Benzene, poly(propene) derivatives

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

9 October 2007 to 27 November 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

yes

Remarks:

(see below)

Principles of method if other than guideline:

In view of the difficulties associated with the evaluation of aquatic toxicity of poorly water soluble materials, a modification of the standard method for the preparation of aqueous media was performed. An approach endorsed by several regulatory authorities (ECETOC, OECD) is to expose organisms to a saturated solution of the test material in cases where the test material is of high purity and poorly soluble in water and in the permitted auxiliary solvents and surfactants. Using this approach, a saturated solution was prepared by stirring an excess (100 mg/L) of test material in dechlorinated tap water for 24 hours and then removing the undissolved test material by filtration through a pre-conditioned filter (0.2 µm) to give a 100% v/v saturated solution.

GLP compliance:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): AL305B
- Physical state: clear, colourless, slightly viscous liquid
- Lot/batch No.: TS07003
- Analytical purity: 100%
- Expiration date of the lot/batch: 24 May 2007
- Storage condition of test material: room temperature, in the dark

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
100% v/v saturated solution

- Sampling method:
not specified

- Sample storage conditions before analysis:
samples were pretreated and analysed almost immediately after collect and pooling. Duplicate samples were taken and stored at approximately -20ºC for further analysis if necessary.

Test solutions

Vehicle:

no

Details on test solutions:

1100 mg of test material was dispersed in 11 litres of reconstituted water with the aid of propeller stirring at approximately 1500 rpm at a temperature of 21 degrees C for a period of 24 hours. After 24 hours, the stirring was stopped and the undissolved test material removed by filtration (0.2 um Sartopore filter, first approximate 1 litres discarded in order to pre-condition the filter) to give 100% v/v saturated solution. The concentration and stability of the test material in the test preparations were verified by chemical analysis at 0 and 48 hours.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: in-house laboratory cultures
- Age at study initiation (mean and range, SD): Less than 24 hours old
- Weight at study initiation (mean and range, SD): Not stated
- Length at study initiation (length definition, mean, range and SD): Not stated
- Valve height at study initiation, for shell deposition study (mean and range, SD): Not applicable
- Peripheral shell growth removed prior to test initiation: Not applicable
- Method of breeding: Parthogenesis
- Feeding during test: None during exposure

ACCLIMATION
- Acclimation period: None
- Acclimation conditions (same as test or not): Not applicable
- Type and amount of food: Not applicable
- Feeding frequency: Not applicable
- Health during acclimation (any mortality observed): Not applicable

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

Not applicable

Test conditions

Hardness:

Theoretical total hardness of 250 mg/L as CaCO3

Test temperature:

Maintained at approx. 21 degrees C throughout the test

pH:

7.8-8.1

Dissolved oxygen:

8.2-9.0 mg O2/l

Salinity:

Not stated

Nominal and measured concentrations:

100% v/v saturated solution

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): covered to reduce evaporation
- Material, size, headspace, fill volume: 250 mL glass jars containing approximately 250 mL test preparation
- Aeration: None
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Not applicable
- 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): Not applicable
- Biomass loading rate: Not stated

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water used to maintain stock animals
- Total organic carbon: Not stated
- Particulate matter: Not stated
- Metals: Not stated
- Pesticides: Not stated
- Chlorine: Not stated
- Alkalinity: Not stated
- Ca/mg ratio: Not stated
- Conductivity: Not stated
- Culture medium different from test medium: Not stated
- Intervals of water quality measurement: Not stated

OTHER TEST CONDITIONS
- Adjustment of pH: Not required
- Photoperiod: 16 h light/8 h dark
- Light intensity: Not stated

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Any immobilisation or adverse reactions in 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 immobilised if they were unable to swim for approximatelty 15 seconds after gentle agitation.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

> 100 other: % V/V

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 other: % v/v

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

There was no immobilisation in 20 daphnids exposed to a test concentration of 100% v/v saturated solution for a period of 48 hours. Inspection of the immobilisation data gave the following results:

Time (h) EC50 (% v/v saturated solution) 95% confidence limits (% v/v saturated solution)
24 > 100 -
48 > 100 -

The NOEC after 24 and 48 hours exposure was > 100% v/v saturated solution based upon zero immobilisation at this concentration.

The test concentration of 100% v/v saturated solution was the highest attainable test concentration that could be prepared due to the limited solubility of the test material in water.

Results with reference substance (positive control):

not applicable.

Reported statistics and error estimates:

Not stated

Any other information on results incl. tables

Table 1. Cumulative Immobilisation Data in the Definitive Test.

Nominal concentration (% v/v saturated solution		Cumulative Immobilised Daphnia (initial population: 5 per replicate)
		24 hours			48 hours
		No. per replicate	Total	%	No. per replicate	Total	%
Control	R1	0	0	0	0	0	0
	R2	0			0
	R3	0			0
	R4	0			0
100	R1	0	0	0	0	0	0
	R2	0			0
	R3	0			0
	R4	0			0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

the validation criteria were fully fulfilled (as above).

Conclusions:

The acute toxicity of the test material to Daphnia magna has an 48-h EC50 >100% v/v saturated solution with the corresponding NOEC of 100% v/v saturated solution.

Executive summary:

Introduction.

A study was performed to assess the acute toxicity of the test material to Daphnia magna.The method followed that described in the OECD Guidelines for Testing of Chemicals (April 2004) No 202,“Daphnia sp, Acute Immobilisation Test” referenced as Method C.2 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC), US Code of Federal Regulations, Title 40, Part 797, Section 1300 and US EPA Draft Ecological Effects Test Guidelines OPPTS 850.1010.

 

Methods.

Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to a saturated solution of the test material for 48 hours at a temperature of approximately 21°C under static test conditions. The test material solution was prepared by stirring an excess (100 mg/l) of test material via a propeller stirrer in reconstituted water at approximately 1500 rpm at a temperature of 21°C for 24 hours prior to removing any undissolved test material by filtration (0.2 µm Sartopore filter, first approximate 1 litre discarded in order to pre-condition the filter) to produce a saturated solution. Immobilisation and any adverse reactions to exposure were recorded after 24 and 48 hours.

 

Results.

The 48-Hour EC50 for the test material to Daphnia magna based on nominal test concentrations was greater than 100% v/v saturated solution and correspondingly the No Observed Effect Concentration was 100% v/v saturated solution. Analysis of the test preparations at 0 and 48 hours showed the measured concentrations to be less than the limit of quantitation of the analytical method. This does not infer that no test material was in solution but that the dissolved concentration (i.e. bioavailable to the test organisms) was below the limit of quantitation which was assessed down to 0.0045 mg/l.

This study showed that there were no toxic effects at saturation.