Aluminium tributanolate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Upon contact with water or moisture (e.g. within mucous membranes) aluminium tributanolate hydrolyses immediately to butan-1ol and aluminium 3+ cations (as hydroxide and oxyhydroxide). Hence, toxicity is determined by the toxicity of these two species

Butanol

Two concentrations of 1-butanol (3000 and 6000 ppm) were administered by inhalation to separate groups of 15 pregnant Sprague-Dawley rats for 7 hr per day throughout gestation; 18 male rats were similarly exposed for 7 hr per day for 6 weeks, and mated to unexposed females (Nelson 1989). Litters were culled to 4 female and 4 male pups and fostered to untreated controls. From days 10-90, offspring were tested as follows: a) ascent on a wire mesh screen, b) rotorod, c) open field and photoelectrically-monitored activity, d) running wheel, e) avoidance conditioning, and f) operant conditioning. Additionally, brains from 10 offspring at 21 days of age were dissected into cerebrum, cerebellum, brainstem, and midbrain. Each sample was assayed for protein and the neurotransmitters acetylcholine, dopamine, norepinephrine, serotonin, met-enkephalin, beta-endorphin, and substance P. Overall, there were few behavioral or neurochemical alterations detected in the offspring following maternal or paternal exposure to either 3000 or 6000 ppm 1-butanol. No effects on maternal and parental animals became apparent. Therefore a NOAEC of 6000 ppm (18.5 mg/L) can be derived from this study.

11-17 female rats were given aqueous solutions (in drinking water) of n-butanol containing 0.24; 0.8 and 4% n-butanol (0.3; 1.0 and 5.0 g/kg bw/day) during 8 weeks before mating, maximum 3 weeks during mating and until gestation day 20. During the first 8 weeks oestrus cycle was assessed and thereafter females were mated with untreated males. The day of detection of spermatozoa in the vaginal smears was assumed to be day 0 of gestation. An assessment of teratogenicity, embryo- and fetotoxicity was performed (Sitarek 1994).
No treatment related effects were found on mortality, clinical signs, body weight (gain), food consumption, haeamatocrit and Hb, organ weights and oestrus cycle. Pregnancy rate, number of corpora lutea, resorptions, pre-and post-implantation loss and placental weights were unaffected by treatment.
No treatment related effects on number of live pups, pup body weight. At 5000 mg/kg bw the crown-rump length was decreased. Developmental effects were reported in all 3 dose groups. Limited skeletal effects were reported without a relation with dose (extra 14th rib and wavy ribs) CNS defects included dilation of either the subarachnoid space or lateral and/or third ventricles of the brain, or external or internal hydrocephalus. Based on the absence of a dose response, it appears probable that the observations in the low dose group were not treatment related.

Aluminium

Swiss Webster mice received diets containing 7 (control), 100, 500, or 1000 microg aluminum (Al)/g (<1, 10, 50 and 100 mg/kg bw as Al) throughout development (conception to 35 days of age) and were tested behaviorally as adults (>90 days of age) (Golub 2001).

The basal diet contained the same percent of recommended dietary amounts of phosphate, calcium, iron, magnesium, and zinc as young women usually consume. These "realistic" dietary conditions led to 12--15% growth retardation at 100 mg/kg bw at the time of testing (also observed during lactation at 50 mg/kg bw). No additional general toxic effects on dams and studied gestational parameters (number of animals completing pregnancy, gestation length, and litter size at birth) were reported.
Female offspring was evaluated in a cognitive task (Morris water maze) at 3 months of age and males were evaluated in a motor test battery at 5 months of age. At 100 mg/kg bw decreased learning abilities in a Morris maze (low latency) was seen in females and males at this dose showed significantly lower hindlimb grip strength than controls. Subtle influences of Al on rotarod and wire suspension tests were also noted. The data suggest that developmental Al exposure under normal, but less than optimal, dietary conditions can lead to subtle but long-term effects on growth and brain function in adulthood. A NOAEL of 10 mg/kg bw can be derived from this study.

In a two-generation reproductive toxicity study, male and female rats were given aluminium sulfate (AS) in drinking water at 0, 120, 600 or 3000 ppm (Hirata Kozumi) 2011). AS reduced water consumption in all treatment groups, and body weight was transiently decreased in the 3000 ppm group. In the F1 and F2 pups, preweaning body weight gain was inhibited at 3000 ppm, and the liver and spleen weight was decreased at weaning. At this dose, vaginal opening was slightly delayed. There were no compound-related changes in other reproductive/developmental parameters, including developmental neurobehavioral endpoints. The data indicated that the NOAEL of AS in this two-generation study is 600 ppm for parental systemic toxicity and reproductive/developmental toxicity. The total ingested dose of aluminium from drinking water and food (standard rat diet, containing 25-29 ppm of aluminium) combined for this 600 ppm group was calculated to be 8.06 mg Al/kg bw/day.

 

Link to relevant study records

Reference

Reference 1

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2008-2009

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

according to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

GLP compliance:

yes

Remarks:

The publication describes a study performed under GLP at the Safety Research Institute Kanto Chemical Compounds Co. Ltd, Sapporo Japan

Limit test:

no

Specific details on test material used for the study:

lot: 007X1828
purity: 98.5%
storange of the sample: cool and dark in sealed container

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Atsugu Breeding Center, Charles River Laboratories Japan Inc. Yokohama, Japan
- Females: no data
- Age at study initiation: (P) 5 weeks
- Weight at study initiation: (P) Males: ca 180 g; Females: ca 120 g; (F1) Males: 80 g; Females: 70 g Values taken from figures in the publication)
- Fasting period before study: no
- Housing:
- Diet standard rat diet (CRF-1 Oriental Yeast Co. Ltd, Tokyo, Japan containing 25-29 ug/g Al) ad libitum
- Water: ionexchanged water containing the substance ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-25 °C
- Humidity (%): 35-59 %
- Air changes (per hr): 10-15/h
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: drinking water

Details on exposure:

WATER PREPARATION
- Rate of preparation of diet (frequency): every 6 days
- Storage temperature of water: cooled
- FRequency based on stability for 4 days at room temperature and 6 days refrigerated (tested at 0.1, 0.6 and 15 mg/mL)

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 2 weeks
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
- After 2 weeks
- Further matings after two unsuccessful attempts: not indicated

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

In first and last preparation and every three months by HPLC (97.5-106.3% of nominal)
Drinking water was below the LOQ (0.5 ug/mL)

Duration of treatment / exposure:

P: 10 weeks and during mating gestation and lactation (parental males were necropsied at the same time as females)
F1: after weaning until mating and during mating gestation and lactation
Necropsy for non-selected F1 and F2 pups on day 26 after weaning

Frequency of treatment:

continuously

Details on study schedule:

- F1 parental animals not mated until unknown] weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were 21-25 days of age.

Dose / conc.:

0.12 mg/L drinking water

Dose / conc.:

0.6 mg/L drinking water

Dose / conc.:

3 mg/L drinking water

No. of animals per sex per dose:

24 males and 24 females for mating in P and F1 generation

Control animals:

other: ion exchanged drinking water

Details on study design:

- Dose selection rationale: based on a pre-test during 6-8 weeks, mating gestation and until day 4 post partum (at 1, 3, 10 and 30 mg/L in drinking water, NOAEL 3 mg/L)

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: weekly (females during gestation on day 0, 7, 14 and 20; during lactation on day 0, 7, 14 and 21)

FOOD CONSUMPTION : Yes
- Time schedule for examinations:weekly (females during gestation on day 0, 7, 14 and 20; during lactation on day 0, 7, 14 and 21)

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: twice weekly females during gestation on day 0,4, 7, 11, 14, 17 and 20; during lactation on day 0, 4, 7, 11, 14, 17, 19 and 21)
- Calculation intake: based on mean values for body weight and water intake per group


OTHER:

Oestrous cyclicity (parental animals):

2 weeks pre-mating and during cohabitation (4-6 days oestrus cycle considered as normal)

Sperm parameters (parental animals):

Parameters examined in all adult male parental generations:
testis weight, epididymis weight, sperm count in testes, sperm count in epididymides, enumeration of cauda epididymal sperm reserve, sperm motility, sperm morphology

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes to 4 males and 4 females
excess pups were killed and necropsied (gross external and internal observations)

PARAMETERS EXAMINED
The following parameters were examined in F1 / F2 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, body weight, physical or behavioural abnormalities, pinna unfolding, anogenital distance (AGD)(
In one male + one female pup per litter:onset and completion of incisor eruption, eye opening, perpetual separation and vaginal opening

GROSS EXAMINATION OF DEAD PUPS: yes, for external and internal abnormalities

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: no

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: no

ASSESSMENT OF BEHAVIOURAL EFFECTS:
On day 5, 8 and 18 in one male + one female pup per litter: surface rightening reflex, negative geitaxis, mid-air rightening reflex
After 4 weeks in 10/sex/group: motoractivity and swim test (T-maze)

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals at the same time as the females]
- Maternal animals: All surviving animals depending on oestrus cycle status after weaning

GROSS NECROPSY
- Gross necropsy in all animals consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera; females counts of implementation sites; males testis and epididymides preservation:

HISTOPATHOLOGY / ORGAN WEIGHTS:
From all animals: weights of brain, pituitary, thyroids, thymus, liver, kidneys, spleen, adrenals, testes, epididymides,, seminal vessels, prostate, uterus, ovaries
From control and high dose (+ all females with abnormalities + non copulating animals + animals with abnormalities ate the sex organs): histopathology of testes, epididymides,, seminal vessels, coagulating gland, ventral prostate, uterus, ovaries and vagina
From 10 females from control and high dose group: count of premordial follicles in the right ovary

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at 26 days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations

HISTOPATHOLOGY / ORGAN WEIGTHS
Organ weight of for 1 male + 1 female F1/F2 weanling/litter: brain, thymus, liver, kidneys, spleen, adrenals, testes, epididymides,, seminal vessels, prostate, uterus, ovaries
Microscopic examination of 10 males + females in control and high dose group of spleen and liver

Statistics:

Bartlett's test, Dunnett's test, Kruskal-Wallis test, Mann-Whitney's U-test, Fisher exact test, Student t-test (f-test)

Reproductive indices:

Copulation Index; Fertility Index; Gestation Index; Delivery Index

Offspring viability indices:

Viability Index on post-natal day 0, 4 and 21

Clinical signs:

no effects observed

Mortality:

mortality observed, non-treatment-related

Description (incidence):

P0: 1 female at 0.6 mg/L (mass)

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Decreased body weight at 3 mg/L during week 2 and 3 in males = females

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Males: sign decreased at 0.6 and 3 mg/L during week 1; sign decreased during week 8 and 13-14 at 3 mg/L
Females: sign decreased at 3 mg/L during week 1; sign decreased during week 3 of lactation at 0.6 and 3 mg/L

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

effects observed, treatment-related

Description (incidence and severity):

significantly decreased in all dose groups (related to the acidity of the formulated water)

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No treatment related effects

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

No treatment related effects

Reproductive function: sperm measures:

effects observed, treatment-related

Description (incidence and severity):

at 3 mg/L sign decrease in cauda epidydimal sperm (control 268E06/cauda; treated 254E06/cauda)

Reproductive performance:

no effects observed

Description (incidence and severity):

Copulation Index (%)
Males: 91.7, 91.7, 100. 91.7 at 0, 0.12,0.6 and 3 mg/L
Females:95.8, 100, 100, 100 at 0, 0.12,0.6 and 3 mg/L
Fertility Index (%):
Males: 95.5, 90,9. 100, 95.5 at 0, 0.12,0.6 and 3 mg/L
Females: 95.7, 91.7, 100, 91.8 at 0, 0.12,0.6 and 3 mg/L

Gestation Index (%): 100, 95.5, 95.7, 95.7 at 0, 0.12,0.6 and 3 mg/L
Delivery Index (%): 94.3, 88.6, 90.7, 92.0 at 0, 0.12,0.6 and 3 mg/L

Dose descriptor:

NOAEL

Effect level:

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

Based on:

element

Sex:

male

Basis for effect level:

food consumption and compound intake

water consumption and compound intake

organ weights and organ / body weight ratios

reproductive function (sperm measures)

Dose descriptor:

NOAEL

Effect level:

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

Based on:

element

Sex:

female

Basis for effect level:

food consumption and compound intake

water consumption and compound intake

Remarks on result:

other: related to the acidity of the drinking water

Critical effects observed:

yes

Lowest effective dose / conc.:

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

System:

male reproductive system

Organ:

cauda epididymis

Treatment related:

no

Dose response relationship:

no

Relevant for humans:

no

Clinical signs:

no effects observed

Mortality:

mortality observed, non-treatment-related

Description (incidence):

F1: 1 male at 0.12 mg/L and 1 male at 3 mg/L

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No significant difference with controls
Females at 3 mg/L showed increased body weight during week 6-8

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Males: sign decreased during week 10 at 0.6 and 3 mg/L
Females: sign decreased during week 3 of lactation at 0. and 3 mg/L
Incidental increases were also reported

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

effects observed, treatment-related

Description (incidence and severity):

Males: sign decreased at 0.6 and 3 mg/L; sign decreased during week 3-6, 8 and 10 at 0.12 mg/L
Females: sign decreased at 3 mg/L; sign decreased during week 10 of dosing and 3 of lactation at 0.6 mg/L; sign decreased during week 9-10 at 0.12 mg/L
Effects related to the acidity of the formulations

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No difference with control animals

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No difference with control animals.

Gross pathological findings:

no effects observed

Description (incidence and severity):

No treatment related effects

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No treatment related effects

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

No treatment related effects

Reproductive function: sperm measures:

no effects observed

Description (incidence and severity):

No treatment related effects

Reproductive performance:

no effects observed

Description (incidence and severity):

Copulation Index (%)
Males: 95.8. 91.3, 95.8, 87.5 at 0, 0.12,0.6 and 3 mg/L
Females:100, 95.8, 100, 95.8 at 0, 0.12,0.6 and 3 mg/L
Fertility Index (%):
Males:91.3, 81.0, 91.3, 95.2 at 0, 0.12,0.6 and 3 mg/L
Females: 91.7, 82.6, 91.7, 91.3 at 0, 0.12,0.6 and 3 mg/L

Gestation Index (%): 100, 94.7, 100, 100 at 0, 0.12,0.6 and 3 mg/L
Delivery Index (%): 94.0, 87.5, 91.4, 94.6 at 0, 0.12,0.6 and 3 mg/L

Dose descriptor:

NOAEL

Effect level:

>= 9.78 - <= 14 mg/kg bw/day (actual dose received)

Based on:

element

Sex:

male/female

Basis for effect level:

food consumption and compound intake

water consumption and compound intake

Remarks on result:

other: No treatment related effects

Critical effects observed:

no

Clinical signs:

no effects observed

Description (incidence and severity):

see P1

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

see P1
Viability Index:
post natal day 0 (%): 100, 99.3, 99.7 and 99,5 at 0, 0.12, 0.6 and 3 mg/L
post natal day 4 (%): 98.7, 95.2, 98.8 and 98.0 at 0, 0.12, 0.6 and 3 mg/L
postnatal day 21 (%): 99.4, 100, 100 and 99.4 at 0, 0.12, 0.6 and 3 mg/L

Body weight and weight changes:

no effects observed

Description (incidence and severity):

see P1
post natal day 21: at 3 mg/L sign decreased body weight in both sexes

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

see P1

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

effects observed, treatment-related

Description (incidence and severity):

see P1

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

see P1
sign decreased abs thymus and spleen weight in both sexes at 3 mg/L
sign decreased abs kidney, testes and epididymus weight in males at 3 mg/L
sign decreased abs uterus weight in females at 0.6 and 3 mg/L
sign increased rel brain weight in both sexes at 3 mg/L

Gross pathological findings:

no effects observed

Description (incidence and severity):

see P1
No treatment related effects

Histopathological findings:

no effects observed

Description (incidence and severity):

see P1
No treatment elated effects in liver and spleen

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Delayed vaginal openeing in females at 3 mg/L
No other treatment related developmental effects

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No treatment related behavioural effects (no dose response)

Developmental immunotoxicity:

not examined

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Based on:

element

Sex:

female

Basis for effect level:

body weight and weight gain

organ weights and organ / body weight ratios

other: developmental toxicity (vaginal opening delayed)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

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

Based on:

element

Sex:

male

Basis for effect level:

body weight and weight gain

organ weights and organ / body weight ratios

Clinical signs:

not specified

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

Viability Index:
post natal day 0 (%): 99.7, 99.6, 99.4 and 99.7 at 0, 0.12, 0.6 and 3 mg/L
post natal day 4 (%): 94.7, 98.1, 99.1and 99.0 at 0, 0.12, 0.6 and 3 mg/L
postnatal day 21 (%): 100, 98.6, 100 and 100 at 0, 0.12, 0.6 and 3 mg/L

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

post natal day 21: at 3 mg/L sign decreased body weight in females

Food consumption and compound intake (if feeding study):

not specified

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

sign decreased abs spleen, abs thymus and rel brain weight in males at 3 mg/L
sign decreased abs liver and epididymus weight in males at 3 mg/L
sign decreased abs spleen, ovary and uterus weight in females at 3 mg/L
sign decreased abs/rel liver weight in females at 3 mg/L
sign increased rel brain weight in females at 3 mg/L

Gross pathological findings:

no effects observed

Description (incidence and severity):

No treatment related effects

Histopathological findings:

no effects observed

Description (incidence and severity):

No treatment related effects in liver and spleen

Other effects:

no effects observed

Description (incidence and severity):

No treatment related developmental effects

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No treatment related behavioural effects

Developmental immunotoxicity:

not examined

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

>= 9.78 - <= 14 mg/kg bw/day (actual dose received)

Based on:

element

Sex:

male/female

Basis for effect level:

body weight and weight gain

organ weights and organ / body weight ratios

Reproductive effects observed:

yes

Lowest effective dose / conc.:

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

Treatment related:

yes

Relation to other toxic effects:

reproductive effects as a secondary non-specific consequence of other toxic effects

Dose response relationship:

yes

Relevant for humans:

yes

Conclusions:

In parental animals the most prominent effect of the substance was the decreased water intake (most likely due to the acidity of the formulation). THis has lead to effects on food intake and body weight at 3 mg/L. The NOAEL for parental toxicity is therefore set at 8.06 mg/kg bw in a worst case approch
Effects on sperm parameters were seen in parental males (P0) at 3 mg/L. No effects on reproduction were seen in the P0 and P1 generation. The NOAEL for reproduction toxicity is set at 8.06 mg/kg bw in a worst case approch
In the offspring (F1 and F2) effects were limited to a lowered body weight and some changes in organ weights at 3 mg/L. Delayed valginal opening was observed in F1 females at 3 mg/L. The NOAEL for developmental effects is therefore set at 8.06 mg/kg bw.

Executive summary:

In a two-generation reproductive toxicity study, male and female rats were given aluminium sulfate (AS) in drinking water at 0, 120, 600 or 3000 ppm. AS reduced water consumption in all treatment groups, and body weight was transiently decreased in the 3000 ppm group. In the F1 and F2 pups, preweaning body weight gain was inhibited at 3000 ppm, and the liver and spleen weight was decreased at weaning. At this dose, vaginal opening was slightly delayed. There were no compound-related changes in other reproductive/developmental parameters, including developmental neurobehavioral endpoints. The data indicated that the NOAEL of AS in this two-generation study is 600 ppm for parental systemic toxicity and reproductive/developmental toxicity. The total ingested dose of aluminium from drinking water and food (standard rat diet, containing 25-29 ppm of aluminium) combined for this 600 ppm group was calculated to be 8.06 mg Al/kg bw/day.

Reference 2

Endpoint:

fertility, other

Remarks:

non-guideline study

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

Males were exposed only 6 weeks prior to mating compared to the 10 weeks recommended by the guidelines 415 and 416 but sufficiently long enough if compared to the screening test (guideline 421) which recommends only 2 weeks of exposure prior to mating. Older male rats were used as can be extrapolated from the initial body weights whereas younger rats are recommended in the guidelines.

Qualifier:

no guideline followed

Principles of method if other than guideline:

Two concentrations of butan-1-ol (3000 and 6000 ppm) were administered by inhalation to separate groups of 15 pregnant Sprague-Dawley rats for 7 hr per day throughout gestation ; 18 male rats were similarly exposed for 7 hr per day for 6 weeks, and mated to unexposed females.

GLP compliance:

not specified

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: (P) Males: mean: 429-512 g; Females: mean: no data


ENVIRONMENTAL CONDITIONS
no data

Route of administration:

inhalation: vapour

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: air

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentrations measured in the exposure chambers approximated the target concentrations of 3000 and 6000 ppm. Mean (±s.d.) 1-butanol concentrations were 3010 (±50) and 6000 (±80) ppm, and results of periodic confirmatory charcoal tube samples were 3000 (± 90) and 5960 (± 110) ppm, respectively .

Duration of treatment / exposure:

females: Exposure period: day 1 - 20 of gestation
males: 6 weeks before mating

Frequency of treatment:

7 h/d

Dose / conc.:

3 000 ppm

Remarks:

nominal

Dose / conc.:

6 000 ppm

Remarks:

nominal

No. of animals per sex per dose:

15 females per dose
18 males per dose

Control animals:

yes

Statistics:

Data were analyzed using multivariate analysis of variance (MANOVA) on tests with multiple dependent measures, followed by analysis of variance (ANOVA) on each dependent variable if a significant MANOVA was observed. If only one dependent variable was obtained, ANOVA was used, followed by the Tukey Range Test to determine which cell means differed from one another.

Dose descriptor:

NOAEL

Effect level:

18.5 mg/L air

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effects observed

Critical effects observed:

no

No significant effects on ascent on a wire mesh screen, rotorod, open-field and operant conditioning. A significant decrease of photoelectrically-monitored activity was observed in 3000 ppm female offspring of exposed males. In avoidance conditiong the reaction was increased from day 34 until day 60 in offspring of males exposed at 6000 ppm. In total 4 of the 78 tests run gave effects that were significantly different from controls

Neurochemical analyses gave effects in 4 out of 62 measurements without a relation with exposure of parental animals.

Dose descriptor:

other: NOAEL developmental neurotoxicity

Generation:

F1

Effect level:

18.5 mg/L air

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effects observed

Critical effects observed:

no

Reproductive effects observed:

no

No general maternal or paternal toxicity was reported. Paternal exposure had no detectable effect on pregnancy rate in non-exposed females; maternal exposure was also without detectable effect on pregnancy rate. Although the study was designed to provide animals for postnatal assessment of developmental neurotoxicity, the lack of effect on pregnancy rate following either maternal or paternal exposure suggested that butan-1 -ol had no effect on fertility up to 6000 ppm. In the 6000 ppm group, 4 of the 78 (5%) behavioural measures, and 4 of the 64 (6%) neurochemical measures differed from those of controls.

There was no discernible pattern of effects.

Conclusions:

No effects were found at 18.5 mg/L related to parental toxicity or behavioural effects in their off-spring

Executive summary:

Two concentrations of 1-butanol (3000 and 6000 ppm) were administered by inhalation to separate groups of 15 pregnant Sprague-Dawley rats for 7 hr per day throughout gestation; 18 male rats were similarly exposed for 7 hr per day for 6 weeks, and mated to unexposed females. Litters were culled to 4 female and 4 male pups and fostered to untreated controls. From days 10-90, offspring were tested as follows: a) ascent on a wire mesh screen, b) rotorod, c) open field and photoelectrically-monitored activity, d) running wheel, e) avoidance conditioning, and f) operant conditioning. Additionally, brains from 10 offspring at 21 days of age were dissected into cerebrum, cerebellum, brainstem, and midbrain. Each sample was assayed for protein and the neurotransmitters acetylcholine, dopamine, norepinephrine, serotonin, met-enkephalin, beta-endorphin, and substance P. Overall, there were few behavioral or neurochemical alterations detected in the offspring following maternal or paternal exposure to either 3000 or 6000 ppm 1-butanol. No effects on maternal and parental animals became apparent. Therefore a NOAEC of 6000 ppm (18.5 mg/L) can be derived from this study.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

8.04 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

two generation study on Aluminium sulfate in drinking water

Effect on fertility: via inhalation route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

18 500 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

study on butan-1-ol the most volatile hydrolysis product

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Description of key information

Upon contact with water or moisture (e.g. within mucous membranes) aluminium tributanolate hydrolyses immediately to butan-1ol and aluminium 3+ cations (as hydroxide and oxyhydroxide). Hence, toxicity is determined by the toxicity of these two species

Butanol

Pregnant rats were given drinking water containing 1-butanol at 0.2%, 1.0% or 5.0% (316, 1454 or 5654 mg/kg/day) on days 0–20 of pregnancy (Ema 2005). The rats were sacrificed on day 20 of pregnancy and both the dams and fetuses were examined. No effects in maternal animals on mortality or pregnancy rates. The body weight gain was significantly reduced at 5.0%. Food consumption on days 0–7, days 7–14, days 14–20 and days 0–20 of pregnancy was significantly lower in the 1.0% and 5.0% groups than the control group. The water consumption on days 0–7 at 1.0 and 5.0% and on days 7–14, days 14–20 and days 0–20 at 5.0% was significantly decreased. No significant increase in the incidence of pre- and post-implantation embryonic loss was observed in any groups treated with 1-butanol. Fetal weight was significantly lowered at 5.0%. Although a significant increase in the incidence of fetuses with skeletal variations and decreased degree of ossification was found at 5.0%, no increase in the incidence of fetuses with external, skeletal and internal abnormalities was detected in any groups treated with 1-butanol. The data demonstrate that 1-butanol is developmental toxic only at maternal toxic doses. No evidence for teratogenicity of 1-butanol was noted in rats. Based on the significant decreases in maternal body weight gain and fetal weight, it is concluded that the no observed adverse effect levels (NOAELs) of 1-butanol for both dams and fetuses are 1.0% (1454 mg/kg/day) in rats.

Groups of approximately 15 Sprague-Dawley rats were exposed at 8000, 6000, 3500, or 0 ppm 1-butanol for 7 hr/day on Gestation Days 1-19 (sperm = 0) (Nelson 1991, Nelson 1992). Dams were sacrificed on Gestation Day 20, and fetuses were individually weighed, tagged, and examined for external malformations. One-half of the fetuses were stained and examined for skeletal abnormalities, and the other half were examined for visceral defects using the Wilson technique. The highest and intermediate concentration was maternally toxic, as manifest by reduced weight gain, feed intake and narcosis (at 8000 ppm). At a maternally toxic dose a dose-dependent reduction in fetal weights was observed and the only teratogenicity observed was a slight increase in skeletal malformations (primarily rudimentary cervical ribs), seen with the highest concentration of 1-butanol. Thus, although teratogenicity was observed at 8000 ppm 1-butanol (24.7 mg/L), and developmental toxicity was observed at the intermediate dose level 6000 ppm, these effects were seen in presence of maternal toxicity. The NOAEL is therefore 3500 ppm (10.8 mg/L) for maternal and developmental effects

Aluminium

A group of pregnant Swiss mice was given by gavage daily doses of Al(OH)3 (166 mg/kg) on gestational days 6-15 (Colomina 1992). Caesarean sections were performed on gestation day 18, and live fetuses were sexed, weighed and examined for morphological defects. No maternal toxicity was observed. The reproductive data did not show embryotoxic effects. 

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Qualifier:

according to guideline

Guideline:

other: Ministry of Health and Welfare, Japan; Guidelines for Toxicity Studies of Drugs

Principles of method if other than guideline:

Pregnant rats were given drinking water containing 1-butanol at 0.2%, 1.0% or 5.0% (316, 1454 or 5654 mg/kg/day) on days 0–20 of pregnancy. The rats were sacrificed on day 20 of pregnancy and both the dams and fetuses were examined.

GLP compliance:

yes

Species:

rat

Strain:

Crj: CD(SD)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Tsukuba Breeding Center
- Age at study initiation: males: 10 weeks; females: 9 weeks
- Weight at study initiation: females: 217-273 g
- Housing: individually
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 7 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-25 °C
- Humidity (%): 40-70 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: drinking water

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
1-butanol was mixed with water to the according target concentrations. The stability of formulations in a dark and cool place under airtight conditions has been confirmed for up to 3 days. During use, the formulations were maintained under such conditions for no more than 3 days and were 95.7–103.5% of the target concentration.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The concentrations were determined analytically and were 95.7–103.5% of the target concentration.

Details on mating procedure:

- Impregnation procedure: [cohoused]
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: over night
- Verification of same strain and source of both sexes: [yes]
- Proof of pregnancy: [sperm in vaginal smear] referred to as [day 0] of pregnancy

Duration of treatment / exposure:

day 0 through day 20 of pregnancy

Frequency of treatment:

continuous through drinking water

Duration of test:

until day 20 of pregnancy

Dose / conc.:

0.2 other: % in drinking water

Remarks:

316 mg/kg bw

Dose / conc.:

1 other: % in drinking water

Remarks:

1454 mg/kg bw

Dose / conc.:

5 other: % in drinking water

Remarks:

5654 mg/kg bw

No. of animals per sex per dose:

20 females per dose

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The dosage levels were determined based on the results of our range-finding study in which administration of 1-butanol in the drinking water on days 0–20 of pregnancy caused decreases in maternal body weight gain and food and water consumption and tended to reduce in fetal weight at 4% and 7% in rats.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: not specified



DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: not not specified


BODY WEIGHT: Yes
- Time schedule for examinations: once daily


FOOD CONSUMPTION: Yes
- Time schedule for examinations: every 3 or 4 days


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 10
- Organs examined: not specified

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- placental weight

Fetal examinations:

- External examinations: Yes: [all per litter] including fetal weight and crown-rumplength
- Soft tissue examinations: Yes: [half per litter]
- Skeletal examinations: Yes: [half per litter]
- Head examinations: Yes: [half per litter]

Statistics:

The statistical analysis of fetuses was carried out using the litter as the experimental unit. The initial body weight, body weight gain and food and water consumption of pregnant rats, numbers of corpora lutea, implantations and live fetuses per litter, fetal weight and crown-rump length and placental weight were analyzed with Bartlett's test for homogeneity of variance at the 5% level of significance.
If it was homogeneous, the data were analyzed using Dunnett's multiple comparison test to compare the mean of the control group with that of each dosage group, and if it was not homogeneous, the mean rank of the 1-butanol-treated groups was compared with that of the control group with the Dunnett type test. The Dunnett type test was used for the incidences of pre- and postimplantation embryonic loss and fetal anomalies and sex ratio of fetuses to compare the mean rank of groups treated with 1-butanol and that of the control group. The incidence of dams with anomalous fetuses was analyzed by Chi-square test or Fisher's exact test. The significance of differences from the control group was estimated at probability levels of 1% and 5%.

Indices:

no data

Historical control data:

International Genetic Standard (Crj: CD (SD) IGS) rats were used throughout this study. This strain was chosen because it is most commonly used in reproductive and developmental toxicity studies and historical control data are available.

Clinical signs:

not specified

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

sign decreased at 5% during day 0-7 and over the whole pregnancy

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

sign decreased at 2 and 5 % over the whole pregnancy (DR)

Food efficiency:

not examined

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

effects observed, treatment-related

Description (incidence and severity):

sign decreased at 5% over the whole pregnancy, sign decreased during day 0-7 at 2%

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

no effects observed

Early or late resorptions:

no effects observed

Dead fetuses:

no effects observed

Changes in pregnancy duration:

not examined

Changes in number of pregnant:

no effects observed

Other effects:

no effects observed

Description (incidence and severity):

placental weights were similar for all treatment groups and controls

Details on maternal toxic effects:

Details on maternal toxic effects:
No death was found in female rats of any group. All females in all groups became pregnant. The body weight gains on days 0–7 of pregnancy were significantly reduced at 5.0%. The body weight gain during the whole period of pregnancy was also significantly decreased at 5.0%. No significant decrease in the body weight gain was noted at 0.2 or 1.0, except for a transient decrease on days 0–2 of pregnancy at 1.0%. The food consumption on days 0–7, days 7–14, days 14–20 and days 0–20 of pregnancy was significantly lower in the 1.0% and 5.0% groups than the control group. The water consumption on days 0–7 at 1.0 and 5.0% and on days 7–14, days 14–20 and days 0–20 at 5.0% was significantly decreased. The mean daily intakes of 1-butanol were 316 mg/kg for the 0.2% group, 1454 mg/kg for the 1.0% group and 5654 mg/kg for the 5.0% group.

Dose descriptor:

NOAEL

Effect level:

1 454 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Fetal body weight changes:

effects observed, treatment-related

Description (incidence and severity):

signi decreased at 5%

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

effects observed, treatment-related

Description (incidence and severity):

the number of litters and litter size was unaffected, the litterweight was decreased (due to lower pup weights)

Changes in postnatal survival:

not examined

External malformations:

effects observed, non-treatment-related

Description (incidence and severity):

One fetus with spina bifida in the control group and one fetus with thread-like tail and anal atresia in the 0.2% group were observed.

Skeletal malformations:

no effects observed

Description (incidence and severity):

Skeletal examination revealed one fetus with supernumerary thoracic vertebral bodies and malpositioned thoracic vertebrae at 1.0%. Although the total number of fetuses with skeletal variations was significantly increased at 5.0%, the number of fetuses with individual skeletal variations was not significantly increased, except for fetuses with short supernumerary ribs at 5.0%. A significantly lower number of forepaw proximal phalanges was observed at 5.0%

Visceral malformations:

no effects observed

Description (incidence and severity):

Membranous ventricular septum defect occurred in one fetus of the control and 0.2% groups and 3 fetuses in 3 dams of the 5.0% group. One fetus with a double aorta in the control group and one fetus with a left umbilical artery in the control and 2.0% groups were observed. Thymic remnants in the neck were found in 4–11 fetuses of the control and groups treated with 1-butanol.

Details on embryotoxic / teratogenic effects:

Details on embryotoxic / teratogenic effects:
No litters totally resorbed were found in any group. No effects of the administration of 1-butanol were observed on the numbers of corpora lutea, implantations, pre- or postimplantation loss, resorptions or dead or live fetuses or sex ratio of live fetuses. The body weights of male and female fetuses were significantly lower in the 5.0% group than in the control group. There was no significant difference in the crown-rump length of male and female fetuses or placental weight between the control and groups treated with 1-butanol.
One fetus with spina bifida in the control group and one fetus with thread-like tail and anal atresia in the 0.2% group were observed. Skeletal examination revealed one fetus with supernumerary thoracic vertebral bodies and malpositioned thoracic vertebrae at 1.0%. Although the total number of fetuses with skeletal variations was significantly increased at 5.0%, the number of fetuses with individual skeletal variations was not significantly increased, except for fetuses with short supernumerary ribs at 5.0%. A significantly lower number of forepaw proximal phalanges was observed at 5.0%. Membranous ventricular septum defect occurred in one fetus of the control and 0.2% groups and 3 fetuses in 3 dams of the 5.0% group. One fetus with a double aorta in the control group and one fetus with a left umbilical artery in the control and 2.0% groups were observed. Thymic remnants in the neck were found in 4–11 fetuses of the control and groups treated with 1-butanol.
However, there was no significant difference in the incidence of fetuses with internal abnormalities between the control and groups treated with 1-butanol.

Dose descriptor:

NOAEL

Effect level:

5 654 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Dose descriptor:

NOAEL

Effect level:

1 454 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: fetotoxicity

Abnormalities:

not specified

Developmental effects observed:

yes

Lowest effective dose / conc.:

5 654 mg/kg bw/day (nominal)

Treatment related:

yes

Relation to maternal toxicity:

developmental effects as a secondary non-specific consequence of maternal toxicity effects

Dose response relationship:

yes

Relevant for humans:

not specified

Maternal findings in rats given 1-butanol on days 0–20 of pregnancy

Dose (%)	0 (Control)	0.2	1.0	5.0
No. of rats	20	20	20	20
No. of pregnant rats	20	20	20	20
No. of dead rats	0	0	0	0
Initial body weight	245 ± 14	247 ± 13	245 ± 11	244±12
Body weight gain during pregnancy (g)^a
Days 0–7	44 ± 7	45 ± 7	40 ± 6	20 ± 28**
Days 7–14	40±6	41±5	41±7	42±10
Days 14–20	78±14	82 ± 8	84 ± 7	75 ± 11
Days 0–20	162 ± 19	168 ± 16	165 ± 15	146 ± 16**
Food consumption during pregnancy (g)^a
Days 0–7	179 ± 12	180 ± 16	164 ± 12*	138 ± 21**
Days 7–14	193 ± 14	194 ± 17	177 ± 14**	160 ± 11**
Days 14–20	176 ± 14	175 ± 15	161 ± 12**	143 ± 11**
Days 0–20	548 ± 38	548 ± 46	503 ± 34**	441 ± 34**
Water consumption during pregnancy (ml)^a
Days 0–7	284 ± 28	305 ± 37	258 ± 29*	175 ± 34**
Days 7–14	318 ± 35	337 ± 48	299 ± 40	239 ± 80**
Days 14–20	328 ± 47	342 ± 47	334 ± 46	256 ± 85**
Days 0–20	930 ± 105	983 ± 126	890 ± 106	669 ± 182**
Mean daily intakes of 1-butanol (mg/kg)^a	0	316 ± 30	1454 ± 186	5654 ± 1402

** Significantly different from the control, *P< 0.05 and **P< 0.01.^aValues are given as the mean ± SD.

 

Reproductive findings in rats given 1-butanol on days 0–20 of pregnancy

Dose (%)	0 (Control)	0.2	1.0	5.0
No. of litters	20	20	20	20
No. of litters totally resorbed	0	0	0	0
No. of corpora lutea per litter^a	16.4 ± 3.6	16.7 ± 3.0^d	16.1 ± 2.1	16.3 ± 2.6
No. of implantations per litter^a	14.3 ± 2.8	15.1 ± 1.7	15.2 ± 1.2	14.7 ± 2.5
% Preimplantation loss per litter^b	9.0	9.0d	4.4	9.2
% Postimplantation loss per litter^c	6.0	5.4	3.7	8.0
No. of live fetuses per litter^a	13.4 ± 2.6	14.3 ± 1.4	14.7 ± 1.5	13.5 ± 2.5
Sex ratio of live fetuses (male/female)	128/139	145/140	149/144	131/139
Body weight of live fetuses (g)^a
Male	4.18±0.27	4.00±0.24	4.04±0.25	3.83±0.18**
Female	3.97 ± 0.25	3.86 ± 0.20	3.83 ± 0.16	3.59 ± 0.17**
Fetal crown-rump length (mm)^a
Male	40.5 ± 1.2	40.3 ± 1.4	40.2 ± 1.2	39.7 ± 1.3
Female	39.4 ± 1.2	39.4 ± 1.2	39.3 ± 1.1	38.5 ± 1.4
Placental weight (g)
Male	0.50 ± 0.05	0.49 ± 0.05	0.48 ± 0.06	0.50 ± 0.06
Female	0.49 ± 0.05	0.48 ± 0.05	0.47 ± 0.05	0.49 ± 0.06

** Significantly different from the control, *P< 0.01

^aValues are given as the mean ± SD.

^b(No. of preimplantation embryonic loss/no. of corpora lutea)x100.

c(No. of resorptions and dead fetuses/no. implantations)x100.

^dValue was obtained from 19 pregnant rats.

 

Morphological examinations in fetuses of rats given 1-butanol on days 0–20 of pregnancy

Dose (%)	0 (Control)	0.2	1.0	5.0
External examination
Total no. of fetuses (litters) examined	267(20)	285(20)	293(20)	270(20)
Total no. of fetuses (litters) with abnormalities	1 (1)	1 (1)	0	0
Spina bifida	1 (1)	0	0	0
Thread-like tail and anal atresia	0	1 (1)	0	0
Skeletal examination
Total no. of fetuses (litters) examined	139(20)	147(20)	152(20)	140(20)
Total no. of fetuses (litters) with abnormalities	0	0	1 (1)	0
Supernumerary of thoracic vertebral bodies and malpositioned thoracic vertebrae	0	0	1 (1)	0
Total no. of fetuses (litters) with variations	28(11)	23(12)	52(17)	69 (20)**
Bipartite ossification of thoracic centra	1 (1)	1 (1)	1 (1)	7(5)
Dumbbell ossification of thoracic centra	0	1 (1)	2(2)	3 (3)
Bipartite ossification of lumbar centra	0	0	0	2(2)
Supernumerary lumbar vertebrae	4(1)	1 (1)	5 (3)	5 (2)
Lumbarization	0	0	1 (1)	1 (1)
Bipartite ossification of sternebrae	1 (1)	1 (1)	1 (1)	1 (1)
Misaligned sternebrae	0	0	0	1 (1)
Cervical ribs	2(2)	3(3)	3 (3)	7(5)
Full supernumerary ribs	5(2)	1 (1)	10 (5)	9(5)
Short supernumerary ribs	20(10)	18 (9)	43(16)	55 (19)**
Wavy ribs	0	0	0	1 (1)
Degree of ossification^a
No. of sacral and caudal vertebrae	8.4 ± 0.5	8.4 ± 0.4	8.3 ± 0.5	8.1 ± 0.3
No. of sternebrae	5.9 ± 0.2	5.8 ± 0.2	5.8 ± 0.2	5.8 ± 0.2
No. of forepaw proximal phalanges	1.6 ± 1.3	1.6 ± 0.9	1.2 ± 1.1	0.3 ± 0.4**
Internal examination
Total no. of fetuses (litters) examined	128(20)	138(20)	141 (20)	130(20)
Total no. of fetuses (litters) with abnormalities	7(6)	9(6)	11 (8)	14(9)
Membranous ventricular septum defect	1 (1)	1 (1)	0	3 (3)
Double aorta	1(1)	0	0	0
Left umbilical artery	1 (1)	0	1 (1)	0
Thymic remnant in neck	4(4)	8 (5)	10 (8)	11 (8)

^aValues are given as the mean ± SD.

** Significantly different from the control, *P< 0.01

 

Conclusions:

Summary:
A significant decrease in maternal body weight gain accompanied by reduced food and water consumption was found at 5.0%. No significant increase in the incidence of pre- and postimplantation embryonic loss was observed in any groups treated with 1-butanol. Fetal weight was significantly lowered at 5.0%. Although a significant increase in the incidence of fetuses with skeletal variations and decreased degree of ossification was found at 5.0%, no increase in the incidence of fetuses with external, skeletal and internal abnormalities was detected in any groups treated with 1-butanol. The data demonstrate that 1-butanol is developmental toxic only at maternal toxic doses. No evidence for teratogenicity of 1-butanol was noted in rats. Based on the significant decreases in maternal body weight gain and fetal weight, it is concluded that the no observed adverse effect levels (NOAELs) of 1-butanol for both dams and fetuses are 1.0% (1454 mg/kg/day) in rats.

Reference 2

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

no

Limit test:

yes

Species:

mouse

Strain:

Swiss

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Interfauna Iberica, Barcelona, Spain
- Strain: CD-1
- Weight at study initiation: 28-32 g
- Fasting period before study: no data
- Housing: individually in solid-bottom polycarbonate cages
- Diet: Panlab rodent chow (Barcelona) ad libitum
- Wate: ad libitum
- Acclimation period: 7 days (prior to mating 1 male/2 females)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): 40-60%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light):12/12

Route of administration:

oral: gavage

Vehicle:

not specified

Remarks:

no information on vehicle

Analytical verification of doses or concentrations:

no

Details on mating procedure:

- M/F ratio per cage: 1/2
- Length of cohabitation: no data
- Proof of pregnancy: vaginal plug (designated as day 0 of gestation)

Duration of treatment / exposure:

day 6-15 of gestation

Frequency of treatment:

daily

Duration of test:

18 days (dams were sacrified at day 18)

Dose / conc.:

166 mg/kg bw/day (nominal)

Remarks:

as Al(OH)3

Dose / conc.:

57.5 mg/kg bw/day (nominal)

Remarks:

as Al

No. of animals per sex per dose:

11 females in treatment group and 13 in controls

Control animals:

other: water treated

Maternal examinations:

BODY WEIGHT: Yes on day 0, 6, 9, 12, 15 and 18

FOOD CONSUMPTION: Yes on day 0, 6, 9, 12, 15 and 18

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day18
- Organs examined: weight and luminium concentration of liver, bone (femur), brain and kidney

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: No
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

- External examinations: No data
- Soft tissue examinations: Yes: ca 1/3 --> control 54/192, treatment 40/130
- Skeletal examinations: Yes: remaining --> control 74/192, treatment 55/130
- Head examinations: No
Total Al/fetus in treatment group

Statistics:

ANOVA with Duncan's test, Kruskal-Wallis, chi-square test

Clinical signs:

not specified

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Body weight gain between treated and control animals did not differ significantly

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No differences between treated mice and controls

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

not specified

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No differences in utertus weight, abs/rel liver and abs/rel kidney weight

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Aluminium concentrations in liver, kidney and brain did not differ significantly.
Aluminium concentrations in bone were significantly increased

Number of abortions:

no effects observed

Description (incidence and severity):

none observed

Pre- and post-implantation loss:

no effects observed

Description (incidence and severity):

post-implementation loss: 4.45% and 6.29% in controls and treated mice

Total litter losses by resorption:

no effects observed

Description (incidence and severity):

none

Early or late resorptions:

no effects observed

Description (incidence and severity):

early resorptions: 0.66% and 0.80 % in controls and treated mice

Dead fetuses:

no effects observed

Description (incidence and severity):

none in controls and treated mice

Changes in pregnancy duration:

not specified

Changes in number of pregnant:

no effects observed

Description (incidence and severity):

all females evaluated were pregnant

Key result

Dose descriptor:

NOAEL

Effect level:

166 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: no effects observed

Key result

Dose descriptor:

NOAEL

Effect level:

57.5 mg/kg bw/day (nominal)

Based on:

element

Basis for effect level:

other: no effects observed

Abnormalities:

no effects observed

Fetal body weight changes:

no effects observed

Description (incidence and severity):

1.24 g and 1.26 g in controls and treated mice

Reduction in number of live offspring:

no effects observed

Description (incidence and severity):

no dead foetuses

Changes in sex ratio:

no effects observed

Description (incidence and severity):

M/F ratio: 0.88 and 0.93 in controls and treated mice

Changes in litter size and weights:

no effects observed

Description (incidence and severity):

14.7 and 11.7 in controls and treated mice

Changes in postnatal survival:

not examined

External malformations:

not specified

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

assymetric sternebrae in 3 fetuses in 2 litters and in 4 fetuses in 3 litters in controls and treated mice

Visceral malformations:

no effects observed

Description (incidence and severity):

no effects

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Aluminium concentration in treated pups: 0.18 ug/g wet weight

Details on embryotoxic / teratogenic effects:

None observed

Key result

Dose descriptor:

NOAEL

Effect level:

166 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effects obeserved

Key result

Dose descriptor:

NOAEL

Effect level:

57.5 mg/kg bw/day (nominal)

Based on:

element

Sex:

male/female

Basis for effect level:

other: no effects obeserved

Abnormalities:

no effects observed

Developmental effects observed:

no

Treatment related:

no

Conclusions:

The NOAEC for maternal and developmental toxicity is 166 mg/kg bw as Al(OH)3 and 57.5 as Al

Executive summary:

The influence of lactate on the potential developmental toxicity of high doses of aluminum (57.5 mg/kg/day) was evaluated. Three groups of pregnant Swiss mice were given by gavage daily doses of Al(OH)3 (166 mg/kg), aluminum lactate (627 mg/kg), or Al(OH)3 (166 mg/kg) concurrent with lactic acid (570 mg/kg) on gestational days 6-15. An additional group of pregnant mice received lactic acid (570 mg/kg) during the same period. Cesarean sections were performed on gestation day 18, and live fetuses were sexed, weighed and examined for morphological defects. Maternal toxicity was observed in the groups treated with aluminum lactate, and Al(OH)3 concurrent with lactic acid. The reproductive data did not show embryotoxic effects in any group, whereas fetal body weight was significantly reduced in the aluminum lactate group. In this group, morphological changes included cleft palate and an increased incidence of parietals with delayed ossification. Although not statistically significant, the incidence of skeletal variations was also increased in the group given Al(OH)3 concurrent with lactic acid. Taken together the present data, as well as the results of previous studies strongly suggest that the consumption of high doses of aluminum-containing compounds should be avoided during pregnancy.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

166 mg/kg bw/day

Study duration:

subacute

Species:

mouse

Quality of whole database:

study on aluminium hydroxide

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

10 800 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

Study on butan-1-ol the most volatile hydrolysis product

Mode of Action Analysis / Human Relevance Framework

Upon contact with water or moisture (e.g. within mucous membranes) aluminium tributanolate hydrolyses immediately to butan-1ol and aluminium 3+ cations (as hydroxide and oxyhydroxide). Hence, toxicity is determined by the toxicity of these two species.

For risk assessment purposes the lowest derived occupational exposure limit for butan-1-ol (61 mg/m3) will be used as starting point, as inhalative absorption of butan-1-ol is expected to be >100 times higher than that of aluminium hydroxide. For dermal toxicity the NOAEL (125 mg/kg bw) from the oral repeated dose study on butan-1-ol will be used as starting point, also based on the large diffecrence is absorption to be expected between butan-1-ol and aluminium (III) compounds.

Justification for classification or non-classification

Based on the absence of effects on reproduction and development for both butan-1-ol and Al3+ compounds, it can be concluded that aluminium tributanolate does not require classification for reproductibe toxicity according to CLP (Regulation EC No 1282/2008).