Aluminium ammonium bis(sulphate)

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

11 September 2006 - 03 November 2006

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study was conducted according to OECD guideline 422 and under GLP conditions.Data from Aluminium chloride are relevant for ammonium alum considering the dissociation of both salts as aluminium ions (and chloride or sulfate and ammonium ions) after oral administration.

Justification for data waiving:

other:

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

Principles of method if other than guideline:

Not relevant

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Aluminum chloride, basic

EC Number:

215-477-2

EC Name:

Aluminum chloride, basic

Cas Number:

1327-41-9

IUPAC Name:

aluminum trichloride

Constituent 2

Reference substance name:

Aluminium chloride basic

IUPAC Name:

Aluminium chloride basic

Details on test material:

- Name of test material (as cited in study report): Aluminium chloride basic
- Substance type: Aqueous solution
- Physical state: Liquid
- Analytical purity: No data
- Impurities (identity and concentrations): Confidential information
- Composition of test material, percentage of components: Confidential information
- Lot/batch No.: Confidential information
- Expiration date of the lot/batch: Confidential information
- Stability under test conditions: At least 48 hours
- Storage condition of test material: At room temperature in the dark

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation:
Males: approx. 9 weeks
Females: approx. 11 weeks
- Weight at study initiation: All animals within +/- 20% of the sex mean
- Fasting period before study:
- Housing:
Pre-mating: groups of 5 animals/sex/cage
Mating: 1 male, 1 female
Post-mating: males in groups of 5 animals/sex/cage, females individually
Lactation: offspring together with dam
- Diet (e.g. ad libitum): Ad libitum, pelleted rodent diet
- Water (e.g. ad libitum): Ad libitum, tap-water
- Acclimation period: 4 days prior to start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.9-23.4
- Humidity (%): 37-95
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Formulations (w/w) were prepared within 4 hours prior to dosing and were homogenised to a visually acceptable level. Adjustment was made for specific gravity of the test substance. No adjustment was made for specific gravity of the vehicle and formulation.

VEHICLE
- Justification for use and choice of vehicle (if other than water): Water
- Concentration in vehicle: ?
- Amount of vehicle (if gavage): 5 ml/kg bw

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Formulated samples were analysed using ICP-MS

Duration of treatment / exposure:

Males: 28 days
Females: 37 to 53 days (because of developmental purpose of study)

Frequency of treatment:

Once daily, 7 days a week

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
40, 200, 1000 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 2

Remarks:

Doses / Concentrations:
= 0, 3.6, 18 and 90 mg/kg bw/day Aluminium
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent no treatment

Details on study design:

- Dose selection rationale: Based on the results of a dose range finding study (NOTOX project 473524)
- Rationale for animal assignment (if not random): Random
- Section schedule rationale (if not random): Not data

Positive control:

Not relevant

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily
- Cage side observations checked: Mortality/viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily

BODY WEIGHT: Yes
- Time schedule for examinations: At start, once weekly and at death (females also at: gestation days 0, 4, 7, 11, 17, 20, lactation day 1, 4)

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, weekly
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: No, not recorded, only subjective appraisal

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Immediately prior to scheduled post mortem examination
- Anaesthetic used for blood collection: Yes (Isoflurane)
- Animals fasted: Yes, max. 20 hours overnight
- How many animals: 5 males and 5 females randomly selected from each group
- Parameters checked: see "Any other information on materials and methods incl. tables"

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Immediately prior to scheduled post mortem examination
- Animals fasted: Yes, max. 20 hours overnight
- How many animals: 5 males and 5 females randomly selected from each group
- Parameters checked: see "Any other information on materials and methods incl. tables"

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Males during week 4, females during lactation
- Dose groups that were examined: 5 males and 5 females, randomly selected from all groups
- Battery of functions tested: hearing ability / pupillary reflex / static righting reflex / grip strength / motor activity test

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, of all organs of all animals
HISTOPATHOLOGY: Yes, examination of following organs from 5 animals/sex/group: see "Any other information on materials and methods incl. tables"

Other examinations:

Not relevant

Statistics:

- Variables with normal distribution: Dunnett-test based on a pooled variance estimate was applied for the comparison of the treated groups and the control groups for each sex
- The steel-test was applied if the data was not assumed to follow a normal distribution
- The Fisher Exact-test was applied to frequency data

All tests were two-sided and in all cases p<0.05 was accepted as the lowest level of significance.
No statistical analysis performed on histopathology findings.
Test statistics were calculated on the basis of exact values for means and pooled variances.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

No clinical signs noted. Functional observation battery: all parameters normal in all animals None related to test substance One female sacrificed in extremis due to parturition difficulties on day 22 p.c. Cause of death could not be established

Mortality:

no mortality observed

Description (incidence):

No clinical signs noted. Functional observation battery: all parameters normal in all animals None related to test substance One female sacrificed in extremis due to parturition difficulties on day 22 p.c. Cause of death could not be established

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Slightly lower mean bw for females at 1000 mg/kg bw/d at day 8 with slight weight loss for three of these females. BW recovered during the following mating and post-coital period. In the other groups no deviations compared to controls

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Mean food consumption of females at 1000 mg/kg bw/d slightly lower than controls. Otherwise no deviations compared to controls

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Description (incidence and severity):

Not performed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Minor, but statistically significant lowering of MCHC in M and F of 1000 mg/kg bw/g group Minor but statistically significant higer plateled count in males at 1000 mg/kg bw

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Males at 1000 mg/kg bw/d: statistically significant - lower ALP activity - lower albumin levels - higher Potassium levels - higher inorganic phospate levels no deviations in females or in M?F at 40 & 200 mg/kg/d

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Minor changes in brain weight of females at 1000 mg/kg and kidney weight of females at 200 mg/kg were not considered to be relevant or substance-related.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Red foci on glandular mucosa of the stomach in 5/10 males at 1000 mg/kg/day, with thickening of glandular mucosa or limiting ridge in two of these five. No other relevant abnormalities were seen except coagulum in the caecum and enlargement of liver and s

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Mild to moderate subacute inflammation of the glandualr mucosa and minimal to moderate superficial easinophilic spheroids in alle xamined animals of both sexes at 1000 mg/kg bw/d

Histopathological findings: neoplastic:

not examined

Details on results:

BODY WEIGHT AND WEIGHT GAIN
Slightly lower mean bw for females at 1000 mg/kg bw/d at day 8 with slight weight loss for three of these females. BW recovered during the following mating and post-coital period. In the other groups no deviations compared to controls

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Mean food consumption of females at 1000 mg/kg bw/d slightly lower than controls. Otherwise no deviations compared to controls

HAEMATOLOGY
Minor, but statistically significant lowering of MCHC in M and F of 1000 mg/kg bw/g group
Minor, but statistically significant higher plateled count in males at 1000 mg/kg bw

CLINICAL CHEMISTRY
Males at 1000 mg/kg bw/d: statistically significant
- lower ALP activity
- lower albumin levels
- higher Potassium levels
- higher inorganic phospate levels
no deviations in females or in M/F at 40 & 200 mg/kg/d

GROSS PATHOLOGY
Red foci on glandular mucosa of the stomach in 5/10 males at 1000 mg/kg/day, with thickening of glandular mucosa or limiting ridge in two of these five. No other relevant abnormalities were seen except coagulum in the caecum and enlargement of liver and spleen in the animal sacrificed in extremis.

HISTOPATHOLOGY: NON-NEOPLASTIC
Mild to moderate subacute inflammation of the glandular mucosa and minimal to moderate superficial easinophilic spheroids in alle examined animals of both sexes at 1000 mg/kg bw/day

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

200 mg/kg bw/day (actual dose received)

Sex:

male

Basis for effect level:

other: Local effects (stomach) at 1000 mg/kg bw/day

2

Dose descriptor:

NOAEL

Effect level:

18 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Remarks:

Al3+

Sex:

male

Basis for effect level:

other: Local effects (stomach) at 90 mg Al3+/kg bw/day

3

Dose descriptor:

LOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: Local effects (stomach) at 90 mg Al3+/kg bw/day

4

Dose descriptor:

LOAEL

Effect level:

90 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Remarks:

Al3+

Sex:

male

Basis for effect level:

other: Local effects (stomach) at 90 mg Al3+/kg bw/day

5

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Remarks:

Al chloride basic

Sex:

male

Basis for effect level:

other: Systemic effects: haematology, clinical chemistry, gross pathology, histopathology

6

Dose descriptor:

NOAEL

Effect level:

90 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Remarks:

Al3+

Sex:

male

Basis for effect level:

other: Systemic effects: haematology, clinical chemistry, gross pathology, histopathology

7

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Sex:

female

Basis for effect level:

other: Local and systemic (HISTOPATHOLOGY EFFECTS at 1000 mg/kg/day)

8

Dose descriptor:

NOAEL

Effect level:

90 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Remarks:

Al3+

Sex:

female

Basis for effect level:

other: Local and systemic (HISTOPATHOLOGY EFFECTS at 90 mg Al3+/kg bw/day)

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Not relevant

Applicant's summary and conclusion

Conclusions:

In a combined repeated dose / reproductive screening study (OECD 422), administration of Aluminium chloride basic by oral gavage to male and female rats at dose levels of 20, 200 or 1000 mg /kg/d (equivalent to 3.6, 18 and 90 mg/kg bw/d) was studied. No toxic effects were observed in females at any dose. Therefore, the overall NOAEL for female rats was established to be 1000 mg/kg bw/day(equivalent to 90 mg/kg bw/d). For males the NOAEL for local effects was established to be 200 mg/kg bw/day (equivalent to 18 mg/kg bw/d) and for systemic toxicity 1000 mg/kg bw/day (equivalent to 90 mg/kg bw/d).

Executive summary:

A combined repeated dose / reproductive screening study (OECD 422), studied the administration of Aluminium chloride basic by oral gavage to male and female Wistar rats at dose levels of 20, 200 or 1000 mg /kg bw/d (equivalent to 3.6, 18 and 90 mg Al³⁺/kg bw/d). 10 animals/sex/group were used. Males were exposed for 28 days, females between 37 -53 days (because of developmental toxicity purpose of study). Detailed clinical signs, body weight, food consumption, water consumption (subjective appraisal), haematology, clinical chemistry, neurobehaviour and gross and histopathology were studied.

Slightly lower mean bodyweight was observed for females at the 1000 mg/kg bw/day

(equivalent to 90 mg Al³⁺/kg bw/d)

group at day 8, with slight weight loss for three of these females. Bodyweight recovered during the following mating and post-coital period. In the other groups no deviations were observed as compared to controls.

Mean food consumption of females at 1000 mg/kg bw/day (equivalent to 90 mg Al³⁺/kg bw/d) was slightly lower than controls. Otherwise no deviations were observed as compared to controls.

Minor, but statistically significant lowering of MCHC was observed in males and females of the 1000 mg/kg bw/day group (equivalent to 90 mg Al³⁺/kg bw/d).

Minor, but statistically significant higher plateled count was observed in males at a dose of 1000 mg/kg bw/d

(equivalent to 90 mg Al³⁺/kg bw/d).

Males at 1000 mg/kg bw/day had statistically significant lower ALP activity, lower albumin levels, higher Potassium levels, higher inorganic phospate levels. No deviations were observed in females or in males and females at a dose of 40 and 200 mg/kg bw/d

(equivalent to 3.6 and 18 mg Al^3+/kg bw/d).

.

Red foci on glandular mucosa of the stomach were observed in 5 of 10 males at a dose of 1000 mg/kg bw/day

(equivalent to 90 mg Al³⁺/kg bw/d)

, with thickening of glandular mucosa or limiting ridge in two of these five. No other relevant abnormalities were seen.

Mild to moderate subacute inflammation of the glandular mucosa and minimal to moderate superficial easinophilic spheroids was observed in all examined animals of both sexes at 1000 mg/kg bw/day

(equivalent to 90 mg Al³⁺/kg bw/d).

Based on these results, the overall NOAEL for female rats was established to be 1000 mg/kg bw/day

(equivalent to 90 mg Al³⁺/kg bw/d).

For males the NOAEL for local effects was established to be 200 mg/kg bw/day and for systemic toxicity 1000 mg/kg bw/day (equivalent to 90 mg Al³⁺/kg bw/d ). In a combined repeated dose / reproductive screening study, administration of Aluminium chloride basic by oral gavage to male and female rats at dose levels of 20, 200 or 1000 mg /kg bw/d,

From two weeks prior to mating to at least 3 days of lactation (females) or 28 days (males) revealed no toxicity in females at any dose.

The NOAEL for local (stomach) toxicity in males is 200 mg/kg bw/d (equivalent to 18 mg Al³⁺/kg bw/d).

The LOAEL for local (stomach) toxicity in males is 1000 mg/kg bw/d (equivalent to 90 mg Al³⁺/kg bw/d).

The NOAEL for systemic toxicity in males is 1000 mg/kg bw/d (equivalent to 90 mg/kg bw/d Al³⁺/kg bw/d).

The NOAEL for local and systemic toxicity in females is 1000 mg/kg bw/d (equivalent to 90 mg/kg bw/d Al³⁺).