Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

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

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
EU Method C.3 (Algal Inhibition test)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
474-080-4
EC Name:
-
Cas Number:
76801-93-9
Molecular formula:
C14H18I3N3O6
IUPAC Name:
5-amino-N1,N3-bis(2,3-dihydroxypropyl)-2,4,6-triiodobenzene-1,3-dicarboxamide
impurity 1
Reference substance name:
Unknown impurities
Molecular formula:
Not available
IUPAC Name:
Unknown impurities
Test material form:
solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 00104016
- Expiration date of the lot/batch: 12/2020

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in cool place. Keep container tightly closed in a dry and well-ventilated place.
- Stability under test conditions: Stable

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
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 (72 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.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The stock solutions of the test item were prepared in the test medium. 50 mg of the test item was weighed into 500 mL of the test medium for the preliminary and limit test. The stock solutions were ultrasonicated for 10 minutes. The concentrations of test solutions used in the preliminary test were obtained by dilution of the stock solution with test medium.

Differential loading: The preliminary test was performed in a range of the test item nominal concentrations 1 – 100 mg·L-1. The concentration of 100 mg·L-1 was tested in the limit test.
Controls - Preliminary test: The test contained three parallel series of the test item concentrations and three controls without the test item.
Controls - Limit test: The test contained six parallel series of the test item concentration and six controls without the test item.

The water with conductivity smaller than 5 μS·cm-1 was used for the preparation of solutions.
The volume of the test solution was 50 mL in each testing flask. The initial density of 5 000 cells per mL was used in the test.

Test organisms

Test organisms (species):
Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
Details on test organisms:
TEST ORGANISM
- Common name: green freshwater algae Desmodesmus subspicatus
- Strain: Desmodesmus subspicatus Brinkmann 1953/SAG 86.81
- Source: the collection of autotrophic organisms of The Botanic Institute of the Czech Academy of Science, Třeboň on date 8.11.2019
- Method of cultivation: The strain culture was preinoculated from the stock solution and cultivated in flasks with the test medium on indirect daylight at laboratory temperature. Algae inoculum for the test was sampled from exponentially growing inoculum culture.
- Age of inoculum (at test initiation): The strain culture was always set to pre-culturing of cells for 3-4 days before the start of the test. Inoculum culture was kept 3-4 days under conditions at which the test was performed. After this period the culture was in the state of the exponential growth and had the cell density suitable for the performance of the test. The cell density of the pre-culture was measured just before the start of the test and the needed inoculum volume was calculated.

ACCLIMATION
- Acclimation period: 3-4 days
- Culturing media and conditions (same as test or not): same as test

Study design

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

Test conditions

Test temperature:
22.0 – 23.0 °C
pH:
8.1 (test medium)
Conductivity:
1.84 µS·cm-1 (deionized water)
Nominal and measured concentrations:
Limit test:
nominal concentration: 100 mg·L-1
measured concentration: 99 mg.L-1 (0 h), 96.5 mg.L-1 (72 h)
measured concentration: 101 mg.L-1 (0 h), 99.2 mg.L-1 (72 h) without algae
Details on test conditions:
Study design: The preliminary test was performed in a range of the test item nominal concentrations 1 – 100 mg·L-1. Based on low toxicity of the test item found in the preliminary test, the limit test with the test item nominal concentration 100 mg·L-1 was performed subsequently.

Limit test
Stock solution of the test item: 100.04 mg·L-1
Test nominal concentration: 100 mg·L-1
Conductivity of deionized water: 1.54 µS·cm-1
pH of the test medium: 8.0
Volume of inoculum algae culture: 1.17 mL in 500 mL mixture
Lighting during the test: 7 490 – 7 495 lux
Temperature during the test: 22.0 – 22.5 °C
Inicial density: 5000 cells per mL

The volume of the test solution was 50 mL in each testing flask. The flasks were placed on a shaker under the lighting ramp and were incubated under continuous illumination and shaking for 72 hours. At the beginning and the end of the test the pH of the test mixtures was measured. The light intensity and temperature were measured every 24 hours. The density of algae culture was evaluated microscopically at 24, 48 and 72 hours. The cell density was measured by direct counting of living cells in Burker´s counting chamber. The growth rates () and the yield (Y) and subsequently percentage reduction of growth rate and percentage inhibition of yield were calculated.
Reference substance (positive control):
yes
Remarks:
potassium dichromate

Results and discussion

Effect concentrations
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Details on results:
- Exponential growth in the control (for algal test): yes
Results with reference substance (positive control):
- Results with reference substance valid? Yes
The value EC50 for inhibition of growth rate (72h – ErC50) obtained from our last reference test meets the calculated range from the interlaboratory test. The sensitivity of test species and proficiency of our laboratory in test performance was proved.

Reference test: 72 hour – ErC50 = 0.88 mg·L-1 (95% confidence limit: 0.69 – 1.11 mg·L-1)
Interlaboratory test: 72 hour – ErC50 = 0.57 – 1.33 mg·L-1

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The inhibition of growth rate was 0.4 % and inhibition of yield was 1.4 % in the limit test. Therefore exact values of ErC50 and EyC50 could not be calculated and the value of EC is given in the form of a range.
Final values
72 hour – ErC50 > 100 mg·L-1 (nominal concentration)
72 hour – EyC50 ˃ 100 mg·L-1 (nominal concentration)
Executive summary:

The test item, ATIBA-A, was tested for growth inhibition on algae Desmodesmus subspicatus.

The test was performed according to method C.3. - Freshwater Algae and Cyanobacteria, Growth Inhibition Test, Commission Regulation (EU) No.2016/266.

The preliminary test was performed in a range of the test item nominal concentrations

1 – 100 mg·L-1.

The inhibition of growth rate was 1.1 % in the highest concentration and inhibition of yield was 3.9 % in the preliminary test. Based on no toxicity of the test item found in the preliminary test, the limit test was performed subsequently.

Samples for analytical determination were taken at the beginning and at the end of the test. The analytical results showed, that the test item was sufficiently stable in test medium in the conditions of the test, therefore the limit test was performed as static one.

The concentration of 100 mg·L-1 was tested in the limit test. The inhibition of growth rate was 0.4 % and inhibition of yield was 1.4 %.

Samples for analytical determination were taken at the beginning and at the end of the test.

There is stated in the guideline that, if evidence is available to demonstrate that the concentration of the test item 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.

In this study concentration of the test item has been satisfactorily maintained within ± 20 percent of the nominal concentration throughout the test.

The nominal concentrations were used for all evaluations and results.

Test results:

72 hour – ErC50>100 mg·L-1(nominal concentration)

72 hourEyC50˃ 100mg·L-1(nominal concentration)