Aluminium sulphate

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 September 2006 - 03 November 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was conducted according to OECD guideline 422 and under GLP conditions.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 September 2006 - 03 November 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study was conducted according to OECD guideline 422 and under GLP conditions, performed on an analogue substance

Reason / purpose for cross-reference:

reference to same study

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

Specific details on test material used for the study:

- Stability under test conditions: At least 48 hours

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:
Males: mean 266-269 g, SD 6.6-12.2
Females: mean 237-240, SD 7.0-10.7
- Housing: According to guideline
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 (guideline: max. 70%, but no effects expected)
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

other: Not relevant

Vehicle:

other: Water (Milli-U)

Details on 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

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: max. 14 days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: Individually
- Any other deviations from standard protocol: Not relevant

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 (14 days prior to and 14 days during mating)
Females: 28 days (14 days prior to and 14 days during mating) + 9-25 days (during gestation and lactation)

Frequency of treatment:

Once daily, 7 days a week

Details on study schedule:

Not relevant

Dose / conc.:

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

Remarks:

3.6 mg Al3+/kg bw/day

Dose / conc.:

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

Remarks:

18 mg Al 3+/kg bw/day

Dose / conc.:

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

Remarks:

90 mg Al 3+/kg bw/day

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): No data

Positive control:

Not relevant

Parental animals: Observations and examinations:

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 study start, once weekly and at death (females: gestation days 0, 4,7, 11, 17, 20, lactation day 1, 4)

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes, weekly

WATER CONSUMPTION: Subjective appraisal but not recorded

OTHER:
FUNCTIONAL OBSERVATIONS:
Males during week 4, females during lactation; 5 M and5F, randomly selected from all groups:
Hearing ability, pupillary reflex, static righting reflex, grip strength, motor activity test (recorded 12 hrs overnight)

REPRODUCTIVE BEHAVIOUR:
Male number paired with, mating data, confirmation of pregnancy, delivery day

Oestrous cyclicity (parental animals):

Not performed

Sperm parameters (parental animals):

No data

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: Not applicable

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
numbers of live and dead pups( daily)
individual bw on day 1 and 4 of lactation
sex of all pups on day 1 and day 4 by assessment of ano-genital distance
physical or behavioural abnormalities daily

GROSS EXAMINATION OF DEAD PUPS:
Yes, sexed and externally examined if practically possible. The stomach was examined for the presence of milk.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals on day 29 of study
- Maternal animals: All surviving animals on day 4 post partum

GROSS NECROPSY
- Gross necropsy consisted of macroscopic examination of all organs of all animals.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [1] (see "Any other information on materials and methods incl. tables) were prepared for microscopic examination and weighed, respectively.

Postmortem examinations (offspring):

SACRIFICE
The F1 offspring was sacrificed at 4 days of age.

GROSS NECROPSY
All offspring sexed and externally examined. Stomach examined for the presence of milk
All macroscopic abnormalities recorded, and defects or cause of death evaluated. Any abnormal pup preserved in 10% buffered formaldehyde

Statistics:

Dunnet -Test (Dunnet, 1955) based on pooled variable estimate for variables assumed to have a normal distribution
Sett-test (Miller, 1981) for variables asssumed not to follow a normal distribution
Student’s T- test for pup organ weights
Fisher exact test for frequency data. All tests two-sided, P<0.05

Reproductive indices:

Percentage mating
Fertility index
Conception rate
Gestation index

Offspring viability indices:

Percentage live male & female pups
Percentage post-natal loss
Viability index

Clinical signs:

no effects observed

Description (incidence and severity):

Observations: Mortality / viability at least twice daily Detailed clinical signs at least once daily Body weights at start, once weekly and at death (females: gestation days 0, 4,7, 11, 17, 20, lactation day 1, 4 food consumtion weekly water consumption

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

See RS on 28-day repeated dose part of study

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

See RS on 28-day repeated dose part of study

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No histopatological abnormalities were obsurved

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Test substance intake: Dose volume 5 ml/kg bw. 0, 40, 200, 1000 mg/kg/day of the test substance solution, equal to 0, 3.6, 18, 90 mg/kg bw/ d of Al3+

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

effects observed, treatment-related

Description (incidence and severity):

M/F ratio : 1/1 per cage Maximum of 14 days, separation once mating occurred (i.e evidence of sperm in vaginal lavage or intravaginal copulatory plug) Proof of pregnancy: littering and post-mortem examination of reproductive organs

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

% pregnant per dose level: see under details on results

For Food consumption , Clinical signs, Haematological findings and duration, Clinical Biochemistry findings, Gross pathology : See RS on 28-day repeated dose part of study


Body weight (parenteral animals)
Slightly lower mean bw for females at 1000 mg/kg bw/d (90 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

Parameters assessment during study (maternal and fetal)
Clinical signs as above
Percentage mating
Fertility index
Conception rate
Gestation index
Percentage live male & female pups
Percentage post-natal loss
Viability index

% pregnant per dose level:
Dose (mg/kg bw/d) 0 40 200 1000
% pregnant 100 90 90 90

Number aborting:
None

Number of resorptions, early/late if available:
Number of implantations:
Pre and post implantation loss, if available; Number of corpora lutea
Corpora lutea:
no significant differences between exposed and controls
Mean (SD):
Dose (mg/kg bw/d) 0 40 200 1000
Corpora lutea 18.5 (2.01) 18.2 (7.94) 15.0 (5.40) 16.9 (1.91)
Implantation sites:
no significant differences between exposed and controls
Mean (SD):
Dose (mg/kg bw/d) 0 40 200 1000
Inplantation sitea 16.6 (2.55) 15.7 (5.72) 14.0 (5.35); 16.2 (1.99)
Litter size:
no significant differences between exposed and controls
Mean (SD):
Dose (mg/kg bw/d) 0 40 200 1000
Litter size 15.1 (3.1) 14.0 (5.1) 14.7
(2.7) 14.8
(1.9)

Duration of Pregnancy:
No difference between dose groups and controls
Mean (SD):
Dose (mg/kg bw/d) 0 40 200 1000
MF ratio 21.9 (0.3) 21.9
(0.6) 21.9
(0.3) 21.8
(0.4)

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No abnormalities at any dose level

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

act. ingr.

Remarks:

Aluminium (Al 3+)

Sex:

female

Basis for effect level:

other: No abnormalities at any dose level

Clinical signs:

not specified

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

No difference between exposed and controls Alive / dead (nrs per group): Dose (mg/kg bw/d) 0 40 200 1000 Live /dead 136/0 126/12 132/1 148/0

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Litter size: no significant differences between exposed and controls Mean (SD): Dose (mg/kg bw/d) 0 40 200 1000 Litter size 15.1 (3.1) 14.0 (5.1) 14.7 (2.7) 14.8 (1.9) Litter weights: No difference between exposed and controls Lactation day 1; M

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Description (incidence and severity):

Grossly visible abnormalities, external, soft tissue and skeletal abnormalities : No abnormalities observed either in controls or in exposed animals ouside the normal biological variation for race/strain No behavioural abnormalities in pups observed

Histopathological findings:

not examined

Sex ratio
No difference between exposed and controls
M / F ratio:
Dose (mg/kg bw/d) 0 40 200 1000
MF ratio 1.1 0.84 0.99 0.92


Postnatal growth
No difference between exposed and controls
Postnatal weight, Lactation day 4; Mean (SD):
Dose (mg/kg bw/d) 0 40 200 1000
Weight (gr) 10.5 (1.5) 10.3 (0.9) 10.5 (1.4) 10.4 (0.6)

Postnatal survival
No difference between exposed and controls
Post-natal pup deaths:
Dose (mg/kg bw /d 0 40 200 1000
Nr. Of dead pups 1 2 0 3

Organs examined at necropsy (macroscopic and microscopic)
Pups:
All offspring sexed and externally examined. Stomach examined for the presence of milk
All macroscopic abnormalities recorded, and defects or cause of death evaluated. Any abnormal pup preserved in 10% buffered formaldehyde

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: No abnormalities at any dose level

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Based on:

act. ingr.

Remarks:

Aluminium (Al 3+)

Sex:

male/female

Basis for effect level:

other: No abnormalities at any dose level

Reproductive effects observed:

not specified

1 female of the control group sacrified in extremis at day 22 of pregnancy due to parturition difficulties

Conclusions:

This study revealed no maternal toxicity at any dose and no reproductive, breeding or developmental toxicity at any dose from two weeks prior to mating to at least 3 days of lactation (females). Therefore, a NOAEL for maternal local and systemic toxicity of 1000 mg/kg bw/d (equivalent to 90 mg/kg/d Al3+) was established. For reproductive and developmental toxicity the NOAEL was 1000 mg/kg bw /d (equivalent to 90 mg/kg/d Al3+).

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 40, 200 or 1000 mg /kg bw/day (equivalent to 3.6, 18 or 90 mg/kg bw/day Al3 +). 10 animals/sex/group were used. Males were exposed for 28 days, females between 37 -53 days, i.e. during 2 weeks prior to mating, during mating, during postcoitum, and during at least 3 days of lactation. Litter was not exposed.

There were no treatment-related effects found regarding to mortality, clinical signs, functional observations, histopathology, organ weigths, reproduction, breeding data and pup development. No reproduction, breeding and developmental toxicity was observed for treatment up to 1000 mg/kg bw/day of aluminium chloride basic (equivalent to 90 mg/kg bw/day Al3+).

Based on these results, the No Observed Adverse Effect Level for maternal and developmental toxicity was established to be 1000 mg/kg bw/day for Aluminium chloride basic (equivalent to 90 mg/kg bw/day Al3+)

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 maternal toxicity at any dose and no reproductive, breeding or developmental toxicity at any dose.

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

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

The NOAEL for reproductive and developmental toxicity is 1000 mg/kg bw /d (equivalent to 90 mg/kg/d Al³⁺)

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 applicable.

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

Molecular formula:

General formula: Al(OH)x(Cl)(3-x), with x ranging from > 0 to 2.3 and typically being > 0.5.

IUPAC Name:

Aluminum Chloride, basic

Test material form:

solid - liquid: aqueous solution

Details on test material:

- Name of test material (as cited in study report): Aluminium chloride basic
- Molecular formula: Al(OH)1.3Cl1.7 (in aqueous solution)
- Molecular weight: 109.36
- CAS Number: 1327-41-9
- Substance type: Aqueous solution
- Physical state: Yellow liquid
- Storage condition of test material: At room temperature in the dark

Specific details on test material used for the study:

- Stability under test conditions: At least 48 hours

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
- 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

Dose / conc.:

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

Remarks:

= 3.6 mg/kg bw/day Aluminium

Doses / concentrations 2

Dose / conc.:

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

Remarks:

= 18 mg/kg bw/day Aluminium

Doses / concentrations 3

Dose / conc.:

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

Remarks:

= 90 mg/kg bw/day Aluminium

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): No 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

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:

Al 3+

Sex:

male

Basis for effect level:

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

4

Dose descriptor:

LOAEL

Effect level:

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

Based on:

act. ingr.

Remarks:

Al 3+

Sex:

male

Basis for effect level:

other: Local effects (stomach) at 90 mg Al/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:

Al 3+

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:

Al 3+

Sex:

female

Basis for effect level:

other: Local and systemic (HISTOPATHOLOGY EFFECTS at 90 mg Al/kg/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 aluminium chloride (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/d (equivalent to 3.6, 18 and 90 mg/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/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 control.

Mean food consumption of females at 1000 mg/kg bw/day (equivalent to 90 mg/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 (equivalent to 90 mg/kg bw/d)

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

(equivalent to 90 mg/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 & 200 mg/kg bw/d (equivalent to 3.6 and 18 mg/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/day (equivalent to 90 mg/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/kg/d Al 3 +).

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

(equivalent to 90 mg/kg/d Al 3 +).

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/kg/d Al 3 +). 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/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⁺⁾

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

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

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