4-poly(propyl), benzene sulfonic acid

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

7 August 2007 to 20 December 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:

yes

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Principles of method if other than guideline:

Due to a technical oversight, particulate maner, pH, hardness, alkalinity, conductivity and Total Organic Carbon analysis of the diluent water was not performed at the start of the definitive test. Given that the control daphnids showed no adverse effects of exposure, the diluent water was considered not to contain any deficiencies in the above water quality parameters that would affect the outcome or integrity of the study.

GLP compliance:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

Sponsor's identification: AS305BD
Description: dark brown, viscous liquid
Lot number: TS07002
Analytical purity: 100%
Storage conditions: room temperature in the dark

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The recovery values for the pre-study samples were observed to be low. Therefore, in order to increase detection and improve accuracy a larger volume of sample was taken for the definitive test. A Strata C8 (500 mg/ 3 mL) solid phase extraction (SPE) cartridge was sequentially preconditioned with methanol and water. A volume (500 mL) of test sample was eluted through the cartridge and the cartridge dried on the SPE manifold. The test material was eluted from the cartridge with methanol (10 mL).

Water samples were taken from the control (replicates R1 - R4 pooled) and the 100% v/v saturated solution test group (each replicate analysed separately) at 0 and 48 hours for quantitative analysis.
Duplicate samples were taken and stored at approximately -20°C for further analysis if necessary.

Test solutions

Vehicle:

no

Details on test solutions:

An amount of test material (2250 mg) was dispersed in 22.5 Iitres of dechlorinated tap water with the aid of propeller stirring at approximately 1500 rpm at a temperature of approximately 21°C for a period of 24 hours. After 24 hours the stirring was stopped and the undissolved test material removed by filtration (0.2 µm Sartopore filter, first approximate 1 litre discarded in order to precondition the filter) to give the 100% v/v saturated solution.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: in-house laboratory cultures
- Age at study initiation (mean and range, SD): less than 24 hours old
- Method of breeding: parthenogenesis
- Feeding during test: no


ACCLIMATION
not applicable to laboratory bred Daphnia.

QUARANTINE (wild caught)
not applicable to laboratory bred Daphnia.

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:

The reconstituted water has an approximate theoretical total hardness of 250 mg/l as CaCO3. Due to a technical oversight water hardness was not tested at the beginning of the definitive test.

Test temperature:

20°C

pH:

7.1-8.0
Due to a technical oversight pH was not tested at the beginning of the definitive test.

Dissolved oxygen:

8.8-9.0 mg O2/L

Salinity:

Not stated

Nominal and measured concentrations:

Following a range-finding tests, a 'limit test' was conducted at a concentration of 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
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- Stock Solutions:
a) CaCI,.2H,O 11.76 g/L
b) MgSO,.7H,O 4.93 g/L
c) NaHCO, 2.59 g/L
d) KCI 0.23 g/L

- Preparation:
An aliquot (25 mL) of each of solutions a-d was added to each litre (final volume) of deionised water with a conductivity of <5 µS cm-I. 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 mgIL as CaCO3.

- Total organic carbon: Due to a technical oversight TOC was not measured at the start of the definitive test.
- Particulate matter: Due to a technical oversight particluate matter was not measured at the start of the definitive test.
- Pesticides: not reported.
- Chlorine: not reported.
- Alkalinity: Due to a technical oversight alkalinity was not measured at the start of the definitive test.
- Conductivity: Due to a technical oversight conductivity was not measured at the start of the definitive test.
- Culture medium different from test medium: no
- Intervals of water quality measurement: 24 hours

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16 hours light / 8 hours dark (20 minute dawn/dusk period).

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Immobilisation:
The criterion of effect used was that Daphnia were considered to be immobilised if they were unable to swim for approximately 15 seconds after gentle agitation. Any immobilisation or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: 100% v/v saturated solution preparation was used as the substance is highly insoluble in water (<0.01mg/l).

- Range finding study
- Test concentrations: 1.0, 10, 100% v/v saturated solution.
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 other: % v/v

Nominal / measured:

nominal

Conc. based on:

other: Saturated

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:

other: Saturated

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. 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:

An estimate of the EC50 values was given by inspection of the immobilisation data.

Any other information on results incl. tables

No significant immobilisation was observed at any test concentration throughout the test. A single daphnid was observed to be immobile after 24 and 48 hours exposure however, this was considered to be due to natural causes and not a toxic effect as no immobilisation was observed at the higher test concentrations throughout the test. Based on this information, a single test concentration of four replicates, of 100% v/v saturated solution was selected for the definitive test. This experimental design conforms to a "Limit test" to confirm that at the highest attainable test concentration of 100% v/v saturated solution, no immobilisation or adverse reactions to exposure were observed.

Table 1. Cummulative Immobilisation data in the range-finding test.

Nominal concentration (% v/v saturated solution)	Cumulative immobilised Daphnia (initial population: 10 per replicate)
	24 hours	48 hours
Control	0	0
1	1*	1*
10	0	0
100	0	0

* Immobilisation considered to be due to natural causes and not a toxic effect as no immobilisation was observed at the higher tier concentrations.

Table 2. Cummulative immobilisation data in the definitive test.

Nominal concentration (% v/v saturated solution)		Cumulative immobilised Daphnia (initial population: 10 per replicate)
		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:

validity criteria (as stated above) fulfilled.

Conclusions:

The acute toxicity of the test material to Daphnia magna gave a 48-hour EC50 of >100% v/v saturated solution and a 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 20°C under static test conditions. The test material solution was prepared by stirring an excess (100 mg/L) of test material via propeller stirrer in reconstituted water at approximately 1500 rpm at a temperature of approximately 21°C for 24 hours prior to removing any undissolved test material by filtration (0.2µm 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.

The chromatograms for the saturated solutions of the test material showed two sets of peaks, one set at approximately 4 to 6 minutes and one set at 8 to 12 minutes. The peaks at 8 to 12 minutes were consistent with the test material standard and were considered to be the test material AS305BD. It was considered, and agreed with the Sponsor, that the peaks at 4 to 6 minutes were consistent with those of AL305B a starting component in the production of the test material AS305BD. The test preparations were therefore analysed for both the notifiable material AS305BD and unreacted AL305B present. Analysis of the test preparations at 0 and 48 hours showed the measured concentrations for AS305BD and AL305B 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 0.019 and 0.0045 mg/L respectively.

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