Oxoaluminium, C16-C18-alkyl esters

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

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:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

See test material information

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.

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 (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

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

The water hardness ranged from 106 to 136 mg/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: 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

Key result

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)

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.

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

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.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Read across data

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH

In accordance with the Regulation (EC) No 1907/2006, Annex XI, section 1.5, read-across to aluminum, benzoate C16-18-fatty acids complexes has been used to fulfil REACH information requirements where appropriate and is justified by the chemical structures and common physiological active moieties of the substances. The chemical structures of the target and read-across substances are very closely aligned. The aluminium cation, a long chain fatty acid, and the –Al=O (-AlOH in aqueous solution) moieties are identical in both substances. The key difference is that read-across substance contains a benzoate moiety linked to the aluminium cation, which is absent from the target substance. Benzoic acid and benzoates have been well characterized (eco)toxicologically, but in this case generating experimental data on the aluminium salt containing benzoate would be expected to demonstrate a ‘worst case’ hazard profile when compared to the target substance. Since no intrinsic toxicity could be demonstrated from any of the Annex VII or VIII endpoints with the benzoate-containing aluminium salt, then these results can be read across to the target substance without restriction.

The substances consist of aluminium salts of fatty acids, with the source substance having additionally a benzoate moiety linked to the aluminium cation as well. As such, both substances have common functional groups and as the fatty acid moieties are considered not to be hazardous, since they are an endogenous part of living cells and are a dietary requirement for many organisms, the toxicity of the substances will be driven by the presence of the aluminium species (and additionally the benzoate for the source substance). The substances have common starting materials and methods of manufacture, principally the reaction of aluminium with organic acids, with the target substance often being used as a pre-cursor reacted with benzoic acid in the manufacture of the source substance. The substances have common breakdown products and, in the environment, the substances will dissociate and biodegrade into inorganic aluminium species and fatty acids then carbon dioxide and water.

2. CHEMICAL(S)

Source chemical: Aluminum, benzoate, C16-18 fatty acids complexes (EC: 303-385-6, CAS: 94166-87-7)

See robust study summaries for further details on the identity of the tested substances and IUCLID dataset for further information on the substance identity and the data to support the read across justification.

3. ANALOGUE APPROACH JUSTIFICATION
The literature search identified no ecotoxicological data for the substance and no experimental ecotoxicity data are available for the substance. The hazard testing was conducted on the substance prepared as a 50% w.w. concentration in pharmaceutical white oil. The presence of the oil restricts the solubility of the substance and reduces its bioavailability to the aquatic environment therefore, no testing was carried out on the substance as no meaningful results would be achieved. Instead, in order to provide a worst-case scenario for the aquatic ecotoxicity potential for the substance, data have been read across from a structural analogue which was tested in in isolated form (i.e. extracted from base oil).

No novel aquatic ecotoxicity testing on the substance was carried out but novel proprietary data were read across from aluminum, benzoate C16-18-fatty acids complexes. This substance is considered suitable for read-across as it contains a fatty acid moiety coordinated to an aluminium atom. The chemical structures of the target and read-across substances are very closely aligned; both substances consist of aluminium salts of fatty acids. The aluminium cation, a long chain fatty acid, and the –Al=O (-AlOH in aqueous solution) moieties are identical in both substances.

The fatty acids present in both substances are the same, consisting of a mixture of C16 and C18 chain lengths at approximately a 1:2 ratio. The C16 and C18 fatty acid moieties are derived from natural fatty materials, or substances which are chemically indistinguishable from natural fatty acids. The fatty acid moieties are considered not to be hazardous to humans as they are natural constituents of the human body and essential components of a balanced human nutrition. REACH Annex V, Entry 9, groups fatty acids and their potassium, sodium, calcium and magnesium salts, including C6 to C24, predominantly even-numbered, unbranched, saturated or unsaturated aliphatic monocarboxylic acids. Provided that they are obtained from natural sources and are not chemically modified, the substances included in REACH Annex V, Entry 9 are exempt from registration, unless they are classified as dangerous (except for flammability, skin irritation or eye irritation) or they meet the criteria for PBT/vPvB substances.

Fatty acids are an endogenous part of living cells and are an essential dietary requirement for many organisms. They are absorbed, digested and transported in animals and humans. When taken up by tissues they can either be stored as triglycerides or can be oxidised via the ß-oxidation and tricarboxylic acid pathways. The ß-oxidation uses a mitochondrial enzyme complex for a series of oxidation and hydration reactions, resulting in a cleavage of acetate groups as acetyl CoA. Acetyl CoA is used mainly to provide energy but also to provide precursors for numerous biochemical reactions. Alternative minor oxidation pathways can be found in the liver and kidney (ω-oxidation and ω-1 oxidation) and in peroxisomes for ß-methyl branched fatty acids (α-oxidation). The metabolic products can then be incorporated for example into membrane phospholipids.

The read across substance also contains a benzoate moiety linked to the aluminium cation, which is absent from the target substance. Although it also contains a coordinated benzoate ion, under environmental relevant conditions the benzoate ion has a LC or EC50s of > 100 mg/L for fish (OECD SIDS 2001) and therefore does not contribute any additional toxicity to the substance. Benzoic acid and benzoates have been well characterized (eco)toxicologically, but in this case generating experimental data on the aluminium salt containing benzoate would be expected to demonstrate a ‘worst case’ hazard profile when compared to the target substance as this substance is tested in isolated form (i.e. outside of the base oil). Since no intrinsic toxicity could be demonstrated from any of the Annex VII or VIII endpoints with the benzoate-containing aluminium salt, then these results can be read across to the target substance without restriction.

Comparison of the data for the two substances indicates that they are expected to have similar properties. Neither the target or read-across substance meets the criteria for classification for physico-chemical, environmental or human health endpoints, based on the available data.

On the basis of the physico-chemical results, the substances are not flammable and have similar densities. The low vapour pressure results indicates that hazards associated with the atmospheric compartment or inhalation routes of toxicity are not expected to be relevant. The substances show similar water solubility, without surface active properties, indicating that they are likely to have similar behaviour in the aquatic environment.

Although the read-across substance met the criteria for ready biodegradability and the target substance did not, neither substance was inhibitory to micro-organisms at the concentration tested. The difference in biodegradation results is expected to derive from the presence of the base oil in the target substance sample, which is designed to minimise leaching of the grease thickener, and therefore less of the grease thickener would have been available for degradation by the micro-organisms.

There are no results available for the ecotoxicity of the target substance and therefore comparison of the effect concentrations against the read-across substance is not possible. However, leaching studies on grease thickeners in base oils have been used to assess the potential bioavailability of the grease components. The bioavailability potential of the water accommodated fractions (WAFs) of metal (lithium and calcium) soap complex based grease thickeners was assessed using a solid-phase micro-extraction (SPME) method combined with gas chromatography (GC). This approach was complemented with metal ion analysis to determine whether the metal leaches out of the base grease during WAF preparation and the ecotoxicity of WAFs was also monitored using an in vitro Microtox assay. The SPME-GC data confirmed that there was negligible leaching of the thickeners from base oils in the samples tested, with measurements for calcium and lithium below the limit of detection (<0.1 mg/L) and the screening ecotoxicity data also showed a lack of toxicity of the greases.

The results of the bioavailability potential of the WAFs, the metal ion analysis and the screening ecotoxicity of lithium and calcium based complexes have been read across to aluminium based thickeners. All of these metal salts of fatty acids are expected to behave in a very similar manner when entrained within a grease matrix, with high temperature stability indicating that the thickener structure is robust and resistant to diffusion out of the oil. Dissolution of grease thickeners from grease into water is very unlikely as the thickeners are poorly water soluble and the thickeners are embedded in the hydrophobic grease matrix and thus unlikely to leach out. Therefore, although there are no data on the ecotoxicity of the target substance, no effects are expected based on the lack of bioavailability of the thickener.

These data on the potential for leaching of other metal salt complex based grease thickeners have been read across to both the target and read across substances. On the basis of these results, it is expected that neither the target nor the read across substance would leach from the base oil in which they are typically marketed and therefore neither substance would be bioavailable. Thus, reading across data from the source substance tested in its isolated form is considered robust as it provides a worst-case conclusion for the target substance which is only manufactured in an inert carrier, typically base oil. In order to provide further evidence for the lack of bioavailability, it is proposed to undertake leaching studies on the target and read-across substances themselves. Dependent on the results, the two studies would then be used to show the similarity in the bioavailability of the two substances and provide further weight of evidence for the read-across approach.

The available mammalian toxicity data show that neither the target nor read-across substance would be classified as irritating to skin or eyes and would not be classified for acute oral toxicity, with LD50 values of >2000 mg/kg. Although no other data are available for comparison of the potential mammalian toxicity of the two substances, the target and read-across substances are expected to behave in a very similar manner. As grease thickeners are entrained within grease matrices which are robust and resistant to diffusion out of the oil, neither substance is expected to be bioavailable. In order to provide further evidence for the lack of bioavailability, it is proposed to undertake leaching studies in fed state simulated intestinal fluid (FeSSIF) on the target and read-across substances. Dependent on the results, the two studies would then be used to show the similarity in the bioavailability of the two substances and provide further weight of evidence for the read-across approach.

For the ecotoxicity of the substances, read across from the source to the target substance is considered justified as both substances have very low solubility in water, would not leach when in situ in base oil during use as grease thickeners and are not expected to be bioavailable. The substances, in the environment, would dissociate and degrade into inorganic aluminium species and firstly, fatty acids (plus benzoic acid for the source substance), then ultimately carbon dioxide and water. As the fatty acid components are biodegradable and are not expected to be hazardous (and the benzoate component of the source substance is not expected to be hazardous), the toxicity is expected to be driven by the aluminium component, so would be the same in both the source and target substances. As such, read across from the source substance, tested in its isolated form, is considered to provide a worst-case scenario for the target substance, which would have even more limited bioavailability as it is manufactured in situ in an inert carrier, typically base oil.

4. DATA

T = target substance (tests were undertaken on a sample prepared as a 50% w.w. concentration in medicinal white oil unless otherwise indicated)
RA = read-across substance

- State: Liquid (T), Solid (RA)
- Melting point: 21°C (T), 224°C (RA)
- Relative density: 0.933 (T), 1.08 (RA)
- Vapour pressure: 0.00015 Pa (T), 0.000044 Pa (RA)
- Surface tension: 72.5 mN/m (T), 72.6 mN/m (RA)
- Water solubility: ≤0.00015 g/L (T), ≤0.00026 g/L (RA)
- Flash-point: 159°C (T), No data available for RA
- Flammability: No data available for T, Not flammable (RA)
- Self-ignition temperature: 374°C (T), 383°C (RA)
- Viscosity: 174.3 mm2/s at 100°C (T), No data available for RA
- Biodegradation: Not readily biodegradable (31%) (T), Readily biodegradable (79%) (RA)
- Acute aquatic invertebrates: No data available for T, EL50 (48 h): > 100 mg/L (RA)
- Algae: No data available for T, EL50 (72 h): > 100 mg/L and NOELR (72 h): 100 mg/L (RA)
- Aquatic microorganisms: NOEC (28 d): 6.7 mg/L (T), NOEC (28 d): 15.4 mg/L (RA)
- Acute fish: No data available for T, LL50 (96 h): > 100 mg/L (RA)
- Skin irritation: Not irritating (T), Not irritating (RA)
- Eye irritation: Not classified (T). Not classified (RA)
- Skin sensitisation: No data available for T, Not sensitising (RA)
- In vitro gene mutation in bacteria: No data available for T, Negative (RA)
- Acute toxicity, oral route: LD50: > 2000 mg/kg (T, test undertaken on solid (isolated) form of the substance), LD50 >2000 mg/kg (RA)
- Acute toxicity, dermal route: No data available for T, LD50 >2000 mg/kg (RA)
- In vitro cytogenicity: No data available for T, Negative (RA)
- In vitro gene mutation in mammalian cells: No data available for T, Negative (RA)
- Short-term repeated dose toxicity, oral route: No data available for T, NOAEL: > 225 mg/kg (RA)
- Reproductive toxicity: No data available for T, NOAEL (P): > 225 mg/kg (RA)
- Developmental toxicity: No data available for T, NOAEL (F1): > 225 mg/kg (RA)

Reason / purpose for cross-reference:

read-across source

Key result

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)

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.

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

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 hazard testing was conducted on the substance prepared as a 50% w.w. concentration in pharmaceutical white oil.The presence of the oil restricts the solubility of the substance and reduces its bioavailability to the aquatic environment therefore, no testing was carried out on the substanceas no meaningful results would be achieved.Instead proprietary data are read across from aluminum, benzoate C16-18-fatty acids complexes. This substance is considered suitable for read-across as it contains a fatty acid moiety coordinated to an aluminium atom. Although it also contains a coordinated benzoate ion, under environmental relevant conditions the benzoate ion has a 96 hour LC50 for fish of > 100 mg/L (OECD SIDS 2001) and therefore does not contribute any additional toxicity to the substance. Aluminum, benzoate C16-18-fatty acids complexes was tested in the form of an isolated solid and showed no toxic effects at a water accommodated fraction loading rate of 100 mg/L (Harlan 2013). Therefore, the 96 hour LL50 is determined to be > 100 mg/L (WAF).

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.

Description of key information

Based on read across, the acute toxicity to the freshwater fish rainbow trout (Oncorhynchus mykiss) is expected to show no effects at a water accommodated fraction loading rate of 100 mg/L. Therefore, the 96 hour LL50 is determined to be > 100 mg/L (WAF).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

The hazard testing was conducted on the substance prepared as a 50% w.w. concentration in pharmaceutical white oil.The presence of the oil restricts the solubility of the substance and reduces its bioavailability to the aquatic environment therefore, no testing was carried out on the substance as no meaningful results would be achieved. Instead proprietary data are read across from aluminum, benzoate C16-18-fatty acids complexes. This substance is considered suitable for read-across as it contains a fatty acid moiety coordinated to an aluminium atom. Although it also contains a coordinated benzoate ion, under environmental relevant conditions the benzoate ion has a 96 hour LC50 for fish of > 100 mg/L (OECD SIDS 2001) and therefore does not contribute any additional toxicity to the substance. Aluminum, benzoate C16-18-fatty acids complexes was tested in the form of an isolated solid and showed no toxic effects at a water accommodated fraction loading rate of 100 mg/L (Harlan 2013). Therefore, the 96 hour LL50 is determined to be > 100 mg/L (WAF).

 

The acute toxicity to rainbow trout (Oncorhynchus mykiss) was determined in GLP-compliant, semi-static limit test (Harlan 2013) following OECD guideline 203. As no effects were observed in the Daphnia and algae toxicity tests or the preliminary range finding test for acute toxicity to rainbow trout, the definitive toxicity to fish test was conducted as a limit test at a nominal loading rate of 100 mg/L alongside a blank control. Seven fish were exposed to a water accommodated fraction (WAF) of aluminum, benzoate C16-18-fatty acids complexes for 96 hours in standard ecotoxicity media adjusted to a hardness of approximately 150 mg/L CaCO₃. Observations of mortalities or abnormalities of behaviour or appearance of the fish were made at 3, 6, 24, 48, 72 and 96 hours and the test solutions analysed for aluminium content and total organic carbon at test initiation and termination. As no toxicity effects were observed, the 96 hour LL50 is determined to be > 100 mg/L (WAF).

Reference

OECD (2001). SIDS, Benzoate, SIDS Initial Assessment Report for 13th SIAM, Bern 7^th– 9^thNovember 2001, UNEP Publications.