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Administrative data

Description of key information

An oral repeated dose toxicity study in rats is available for the read across substance aluminum, benzoate C16-18-fatty acids complexes, incorporating reprotoxicity screening conducted according to OECD 422. 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
Reference
Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant, guideline study, available as an unpublished report.
Reason / purpose:
reference to same study
Qualifier:
according 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: approx 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 deg C
- humidity: 55+/- 15%
- photoperiod: 12h light/12h 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 (if other than water): 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. (if required): 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 degC 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.
Remarks:
Doses / Concentrations:
0, 375, 750, 1500 mg/kg bw/day
Basis:
other: nominal per unit bodyweight
Remarks:
Doses / Concentrations:
0, 56.3, 113, 225 mg/kg bw/day
Basis:
other: expressed as 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 (if not random): 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: Yes
- Time schedule: multiple occasions during each day for morbidity and mortality
- Cage side observations recorded

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

BODY WEIGHT: Yes
- 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:
- 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:
Water intake was observed daily by visual inspection of water bottles for any overt changes.

FOOD EFFICIENCY:
- 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.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- 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: Yes
- 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: Yes
- 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
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


HISTORICAL CONTROL DATA (if applicable)

OTHER FINDINGSThe 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)
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; histopathology 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 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 conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
225 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
As no data was available for the repeat dose oral toxicity of (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium, this endpoints was read across from aluminum, benzoate C16-18-fatty acids complexes. 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 toxicity 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.

Aluminum, benzoate C16-18-fatty acids complexes (CAS No. 94166-87-7, EC No. 303-385-6) is considered suitable for read-across to (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium as it is structurally similar being an approximate 2:1 mixture of (benzoato-O,O')hydroxy(octadecanoato-O,O')aluminium and (benzoato-O,O')hydroxy(hexadecanoato-O,O')aluminium. The only significant difference is the presence of about 30% of the aluminium complex containing the saturated C16-fatty acid, hexadecanoate in place of the saturated C18-fatty acid, octadecanoate. This small difference in composition is expected to have no adverse influence on repeat dose oral toxicity.

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

The read across study was performed on the test substance manufactured in situ in medicinal grade white oil. A key toxicity and reproductive toxicity screen, using the OECD 422 study design, was conducted in rats on aluminum, benzoate C16-18 fatty acid complexes via oral gavage administration. The test material was administered at dose levels of 0, 375, 750 and 1500 mg/kg bw/day nominal in white oil, equating to 56.3, 113 and 225 mg/kg bw/day Active Ingredient. 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 bw/day (225 mg/kg bw/day Active Ingredient).


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

Justification for classification or non-classification

Not classified for STOT systemic. No toxicologically significant adverse effects observed.