Oxo(propan-2-olato)aluminium

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24 February 2013 to 21 March 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant, guideline study, available as an unpublished report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Aluminum, benzoate C16-18-fatty acids complexes

EC Number:

303-385-6

EC Name:

Aluminum, benzoate C16-18-fatty acids complexes

Cas Number:

94166-87-7

Molecular formula:

C23H37AlO5, C25H41AlO5

IUPAC Name:

Aluminum, benzoate C16-18-fatty acids complexes

Test material form:

other: Solid

Details on test material:

- Physical state: Pale Yellow Solid
- Purity: Not applicable - UVCB
- Substance identity: Aluminum, benzoate C16-18-fatty acids complexes
- Batch number: 11074091 + Benzoic acid
- Carbon Content: 65.1%
- Analysis code: A118/99
- Date received: 07 June 2012
- Expiration date: 01 July 2013
- Storage of test material: Room temperature in the dark

Specific details on test material used for the study:

The substance is the Source of the read-across.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Aluminium concentration: Samples taken at 0 and 72 hours (fresh media) and at 24 and 96 hours (old media) from the control and treatment group.
- Total organic carbon: Samples were collected at 0, 48 and 72 (fresh media) and 24, 72 and 96 hours (old media) from the control and treatment group.
- Duplicate samples: Duplicate samples were taken at 24 (fresh media), 48 and 72 hours (old and fresh media) and stored at approximately -20°C for analysis if necessary.

Test solutions

Vehicle:

no

Details on test solutions:

- Preparation: 200 mg aluminum, benzoate C16-18-fatty acids complexes was added to the surface of 2 L of dichlorinated laboratory tap water, to give a loading rate of 100 mg/L, and stirred using a magnetic stirrer at a rate that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixture allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. A glass wool plug was inserted into the opposite end of the tubing and the WAFs removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the WAF.
- Test water: The test water used for the range finding and definitive tests was the same as that used to maintain the stock fish. Dechlorinated laboratory tap water was partly softened and heated to the required temperature.
- Controls: Test water only.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present.


- Test water: The test water used for the range finding and definitive tests was the same as that used to maintain the stock fish. Dechlorinated laboratory tap water was partly softened and heated to the required temperature.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

- Common name: Rainbow trout
- Source: Brow Well Fisheries Ltd., Hebden, Skipton, Yorkshire, UK
- Acclimatisation: Fish acclimatized to test conditions for 12 days
- Feeding during acclimatisation: Fish fed commercial trout pellets daily up to 24 hours prior to test initiation
- Culture conditions: Fish were maintained in a glass fibre tank with a single pass water renewal system under 16 hours light to 8 hours dark with 20 minute transitions. The water was kept at approximately 14°C with at least 10.5 mg O2/L. There were no mortalities in the 7 days prior to test initiation.
- Mean standard length: 4.4 cm (sd = 0.2) at test termination
- Mean weight: 1.10 g (sd = 0.10.) at test termination
- Loading rate: 0.35 g bw/L

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

None

Test conditions

Hardness:

The water hardness ranged from 106 to 136 ml/L as CaCO3 for the duration of the test.

Test temperature:

Vessels were maintained at 14 to 15°C in a temperature controlled room.

pH:

The pH ranged from 7.7 to 7.9 in fresh media and from 8.0 to 8.2 in old media.

Dissolved oxygen:

The dissolved oxygen concentration ranged from 10.5 to 10.7 mg O2/L (104-106 % air saturation value) in fresh media and 9.7 to 10.0 mg O2/L (94-97 % air saturation value) in old media. The oxygen concentration in some of the test vessels was observed to have an air saturation value in excess of 100 %. This was considered to be due to the presence of microscopic air bubbles in the media super saturating the dilutent and was considered not to have had an impact on the outcome or the integrity of the study as no adverse effects were observed in the control group.

Salinity:

Not applicable

Nominal and measured concentrations:

- Nominal concentration: 100 mg/L loading rate WAF
- Basis for concentration: Based on the results of the Acute Toxicity to Daphnia magna test (Harlan Study Number: 41207981) which gave an EC50 value of greater than 100 mg/L loading rate WAF. Therefore this test was conducted at a single loading rate of 100 mg/L loading rate WAF.
- Measured aluminium concentrations: < LOQ to 0.0183 mg/L for the control and < LOQ to 0.0135 mg/L for the treatment group. The limit of quantitation (LOQ) was determined to be 0.0050 mg/L aluminium.
- Measured total organic carbon: Less than the control to 1.19 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: Covered 20 L glass exposure vessel
- Aeration: Vessels were aerated via narrow bore glass tubes
- Renewal rate of test solution (frequency/flow rate): Daily renewal
- No. of organisms per vessel: 7
- No. of vessels per control (replicates): 7
- Number of replicates: 1 test vessel and 1 control
- Biomass loading rate: 0.35 g bodyweight/L
- Feeding: None during test

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated laboratory tap water
- Total organic carbon: 0.920 mg C/L
- Culture medium different from test medium: No
- Intervals of water quality measurement: Water temperature, dissolved oxygen and pH recorded daily. Water hardness measured at test start and and after each renewal.

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light to 8 hours dark with 20 minute transition period

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Fish were inspected at 3, 6, 24, 48, 72 and 96 hours after test initiation for mortalities and sub-lethal effects. The criteria of death were taken to be the absence of both respiratory movement and response to physical stimulation.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: Water Accommodated Fraction

Basis for effect:

mortality (fish)

Details on results:

- Results: No mortalities or sub-lethal effects were observed in limit test conducted at 100 mg/L (WAF). Therefore the NOELR was set at 100 mg/L (WAF) and the LL50 was considered to be greater than this value. It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L. Given that toxicity cannot be attributed to a single component or mixture of components but to aluminum, benzoate C16-18-fatty acids complexes as a whole, the results were based on nominal loading rates only.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

- Effect concentration: An LL50 could not be determined because no effects were observed in the single concentration tested. An estimate of the LL50 (lethal loading rate) was given by inspection of the mortality data.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Total Organic Carbon measured in the Control and 100 mg/L loading rate WAF test concentration at 48 hours was observed to be higher than was expected. This was not considered to impact the validity of the result as no mortality was observed for the duration of the test.

Cumulative Mortality Data in the Definitive Test

Nominal Load Rate (mg/L)	Cumulative Mortality (Initial Population = 7)						% Mortality
	3 Hours	6 Hours	24 Hours	48 Hours	72 Hours	96 Hours	96 Hours
Control	0	0	0	0	0	0	0
100	0	0	0	0	0	0	0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Confirmation of the test concentrations is not applicable as the test is being conducted on WAFs

Conclusions:

The acute toxicity of aluminum, benzoate C16-18-fatty acids complexes to the freshwater fish rainbow trout (Oncorhynchus mykiss) showed no effects at a water accommodated fraction (WAF) nominal loading rate of 100 mg/L. Therefore, the 96 hour LL50 is determined to be > 100 mg/L (WAF).

Executive summary:

The acute toxicity of aluminum, benzoate C16-18-fatty acids complexes to the freshwater fish, rainbow trout (Oncorhynchus mykiss) showed no effects at a water accommodated fraction (WAF) nominal loading rate of 100 mg/L. Therefore, the 96 hour LL50 is determined to be > 100 mg/L (WAF). The acute toxicity of aluminum, benzoate C16-18-fatty acids to fish was determined in a GLP-compliant, limit test (Harlan 2013) following OECD guideline 203. Seven fish were exposed to a single concentration of the test item for 96 hours in a semi-static freshwater study and any effects were recorded. The study is considered reliable and relevant for use for this endpoint.