Benzoic acid, 4-[2-[2-oxo-1-[(phenylamino)carbonyl]propyl]diazenyl]-

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Study, conducted according to all applicable test guidelines with minor or no deviations. The study is well documented.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

4-[(1E)-2-[2-oxo-1-(phenylcarbamoyl)propyl]diazen-1-yl]benzoic acid

EC Number:

603-014-0

Cas Number:

124772-04-9

Molecular formula:

C17 H15 N3 O4

IUPAC Name:

4-[(1E)-2-[2-oxo-1-(phenylcarbamoyl)propyl]diazen-1-yl]benzoic acid

Sampling and analysis

Analytical monitoring:

yes

Test organisms

Test organisms (species):

other: Daphnia magna STRAUS (Clone 5)

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: STRAUS (Clone 5)
- Source: Institut fur Wasser-, Boden- und Lufthygiene (WaBoLu)
- Method of breeding: In 2-3 L glass vessels with approximate 1.8 L culture medium, at 21 °C (temperatures of 20 - 25 °C are tolerated), in an incubator, 16 h illumination, illumination strength max. 20 µE/m2*s.
- Culture medium: Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaC03/L

Feeding during test:
- Food type: mix of Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of > 106 cells/ml, ad libitum.
- Frequency: At least 5x weekly

Age of test organisms: 2 - 24 h old
Acclimation: Ca. 2 h in dilution water
Test temperature: 18-22°C, ± 1 X constant
Illumination: Diffuse light, illumination range max. 20 µE • m-2 • s-1
Photoperiod: 16/8 h light/dark cycle
Feeding: The daphnids were not fed during the study.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

259 [mg CaC03/L]

Test temperature:

20.8°C

pH:

7.67

Dissolved oxygen:

8.13 mg/l

Salinity:

Conductivity 667 [µS/cm]

Details on test conditions:

Test duration: 48 h
Test vessels: Glass beakers (5 cm ID x 8 cm H), 50 mL
Volume of the study medium: 20 mL

Dilution water: According to EC 92/69 L383A C.2. (Annex)
Component Concentration [mg/L] in dilution water
KCL 5.76
NaHC03 64.8
CaCb•2 H20 294
MgSOv 7H20 123

Number of study organisms: 20 animals, divided into 4 (2*) parallel samples, per concentration and each with 5 (10*) animals
control

Number of animals per test vessel: 5(10*)
Number of parallel samples per concentration: 4 (2*)

*The information in brackets refer to the test of the reference item

Reference substance (positive control):

yes

Remarks:

Potassium dichromate p.a. (MERCK)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

0.42 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: Carbon content and DOC-Analysis

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

0.35 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: Carbon content and DOC-Analysis

Basis for effect:

mobility

Effect concentrations 3

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

0.51 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: Carbon content and DOC-Analysis

Basis for effect:

mobility

Remarks on result:

other: 95% CL = 0.48-0.54

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.54 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: Carbon content and DOC-Analysis

Basis for effect:

mobility

Remarks on result:

other: 95% CL=0.49-0.60

Effect concentrations 5

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

2.42 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: Carbon content and DOC-Analysis

Basis for effect:

mobility

Effect concentrations 6

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

0.61 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

other: Carbon content and DOC-Analysis

Basis for effect:

mobility

Effect concentrations 7

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.3 mg/L

Nominal / measured:

estimated

Basis for effect:

mobility

Effect concentrations 8

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

0.61 mg/L

Nominal / measured:

estimated

Basis for effect:

mobility

Details on results:

Results of the Preliminary Tests
The percentage immobility was determined in all test item and control groups after 24 and 48 hours of exposure under static conditions. The immobilisation rates are given in Table 2.
Table 2: Immobilisation Rates of the Preliminary Range Finding Tests [%] (n = 20)

Dilutions of the saturated solution IMMOBILISATION [%]
#) 24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
1:1 100 100 100 100 100 100
1:10 100 80 90 100 100 100
1:100 0 0 0 0 0 0
1:100 0 0 0 0 0 0
Control 0 0 0 0 0 0
MV: Mean value


Results of the Definitive Test
The percentage immobility was determined in ail test item and control groups after 24 h and 48 h. The immobilisation rates in % after 24 h and 48 h of exposure under static conditions are given in Table 3.
Table 3: Percentage of Daphnids Incapable of Swimming after 24 and 48 h of Exposure (n = 20)

Dilutions of Calculated* IMMOBILISATION [%]
the saturated test
solution concentration 24 h 48 h
[mg/L] Replicates Replicates
1. 2. 3. 4. MV 1. 2. 3. 4. MV
1:10 2.42 100 100 100 100 100 100 100 100 100 100
1:20 1.21 100 100 60 80 85 100 100 80 100 95
1:40 0.61 100 80 100 80 90 100 100 100 100 100
1:80 0.30 0 0 0 0 0 0 0 0 0 0
1:160 0.15 0 0 0 0 0 0 0 0 0 0
Control 0 0 0 0 0 0 0 0 0 0

MV Mean value
*) Calculated based on carbon content (ThTOC= 0.628 mg/mg) and DOC analysis
#) Saturated solution: The maximum dissolved concentration of the test item that can be achieved under the test conditions in the test medium, ace. to OECD Series, No. 23 (2000)6.

The 24 and 48 h EC10- and EC50-values were calculated by probit analysis.

Results with reference substance (positive control):

The percentage immobility for the reference item was determined after 24 h. The EC100-value was determined directly from the test results. The EC10-and EC50-with 95 % confidence interval (CI) was determined in a probability network by interpolation according to standard procedures. EC-values after 24 h of the reference item in mg/L:
EC10 :1.08
EC50 : 1.47 (CI 1.36- 1.58)
EC100 : 3.20
The EC50-value of the reference item potassium dichromaie after 24 h is within the prescribed concentration range of 1.0 - 2.5 mg/L of quality criteria according to AQS: DIN Guideline 38412L30.

Reported statistics and error estimates:

EC100-values were deduced directly from the dose-response-relationship after 24 and 48 h. There was no mathematical calculation.
The EC10- and EC50-values were calculated via probit analysis. Probits according to WEBER (1986).
The concentration effect relationship is shown graphically in a probability network. Calculation of the confidence intervals was carried out using standard procedures according to BREITIG & TUMPLING (1982).

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

According to the criteria of the EU regulation 1272/2008 the substance must be classified as Aquat. Acut 1, H400