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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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

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:
2013-12-17 to 2014-12-20
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well performed OECD study with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
N,N'-(3,3'-dimethylbiphenyl-4,4'-ylene)di(acetoacetamide)
EC Number:
202-111-1
EC Name:
N,N'-(3,3'-dimethylbiphenyl-4,4'-ylene)di(acetoacetamide)
Cas Number:
91-96-3
Molecular formula:
C22H24N2O4
IUPAC Name:
N,N'-(3,3'-dimethylbiphenyl-4,4'-diyl)bis(3-oxobutanamide)
Details on test material:
- Name of test material (as cited in study report): Naphtol AS-G trocken
- Molecular formula: C22H24N2O4
- Molecular weight: 380.45 g/mol
- Physical state: Grey powder
- Purity of main component: 99.1 % (w/w) (certified)
- Purity test date: 2013-06-03
- Expiration date of the lot/batch: 2013-06-03
- Stability under test conditions: Not applicable
- Storage condition of test material: Room temperature, protected from light, in original container
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):

OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: not readily biodegradable
- Other:

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: A limit concentration of 1.80 mg/L (nominal) was tested, which corresponds to a geometric mean measured test item concentration of 1.54 mg/L. The limit concentration is based on the maximum solubility level of the test item in water, which was specified by the sponsor to be 1.80 mg/L, and the results of a non GLP preliminary range finding test conducted at the test facility under static conditions. In the non GLP preliminary range finding test, no effects were observed at test item concentrations around the limit of water solubility of the test item up to a loading level of 10 mg test item/L.

- Sampling method:
The limit concentration and the control were analytically verified via HPLC-DAD at the start (0 h) and at the end of the exposure (48 h) as specified below. At the start of the exposure (0 h), samples were taken immediately after preparation of the limit concentration and analysed.
At the end of the exposure (48 h), samples were taken from additional replicates. The additional replicates were prepared with test media, but without daphnids, and incubated under test conditions until sampling. The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All original samples were stored at room temperature. Prepared samples were stored at room temperature until analysis.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:A stock solution (1.80 mg/L of the test item were weighed out) was prepared with dilution water one day prior to the start of the exposure phase. The stock solution was stirred with approximately 1100 rpm for 24 h at room temperature.


- Eluate: Dilution water
- Differential loading: A limit concentration of 1.80 mg/L (nominal) was tested, which corresponds to a geometric mean measured test item concentration of 1.54 mg/L. The limit concentration is based on the maximum solubility level of the test item in water, which was specified by the sponsor to be 1.80 mg/L, and the results of a non GLP preliminary range finding test conducted at the test facility under static conditions. In the non GLP preliminary range finding test, no effects were observed at test item concentrations around the limit of water solubility of the test item up to a loading level of 10 mg test item/L.

- Controls: Dilution water without test item exposed under the same conditions as the test groups.

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, light intensity of
max. 20 µExm-2 x s-1; culture feeding: at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus with an algae cell density of > 10^6 cells/mL.
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: at least 2 h in dilution water
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

DILUTION WATER:
Dilution water ISO test water, according to OECD 202, Annex 3:
Component Concentration [mg/L]
KCl 5.76
NaHCO3 64.8
CaCl2  2 H2O 294
MgSO4  7 H2O 123




Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
No

Test conditions

Hardness:
Dilution water at test start: 240 (mg CaCO3/L)
Test temperature:
At test start: 21.4 °C (Water)
pH:
Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate of the limit concentration and the control)

Geometric mean measured
test item
concentration
[mg/L] pH-Value
1.54 7.28
Control 7.60

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Geometric mean measured
test item
concentration
[mg/L] pH-Values
Replicates
1 2 3 4
1.54 7.33 7.35 7.37 7.39
Control 7.19 7.23 7.28 7.30

Dissolved oxygen:
Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate of the limit concentration and the control)

Geometric mean measured
test item
concentration
[mg/L] Dissolved O2-Concentration
1.54 7.65
Control 8.51

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Geometric mean measured
test item
concentration
[mg/L] Dissolved O2-Concentration
Replicates
1 2 3 4
1.54 8.16 8.15 8.16 8.22
Control 8.17 8.04 8.12 8.20

Salinity:
Not measured, freshwater
Nominal and measured concentrations:
Please refer to "Any other information on materials and methods"
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): Static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start: 669 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and at test end


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
In the non GLP preliminary renge finding test, no effects were observed at test item concentrations around the limit of water solubility of the test item up to a loading level of 10 mg test item/L.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrations
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
1.54 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Details on results:
- Mortality of control: 0 %
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- EC50/LC50: 1.65 (CI 1.60 - 1.70) mg/L
Reported statistics and error estimates:
The effects of the limit concentration of 1.80 mg/L were deduced directly from the observed immobilisation rates.


Any other information on results incl. tables

The percentage immobility, determined in the geometric mean measured concentration of 1.54 mg/L of the test item Naphtol AS-G trocken (limit concentration) and the control after 24 and 48 h under static conditions, is presented in the table below. There was no effect observed at the limit concentration and the control.

Immobilisation Rates of the Definitive Test after 24 and 48 hours of Exposure

 (n = 20, divided into 4 replicates with 5 daphnids each)

Geometric mean measured

test item concentration

 

[mg/L]

IMMOBILISATION [%]

24 h

48 h

Replicates

Replicates

1.

2.

3.

4.

MV

1.

2.

3.

4.

MV

1.54

0

0

0

0

0

0

0

0

0

0

Control

0

0

0

0

0

0

0

0

0

0

MV   = mean value

 

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
At the geometric mean measured concentration of 1.54 mg/L of the test item Naphtol AS-G trocken, representing the maximum solubility of the test item in ISO test water (according to OECD 202, Annex 3), no effects were observed on Daphnia magna.
Executive summary:

In the acute immobilisation test with Daphnia magna(STRAUS), the effects of the limit concentration of 1.80 mg/L of the test item Naphtol AS-G trocken (batch number:95) were determined according to OECD 202 (2004).The limit test was conducted under static conditions over a period of 48 hours.

The test item is a grey powder and has a water solubility of 1.8 mg/L. Therefore, a limit concentration of 1.80 mg/L was chosen representing the maximum water solubility of the test item.Twenty daphnids were exposed to the limit concentrationand the control.

The test item Naphtol AS-G trocken was analytically verified via HPLC-DAD at the start of the exposure (0 h) and at the end of the exposure (48 h) in the limit concentration and in the control.

The measured concentration of the test item in the limit concentration was 1.89 mg/L at the start of the exposure (0 h) and 1.25 mg/L at the end of the exposure (48 h). The geometric mean measured test item concentration was calculated to be 1.54 mg/L.

The geometric mean measured test item concentration is close to the water solubility and therefore regarded to be the maximum solubility level in the used ISO test water. The observed decrease of the test item concentration at the end of the exposure (after 48 h) is regarded to be caused by separation, because in a non GLP preliminary range finding test the measured concentrations at the end of the exposure (48 h) remained stable within±10 % of the initially measured concentrations. It can be concluded, that also no effects will occur at the absolute solubility of the test item in water of 1.80 mg/L, because no effects were observed at test item concentrations up to 10 mg/L in a preliminary test. The effect values given are based on the geometric mean measured concentrations of the test item Naphtol AS-G trocken.

Water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guidelines were fulfilled.

EC-Values, NOEC and LOEC

(based on the geometric mean measured concentration of the test item)

NaphtolAS-G trocken

 

Test

duration

[h]

Geometric mean measured

test item concentration

[mg/L]

Confidence interval
p = 95%

[mg/L]

EC10/ EC50/ EC100

24

> 1.54

Not applicable

48

> 1.54

Not applicable

NOEC

48

  1.54

---

LOEC

48

> 1.54

---

At the geometric mean measured concentration of 1.54 mg/L of the test item
Naphtol AS-G trocken, representing the maximum solubility of the test item in ISO test water(according to OECD 202, Annex 3), no effects were observed on Daphnia magna.