Aluminum, benzoate C16-18 fatty acids complexes - high aluminium

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 September 2017 to 13 October 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The acute toxicity to Daphnia study was undertaken as a bridging study to demonstrate equivalence between the Target substance (XP 453) and the Source Substance (MOL-LUB) for the read-across of other data. Both Target and Source Substances correspond to EC 303-385-6 (previously 272-365-6); CAS 94166-87-7 (previously 68815-27-0), but differ in method of manufacture and ratio of components.
A full discussion of read-across and supporting data is given in the attached justification in the endpoint for short-term toxicity to fish.

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

24 January 2013 to 25 February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-compliant, guideline study, available as an unpublished report.

Justification for type of information:

The acute toxicity to Daphnia study was undertaken as a bridging study to demonstrate equivalence between the Target substance (XP 453) and the Source Substance (MOL-LUB) for the read-across of other data. Both Target and Source Substances correspond to EC 303-385-6 (previously 272-365-6); CAS 94166-87-7 (previously 68815-27-0), but differ in method of manufacture and ratio of components.
A full discussion of read-across and supporting data is given in the attached justification in the endpoint for short-term toxicity to fish.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

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

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 (e.g. precipitate, surface film, etc): 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:

21 ºC to 23 ºC

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: 0.0.012 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 (delete if not applicable): 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 (with observation intervals if applicable) : 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

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 ImmobilizedDaphnia(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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

April 13, 2004.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium

EC Number:

259-105-7

EC Name:

(benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium

Cas Number:

54326-11-3

Molecular formula:

C25H41AlO5

IUPAC Name:

aluminum hydroxide benzoate stearate

Constituent 2

Reference substance name:

aluminum hydroxide benzoate palmitate

Molecular formula:

C23H37AlO5

IUPAC Name:

aluminum hydroxide benzoate palmitate

Constituent 3

Reference substance name:

Aluminium benzoate salts of other fatty acids

IUPAC Name:

Aluminium benzoate salts of other fatty acids

Constituent 4

Reference substance name:

Aluminium fatty acid salts

Cas Number:

Not available.

Molecular formula:

Not available.

IUPAC Name:

Aluminium fatty acid salts

Test material form:

solid: particulate/powder

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The concentration of XP 453, based on percent carbon, was calculated in fresh parent solution samples collected at initiation and from composited spent replicate solutions at 48 hours (termination) of the definitive test.

Test solutions

Vehicle:

no

Details on test solutions:

A definitive limit test was performed from 11 to 13 October 2017 at nominal concentrations of 0 (control) and 100 mg XP 453 total product (TP)/L. The test solution was prepared as a water accommodated fraction (WAF). At initiation, 0.20000 g of the XP 453 test substance was weighed into a glass scintillation vial and rinsed into a glass carboy containing 2.0-L of dilution water to prepare the 100 mg TP/L test substance solution. The control carboy contained only dilution water. A magnetic stir bar was added to each carboy. The contents of each carboy were stirred for approximately 24 hours, with a vortex no greater than approximately 25% of the solution depth, at room temperature while shielded from light. At the start of the stirring period, each carboy was sealed with a silicone stopper to prevent evaporation and contamination. At the end of the 24 hour stirring period, the solutions were removed from the stir plates and allowed to settle for approximately 30 to 60 minutes. After the settling period, approximately 1.5 L of each solution was collected and passed through glass wool packed into a glass funnel. The filtered solutions were then collected and used to prepare the control and 100 mg TP/L replicate test treatment solutions.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Five neonates (<24-hours old) were added to each of four test chambers per treatment at the start of the test. Daphnids were impartially added to a set of labeled containers with each container representing one treatment replicate. Each container was then randomly assigned to a treatment replicate by random number generator. The individuals within each container were then transferred into the corresponding test chamber below the surface of the solution using a wide bore glass pipette. The daphnids were observed for immobility and sublethal effects at approximately 24 and 48 hours after test initiation.

Study design

Test type:

static

Water media type:

freshwater

Remarks:

The dilution water was a moderately hard freshwater prepared by blending naturally hard well water with well water that was demineralized by reverse osmosis (RO).

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

146 mg CaCO3/L

Test temperature:

20 °C

pH:

8.6

Dissolved oxygen:

8.3 to 8.7 mg/L (95 to 100% saturation)

Conductivity:

353 μS/cm

Nominal and measured concentrations:

Nominal, based on water-accommodated fractions (WAF): 0 (control) and 100 mg total product per litre.

Details on test conditions:

A definitive limit test was performed at nominal concentrations of 0 (control) and 100 mg XP 453 total product (TP)/L. The test solution was prepared as a water accommodated fraction (WAF). At initiation, 0.20000 g of the XP 453 test substance was weighed into a glass scintillation vial and rinsed into a glass carboy containing 2.0-L of dilution water to prepare the 100 mg TP/L test substance solution. The control carboy contained only dilution water. A magnetic stir bar was added to each carboy. The contents of each carboy were stirred for approximately 24 hours, with a vortex no greater than approximately 25% of the solution depth, at room temperature while shielded from light. At the start of the stirring period, each carboy was sealed with a silicone stopper to prevent evaporation and contamination. At the end of the 24 hour stirring period, the solutions were removed from the stir plates and allowed to settle for approximately 30 to 60 minutes. After the settling period, approximately 1.5 L of each solution was collected and passed through glass wool packed into a glass funnel. The filtered solutions were then collected and used to prepare the control and 100 mg TP/L replicate test treatment solutions.
All test chambers were labeled with the study number, treatment, and replicate. The test chambers were grouped by treatment in a water bath. No aeration was provided to any test chamber during the test.
Five neonates (<24-hours old) were added to each of four test chambers per treatment at the start of the test. Daphnids were impartially added to a set of labeled containers with each container representing one treatment replicate. Each container was then randomly assigned to a treatment replicate by random number generator. The individuals within each container were then transferred into the corresponding test chamber below the surface of the solution using a wide bore glass pipette. The daphnids were observed for immobility and sublethal effects at approximately 24 and 48 hours after test initiation.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Details on results:

No sublethal effects were noted during the definitive test.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 24- and 48-hour EC50 values were estimated to be >100 mg TP/L, the only concentration tested. No sublethal effects were noted during the definitive test. The 48 hour no effect concentration was 100 mg TP/L.