Aluminium triisopropanolate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Aluminium triisopropylate hydrolyses immediately to isopropanol and aluminium(III) species (hydroxide / oxyhydroxide). Therefore, effects on fertility by oral exposure can be assessed by assessing the oral fertility effects of aluminium 3+ cationic compounds and isopropanol.

 

The reproductive toxicity of isopropanol was assessed in a GLP-compliant study equivalent to the OECD test guideline 415 (one-generation reproduction toxicity study). Wistar rats were administered IPA at concentrations of 0 (tap water), 1.25, or 2.0% in drinking water (Publication by Faber, 2008). 
There were no mortalities, abortions, or early deliveries reported in the F0 generation. Adult male food consumption was decreased in all dose groups compared to controls. This corresponded with decreased body weights in the 2.0% dose group, and a transient slight decrease in body weight in animals treated with 0.5 and 1.0% isopropanol. Water intake was decreased in male rats treated with isopropanol; however, intake returned to normal levels in the 0.5% group. Food consumption also was decreased in adult females treated with 1.0 and 2.0% isopropanol; however, decreases noted in the 1.0% group had recovered by the second day of gestation. Body weights in isopropanol-treated adult females were lower than those reported for the control group at the start of gestation, but had recovered during gestation except in the 2.0% group. Following parturition, body weights for all dose groups were initially similar to controls; however, decreased body weights were noted in the 2.0% dose group on and after post-natal day 4. Water consumption was initially decreased in females treated with 1.0 and 2.0% isopropanol, but had returned to control levels in the 1.0% IPA group by the third day of treatment.
A slight-dose dependent decrease in red blood cells in the 2.0% group adult males and 1.0 and 2.0% adult females was noted. For males, a slight increase in mean cell volume in the mid- and high-dose groups was observed. Increased absolute and relative kidney weights, and relative liver and spleen weights were noted in high-dose F0males. Statistically significant increased absolute liver and kidney weight, and relative liver weight were noted in the 2.0% F0 females. There were no effects of isopropanol exposure on fertility. The number of pups per litter on post-natal day 1 was decreased in the 2.0% group. This increase was attributed to the cannibalism of the pups by the Dam and decreased pup survival, as a decrease in litter size was not observed in the embryotoxicity study. In the embryotoxicity study, increased preimplantation loss, a decrease in mean litter weight, and a decrease in mean fetal body weight was noted in the 2.0% group.
In the F1 Generation, average pup weight was decreased in the 2.0% group on post-natal day 7. Increased mean relative liver weight was reported in F1males and females at the 2.0% dose level. High-dose F1males also had higher relative kidney weights. Slight increases in absolute brain weight and increased relative empty caecum weight were noted in high-dose F1males and females. There were no gross abnormalities noted in the F1generation at necropsy.
Based on a review of the data it is proposed that the NOAEL for parental toxicity be considered to be 347 mg/kg bw/day (based on the lowest calculated parental isopropanol intake for 0.5% isopropanol in drinking water). The proposed NOAEL for reproductive toxicity is 853 mg/kg bw/day (based on the lowest calculated maternal isopropanol intake for 1.0% isopropanol in drinking water) on the basis of effects noted on pre-implantation loss, mean litter and fetal body weight, and fetal survival at the 2.0% dose level.

 

In an oral gavage, GLP, multi-generation study with isopropanol equivalent to the OECD test guideline 416 (Beyer, 1992), a dose of 1000 mg of isopropanol per kg of body weight produced evidence of parental effects, as indicated by increases in absolute and/or relative liver and/or kidney weights compared wtih those of controls. In addition, increased absolute or relative liver weigths were observed in parental animals dosed with 500 mg/kg compared with controls. These effects were not considered to be an adverse effect. However, treatment-related centrilobular hepatocyte hypertrophy was observed in a few high dose P2 male rats. Increased mortality was observed in high dose F1 offspring compared with controls. In addition, high dose offspring of both generations were lighter than controls at several intervals. Therefore, the parental NOAEL (No Observed Adverse Effect Level) was established at 500 mg/kg of isopropanol, while the reproductive NOAEL was established at greater than 1000 mg/kg under the conditions of this study. The offspring toxicity NOAEL was established at 500 mg/kg based on reduced offspring body weights and increased mortality observed at 1000 mg/kg. Reprotoxicity was only observed in the presence of maternal toxicity.

The NOAEL for reproductive effects was 1000 mg/kg bw/day. The NOAEL for offspring toxicity was 500 mg/kg bw/day due to reduced body weights and increased mortality at 1000 mg/kg bw/day.

A GLP-compliant pilot study equivalent to the OECD test guideline 415 in rats served as a range finding study. This study did not identify a NOAEL at isopropanol dose concentrations of 0, 1.25, 2.0, or 2.5% in drinking water (Gaunt, 1986). Decreased adult food and water consumption, decreased adult and pup body weight gain, evidence of embryotoxicity (i.e., fewer live pups, increase in pup mortality, and reduction in pup body weight gain), signs of anaemia, and increased liver and kidney weights were noted. These effects were mainly seen at the 2.0 and 2.5% dose concentrations compared to the control.

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). Aluminium sulfate 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.

In conclusion, neither isopropanol nor soluble aluminium species tested for fertility/developmental toxicity showed significant effects and thus such effects are not expected upon exposure of aluminium tri-isopropylate as described above.

 

For explanation for using surrogate data in a weight of evidence approach see the document in section 13 IUCLID.

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:

one-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented, according to accepted guidelines

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Specified-Pathogen-Free colony (Olac 1976 Ltd., Bicester, Oxon)
- Age at study initiation: Ages of the rats at receipt were 7-8 weeks (males) and 10-11 weeks (females) .
- Housing: Groups of 5 females or 2 males of the same sex were housed in polypropylene cages with stainless steel tops and grid floors.
- Diet (e.g. ad libitum): Free access to Certified Rat and Mouse No. 3 expanded fine ground laboratory feed (Special Diet Services, Witham, Essex, UK)
- Water (e.g. ad libitum): Tap water ad libitum
- Acclimation period: 7–9 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20–24ºC
- Humidity (%): 45%–65%
- Photoperiod (hrs dark / hrs light): 12:12 hr light/dark cycle

Route of administration:

oral: drinking water

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Drinking water formulations were prepared with domestic mains tap water at 2-week intervals for the duration of treatment of the animals. Due to greater water consumption during pregnancy and lactation, drinking water solutions were prepared more frequently during some stages of the reproduction/embryotoxicity study.

Details on mating procedure:

- M/F ratio per cage: 1:3
- Length of cohabitation: Up to 15 days
- Proof of pregnancy: vaginal plug/sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged (how): After mating was confirmed, pregnant females were housed individually in similar cages. Those assigned to the littering phase of the study were given cages with solid floors covered with sawdust and provided with paper nesting material, as necessary.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Prior to use, formulations were analyzed by GLC to confirm concentration of IPA and stability (further details not provided).

Duration of treatment / exposure:

Males (F0) were dosed for 70 days and females (F0) for 21 days after which 2 females from those assigned to the littering phase of the study and 1 female from those assigned to the embryotoxicity phase of the study were housed with 1 male from the same treatment group for up to 15 days.

Male (F0) dosing also occurred during mating and continued until animals were euthanized and female (F0) dosing continued during gestation, during lactation, and continued until animals were euthanized.

Offspring (F1) were dosed during rearing until animals were euthanized.

Frequency of treatment:

Continuous in drinking water.

Remarks:

Doses / Concentrations:
0.5, 1.0, or 2.0%
Basis:
nominal in water

No. of animals per sex per dose:

Each dose group consisted of 10 males and 30 females, of which 10 females were used for the embryotoxicity determinations and the remaining 20 females for the single generation (littering) phase of the study.

Control animals:

yes, concurrent no treatment

Positive control:

None used.

Parental animals: Observations and examinations:

General observations were made each day, with a more thorough examination done once per week. Male body weights were recorded for 3 days prior to and 4 days after the test solutions were first given and then twice weekly throughout the study. Females were weighed daily for 3 days prior to and 4 days after the test solutions were first given and then twice weekly for 3 weeks. During gestation, females were weighed on GD 0 and every day until they littered or were euthanized. During lactation, the weight of the female and the total litter weight were recorded on postnatal days (PNDs) 1, 4, 7, and 14. On PND 21, the female and each of the pups were weighed individually. During the mating period, only male body weights were recorded.

Consumption of feed and water was determined at the same intervals as the body weight measurements except for the females during the postpartum period when intake of feed and water was measured twice weekly.

Dams assigned to the embryotoxicity phase of the study were euthanized on GD 19. The abdominal and thoracic contents were examined for abnormalities. Ovaries were examined and the number of corpora lutea recorded. The uteri were examined and the numbers and locations of viable fetuses, early and late resorptions, total implantations, and pre- and postimplantation losses were recorded.

Males were euthanized on day 126 of the study.

Oestrous cyclicity (parental animals):

Not reported.

Sperm parameters (parental animals):

Not reported.

Litter observations:

Each live fetus was weighed and examined for gross, externally visible abnormalities. The viscera of fetuses that showed evidence of edema in the embryotoxicity study, along with their littermates, were examined by evisceration under a dissecting microscope, and the sex of each fetus was recorded. The remaining females were allowed to litter. Litters were examined on PND 1 for stillborn or abnormal young and then daily for any subsequent deaths. On PND 4, 7, 10, 14, 17, and 21, numbers of survivors and any abnormalities were recorded. On PND 21, pups were weaned and removed from the dams.

Postmortem examinations (parental animals):

Within 21 days of weaning the last litter, each adult animal was fasted overnight (with access to water) and euthanized by exsanguination under barbiturate anesthesia.

During the anesthesia phase, blood was collected from the aorta of each adult rat and analyzed for total erythrocyte and leukocyte counts, hemoglobin concentration, and mean cell volume. Twelve females (5 controls, 3 given 1.0% IPA, 4 given 2.0% IPA) that failed to litter were euthanized 24 days after the last day of pairing. Each rat was examined for general condition and gross abnormalities. Weights of the following organs were recorded: adrenal glands, brain, cecum, gonads, heart, liver, kidney, and spleen. Selective tissues were preserved in 10% neutral buffered formalin. Histopathologic examination was carried out on the following tissues from control and high-dose adult animals: bladder, cervix and uteri, epididymides, ovaries, pituitary, prostate, seminal vesicles, testes, uterine horns, and vagina.

Postmortem examinations (offspring):

Within 21 days of weaning the last litter, one animal of each sex from each litter were fasted overnight (with access to water) and euthanized by exsanguination under barbiturate anesthesia. Approximately 10 days later, the remaining pups (F1) were euthanized by carbon dioxide inhalation and examined for gross, external abnormalities. The liver and kidneys of each animal were weighed and tissue samples preserved in formalin.

Statistics:

Body weights, organ weights, hematological results, feed intakes, water intakes, pup numbers, pup weights, number of preimplantation losses, number of early and late resorptions, total number of postimplantation losses, number of live fetuses, weight of fetuses, and litter weights from the treated and control groups were compared using analysis of variance, the procedure of Least Significant Difference and Student’s t-test. The incidence of abnormalities in adults at necropsy and the incidence of histopathological findings in the treated and control groups were compared using Fisher’s Exact test.

Reproductive indices:

Number of infertile males, length of gestation, number of females with litters.

Offspring viability indices:

Total number of pup survival (%) and pup survival/litter (%).

Clinical signs:

no effects observed

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

effects observed, treatment-related

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

effects observed, treatment-related

There were no deaths, abortions, early deliveries, or dams removed from the study.

Administration of IPA in the drinking water caused an immediate, statistically significant dose-dependent decrease in water intake in the male rats. Although intake returned to normal for males in the 0.5% group, intake was approximately 5%–14% lower during the premating phase for the 1.0% group males and approximately 30% lower from days 7–11 until study termination for the 2.0% group males. For male rats consuming 2.0% IPA in the drinking water, there was a marked decrease in feed intake over the first 4 days of treatment, and feed intake remained significantly less than control values over the remainder of the premating period. Smaller (but still significant) reductions in feed consumption were also seen in the low- and mid-dose groups. The overall mean feed consumption for all three treatment groups was statistically significantly lower than the control. Body weights of male rats receiving 2.0% IPA were 4%–5% lower than control prior to start of treatment. Male rats consuming water containing 0.5% or 1.0% IPA in the drinking water had initial slight reductions in weight gain but recovered after one week of exposure.

An initial decrease in water consumption was observed in 1.0% and 2.0% females during the premating phase. Intakes in the 2.0% group recovered to about 70% of the control value from 50% of the control value on day 1. During lactation, water consumption for the control and two lower doses increased dramatically, whereas intake by the 2.0% group was markedly reduced. For female rats consuming 1.0% or 2.0% IPA in the drinking water, there was a similar initial reduction in feed intake during the premating phase. The 1.0% female group had a return to control values; however, feed intake for the 2.0% group remained significantly lower than control values. Only animals in the 2.0% group continued to consume less feed than the control group, and this was generally within 10% of the control value. Similar reductions in feed intake for 2.0% females were also seen during the postpartum phase, whereas the 0.5% and 1.0% groups were unaffected. During the premating phase, all treated female groups showed an immediate weight loss, with recovery of weight gain after one week of exposure. From PND 4 onward, weights of animals consuming water containing 2.0% IPA were significantly lower.

There were no infertile males in any group and there was no effect of IPA exposure on female fertility. The length of gestation in females consuming water with IPA was comparable to the control values.

The number of pups/litter on gestation day 1 was reduced in the 2.0% group. This decrease in litter size in the 2.0% group was not replicated in the embryotoxicity portion of the study, suggesting an increase in pup mortality during parturition or GD 0, followed by cannibalism of the dead pups by the dam. This explanation is also supported by a decrease in pup survival from postnatal day 1 onward.

There was a slight dose-dependent decrease in red blood cells in males given 2.0% IPA in the drinking water and females in the 1.0% and 2.0% groups. For males only, there was also a slight but statistically significant increase in mean cell volume in the mid- and high-dose groups. There were no treatment-related effects on hematocrit, hemoglobin, or numbers of leucocytes in either sex.

No macroscopic abnormalities were seen at necropsy in females in either the embryotoxicity or one-generation reproductive phases of this study. In addition, no treatment-related histopathological changes were seen in tissues of the reproductive system of parental animals at the highest dose tested.

There was a statistically significant increase in absolute kidney weight and relative kidney, liver, and spleen weights in F0 high-dose group males. For F0 high-dose group females, there was a statistically significant increase in absolute liver and kidney weights and relative liver weights.

The only significant findings from the embryotoxicity study portion of the study was an increase in preimplantation loss, a decrease in mean litter weight, and a decrease in mean fetal body weight in the 2.0% group.

Dose descriptor:

NOAEL

Effect level:

853 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Clinical signs:

effects observed, treatment-related

Mortality / viability:

mortality observed, treatment-related

Body weight and weight changes:

no effects observed

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

A statistically significant increase in the total number of preimplantation losses in animals given 2.0% IPA and a slight but not statistically significant decrease in mean litter and mean fetal weights in this group. The only other finding in this phase of the study was whole body edema seen in 40% of the fetuses in three of eight litters in the high-dose group. No macroscopic abnormalities of the viscera of these fetuses were detected, and the incidence of edema was not related to gender.

There was a statistically significant decrease in postnatal pup survival and in the average pup weight (by PND 7) in the 2.0% group.

F1 generation animals of both sexes showed a significant increase in relative liver weights at all dose levels, with the effects being very highly significant at the 2.0% IPA level. High-dose group males also had a significantly higher relative kidney weight. In the F1 generation group receiving 2.0% IPA, there was a slight but statistically significant decrease in absolute brain weight and increase in relative empty cecum weights in both sexes.

No treatment-related gross abnormalities were observed in the F1 generation animals at necropsy.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

853 mg/kg bw/day

Sex:

male/female

Basis for effect level:

body weight and weight gain

System:

other: fetal body weight

Reproductive effects observed:

not specified

Conclusions:

The fertility of the male or female animals was not effected, even with significant reductions in water and food consumption in the group consuming water with 2.0% IPA. The publication authors state that the weight of evidence suggests that IPA does not affect male mating or fertility at levels up to at least 1000 mg/kg bw/day.
An increase in preimplantation loss, a decrease in mean litter weigth, and a decrease in mean fetal body weight in the 2.0% group were observed.

Executive summary:

Based on a review of this drinking water study, the following maternal and fetal NOAELs are proposed: NOAEL (parental)  F0 (male/female)  0.5% (347 mg/kg bw/d for males and 456, 668, and 1053 mg/kg bw/d for females during pre-mating, gestation, and postpartum phases, respectively). Effects on one or more of the following at 1.0% and/or 2.0% dose levels: food and water intake, body weight, red blood cells, cell volume, and liver, kidney, and spleen weights. NOAEL (reproductive) F0 (female) F0 (male) 1.0% (853, 1330, and 1948 mg/kg bw/d for females during pre-mating, gestation, and postpartum phases, respectively). 2.0 % (1107 mg/kg and 1030.4 bw/day for males during premating phase and days -3 to 126, respectively). Increased pre-implantation loss, decreased mean litter weight, and decreased mean fetal body weight at the 2.0% dose level. IPA does not affect male mating or fertility up to at least 1000 mg/kg bw/day.

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, The NOAEL for the other hydrolysis product isopropanol is 853 mg/kg bw

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

Description of key information

Aluminium tri-isopropylate hydrolyses immediately to isopropanol and aluminium(III) species (hydroxide / oxyhydroxide). Therefore, effects on fertility by oral exposure can be assessed by assessing the oral fertility effects of aluminium hydroxide and isopropanol.

The developmental toxicity of isopropanol was assessed in a GLP, prenatal development toxicity study equivalent to the OECD test guideline 414 (Faber, 2008). Wistar rats were administered 0 (tap water), 0.5, 1.25, or 2.5% isopropanol in drinking water. These dose levels were equivalent to 0, 596, 1242, and 1605 mg/kg bw for the low-, mid-, and high-dose groups, respectively. There were no mortalities reported. Reduced food and water consumption was noted at the 1.25 and 2.5% dose levels. A slight decrease in water consumption was observed in the 0.5% dose group on the first day of dosing, but this did not achieve statistical significance. Body weight loss was noted from gestation days 6 to 8 in the 2.5% dose group, and decreased body weight gain was noted thereafter during the dosing period. Decreased body weight gain was noted in the low dose group on the first day of treatment, and for the first two days of treatment in the mid-dose group. After cessation of treatment, the dams in the 2.5% dose level reported increased weight gain compared to controls. Overall, body weights of the 2.5% dose group were lower than those reported for control animals from gestation day 7 through to termination. There were no effects on embryotoxic parameters. A slight dose-dependent decrease in fetal litter weight was observed. Statistically significant decreased mean fetal weight was noted at the 1.25 and 2.5% dose levels. A statistically significant increase in variations was reported in treated animals, and was indicative of a lower degree of ossification. These changes may have been secondary to decreased water and food consumption and secondary to palatability problems. The authors proposed that fetotoxicity, as manifested by reduced fetal body weights, only occurred at dose levels that also caused maternal toxicity (decreased food and water consumption).
A NOAEL was not reported by the study authors. It is proposed that the NOAEL for maternal toxicity be considered to be 0.5% (596 mg/kg bw/day) due to decreased food and water consumption, and corresponding effects on body weight at higher dose levels. The NOAEL for fetal toxicity is proposed to be 0.5% (596 mg/kg bw/day) on the basis of reduced body weights at higher dose levels.

Isopropanol administered by gavage during major organogenesis in CD (Sprague-Dawley) rats resulted in maternal and developmental toxicity at 800 and 1,200 mg/kg bw/day, with no indication of teratogenicity at any dose tested (GLP-compliant, prenatal development study in rats, equivalent to the OECD Test Guideline 414 (Tyl et al., 1990). The "no observed adverse effect level" (NOAEL) for maternal and developmental toxicity of isopropanol in rats is therefore 400 mg/kg bw/day under the conditions of this study.

In another GLP-compliant developmental toxicity study similar to the OECD Guideline 414 in New Zealand White rabbits (Tyl et al., 1990) administration of isopropanol by gavage resulted in profound maternal toxicity at 480 mg/kg/day but only relatively mild non-specific, transient effects in the 120 and 240 mg/kg/day groups. There was no statistically significant indication of developmental toxicity at any dose. In addition, there was no evidence of teratogenicity at any dose tested. The no-observed-adverse-effect level (NOAEL) for maternal toxicity in rabbits was therefore 240 mg isopropanol/kg/day and the NOAEL for developmental toxicity in rabbits was 480 mg isopropanol/kg/day

 

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. 

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. The NOAEL for the other hydrolysis product isopropanol is 400-596 mg/kg bw

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

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