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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 January 2013 to 25 February 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
See test material information
Analytical monitoring:
yes
Details on sampling:
- Total organic carbon and aluminium analysis: Samples were collected at 0 and 48 hours from the control and pooled treatment replicates and stored at -20°C until analysis. Duplicate samples were taken and stored at approximately -20°C for further analysis if necessary.
Vehicle:
no
Details on test solutions:
- Test water: Reconstituted water comprised of Elendt M7 medium was used.
- Preparation: 200 mg aluminum, benzoate C16-18-fatty acids complexes was added to the surface of 2 L of reconstituted 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 and the WAFs removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the WAF.
- Controls: Test control: test medium only. Positive control: potassium dichromate at concentrations; 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.
- Evidence of undissolved material: Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present.
Test organisms (species):
Daphnia magna
Details on test organisms:
- Age: First instar, less than 24 hours old
- Source: In-house laboratory cultures
- Culture conditions: Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium at approximately 20°C under a 16 hour light to 8 hour dark photoperiod, with 20 minute transitions. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin flake food suspension. Conditions ensured reproduction by parthogenesis.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
None
Hardness:
At the start of the test, the dilutent had a total water hardness of 152 mg/L CaCO3.
Test temperature:
21ºC to 23ºC
pH:
The pH was 8.0 at 0 hours and 7.8 to 8.0 at 48 hours.
Dissolved oxygen:
The dissolved oxygen ranged from 8.3 to 8.6 mg O2/L at 0 hours and 8.5 to 8.8 mg O2/L at 48 hours. The equivalent air saturation values were 97 to 99 % at 0 hours to 96 to 98 % at 48 hours.
Salinity:
Not applicable
Nominal and measured concentrations:
- Preliminary test: The loading rates to be used in the definitive test were determined on the basis of a range finding study at 1.0, 10 and 100 mg/L nominal loading rates showing no effects.
- Nominal concentration: The test was conducted as a limit test with a single nominal loading rate of 100 mg/L (WAF).
- Measured aluminium concentrations: 0.012 mg/L at 0 hours and 0.0057 mg/L at 48 hours.
- Measured total organic carbon (TOC): Total Organic Carbon (TOC) analysis of the test preparation at 0 and 48 hours showed measured concentrations of 0.50 mg C/L and less than the limit of quantitation (assessed to be 1.0 mg C/L for the method employed) respectively.
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass jar
- Type: Closed
- Material, size, headspace, fill volume: 250 mL glass jar containing 200 mL of test preparation.
- 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: Reconstituted water - Elendt M7 medium
- Culture medium different from test medium: No
- Intervals of water quality measurement: Water temperature was recorded daily throughout the test. Dissolved oxygen concentrations and pH were recorded at the start and termination of the test.

OTHER TEST CONDITIONS
- Adjustment of pH: Adjusted to pH 7.8 ± 2 with NaOH or HCl.
- Photoperiod: 16 hours light and 8 hours dark with 20 minutes dawn and dusk transition period.
- Light intensity: 396 to 405 lux

EFFECT PARAMETERS MEASURED: Immoblization or adverse reactions recorded at 24 and 48 hours. Daphnia were considered to be immobilized if they were unable to swim for approximately 15 seconds after gentle agitation.

TEST CONCENTRATIONS
- Range finding study: Yes
- Test concentrations: Limit test at 100 mg test item/L WAF.
Reference substance (positive control):
yes
Remarks:
Potassium dichromate
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction
Basis for effect:
mobility
Details on results:
- Results: There was no immobilisation in 20 daphnids exposed for 48 hours to a 100 mg/L WAF of aluminum, benzoate C16-18-fatty acids complexes 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):
- Reference: Harlan Laboratories Ltd., study number 41203341. Daphnia were exposed to 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L potassium dichromate under similar exposure conditions to the definitive test and the immobilisation data was analysed by the geometric mean method at 24 hours and the trimmed Spearmman-Karber method at 48 hours.
- Results: Based on nominal concentrations, the 48 hour EC50 was 0.45 (95 % confidence limits 0.42-0.48 mg/L). The results were within the normal range for this test item.
Reported statistics and error estimates:
Not reported

Some of the temperatures were measured to be slightly in excess of the 18 to 22ºC given in the study plan. This was considered not to affect the results of the test as no adverse effects of exposure were observed throughout the duration of the test and that the temperatures did not deviate more than ± 1ºC throughout the test.

Table 2 Cumulative Immobilization Data in the Definitive Test

Nominal Loading Rate

(mg/L)

Cumulative Immobilized Daphnia (Initial Population: 5 per Replicate)

24 Hours

48 Hours

No per Replicate

Total

%

No per Replicate

Total

%

Control

R1

4*

4

20

4

4

20

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

R1 – R4 = Replicates 1 to 4

* Whilst 4 daphnids were observed immobilized in this control replicate, this was not thought to impact the validity of the test as this was isolated to one vessel and was considered to be contamination of this vessel rather than being representative of the control group.

Validity criteria fulfilled:
yes
Remarks:
Whilst 4 daphnids were observed immobilized in one control, this was not thought to impact the validity as this was isolated to one vessel and was considered to be contamination of this vessel rather than being representative of the control group.
Conclusions:
The acute toxicity of aluminum, benzoate C16-18-fatty acids complexes to Daphnia showed no effects at a water accommodated fraction nominal loading rate of 100 mg/L. Therefore, the 48 hour EL50 is determined to be > 100 mg/L (WAF).
Executive summary:

The acute toxicity of aluminum, benzoate C16-18-fatty acids complexes to Daphnia magna showed no effects at a water accommodated fraction nominal loading rate of 100 mg/L. Therefore, the 48 hour LC50 is determined to be > 100 mg/L (WAF). The acute toxicity of of the test item to Daphnia magna was determined in a GLP-compliant, limit test (Harlan 2013) following OECD guideline 202. The study is considered reliable and relevant for use for this endpoint.

Endpoint:
short-term toxicity to aquatic invertebrates
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:
read-across source
Key result
Duration:
48 h
Dose descriptor:
EL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction
Basis for effect:
mobility
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction
Basis for effect:
mobility
Details on results:
- Results: There was no immobilisation in 20 daphnids exposed for 48 hours to a 100 mg/L WAF of aluminum, benzoate C16-18-fatty acids complexes 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):
- Reference: Harlan Laboratories Ltd., study number 41203341. Daphnia were exposed to 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L potassium dichromate under similar exposure conditions to the definitive test and the immobilisation data was analysed by the geometric mean method at 24 hours and the trimmed Spearmman-Karber method at 48 hours.
- Results: Based on nominal concentrations, the 48 hour EC50 was 0.45 (95 % confidence limits 0.42-0.48 mg/L). The results were within the normal range for this test item.
Reported statistics and error estimates:
Not reported

Some of the temperatures were measured to be slightly in excess of the 18 to 22ºC given in the study plan. This was considered not to affect the results of the test as no adverse effects of exposure were observed throughout the duration of the test and that the temperatures did not deviate more than ± 1ºC throughout the test.

Table 2 Cumulative Immobilization Data in the Definitive Test

Nominal Loading Rate

(mg/L)

Cumulative Immobilized Daphnia (Initial Population: 5 per Replicate)

24 Hours

48 Hours

No per Replicate

Total

%

No per Replicate

Total

%

Control

R1

4*

4

20

4

4

20

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

R1 – R4 = Replicates 1 to 4

* Whilst 4 daphnids were observed immobilized in this control replicate, this was not thought to impact the validity of the test as this was isolated to one vessel and was considered to be contamination of this vessel rather than being representative of the control group.

Validity criteria fulfilled:
yes
Remarks:
Whilst 4 daphnids were observed immobilized in one control, this was not thought to impact the validity as this was isolated to one vessel and was considered to be contamination of this vessel rather than being representative of the control group.
Conclusions:
The acute toxicity of aluminum, benzoate C16-18-fatty acids complexes to Daphnia showed no effects at a water accommodated fraction nominal loading rate of 100 mg/L. Therefore, the 48 hour EL50 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 48 hour EC50 for Daphnia 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 48 hour EL50 is determined to be > 100 mg/L (WAF).

The acute toxicity of aluminum, benzoate C16-18-fatty acids complexes to Daphnia magna showed no effects at a water accommodated fraction nominal loading rate of 100 mg/L. Therefore, the 48 hour LC50 is determined to be > 100 mg/L (WAF). The acute toxicity of of the test item to Daphnia magna was determined in a GLP-compliant, limit test (Harlan 2013) following OECD guideline 202. The study is considered reliable and relevant for use for this endpoint.

Description of key information

Based on read across, the acute toxicity to Daphnia is expected to show no effects at a water accommodated fraction nominal loading rate of 100 mg/L. Therefore, the 48 hour EL50 is determined to be > 100 mg/L (WAF).

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
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 48 hour EC50 of > 100 mg/L for Daphnia (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 48 hour EL50 is determined to be > 100 mg/L (WAF).

The acute toxicity of aluminum, benzoate C16-18-fatty acids complexes to Daphnia was determined in a GLP-compliant, limit test (Harlan 2013) following OECD guideline 202. As no effects were observed in the preliminary range finding test, the definitive toxicity to Daphnia test was conducted as a limit test at a nominal loading rate of 100 mg/L alongside a blank control. Twenty Daphnia divided into four replicates were exposed to a water accommodated fraction of aluminum, benzoate C16-18-fatty acids complexes for 48 hours in standard ecotoxicity media adjusted to a hardness of approximately 150 mg/L CaCO3. Observations of immobilisation or abnormalities of behaviour or appearance of the Daphnia were made at 24 and 48 hours and the test solution analysed for aluminium content and total organic carbon at test initiation and termination. As no effects were observed, the 48 hour EL50 is determined to be > 100 mg/L (WAF).

Reference

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