4-[(1-amino-9,10-dihydro-4-hydroxy-9,10-dioxo-2-anthryl)oxy]-N-(3-ethoxypropyl)benzenesulphonamide

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:

06 June 2016 - 09 August 2016

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No. of test material: PCR92X140707
- Expiration date of the lot/batch: 14 October 2019

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Approximately +4 °C in the dark

Analytical monitoring:

yes

Details on sampling:

Samples were taken from the control and 100 % v/v saturated solution test group from the freshly prepared bulk test preparation at 0 and 24 h and from the aged pooled replicates at 24 and 48 h for quantitative analysis. All samples were stored frozen prior to analysis. Duplicate samples were taken and stored frozen for further analysis if necessary.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Based on the results of the range-finding test a "limit test" was conducted at a concentration of 100 % v/v saturated solution to confirm that at highest attainable test concentration no immobilization or adverse reactions to exposure were observed. A nominal amount of test item (550 mg) was dispersed in 11 liters of test water with the aid of propeller stirring at approximately 1500 rpm for 48 hours. After 48 hours the stirring was stopped and any undissolved test item was removed by centrifugation at 40000 g to give a 100 % v/v saturated solution. The prepared concentration was inverted several times to ensure adequate mixing and homogeneity.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia Magna
- Source: in-house breeding facility

The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures. Adult daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room maintaining the water temperature at 18 to 22 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Remarks on exposure duration:

Not applicable

Post exposure observation period:

Not applicable

Hardness:

Approximate theoretical hardness = 250 mg/L as CaCO3.

Test temperature:

18 to 22°C with a maximum deviation of 1 °C.
Measured values = 21 - 22 °C

pH:

Reconstituted water - ISO medium was adjusted to pH 7.8 ± 0.2 (adjusted with NaOH or HCl as necessary).
Measured values (fresh and expired media) = 6.9 - 7.4

Dissolved oxygen:

8.2 - 8.8 mg O2/L (measured values)

Salinity:

Not measured.

Conductivity:

Not measured.

Nominal and measured concentrations:

Nominal values = 0 mg/L (control) and100 % saturated solution (refer above for details of test medium preparation).
Measured values: Fresh medium: 0.594 mg/L (0 hours) and 0.675 mg/L (24 hours)
Old medium: 0.158 mg/L (24 hours) and 0.0999 mg/L (48 hours)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers
- Type: covered to reduce evaporation
- Material, size, headspace, fill volume: 150 mL glass beakers with 100 mL fill volume
- Aeration: No
- Type of flow-through: Not applicable
- Renewal rate of test solution: Test solution was replaced after 24 hours
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST WATER PARAMETERS
Reconstituted Water – ISO Medium
Ingredient Final Concentration
(mg/L)
CaCl2.2H2O 294
MgSO4.7H2O 123
NaHCO3 65
KCl 5.8

The reconstituted water had a pH of 7.8 ± 0.2 adjusted (if necessary) with NaOH or HCl and was aerated until the dissolved oxygen concentration was approximately air-saturation value.
The reconstituted water had an approximate theoretical total hardness of 250 mg/L as CaCO3


OTHER TEST CONDITIONS
- Adjustment of pH: pH of the reconstituted water used to prepare the test solutions was adjusted to pH 7.8±0.2 as noted above. pH of test solutions were not adjusted following the introduction of test material.
- Photoperiod: 16 hours light and 8 hours dark, with a 20 minute transition period to mimic dawn and dusk.
- Light intensity: 200 - 1200 lux (light periods)

EFFECT PARAMETERS MEASURED: Any immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that daphnia were considered to be immobilized if they were unable to swim within 15 seconds after gentle agitation.

VEHICLE CONTROL PERFORMED: Not applicable

RANGE-FINDING STUDY
- Test concentrations: A preliminary media preparation trial was conducted and concluded that a dissolved test item concentration of 2.44 mg/L could be achieved using a saturated solution method of preparation. A range finding study was conducted on this basis at nominal concentrations of 1.0, 10, and 100 % saturated solutions.
- Results used to determine the conditions for the definitive study: Yes. No immobilisation of daphnids was observed in the range finding test, so the definitive test was conducted as a limit test using a 100% saturated solution.

Reference substance (positive control):

yes

Remarks:

A positive control was run as a separate study under similar conditions to those used during the definitive test. Daphnids were exposed to a potassium dichromate control. The date of the positive control study was 29 February to 02 March 2016.

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.29 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

No immobilisation or sub-lethal effects were seen in any of the Daphnia in the treated or control groups during the 48-hour exposure period.

Cumulative Immobilization Data and Observations in the Definitive Test

Nominal Concentration (% v/v Saturated Solution)	24 Hours
	Cumulative Immobilized Daphnia (Initial Population: 5 Per Replicate)						Observations
	R1	R2	R3	R4	Total	%	R1	R2	R3	R4
Control	0	0	0	0	0	0	5 N	5 N	5 N	5 N
100	0	0	0	0	0	0	5 N	5 N	5 N	5 N

 

Nominal Concentration (% v/v Saturated Solution)	48 Hours
	Cumulative Immobilized Daphnia (Initial Population: 5 Per Replicate)						Observations
	R1	R2	R3	R4	Total	%	R1	R2	R3	R4
Control	0	0	0	1	1	5	5 N	5 N	5 N	4 N
100	1	0	0	0	1	5	4 N	5 N	5 N	5 N

 

Validity criteria fulfilled:

yes

Conclusions:

The 48-h EC50 value is greater than 0.29 mg/L. The No Observed Effect Concentration was 0.29 mg/L

Executive summary:

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008. Preliminary solubility work conducted indicated that it was not possible to obtain a testable solution of the test item using traditional methods of preparation e.g. ultrasonication and high shear mixing. A preliminary media preparation trial indicated that a dissolved test item concentration of approximately 2.44 mg/L was obtained from a saturated solution method of preparation indicating this to be the limit of water solubility of this item under test conditions. Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to an aqueous solution of the test item at a nominal concentration of 100 % v/v saturated solution for 48 hours at a temperature of 21 to 22 °C under semi-static test conditions. The test item solution was prepared by stirring an excess (50 mg/L) of test item in test water using a propeller stirrer at approximately 1500 rpm for 48 hours. After the stirring period any undissolved test item was removed by centrifugation (40000g, 30 minutes) to produce a 100 % v/v saturated solution of the test item. Chemical analysis of the freshly prepared saturated solution at 0 and 24 h showed measured test concentrations to range from 0.59 to 0.68 mg/L. Analysis of the aged test preparations at 24 and 48 hours showed measured test concentrations had declined, to 0.16 and 0.10 mg/L, respectively (27 % and 15 % of the freshly prepared test concentrations) and hence, it was considered appropriate to calculate the results based on the geometric mean measured test concentrations in order to give a “worst case” analysis of the data. The geometric mean measured test concentration was calculated to be 0.29 mg/L. There was no significant immobilization in 20 daphnids for a period of 48 hours. Exposure of Daphnia magna to the test item gave EC50 values based on the geometric mean measured test concentrations of greater than 0.29 mg/L. The No Observed Effect Concentration was 0.29 mg/L. This study showed that there were no toxic effects at saturation.

Description of key information

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The study was performed OECD Guideline 202 and EU Method C.4. Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to an aqueous solution of the test item at a nominal concentration of 100 % v/v saturated solution for 48 hours at a temperature of 21 to 22 °C under semi-static test conditions. Based on the findings of the study, exposure of Daphnia magna to the test item gave EC₅₀ values based on the geometric mean measured test concentrations of greater than 0.29 mg/L. The No Observed Effect Concentration was 0.29 mg/L. This study showed that there were no toxic effects at saturation.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.29 mg/L