Registration Dossier

Administrative data

Description of key information

Data are read-across from an oral repeated dose toxicity study in rats with aluminum, benzoate C16-18 fatty acid complexes incorporating reprotoxicity screening, and conducted according to OECD 422, in which no adverse effect was seen in any of the toxicological parameters examined at any dose.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose:
reference to same study
Reason / purpose:
reference to same study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Ltd
- Age at study initiation: Approximately 12 weeks
- Weight at study initiation: Males - 322 to 394g; females (nulliparous and nonpregnant) - 195 to 237g
- Housing: Initially in groups of 4 in solid floor propylene cages with softwood bedding. During pairing animals were transferred to polypropylene grid floor cages suspended over trays lined with absorbant paper, one male : one female basis. Following successful mating, males returned to original cages. Mated females housed individually during gestation/lactation in the solid floor cages as for mating.
- Enrichment: Wooden chew blocks and cardboard tunnels.
- Diet: Rodent 2018C Teklad Global Certified Diet, Harlan Laboratories UK Ltd
- Water: Mains drinking water ad libitum
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 ± 2°C
- Humidity: 55 ± 15%
- Photoperiod: 12 h light / 12 h dark
Route of administration:
oral: gavage
Vehicle:
other: MOL WO M 46 Medicinal white oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
VEHICLE
- Justification for use and choice of vehicle: Test material synthesised in the presence of MOL WO M 46 Medicinal white oil. Same white oil used for dilution of test material and as the control vehicle
- Test substance concentration in vehicle: 15%
- Treatment volume: 5 mL/kg bw/day
- Lot/batch no.: 9037038
- Purity: 100%
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentrations of the test material in the vehicle dilutions were determined by Inductively Coupled Plasma Mass Spectroscopy (ICP-MS). The test item formulations were extracted with hexane, evaporated to dryness and re-dissolved in 2% nitric acid. Homogeneity determinations were performed on samples taken from the top, middle and bottom of the container. Stability determinations were performed before and after storage for 13 days at approx +4°C in the dark for 13 days, by IR spectroscopy using a Perkin Elmer Spectrum One Fournier-transform infrared spectrophotometer.
Duration of treatment / exposure:
Males dosed for 42 days and killed on day 43, beginning 14 days prior to mating.
Dosing of females began 14 days before mating, and continued through mating, up to and including day 4 post partum.They were killed on day 5 post partum.
Frequency of treatment:
Daily, once per day
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Basis: Dose (test item in vehicle)
Dose / conc.:
375 mg/kg bw/day (nominal)
Remarks:
Basis: Dose (test item in vehicle)
Dose / conc.:
750 mg/kg bw/day (nominal)
Remarks:
Basis: Dose (test item in vehicle)
Dose / conc.:
1 500 mg/kg bw/day (nominal)
Remarks:
Basis: Dose (test item in vehicle)
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Remarks:
Basis: Concentration of test item (active ingredient)
Dose / conc.:
56.3 mg/kg bw/day (actual dose received)
Remarks:
Basis: Concentration of test item (active ingredient)
Dose / conc.:
113 mg/kg bw/day (actual dose received)
Remarks:
Basis: Concentration of test item (active ingredient)
Dose / conc.:
225 mg/kg bw/day (actual dose received)
Remarks:
Basis: Concentration of test item (active ingredient)
No. of animals per sex per dose:
12 males and 12 females per dose level
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected on the basis of a 14-day dose range finding study where three groups of 3 male and 3 female wistar rats were treated at 375, 750 and 1500 mg/kg bw/day (dosed as supplied, containing 15% active ingredient). A group of 3 males and 3 females received the vehicle (medicinal white oil). No signs of toxicity were observed, and no adverse effects on bodyweight, food consumption, or water consumption. No macroscopic changes were seen at necropsy.
- Rationale for animal assignment: The animals were allocated to dose groups using a randomised procedure based on stratified bodyweights. Group mean bodyweights were then dermined to ensure similarity between the groups.
- Rationale for selecting satellite groups: Not applicable
- Post-exposure recovery period in satellite groups: Not applicable
Positive control:
Not included
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS
- Time schedule: Multiple occasions during each day for morbidity and mortality
- Cage side observations recorded: Yes

DETAILED CLINICAL OBSERVATIONS
- Time schedule: Before dosing, 30 mins, 1 and 5 h after dosing during weekdays; before dosing and 1 h after dosing at weekends

BODY WEIGHT
- Time schedule for examinations: Prior to dosing, then weekly for males until termination, and weekly for females until mating was evident. Then for females bodyweight was recorded on days 0, 7, 14 and 20 post coitum, and on days 1 and 4 post partum.

FOOD CONSUMPTION
- Time schedule for examinations: Food consumption was recorded for each cage of adults and was continued for males after the mating phase. For females showing evidence of mating, food consumption was recorded for the periods covering post coitum Days 0-7, 7-14 and 14-20. For females with live litters, food consumption was recorded on Days 1 and 4 post partum.

WATER CONSUMPTION
- Time schedule for examinations: Water intake was observed daily by visual inspection of water bottles for any overt changes.

FOOD EFFICIENCY
- Time schedule for examinations: Food efficiency (the ratio of body weight change/dietary intake) was calculated retrospectively for males throughout the study period (with the exception of the mating phase) and for females during the pre-pairing phase.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY
- Time schedule for collection of blood: Day 42 for males, day 4 post partum for females
- Anaesthetic used for blood collection: No
- Animals fasted: No
- How many animals: 5 males and 5 females per group
- Parameters checked:
Haemoglobin (Hb)
Erythrocyte count (RBC)
Haematocrit (Hct)
Erythrocyte indices - mean corpuscular haemoglobin (MCH)
- mean corpuscular volume (MCV)
- mean corpuscular haemoglobin concentration (MCHC)
Total leucocyte count (WBC)
Differential leucocyte count - neutrophils (Neut)
- lymphocytes (Lymph)
- monocytes (Mono)
- eosinophils (Eos)
- basophils (Bas)
Platelet count (PLT)
Reticulocyte count (Retic) - Methylene blue stained slides were prepared but reticulocytes were not assessed
Prothrombin time (CT) was assessed by ‘Innovin’ and Activated partial thromboplastin time (APTT) was assessed by ‘Actin FS’ using samples collected into sodium citrate solution (0.11 mol/L).

CLINICAL CHEMISTRY
- Time schedule for collection of blood: Day 42 for males, day 4 post partum for females
- Animals fasted: No
- How many animals: 5 males and 5 females per group
- Parameters checked
Urea
Calcium (Ca++)
Glucose
Inorganic phosphorus (P)
Total protein (Tot.Prot.)
Aspartate aminotransferase (ASAT)
Albumin
Alanine aminotransferase (ALAT)
Albumin/Globulin (A/G) ratio (by calculation)
Alkaline phosphatase (AP)
Sodium (Na+)
Creatinine (Creat)
Potassium (K+)
Total cholesterol (Chol)
Chloride (Cl-)
Total bilirubin (Bili)
Bile acids

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION
- Time schedule for examinations: Prior to start of treatment and weekly intervals thereafter. Functional performance tests performed on 5 selected males and females from each dose level prior to termination, together with an assessment of sensory reactivity to various stimuli.
-Behavioural assessments: Detailed individual clinical observations were performed for each animal using a purpose built arena. This test was developed from the methods used by Irwin (1968) and Moser et al (1988). The scoring system used is outlined in The Key to Scoring System and Explanation for Behavioural Assessments and Sensory Reactivity Tests.
- Functional/performance tests: Motor activity, forelimb/hindlimb grip strength
- Sensory reactivity: Each animal was individually assessed for sensory reactivity to auditory, visual and proprioceptive stimuli. This assessment was developed from the methods employed by Irwin (1968) and Moser et al (1988).
- Dose groups that were examined: Each group
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
Adrenals
Ovaries
Aorta (thoracic)
Pancreas
Bone & bone marrow (femur including stifle joint)
Pituitary
Bone & bone marrow (sternum)
Prostate
Brain (including cerebrum, cerebellum and pons)
Oesophagus
Caecum
Rectum
Coagulating gland
Salivary glands (submaxillary)
Colon
Sciatic nerve
Duodenum
Seminal vesicles
Statistics:
Where appropriate, data transformations were performed using the most suitable method. The homogeneity of variance from mean values was analysed using Bartlett’s test. Intergroup variances were assessed using suitable ANOVA, or if required, ANCOVA with appropriate covariates. Any transformed data were analysed to find the lowest treatment level that showed a significant effect, using the Williams Test for parametric data or the Shirley Test for non-parametric data. If no dose response was found, but the data shows non-homogeneity of means, the data were analysed by a stepwise Dunnett’s (parametric) or Steel (non-parametric) test to determine significant difference from the control group. Where the data were unsuitable for these analyses, pair-wise tests was performed using the Student t-test (parametric) or the Mann-Whitney U test (non-parametric).
Data not analysed by the Provantis data capture system were assessed separately using the SPSS statistical package. Initially, the homogeneity of the data was assessed using Levene’s test. Where Levene’s test was shown to be non-significant (p≥0.05), parametric analysis of the data was applied, incorporating analysis of variance (ANOVA). If this data was shown to be significant, this analysis was followed by pair-wise comparisons using Dunnett’s test. Where Levene’s test was significant, non-parametric analysis of the data was analysed incorporating the Kruskal-Wallis test which if significant, was followed by the Mann-Whitney U test. Dose response relationship was also be investigated by linear regression. Where the data was unsuitable for these analyses, then pair-wise tests were performed using the Student t-test (parametric) or the Mann-Whitney U test (non-parametric).
Due to the preponderance of non-normally distributed data, reproductive parameters (implantation losses, offspring sex ratio and offspring surface righting) were analysed using non-parametric analyses.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
There were no unscheduled deaths and no toxicologically significant clinical observations detected.
BODY WEIGHT AND WEIGHT GAIN
There were no toxicologically significant effects detected on body weight development.
FOOD CONSUMPTION
No adverse effect on food consumption was deteted in treated animals
FOOD EFFICIENCY
No adverse effect on food efficiency was detected in treated animals
WATER CONSUMPTION
No adverse effect on water consumption was detected
OPHTHALMOSCOPIC EXAMINATION
Not examined
HAEMATOLOGY
No toxicologically significant effects were detected
CLINICAL CHEMISTRY
No toxicologically significant effects were detected
URINALYSIS
Not examined
NEUROBEHAVIOUR
There were no toxicologically significant changes in the behavioural paremeters measured, in functional performance or sensory reactivity
ORGAN WEIGHTS
No toxicologically signifcant treatment related trends were detected in the organ weights measured.
GROSS PATHOLOGY
There were no macroscopic abnormalities detected that were considered to be related to treatment
HISTOPATHOLOGY
No treatment related microscopic findings were detected
Key result
Dose descriptor:
NOAEL
Effect level:
> 225 other: mg/kg bw/day Active Ingredient
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects seen on clinical signs; mortality; body weight; food consumption; water consumption; haematology; clinical chemistry; gross pathology; organ weights; histopathlogy at doses up to 1500 mg/kg bw/day (225 mg/kg bw/day Active Ingredient)
Critical effects observed:
not specified
Conclusions:
The subchronic repeated dose oral toxicity of grease containing 15% active ingredient (aluminum, benzoate C16-18-fatty acids complexes) in MOL WO M 46 medicinal white oil to rats gave a NOAEL of 1500 mg/kg bw/day (225 mg/kg bw/day Active Ingredient).
Executive summary:

The subacute repeated dose oral toxicity of grease containing 15% active ingredient (aluminum, benzoate C16-18-fatty acids complexes) in MOL WO M 46 medicinal white oil to rats gave a NOAEL of 1500 mg/kg bw/day (225 mg/kg bw/day Active Ingredient)

The repeated dose oral toxicity of aluminum, benzoate C16 -18 -fatty acids complexes to rats is taken from data presented within an oral (gavage) combined repeat dose toxiciy study with reproduction/developmental toxicity screening test in the rat performed in compliance with, amongst others, the requirements of the OECD Guidelines for Testing of Chemicals No.422 "Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Screening Test" (adopted 22 March 1996). Since the results are taken from a regulatory and GLP compliant study, the data are considered reliable and relevant for use in assessing this endpoint.

Endpoint:
short-term repeated dose 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 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.

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 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% concentration in phamacuetical 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 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 repeated dose oral toxicity of the substances, read across from the source to the target substance is considered justified. Data available for the both substances show that they have an acute oral toxicity LD50 of >2000 mg/kg bw. 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
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
There were no unscheduled deaths and no toxicologically significant clinical observations detected.
BODY WEIGHT AND WEIGHT GAIN
There were no toxicologically significant effects detected on body weight development.
FOOD CONSUMPTION
No adverse effect on food consumption was deteted in treated animals
FOOD EFFICIENCY
No adverse effect on food efficiency was detected in treated animals
WATER CONSUMPTION
No adverse effect on water consumption was detected
OPHTHALMOSCOPIC EXAMINATION
Not examined
HAEMATOLOGY
No toxicologically significant effects were detected
CLINICAL CHEMISTRY
No toxicologically significant effects were detected
URINALYSIS
Not examined
NEUROBEHAVIOUR
There were no toxicologically significant changes in the behavioural paremeters measured, in functional performance or sensory reactivity
ORGAN WEIGHTS
No toxicologically signifcant treatment related trends were detected in the organ weights measured.
GROSS PATHOLOGY
There were no macroscopic abnormalities detected that were considered to be related to treatment
HISTOPATHOLOGY
No treatment related microscopic findings were detected
Key result
Dose descriptor:
NOAEL
Effect level:
> 225 other: mg/kg bw/day Active Ingredient
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects seen on clinical signs; mortality; body weight; food consumption; water consumption; haematology; clinical chemistry; gross pathology; organ weights; histopathlogy at doses up to 1500 mg/kg bw/day (225 mg/kg bw/day Active Ingredient)
Critical effects observed:
not specified
Conclusions:
The subchronic repeated dose oral toxicity of grease containing 15% active ingredient (aluminum, benzoate C16-18-fatty acids complexes) in MOL WO M 46 medicinal white oil to rats gave a NOAEL of 1500 mg/kg bw/day (225 mg/kg bw/day Active Ingredient).
Executive summary:

The subacute repeated dose oral toxicity of grease containing 15% active ingredient (aluminum, benzoate C16-18-fatty acids complexes) in MOL WO M 46 medicinal white oil to rats gave a NOAEL of 1500 mg/kg bw/day (225 mg/kg bw/day Active Ingredient). 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 showed no toxicological effects in an OECD 422 combined repeat dose oral toxicity and reproduvctive/developmental toxicity screen (Harlan 2013). Therefore, aluminium, benzoate C16 -18 fatty acids complexes is considered to have an NOEAL of 1500 mg/kg bw/day (225 mg/kg bw/day active ingredient) and this has been read across to the target substance.

The subchronic repeated dose oral toxicity of grease containing 15% active ingredient (aluminum, benzoate C16-18-fatty acids complexes) in MOL WO M 46 medicinal white oil to rats gave a NOAEL of 1500 mg/kg bw/day (225 mg/kg bw/day Active Ingredient). The repeated dose oral toxicity of aluminum, benzoate C16 -18 -fatty acids complexes to rats is taken from data presented within an oral (gavage) combined repeat dose toxiciy study with reproduction/developmental toxicity screening test in the rat performed in compliance with, amongst others, the requirements of the OECD Guidelines for Testing of Chemicals No.422 "Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Screening Test" (adopted 22 March 1996). Since the results are taken from a regulatory and GLP compliant study, the data are considered reliable and relevant for use in assessing this endpoint.

Endpoint:
sub-chronic toxicity: oral
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
a sub-chronic toxicity study (90 days) does not need to be conducted because the substance is unreactive, insoluble and not inhalable and there is no evidence of absorption and no evidence of toxicity in a 28-day 'limit test' and human exposure is limited
Justification for type of information:
JUSTIFICATION FOR DATA WAIVING
In a regulatory compliant GLP study on the read across substance aluminum, benzoate C16-18 fatty acid complexes, rats were dosed daily by oral gavage for a minimum of 42 days at dose levels of 0, 375, 750 and 1500 mg/kg bw/day (equivalent to 0, 56.3, 113 and 225 mg/kg bw/day Active Ingredient). Since this study was conducted on the substance manufactured in a white oil base, the maximum dose that could be administered was limited by the physico-chemical characteristics of the dosing preparation. Therefore the limit dose of actual aluminum, benzoate C16-18 fatty acid complexes could not be attained. Since no toxicity was observed in the OECD422 study and the maximum dose was well below the limit dose of 1000 mg/kg/day (actual maximum dose achieved was 225 mg/kg bw/day), then the likelihood of identifying any further toxic response from a 90-day study at the same concentrations is extremely low. Thus the Annex IX requirement for a subchronic toxicity study is waived in accordance with Annex XI.

Both the registered substance and aluminium, benzoate, C16-18 fatty acids complexes are used specifically as thickeners in industrial greases and lubricants and as such are manufactured and used solely in situ in base oil. The interactions between the thickener and base oil do not strictly fall under the definitions of a reaction product nor do they act as a simple mixture of components due to matrix effects which occur in the manufacturing/ blending process. Matrix effects should be taken into account when assessing whether adverse effects would be seen in grease products because entrainment of grease thickeners during the manufacturing process as part of the grease matrix severely limits exposure and will have a significant impact on the outcome of any risk assessments. The fact that matrix effects do occur is a point recognised in the OECD Lubricant Emission Scenario document (2004) and it is therefore reasonable to assess the influence the process of manufacturing the thickener in an inert base oil has on factors such as availability, which are critical to assessing the potential risks posed by the grease thickeners when in the grease.

Leaching tests have been undertaken to assess whether the metallic components of the metal-soap and metal-complex-soap grease thickeners remain in base grease(s) and hence determine the degree of availability of the thickeners (see section 4.5 and Appendix 2 of the CSR). The results of the study show no leaching of the thickener from the base oil and a lack of bioavailability of the substance in aqueous media. The same lack of bioavailability is expected in GI fluids as well and, as a further demonstration of this lack of availability, it is proposed to conduct leaching studies on aluminium thickeners in base oil using fed state simulated intestinal fluid (FeSSIF). This approach provides sufficient information to assess the risks associated with the aluminium thickener complexes following exposure of mammals by the oral route. Also, the use of intestinal fluid has been used to evaluate the solubility of drugs for the prediction of oral absorption (Augustijns 2014).

This substance has been registered by a Member of the European REACH Grease Thickeners Consortium (ERGTC). A number of decisions have been made in the dossier with regard to the approach taken for registering the substance including the testing strategy and the justification for waiving certain endpoints. Several of the decisions reflect the technical difficulties of testing the substance and the relevance of data with regard to the potential for exposure, given that the substance typically occurs in situ in base oil. A face to face meeting between the ERGTC and ECHA was held in Helsinki on 8th September 2016 which discussed many of these topics and a copy of the minutes from the meeting are attached to the dossier (See section 13 of IUCLID). Therefore, if there are any queries or concerns which arise when the dossier is reviewed, it is requested that the reviewer discuss these with the ERGTC (ERGTC@wca-consulting.com) as there may be background information and previous discussions between the ERGTC and ECHA which are relevant.
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
225 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Guideline study conducted to GLP

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

A key toxicity and reproductive toxicity screen, using the OECD 422 study design, conducted in rats on aluminum, benzoate C16-18 fatty acid complexes via oral gavage administration has been read across to the registered substance. This study was performed on the test substance manufactured in situ in medicinal grade white oil and the maximum dose that could be administered was limited by the physico-chemical characteristics of the dosing preparation. The test material was administered at dose levels of 0, 375, 750 and 1500 mg/kg/day nominal, equating to 56.3, 113 and 225 mg/kg/day by analysis. There were no treatment-related effects at any dose level on any of the toxicological parameters evaluated in this study. Based on these data, the NOAEL for repeated dose toxicity was 1500 mg/kg/day (225 mg/kg bw/day Active Ingredient). This lack of any systemic toxicity was also observed in the acute oral and dermal toxicity studies, which were conducted on aluminium, benzoate, C16-18 fatty acid complexes isolated from an oil base and re-dissolved in arachis oil, at doses up to 2000 mg/kg bw by either route and in the acute oral toxicity study, which was conducted on the registered substance as a suspension in poly alpha olefin, at doses up to 2000 mg/kg bw.

This substance is considered suitable for read across as it contains a fatty acid moiety coordinated to an aluminium atom. Although it 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 showed no toxicological effects in an OECD 422 combined repeat dose oral toxicity and reproductive/developmental toxicity screen (Harlan 2013). Therefore, aluminium benzoate C16 -18 fatty acids complexes is considered to have a NOAEL of 1500 mg/kg bw/day (225 mg/kg bw/day active ingredient) and this has been read across to the target substance.

The registered substance is manufactured and used solely in situ in an inert carrier. The substance and aluminium, benzoate, C16-18 fatty acids complexes are used specifically as thickeners in industrial greases and lubricants and as such typically occur in situ in base oil. The interactions between the thickener and base oil do not strictly fall under the definitions of a reaction product nor do they act as a simple mixture of components due to matrix effects which occur in the manufacturing/ blending process. Matrix effects should be taken into account when assessing whether adverse effects would be seen in grease products because entrainment of grease thickeners during the manufacturing process as part of the grease matrix severely limits exposure and will have a significant impact on the outcome of any risk assessments. The fact that matrix effects do occur is a point recognised in the OECD Lubricant Emission Scenario document (2004) and it is therefore reasonable to assess the influence the process of manufacturing the thickener in an inert base oil has on factors such as availability, which are critical to assessing the potential risks posed by the grease thickeners when in the grease.

Leaching tests have been undertaken to assess whether the metallic components of the metal-soap and metal-complex-soap grease thickeners remain in base grease(s) and hence determine the degree of availability of the thickeners (see section 4.6 and Appendix 2 of the CSR). The results of the study show no leaching of the thickener from the base oil and a lack of bioavailability of the substance in aqueous media. The same lack of bioavailability is expected in GI fluids as well and, as a further demonstration of this lack of availability, it is proposed to conduct leaching studies on aluminium thickeners in base oil using fed state simulated intestinal fluid (FeSSIF). This approach provides sufficient information to assess the risks associated with the aluminium thickener complexes following exposure of mammals by the oral route. Also, the use of intestinal fluid has been used to evaluate the solubility of drugs for the prediction of oral absorption (Augustijns 2014).

This substance has been registered by a Member of the European REACH Grease Thickeners Consortium (ERGTC). A number of decisions have been made in the dossier with regard to the approach taken for registering the substance including the testing strategy and the justification for waiving certain endpoints. Several of the decisions reflect the technical difficulties of testing the substance and the relevance of data with regard to the potential for exposure, given that the substance typically occurs in situ in base oil. A face to face meeting between the ERGTC and ECHA was held in Helsinki on 8th September 2016 which discussed many of these topics and a copy of the minutes from the meeting are attached to the dossier (See section 13 of IUCLID). Therefore, if there are any queries or concerns which arise when the dossier is reviewed, it is requested that the reviewer discuss these with the ERGTC (ERGTC@wca-consulting.com) as there may be background information and previous discussions between the ERGTC and ECHA which are relevant.

Data taken from the OECD 422 study with aluminium, benzoate C16-18 fatty acid complexes are read across to the registered substance. There were no indications of any target organ toxicity when administered to rats on this study so it can be concluded that no classification for this endpoint is required.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
This screening study provides relevant read-across experimental data on this endpoint from a related substance, aluminum, benzoate C16-18 fatty acid complexes in which a clear NOAEL is identified.

Justification for classification or non-classification

Data taken from the OECD 422 study with aluminium, benzoate C16-18 fatty acid complexes are read across to the registered substance. There were no indications of any target organ toxicity when administered to rats on this study so it can be concluded that no classification for this endpoint is required.