration during the test.

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.