Registration Dossier

Administrative data

Description of key information

The acute oral toxicity of the registered substance to the Wistar strain rat was assessed and determined an LD50 as >2000 mg/kg bodyweight. This study was supported by a study with aluminum, benzoate C16-18-fatty acid complexes also showing an LD50 as >2000 mg/kg bodyweight. Since the level of toxicity by the oral route of administration was similar for both substances, a study with aluminum, benzoate C16-18 fatty acid complexes was taken as indicative of the potential dermal toxicity. There is no evidence of a relevant intrinsic acute oral or dermal toxicity requiring classification or substance specific Risk Management Measures.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 June 2001 to 12 July 2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals approximately 7 weks old
- Weight at study initiation: The bodyweight variation did not exceed ±20% of the sex mean
- Fasting period before study: Food was withheld overnight (for a maximum of 20 hours) prior to dosing until approximately 3-4 hours after administration of the test substance.
- Housing: Group housing of three animals per sex per cage in laballed cages (type IV; height 15 cm) containing purified sawdust as bedding.
- Diet: Free access to standard pelleted laboratory animal diet supplied by Altromin (code VRF 1), Lage, Germany
- Water: Free access to tap water
- Acclimation period: At least five days


ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 3 °C
- Humidity: Relative humidity 30 to 70%
- Air changes: The rate of air exchange was at approximately fifteen changes per hour
- Photoperiod: 12 hours artificial fluorescent light and 12 hours dark per day
Route of administration:
oral: gavage
Vehicle:
other: Poly alpha olefin
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The formulations (w/w) were prepared within 4 hours prior to dosing to give a dose level of 2000mg/kg bodyweight
- Amount of vehicle: Not stated
- Justification for choice of vehicle: The vehicle was selected based on information provided by the sponsor
- Lot/batch no. (if required): Not stated
- Purity: Not stated

MAXIMUM DOSE VOLUME APPLIED: 10mL/kg bodyweight

DOSAGE PREPARATION (if unusual): To dissolve the substance, ethyl acetate was added to the test substance (final concentration in formulation preferably below 30%). After mixing, poly alpha olefin (supplied by sponsor) was added until the requested concentration was reached. This preparation method was selected, in mutual agreement with the sponsor, to facilitate dosing of a homogeneous formulation. It was indicated by the sponsor that by using this method the test substance is stable.

CLASS METHOD (if applicable)
- Ration for the selection of the starting dose: The toxicity of the test substance was assessed by stepwise treatment of groups of three animals. The first group was treated at a dose level of 2000 mg/kg bodyweight. The absence or presence of mortality of animals dosed at one step determined the next step, based on the test procedure defined in the guidelines. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.
Doses:
Three females dosed at 2000 mg/kg bodyweight, followed two days later by three males dosed at 2000 mg/kg bodyweight.
No. of animals per sex per dose:
3 females at 2000 mg/kg bodyweight
3 males at 2000 mg/kg bodyweight
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made on the day of dosing (day 1) and once daily thereafter, until day 15. Mortality/viability checks were made twice daily. Individual bodyweights were recorded on Day 1 (the day of dosing) and on Days 8 and 15.
- Necropsy of survivors performed: Yes
Statistics:
No statistical analysis was performed.
Preliminary study:
Not applicable
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- Lethal results: There were no deaths
Clinical signs:
- Non-lethal results: Lethargy, hunched posture and piloerection were noted among animals between days 1 and 3.
Body weight:
- Bodyweight results: The mean bodyweight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
- Gross pathology results: No abnormalities were noted at macroscopic post mortem examination of the animals.
Other findings:
No data reported

Table1: Clinical Signs

 

Test Day

1

1

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Hours After Treatment

0

2

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Female at 2000 mg/kg

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Animal 1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Behaviour - Lethargy

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Posture – Hunched Posture

-

-

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/Fur/Plumage - Piloerection

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Female at 2000 mg/kg

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Animal 2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Behaviour - Lethargy

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Posture – Hunched Posture

-

1

1

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/Fur/Plumage - Piloerection

-

-

1

1

 

-

-

-

-

-

-

-

-

-

-

-

-

Female at 2000 mg/kg

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Animal 3

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Behaviour - Lethargy

-

2

2

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Posture – Hunched Posture

-

1

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Skin/Fur/Plumage - Piloerection

-

-

1

1

1

-

-

-

-

-

-

-

-

-

-

-

-

Male at 2000 mg/kg

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Animal 1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Behaviour - Lethargy

-

1

.

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Posture – Hunched Posture

-

1

.

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/Fur/Plumage - Piloerection

-

1

.

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Male at 2000 mg/kg

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Animal 2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Behaviour - Lethargy

-

1

.

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Posture – Hunched Posture

-

1

.

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/Fur/Plumage - Piloerection

-

1

.

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Male at 2000 mg/kg

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Animal 3

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Behaviour - Lethargy

-

1

.

-

-

-

-

-

-

-

-

-

-

-

-

-

-

Posture – Hunched Posture

-

1

.

1

-

-

-

-

-

-

-

-

-

-

-

-

-

Skin/Fur/Plumage - Piloerection

-

1

.

1

-

-

-

-

-

-

-

-

-

-

-

-

-

- = sign not observes,. = Observation not made, + = Animal dead
Interpretation of results:
other: Criteria for classification as an acute oral toxicant not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of in both the male and female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.
Executive summary:

Introduction. 

The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat.The acute oral toxicity of the test item to the Wistar strain rat was assessed in a GLP-compliant study following OECD guideline 423 (adopted 1996) andEU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)in a proprietary, experimental study (NOTOX 2013). The study is considered reliable and relevant for use for this endpoint.

Method. 

A group of three fasted females was given a single oral dose of test material, as a suspension in poly alpha olefin, at a dose level of 2000 mg/kg bodyweight. A group of three fasted males were treated in the same way. Clinical signs and bodyweight development were monitored during the study period of 15 days (starting from day of dosing). All animals were subjected to gross necropsy.

Mortality. 

There were no deaths.

 

Clinical Observations.

Lethargy, hunched posture and piloerection were noted among animals between days 1 and 3.

 

Bodyweight. 

The mean bodyweight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.

 

Necropsy. 

No abnormalities were noted at macroscopic post mortem examination of the animals..

 

Conclusion. 

The acute oral median lethal dose (LD50) of in both the male and female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
16 October 2012 and 22 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Oxon, UK
- Age at study initiation: At the start of the study the animals were eight to twelve weeks of age
- Weight at study initiation: The bodyweight variation did not exceed ±20% of the bodyweight of the initially dosed animal
- Fasting period before study: Overnight fast immediately before dosing
- Housing: The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes
- Diet: 2014C Teklad Global Rodent diet supplied by Harlan Laboratorius UK Limited, Oxon, UK, was allowed ad libitum throughout the study
- Water: Free access to mains drinking water
- Acclimation period: At least five days


ENVIRONMENTAL CONDITIONS
- Temperature: 19 to 25°C
- Humidity: Relative humidity 30 to 70%
- Air changes: The rate of air exchange was at least fifteen changes per hour
- Photoperiod: Lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness
- In-life dates: Days 1 to 14
Route of administration:
oral: gavage
Vehicle:
other:
Details on oral exposure:
VEHICLE
- Concentration in vehicle: For the purpose of the study the test item was ground using a mortar and pestle and freshly prepared, as required, as a suspension in arachis oil BP water to give a dose level of 300 or 2000mg/kg bodyweight
- Amount of vehicle: Not stated
- Justification for choice of vehicle: Arachis oil BP was used because the test item did not dissolve/suspend in distilled water
- Lot/batch no. (if required): Not stated
- Purity: Not stated

MAXIMUM DOSE VOLUME APPLIED: 10mL/kg

DOSAGE PREPARATION (if unusual): Not applicable

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: In the absence of data regarding the toxicity of the test material, 300 mg/kg was chosen as the starting dose.

Doses:
Following a sighting test at dose levels of 300 and 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test
material, as a suspension in Arachis oil BP, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored
during the study. All animals were subjected to gross necropsy.
No. of animals per sex per dose:
1 female at 300 mg/kg
1 female at 2000 mg/kg
4 females at 2000 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made ½, 1, 2, and 4 hours after dosing and then daily for fourteen days. Morbidity and mortality checks were made twice daily. Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: Yes
Statistics:
No statistical analysis was performed.
Preliminary study:
- Preliminary results: A sighting test at a dose levels of 300 and 2000 mg/kg showed no signs of toxicity or mortality
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- Lethal results: There were no deaths
Clinical signs:
- Non-lethal results: No signs of systemic toxicity were noted during the observation period.
Body weight:
- Bodyweight results: All animals showed expected gains in bodyweight over the observation period

Gross pathology:
- Gross pathology results: No abnormalities were noted at necropsy
Other findings:
No data reported

Table1              Individual Clinical Observations and Mortality Data -300mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

300

1-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0=     No signs of systemic toxicity

Table 2              Individual Bodyweights and Bodyweight Changes -300mg/kg

Dose Level

mg/kg

Animal Number
and Sex

Bodyweight (g) at Day

Bodyweight Gain (g)
During Week

0

7

14

1

2

300

1-0 Female

180

202

212

22

10

Table 3              Necropsy Findings -300 mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

300

1-0 Female

Killed Day 14

No abnormalities detected

Table4              Individual Clinical Observations and Mortality Data -2000mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0=     No signs of systemic toxicity

Table5              Individual Bodyweights and Bodyweight Changes -2000mg/kg

Dose Level

mg/kg

Animal Number
and Sex

Bodyweight (g) at Day

Bodyweight Gain (g) During Week

0

7

14

1

2

2000

2-0 Female

155

173

193

18

20

3-0 Female

171

201

213

30

12

3-1 Female

172

187

204

15

17

3-2 Female

168

196

215

28

19

3-3 Female

158

176

194

16

18

Table 6              Individual Necropsy Findings-2000mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

2-0 Female

Killed Day 14

No abnormalities detected

3-0 Female

Killed Day 14

No abnormalities detected

3-1 Female

Killed Day 14

No abnormalities detected

3-2 Female

Killed Day 14

No abnormalities detected

3-3 Female

Killed Day 14

No abnormalities detected


Interpretation of results:
other: Criteria for classification as an acute oral toxicant not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexes in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.
Executive summary:

Introduction. 

The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat.The acute oral toxicity of the test item to the Wistar strain rat was assessed in a GLP-compliant study following OECD guideline 420 (adopted 2001) and Method B1 bis Acute Toxicity (Oral) of Commission Regulation (EC) No. 440/2008 in a proprietary, experimental study (Harlan 2013). The study is considered reliable and relevant for use for this endpoint.

Method. 

Following a sighting test at dose levels of 300 mg/kg bodyweight and 2000 mg/kg bodyweight, a further group of four fasted females was given a single oral dose of test material, as a suspension in Arachis oil BP, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study period of 14 days. All animals were subjected to gross necropsy.

Mortality. 

There were no deaths.

 

Clinical Observations.

There were no signs of systemic toxicity noted in the animal treated at a dose levels of 300 mg/kg bodyweight or 2000 mg/kg bodyweight.

 

Bodyweight. 

All animals showed expected gains in bodyweight.

 

Necropsy. 

No abnormalities were noted at necropsy.

 

Conclusion. 

The acute oral median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexes in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

Endpoint:
acute toxicity: oral
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 (oxoaluminium, C16-C18-alkyl esters) and read-across (aluminum, benzoate C16-18-fatty acids complexes) 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.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)

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
Aluminum, benzoate C16-18-fatty acids complexes 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. The fatty acid components of the two substances are therefore expected to be exempt from registration under REACH.

Fatty acids are an endogenous part of every living cell and are an essential dietary requirement. 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.

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 (tested as a 50% w.w. concentration in pharmaceutical white oil) 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 everted gut studies on the target and read-across substances to determine whether the substances are capable of translocation across the intestinal wall. 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 acute oral toxicity of the substances, read across from the source substance to the target substance is considered justified. Data available for the source substance indicate an LD50 of >2000 mg/kg and data on the target substance (tested on the solid, isolated form, stored in the dark under nitrogen until immediately prior to formulation) also gave an LD50 of >2000 mg/kg. The comparable results support the read across from the source to the target substance for this and other mammalian toxicity endpoints.

Both substances would not leach when in situ in base oil during use as grease thickeners and are not expected to be bioavailable. The substances would dissociate into inorganic aluminium species and fatty acids (plus benzoic acid for the source substance), the organic components of which are readily metabolised. As the fatty acid components are essential nutrients to many organisms 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 is considered to provide a worst-case scenario for the target substance.

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
Preliminary study:
- Preliminary results: A sighting test at a dose levels of 300 and 2000 mg/kg showed no signs of toxicity or mortality
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- Lethal results: There were no deaths
Clinical signs:
- Non-lethal results: No signs of systemic toxicity were noted during the observation period.
Body weight:
- Bodyweight results: All animals showed expected gains in bodyweight over the observation period

Gross pathology:
- Gross pathology results: No abnormalities were noted at necropsy
Other findings:
No data reported

Table1              Individual Clinical Observations and Mortality Data -300mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

300

1-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0=     No signs of systemic toxicity

Table 2              Individual Bodyweights and Bodyweight Changes -300mg/kg

Dose Level

mg/kg

Animal Number
and Sex

Bodyweight (g) at Day

Bodyweight Gain (g)
During Week

0

7

14

1

2

300

1-0 Female

180

202

212

22

10

Table 3              Necropsy Findings -300 mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

300

1-0 Female

Killed Day 14

No abnormalities detected

Table4              Individual Clinical Observations and Mortality Data -2000mg/kg

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0=     No signs of systemic toxicity

Table5              Individual Bodyweights and Bodyweight Changes -2000mg/kg

Dose Level

mg/kg

Animal Number
and Sex

Bodyweight (g) at Day

Bodyweight Gain (g) During Week

0

7

14

1

2

2000

2-0 Female

155

173

193

18

20

3-0 Female

171

201

213

30

12

3-1 Female

172

187

204

15

17

3-2 Female

168

196

215

28

19

3-3 Female

158

176

194

16

18

Table 6              Individual Necropsy Findings-2000mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

2-0 Female

Killed Day 14

No abnormalities detected

3-0 Female

Killed Day 14

No abnormalities detected

3-1 Female

Killed Day 14

No abnormalities detected

3-2 Female

Killed Day 14

No abnormalities detected

3-3 Female

Killed Day 14

No abnormalities detected


Interpretation of results:
other: Criteria for classification as an acute oral toxicant not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexes in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.
Executive summary:

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, no toxicological effects were observed and therefore it is concluded that the benzoate ion 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 2000 mg/kg bw (Harlan 2013). Therefore, the LD50 is determined to be > 2000 mg/kg bw.

Introduction. 

The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat.The acute oral toxicity of the test item to the Wistar strain rat was assessed in a GLP-compliant study following OECD guideline 420 (adopted 2001) and Method B1 bis Acute Toxicity (Oral) of Commission Regulation (EC) No. 440/2008 in a proprietary, experimental study (Harlan 2013). The study is considered reliable and relevant for use for this endpoint.

Method. 

Following a sighting test at dose levels of 300 mg/kg bodyweight and 2000 mg/kg bodyweight, a further group of four fasted females was given a single oral dose of test material, as a suspension in Arachis oil BP, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study period of 14 days. All animals were subjected to gross necropsy.

Mortality. 

There were no deaths.

 

Clinical Observations.

There were no signs of systemic toxicity noted in the animal treated at a dose levels of 300 mg/kg bodyweight or 2000 mg/kg bodyweight.

 

Bodyweight. 

All animals showed expected gains in bodyweight.

 

Necropsy. 

No abnormalities were noted at necropsy.

 

Conclusion. 

The acute oral median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexes in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
High. Both studies are GLP compliant.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 October 2012 and 14 November 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source :Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: Eight to twelve weeks of age
- Weight at study initiation: At least 200g
- Housing: The animals were housed in suspended solid floor polypropylene cages furnished with woodflakes. The animals were housed individually during the 24 Hour exposure period and in groups of up to four, by sex, for the remainder of the study.
- Diet: 2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK
- Water: Mains drinking water
- Acclimation period: At least five days

ENVIRONMENTAL CONDITIONS
- Temperature: 19 to 25ºC
- Humidity: Relative humidity 30 to 70%
- Air changes: At least fifteen changes per hour
- Photoperiod: The lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness
Type of coverage:
semiocclusive
Vehicle:
arachis oil
Details on dermal exposure:
TEST SITE
- Area of exposure: Back and flanks of each anomal
- % coverage: Approximately 10% of the total body surface
- Type of wrap if used: Surgical gauze

REMOVAL OF TEST SUBSTANCE
- Washing: Treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test item
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied: 2000 mg/kg bodyweight
- For solids, paste formed: Yes

VEHICLE
- Amount(s) applied: Not reported
- Concentration: Not reported
- Lot/batch no.: Not reported
- Purity: Not reported
Duration of exposure:
24 hours contact period.
Doses:
2000 mg /kg body weight
No. of animals per sex per dose:
In the absence of data suggesting the test item was toxic, one male and one female rat were initially treated with the test item at a dose level of 2000 mg/kg. As no mortalities were noted a further group of animals (four males and four females) was similarly treated with the test item at a dose level of 2000 mg/kg bodyweight to give a total of five males and five females.
Control animals:
not required
Details on study design:
- Duration of observation period following administration: The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days.
- Frequency of observations and weighing: Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs, body weight,organ weights, histopathology. After removal of the dressings and subsequently once daily for fourteen days, the test sites were examined for evidence of primary irritation and scored according to the following scale from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.31

EVALUATION OF SKIN REACTIONS
Erythema and Eschar Formation Value
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to slight eschar formation (injuries in depth) 4
Oedema Formation
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimetre) 3
Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) 4
Statistics:
No statistical analysis was performed.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 95% confidence limits not reported
Mortality:
No deaths occurred during the study.


Clinical signs:
No clinical signs of systemic toxicity were observed during the course of the study.
Body weight:
Animals showed expected gains in bodyweight over the study period, except for three animals which showed bodyweight loss during the first week but expected gain in bodyweight during the second week.
Gross pathology:
No macroscopic findings were recorded at necropsy.
Other findings:
Individual dermal reactions are given in Table 2 and Table 3 in overall remarks section.
Very slight erythema was noted at the test site of three females up to two days after dosing. There were no signs of dermal irritation noted at the test sites of the remaining animals.

Table 1              Individual Clinical Observations and Mortality Data

Dose Level

mg/kg

Animal Number and Sex

Effects Noted After Initiation of Exposure (Hours)

Effects Noted After Initiation of Exposure (Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-1

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0


0 = No signs of systemic toxicity

See Overall remarks for Dermal Reactions Tables 2 and 3.

Table 4              Individual Bodyweights and Weekly Bodyweight Changes

Dose Level mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight Change (g) During Week

0

7

14

1

2

2000

1-0 Male

412

403

419

-9

16

3-0 Male

253

271

291

18

20

3-1 Male

252

265

285

13

20

3-2 Male

260

274

296

14

24

3-3 Male

241

251

266

10

15

2-0 Female

252

245

260

-7

15

4-0 Female

203

206

209

3

3

4-1 Female

210

217

222

7

5

4-2 Female

219

217

229

-2

12

4-3 Female

208

209

218

1

9

Table 5              Individual Necropsy Findings

Dose Level

mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

1-0

Male

Killed Day 14

No abnormalities detected

3-0

Male

Killed Day 14

No abnormalities detected

3-1

Male

Killed Day 14

No abnormalities detected

3-2

Male

Killed Day 14

No abnormalities detected

3-3

Male

Killed Day 14

No abnormalities detected

2-0

Female

Killed Day 14

No abnormalities detected

4-0

Female

Killed Day 14

No abnormalities detected

4-1

Female

Killed Day 14

No abnormalities detected

4-2

Female

Killed Day 14

No abnormalities detected

4-3

Female

Killed Day 14

No abnormalities detected

Interpretation of results:
other: Criteria for classification as an acute dermal toxicant not met
Remarks:
Criteria used for interpretation of results: Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.31
Conclusions:
The acute dermal median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexes in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight.

Executive summary:

Introduction. 

The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat.The acute oral toxicity of the test item to the Wistar strain rat was assessed in a GLP-compliant study following OECD guideline 402 (adopted 1987) and Method B3 Acute Toxicity (Dermal) of Commission Regulation (EC) No. 440/2008 in a proprietary, experimental study (Harlan 2013). The study is considered reliable and relevant for use for this endpoint.

Method. A group of ten animals (five males and five females) was given a single, 24 hour, semi occluded dermal application of the test material to intact skin at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Dermal Irritation.  Very slight erythema was noted at the test site of three females up to two days after dosing. There were no signs of dermal irritation noted at the test sites of the remaining animals.

Bodyweight.  Animals showed expected gains in bodyweight over the study period, except for three animals which showed bodyweight loss during the first week but expected gain in bodyweight during the second week.

Necropsy. No abnormalities were noted at necropsy.

Conclusion. The acute dermal median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexess in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight.

Endpoint:
acute toxicity: dermal
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 (oxoaluminium, C16-C18-alkyl esters) and read-across (aluminum, benzoate C16-18-fatty acids complexes) 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.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)

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
Aluminum, benzoate C16-18-fatty acids complexes 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. The fatty acid components of the two substances are therefore expected to be exempt from registration under REACH.

Fatty acids are an endogenous part of every living cell and are an essential dietary requirement. 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.

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 (tested as a 50% w.w. concentration in pharmaceutical white oil) 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 everted gut studies on the target and read-across substances to determine whether the substances are capable of translocation across the intestinal wall. 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 acute dermal toxicity of the substances, read across from the source substance to the target substance is considered justified. Both substances showed no acute oral toxicity, with LD50 of >2000 mg/kg, and were not corrosive or irritating to skin. Both substances would not leach when in situ in base oil during use as grease thickeners and are not expected to be bioavailable. The substances would dissociate into inorganic aluminium species and fatty acids (plus benzoic acid for the source substance). As the fatty acid components are essential nutrients to many organisms 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 is considered to provide a worst-case scenario for the target substance.

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
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 95% confidence limits not reported
Mortality:
No deaths occurred during the study.


Clinical signs:
No clinical signs of systemic toxicity were observed during the course of the study.
Body weight:
Animals showed expected gains in bodyweight over the study period, except for three animals which showed bodyweight loss during the first week but expected gain in bodyweight during the second week.
Gross pathology:
No macroscopic findings were recorded at necropsy.
Other findings:
Individual dermal reactions are given in Table 2 and Table 3 in overall remarks section.
Very slight erythema was noted at the test site of three females up to two days after dosing. There were no signs of dermal irritation noted at the test sites of the remaining animals.

Table 1              Individual Clinical Observations and Mortality Data

Dose Level

mg/kg

Animal Number and Sex

Effects Noted After Initiation of Exposure (Hours)

Effects Noted After Initiation of Exposure (Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-1

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3

Male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

4-3

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0


0 = No signs of systemic toxicity

See Overall remarks for Dermal Reactions Tables 2 and 3.

Table 4              Individual Bodyweights and Weekly Bodyweight Changes

Dose Level mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight Change (g) During Week

0

7

14

1

2

2000

1-0 Male

412

403

419

-9

16

3-0 Male

253

271

291

18

20

3-1 Male

252

265

285

13

20

3-2 Male

260

274

296

14

24

3-3 Male

241

251

266

10

15

2-0 Female

252

245

260

-7

15

4-0 Female

203

206

209

3

3

4-1 Female

210

217

222

7

5

4-2 Female

219

217

229

-2

12

4-3 Female

208

209

218

1

9

Table 5              Individual Necropsy Findings

Dose Level

mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

1-0

Male

Killed Day 14

No abnormalities detected

3-0

Male

Killed Day 14

No abnormalities detected

3-1

Male

Killed Day 14

No abnormalities detected

3-2

Male

Killed Day 14

No abnormalities detected

3-3

Male

Killed Day 14

No abnormalities detected

2-0

Female

Killed Day 14

No abnormalities detected

4-0

Female

Killed Day 14

No abnormalities detected

4-1

Female

Killed Day 14

No abnormalities detected

4-2

Female

Killed Day 14

No abnormalities detected

4-3

Female

Killed Day 14

No abnormalities detected

Interpretation of results:
other: Criteria for classification as an acute dermal toxicant not met
Remarks:
Criteria used for interpretation of results: Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.31
Conclusions:
The acute dermal median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexes in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight.

Executive summary:

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, no toxicological effects were observed and therefore it is concluded that the benzoate ion 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 2000 mg/kg bw (Harlan 2013). Therefore, the LD50 is determined to be > 2000 mg/kg bw.

Introduction. 

The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat.The acute oral toxicity of the test item to the Wistar strain rat was assessed in a GLP-compliant study following OECD guideline 402 (adopted 1987) and Method B3 Acute Toxicity (Dermal) of Commission Regulation (EC) No. 440/2008 in a proprietary, experimental study (Harlan 2013). The study is considered reliable and relevant for use for this endpoint.

Method. A group of ten animals (five males and five females) was given a single, 24 hour, semi occluded dermal application of the test material to intact skin at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Dermal Irritation.  Very slight erythema was noted at the test site of three females up to two days after dosing. There were no signs of dermal irritation noted at the test sites of the remaining animals.

Bodyweight.  Animals showed expected gains in bodyweight over the study period, except for three animals which showed bodyweight loss during the first week but expected gain in bodyweight during the second week.

Necropsy. No abnormalities were noted at necropsy.

Conclusion. The acute dermal median lethal dose (LD50) of aluminum, benzoate C16-18-fatty acids complexess in the Wistar strain rat was found to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
High. GLP compliant study.

Additional information

In the acute oral toxicity study, there were no deaths or effects on bodyweight and only some minor transient clinical observations made between days 1 and 3. The acute oral median lethal dose (LD50) in both the male and female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight. These data are supported by the acute oral toxicity study with aluminum, benzoate C16-18 fatty acid complexes. The acute dermal toxicity study for aluminum, benzoate C16 -18 fatty acid complexes demonstrates a lack of dermal toxicity associated with the aluminium moiety under the conditions of the test up to the highest dose tested (2000 mg/kg bw in arachis oil BP).


Justification for selection of acute toxicity – oral endpoint
This study is the formal study conducted on the registered substance. It is supported by a similar study conducted on aluminum, benzoate C16-18 fatty acid 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, no toxicological effects were observed and therefore it is concluded that the benzoate ion does not contribute any additional toxicity to the substance. Aluminium, benzoate, C16 -18 fatty acids complexes was tested in the form of an isolated solid and showed no toxic effects at 2000 mg/kg bw (Harlan 2013). Therefore the LD50 is determined to be >2000 mg/kg bw.


Justification for selection of acute toxicity – dermal endpoint
This study was conducted on aluminum, benzoate C16-18 fatty acid complexes and is considered an acceptable read-across substance for the potential dermal toxicity of the aluminium moiety. 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, no toxicological effects were observed and therefore it is concluded that the benzoate ion does not contribute any additional toxicity to the substance. Aluminium, benzoate, C16 -18 fatty acids complexes was tested in the form of an isolated solid and showed no toxic effects at 2000 mg/kg bw (Harlan 2013). Therefore the LD50 is determined to be >2000 mg/kg bw.

Justification for classification or non-classification

There is no evidence of a relevant intrinsic acute oral or dermal toxicity that can be attributed to the registered substance requiring classification or substance specific Risk Management Measures.