Sodium 2-[[2-amino-8-hydroxy-6-[(methylanilino)sulphonyl]-1-naphthyl]azo]-5-(chloroacetamido)benzenesulphonate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 September 2014 to 08 October 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: AT-0027506800
- Expiration date of the lot/batch: January 20, 2019

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (20 ± 5 °C) under dark and dry conditions

Analytical monitoring:

yes

Details on sampling:

Duplicate samples from the freshly prepared test medium of the only test concentration (filtrate) and the control were taken at test start. 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 2 with acetonitrile.One additional sample of the control and the dilution solution were taken at test start and test end without any sample treatment.

Vehicle:

no

Details on test solutions:

Test Concentration: Filtrate of a supersaturated stock solution of nominal 100 mg test item/L and a control.
Control: In the control, test water was used without addition of the test item.
Dosage of Test Item: The test item was not well soluble in test water. To avoid physical effects of undissolved test item on the daphnids, no concentration above the solubility limit of the test item in test water was tested. Therefore, a supersaturated stock solution of nominal 100 mg test item/L was prepared by suspending 51.1 mg of test item in 511 mL test water. The stock solution was heated up to 80 °C and cooled down while stirring for 25 hours in the dark to dissolve as much test item as possible. Then, the undissolved test item was separated by filtration (0.45 µm cellulose nitrate filter) and the filtrate was used as only test concentration. The test medium was prepared just before introduction of the daphnids (= start of the test).

Appearance of the Test Item in Test Medium:
No remarkable observations.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (Straus), clone 5
Age at Test Start: From 6.0 to 22.0 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 reconstituted water of a similar quality with regards to the constituent salts and total hardness as the test water used in the test (see 6.5). 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.
Acclimatisation: For 6.0 hours under test conditions

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Hardness:

Water Hardness: 2.5 mmol/L (= 250 mg/L) as CaCO3

Test temperature:

20 °C at test start;
20 °C at test end

pH:

6.2 to 6.3 at test start;
6.3 to 6.4 at test end; and thus the pH-value did not vary by more than 1.5 units

Dissolved oxygen:

3.6 to 8.0 mg/L at test start;
7.1 to 7.4 mg/L at test end

Nominal and measured concentrations:

Test Concentration: Filtrate of a supersaturated stock solution of nominal 100 mg test item/L and a control.
Control: In the control, test water was used without addition 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 °C at test start;
20 °C at test end
pH-Values: 6.2 to 6.3 at test start;
6.3 to 6.4 at test end; and thus the pH-value did not vary by more than 1.5 units (see Table 6)
Dissolved Oxygen Concentration:
3.6 to 8.0 mg/L at test start;
7.1 to 7.4 mg/L at test end
Light Regime: 16 h light; 8 h dark
Light Intensity: The light intensity was 1250 to 1280 lux (measured once during the test).

Reference substance (positive control):

yes

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

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 in the control and in the only test concentration of nominal 100 mg test item/L.

Results with reference substance (positive control):

Results of the most recent Test with the Reference Item Potassium dichromate performed in July 2014 (88292220)
Nominal Concentration % of immobilised daphnids after
[mg test item/L] 24 hours 48 hours
Control 0 0
0.125 0 0
0.25 0 0
0.5 0 0
1.0 25 70
2.0 95 100
EC50 [mg/L]: 1.225 0.946
95 % CI [mg/L]: 1.043 - 1.451 n.d.
EC20 [mg/L]: 0.956 0.865
95 % CI [mg/L]: 0.734 - 1.113 n.d.
EC10 [mg/L]: 0.840 0.825
95 % CI [mg/L]: 0.594 - 0.995 n.d.
NOEC [mg/L]: 0.5 0.5
LOEC [mg/L]: 1.0 1.0
Values refer to nominal test concentrations; CI: Confidence interval; n.d.: not determinable; NOEC and LOEC were determined directly from the raw data.

In the most recent test with the reference item potassium dichromate performed in July 2014 (study code 88292220) the EC50 after 24 hours was determined to be 1.225 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 EC50, NOEC and the LOEC were determined directly from the raw data.

Influence of FAT 20075/C TE on the Mobility of Daphnia magna
Endpoint [mg test item/L]	24 hours	48 hours
EC50	> 100	> 100
NOEC	>100	> 100
LOEC	> 100	> 100
Values refer to nominal loading rate EC50, NOEC and LOEC were determined directly from the raw data.

Analytical Results

sample description	% of
[mg test item/L]	nominal^1	n
Control	<LOD	4
Filtrate of 100 mg/L	<LOQ	4

¹mean value of all measured samples per treatment group
n: number of analysed samples

Validity criteria fulfilled:

yes

Conclusions:

The 48-h NOEC, LOEC and EC50 of FAT 20075/C to Daphnia magna was determined to be >100 mg test item/L.

Executive summary:

Acute Toxicity of FAT 20075/C TE to Daphnia magna in a Static 48-hour Immobilisation Limit Test was carried out according to OECD guideline 202 and EU method C.2. The purpose of this study was to evaluate the influence of the test item FAT 20075/C TE on the mobility of Daphnia magna. This study encompassed two treatment groups, one control and a filtrate of a supersaturated stock suspension of nominal 100 mg/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. Young daphnids (<24 hours old) were exposed in a static test for 48 hours to a filtrate of a supersaturated stock suspension of nominal 100 mg/L. The mobility of the daphnids was determined in a static 48-hour test by visual observation after 24 and 48 hours. Number of immobile organisms after 24 and 48 hours. Filtrate of a supersaturated stock solution of nominal 100 mg test item/L and a control. Test Conditions in the study was water temperature: 20 °C; pH value: 6.2 to 6.4; dissolved oxygen concentration: 3.6 to 8.0 mg/L; photoperiod: 16 h light - 8 h dark; light intensity: 1250 to 1280 lux. After 48 hours of exposure no immobilisation of the test animals was observed in the control and in the only test concentration which was a filtrate of a supersaturated stock solution of nominal 100 mg test item/L. There were no immobilization of daphnids seen after 48 h.

 

In conclusion, the toxic effect of FAT 20075/C to Daphnia magna was assessed in a static limit test. The 48-h NOEC, LOEC and EC₅₀ of FAT 20075/C was determined to be >100 mg test item/L.

Description of key information

In a key study, acute toxicity of FAT 20075/C TE to Daphnia magna in a static 48-hour immobilisation limit test was carried out according to OECD guideline 202 and EU method C.2. The purpose of this study was to evaluate the influence of the test item FAT 20075/C on the mobility of Daphnia magna. After 48 hours of exposure no immobilisation of the test animals was observed in the control and in the only test concentration which was a filtrate of a supersaturated stock solution of nominal 100 mg test item/L. In conclusion, the toxic effect of FAT 20075/C to Daphnia magna was assessed in a static limit test. The 48-h NOEC, LOEC and EC₅₀ of FAT 20075/C was determined to be >100 mg test item/L. In another supporting study, FAT 20075/B was tested for its acute toxicity to Daphnia magna in a 48-hour static test following OECD test guideline 202. The nominal concentrations of FAT 20075/B tested were: 0.1, 1, 10, 100 mg test item/L. Additionally, a blank control without test item was carried out. Test concentrations were prepared from a stock solution. No daphnids were immobilised in the control and test concentrations of 0.1, 1, 10 and 100 mg/L after an exposure period of 48h. The EC₅₀ value after 48 hours was determined to be greater than 100 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L