Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reliable data on reproductive toxicity are available for the source substances aluminium dihydrogen triphosphate and aluminium sulphate. Aluminium dihydrogen triphosphate was tested in a Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test according to OECD 422 in which a NOAEL of 1000 mg/kg bw/day was defined for fertility. Furthermore, a Two-Generation study was performed with aluminium sulfate. No adverse effects on fertility were noted when testing up to the highest dose level (3000 ppm aluminium sulfate) and hence, a NOAEL ≥ 31.2 mg aluminium/kg bw/day is considered for fertility.

Link to relevant study records

Referenceopen allclose all

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 with acceptable restrictions
Remarks:
limited documentation, differences in pH of drinking water in control and test groups, no analysis of Al content in blood, urine and tissues.
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

The source substance and the target substance are considered to be similar enough to facilitate read-across for the following reasons:
(1) Both substances are inorganic salts containing a trivalent aluminium cation.
(2) Both substances will ultimately dissociate into the common breakdown products of the Al3+ following chemical or biological hydrolysis and dissolution of the ionic bonds.
In accordance with the provisions set out in Annex XI, Section 1.5, the results of the studies used for assessment and read-across are adequate for the purpose of classification and labelling and/or risk assessment; have adequate and reliable coverage of the key parameters addressed in the corresponding test method; cover an exposure duration comparable to or longer than the corresponding test method; and adequate and reliable documentation of the applied method is provided in the technical dossier.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
Deviations:
yes
Remarks:
(limited information of methods, differences in the pH of drinking water in control and test groups)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crl:CD(SD)
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Atsugi Breeding Center, Charles River Laboratories Japan, Inc., Yokohama, Japan
- Age at study initiation: 5 weeks
- Housing: Animals were housed individually, except for acclimation, mating and nursing periods, in suspended wire-mesh cages
From gestational day 17 (GD 17) to postnatal Day 21 (PND 21), individual dams and litters were reared in stainless-steel trays, and using wood chips as bedding (White Flake; Charles River Laboratories Japan, Inc., Yokohama, Japan).
- Diet: CRF-1; Oriental Yeast Co., Ltd., Tokyo, Japan, ad libitum (standard laboratory rodent chow).
- Al content in diet: 25 - 29 ppm (based on atomic absorption spectrometry)
- Water: deionized drinking water, ad libitum
- Al content in water: < 5 µg Al/mL (drinking water of controls)
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 – 25
- Humidity (%): 36 - 59
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: drinking water
Vehicle:
water
Remarks:
ion-exchanged water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Dosing solutions were prepared freshly at least every 6 days and kept in a cool place until serving. Fresh drinking water was replaced at least once every 4 days.
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: up to 2 weeks (until successful mating)
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- After 14 days of unsuccessful pairing, the first male was replaced by another male with proven fertility.
- After successful mating each pregnant female was caged (how): individually except for nursing period
- For F1 mating, cohabitation of siblings was avoided.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The concentrations of aluminium sulphate in drinking water were analysed in the first and last preparations and once every 3 months using high performance liquid chromatography (quantitation limit: 5 µg/mL) (analysed Al concentrations: 97.5 – 106.3% of the target).
Duration of treatment / exposure:
(P) Males: 10 weeks before mating.
(P) Females: 10 weeks before mating, during mating, gestation and lactation (weaning of pups: PND 26).
(F1) Males: starting at PND 21 - 25 (selection of F1 parental animals on PND 21- 25 to equalize mean body weights) until mating, during mating and production of F2 generation, until weaning (on PND 26)
(F1) Females: starting at PND 21 - 25 (selection of F1 parental animals on PND 21- 25 to equalize mean body weights) until mating, during mating, gestation and lactation until weaning (on PND 26)
Frequency of treatment:
daily, 7 days/week
Details on study schedule:
Dams (P) delivered the offspring spontaneously and nursed their pups until PND 26. Litters were normalized on PND 4.
During PND 21 - 25, F1 parental animals were selected (desiganted as Day 0 or dosing) for the F1 generation. Exposure of the F1 weanlings started on the day of selection at the same doses as their parents.
F1-selected rats were necropsied in the same manner as described for F0 rats. The remaining F1 weanlings and all F2 weanlings were necropsied on PND 26.
Dose / conc.:
120 ppm (nominal)
Remarks:
in water
Dose / conc.:
600 ppm (nominal)
Remarks:
in water
Dose / conc.:
3 000 ppm (nominal)
Remarks:
in water
Remarks:
Doses / Concentrations:
P males: 8.6, 41.0 and 188 mg Al2(SO4)3/kg bw/day; P females: 14.4, 71.5 and 316 mg Al2(SO4)3/kg bw/day
Basis:
other: mean dose value calculated based on reported body weight and water consumption (including aluminium from diet and drinking water)
Remarks:
Doses / Concentrations:
P males: 2.96, 8.06 and 31.2 mg Al/kg bw/day; P females: 4.5, 13.5 and 52.0 mg Al/kg bw/day
Basis:
other: total doses of aluminium (food and water combined)
Remarks:
Doses / Concentrations:
F1 males: 10.7, 50.2 and 232 mg Al2(SO4)3/kg bw/day; F1 females: 15.3, 74.2 and 338 mg Al2(SO4)3/kg bw/day
Basis:
other: mean dose value calculated based on reported body weight and water consumption (including aluminium from diet and drinking water)
Remarks:
Doses / Concentrations:
F1 males: 3.55, 9.78 and 38.5 mg Al/kg bw/day; F1 females: 4.72, 14.0 and 55.6 mg Al/kg bw/day
Basis:
other: total doses of aluminium (food and water combined)
No. of animals per sex per dose:
24
Control animals:
other: concurent vehicle (total dose of aluminium via food and water: F0: 1.62 (males) and 2.29 (females) mg Al/kg bw/day; F1: 1.93 (males) and 2.35 (females) mg Al/kg bw/day)
Details on study design:
- Dose selection rationale: based on a range finding study (animals were orally exposed to 1000, 3000, 10,000 and 30,000 ppm aluminium sulphate for 7 weeks).

Results: Sacrifice of high-dose animals after 2 weeks due to water avoidance. Decreased water consumption was observed in the remaining groups. At 3000, 10,000 and 30,000 ppm, reduced food consumption and decreased body weights were observed. At necropsy, thickening of the limiting ridge in the stomach and atrophy of the thymus and spleen were observed at 10,000 ppm. Females exposed to 3000 or 10,000 ppm revealed decreased relative weights of the liver, thymus and spleen. Body weights of pups were decreased at 10,000 ppm on PND 4. Based on these results, 120, 600 and 3000 ppm were selected for the main study.
Parental animals: Observations and examinations:
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice a day

BODY WEIGHT: Yes
- Time schedule for examinations: weekly. Body weights of dams were recorded on GD 0, 7, 14 and 20 and PND 0, 4, 7, 14 and 21

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/day: Yes
- Time schedule for examinations: weekly. Food consumption of dams was recorded on GD 0, 7, 14 and 20 and PND 0, 7, 14 and 21

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: twice a week, and on GD 0, 4, 7, 11, 14, 17 and 20 and PND 0, 4, 7, 11, 14, 17, 19 and 21
Oestrous cyclicity (parental animals):
Estrous cyclicity was tested during the last 2 weeks of the premating and during mating (until evidence of copulation was detected based on daily vaginal lavage).
Repeated estrous cycles of 4 – 6 days were considered normal.
Sperm parameters (parental animals):
Parameters examined in P/F1 male parental generations:
epididymis weight, sperm count in epididymides, sperm motility and sperm morphology
Litter observations:
STANDARDISATION OF LITTERS: Yes
Performed on day 4 postpartum: maximum of 8 pups/litter (4/sex/litter); no adjustment for litters with less than eight pups.

PARAMETERS EXAMINED:
F1 / F2 offspring: number of pups, sex ratio, live birth, postnatal mortality, presence of gross anomalies, weight gain, physical and behavioural abnormalities. Clinical signs of toxicity (daily) and body weight were measured on PND 0, 4, 7, 14 and 21.

GROSS EXAMINATION OF DEAD PUPS: Yes

DEVELOPMENTAL LANDMARKS
Developmental landmarks:
- Pinna unfolding in all F1 and F2 live pups (PND 1 to PND 4).
- Anogenital distance (AGD, measured using calipers on PND 4) in all F1 and F2 pups, normalized value of AGD to body weight, AGD/cube root of the body weight ratio
- Incisor eruption (one male and one female F1 and F2 pup/litter, evaluated on PND 8)
- Eye opening (starting on PND 12).
- Body weight (recorded on the day the criteria were fulfilled).

Neuromotor performance:
- Surface righting reflex, negative geotaxis and mid-air righting reflex (assessed on PND 5, 8 and 18, respectively), for one F1/2 male and female /litter).

Sexual maturation:
- Preputial separation (recorded daily in all F1 males selected as F1 parents, beginning on PND 35)
- Vaginal opening (recorded daily in all F1 females selected as F1 parents beginning on PND 25)
- Body weights (recorded on the day of completion of these pubertal landmarks).

NEUROBEHAVIOURAL EXAMINATIONS
Locomotor activity:
- Spontaneous locomotor activity (measured in 10 male and 10 female F1 rats which were randomly selected from each group at 4 weeks of age) using a multi-channel activity monitoring system (SUPERMEX; Muromachi Kikai Co., Ltd., Tokyo, Japan)


T-maze test:
- Water-filled multiple T-maze test (Biel (1940) (conducted in 10 male and 10 female F1 rats selected randomly from each group at 6 weeks of age)
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: as soon as possible after parturition of their paired females
- Maternal animals: after weaning on PND 26.

GROSS NECROPSY
- External and Internal examination, including the cervical, thoracic, and abdominal viscera
- Organs examined: brain, pituitary, thyroid, thymus, liver, kidneys, spleen, adrenals, testes, epididymides, seminal vesicles (with coagulating glands and their fluids), ventral prostate, uterus and ovaries
In 10 randomly selected F1 females from the control and highest dose groups, the number of primordial follicles was counted.

HISTOPATHOLOGY / ORGAN WEIGHTS
Histopathological evaluations were performed:
- in all animals of the control and highest dose groups;
- in females showing abnormal estrous cycles, abnormal delivery or totally dead pups;
- in females and males without evidence of copulation or insemination;
- in all animals with grossly abnormal reproductive organs.
- Organs examined: testes, epididymides, seminal vesicles, ventral prostate, coagulating gland, ovaries, uterus and vagina
When the highest dose group showed treatment-related changes in some tissues, examination of the same tissues was performed in the next lower dose group.
Postmortem examinations (offspring):
SACRIFICE
Following the adjustment of litter size on PND4, culled pups were euthanized and subjected to gross necropsy. Pups found dead before weaning were necropsied immediately. Not selected F1 pups as parental animals and all F2 pups were sacrificed on PND 26.

HISTOPATHOLOGY / ORGAN WEIGTHS
Organ weights were determined for one male/female F1 and F2 weanling selected from each dam:
- brain, thymus, liver, kidneys, spleen, adrenals, testes, epididymides, ventral prostate, uterus and ovaries
- based on organ weight changes observed in the liver and spleen of the highest dose group in F1 and F2 generations, these tissues were histopathologically examined from 10 male and 10 female F1/F2 weanlings of the control and highest dose groups;
When the highest dose group showed treatment-related changes in some tissues, examination of the same tissues was performed in the next lower dose group.
Statistics:
Parametric data (body weight, food and water consumption, length of the estrous cycle and gestation, precoital interval, the number of implantations and pups born, delivery index, reflex response time, age at sexual maturation, behavioural test parameters, organ weight and sperm parameters) were analysed for homogeinity and distribution by Bartlett’s test.
Pre-weaning pups, body weight, AGD, viability, and age at the completion of developmental landmarks were similarly analysed using the litter as the experimental unit.
When homogeneity of distribution was established, One Way Analysis of Variance followed by Dunnett’s test were performed.
Data without homogeneity were analysed using the Kruskal–Wallis rank sum test followed by Mann Whitney’s U test.
The incidence of parental animals showing clinical signs, autopsy and histopathological findings, the incidence of females with normal estrous cycles, incidence of weanlings with histopathological findings, copulation, fertility and gestation index, neonatal sex ratio and completion rate of negative geotaxis were analysed by Fisher’s exact test.
The Wilcoxon rank sum test was used to analyse the incidence of pups showing clinical signs and necropsy findings, completion rate of pinna unfolding, and the success rate of surface and mid-air righting reflex.
Student’s t-test was used to compare the number of primordial follicles in the control and highest dose group, homogeneity of variance was indicated by the F-test.
A 5% level of probability was used as criterion for significance.
Reproductive indices:
The following reproductive indices were calculated in F0 and F1 parental animals:
- Copulation index (%) (no. of animals with successful copulation/no. of animals paired) ×100;
- Precoital interval (days);
- Fertility index (%) (no. of males that impregnated a female or no. pregnant/no. of animals with successful copulation) × 100;
- Gestation index (%) (no. of females that delivered live pups/no. of pregnant females) × 100;
- Gestation length (days);
- Delivery index (%) (no. of pups delivered/no. of implantations) × 100;
- Estrous cycle in F0 and F1 females
Offspring viability indices:
The following parameters were analysed:

F1 and F2 offspring:
Maternal indices:
No. of litters;
No. of pups delivered;
Sex of all pups;
Sex ratio of pups total (no. of male pups/total no. of pups).

Viability index:
on PND 0 (%) = (no. of live pups on PND 0/no. of pups delivered) × 100;
on PND 4 (%) = (no. of live pups on PND 4/no. of live pups on PND 0) × 100;
on PND 21 (%) = (no. of live pups on PND 21/no. of live pups on PND 4 after cull) × 100.

Individual body weight:
Male and female individual weight during lactation on PND 0, 4, 7, 14 and 21.
Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
3000 ppm: significant reduction in body weight (P males/females in the first 2 -3 weeks of dosing); 600 and 3000 ppm: reduced food consumption
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
3000 ppm: significant reduction in body weight (P males/females in the first 2 -3 weeks of dosing); 600 and 3000 ppm: reduced food consumption
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
Description (incidence and severity):
Test substance intake: 120, 600 and 3000 ppm: significant decrease in water consumption (P males and females)
Reproductive function: oestrous cycle:
effects observed, treatment-related
Description (incidence and severity):
persistent diestrus (control and test animals, non-adverse)
Reproductive function: sperm measures:
effects observed, treatment-related
Description (incidence and severity):
3000 ppm: decreased number of cauda epdididymal sperm (non-adverse)
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No clinical signs of toxicty were observed in the P generation. One unscheduled death was determined in the P mid-dose group at 2 weeks of gestation, the respective dam showed a subcutaneous mass in the abdominal region starting 5 weeks of dosing.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Significantly decreased body weights were observed in high-dose males and females in the first 2 - 3 weeks of dosing without showing a direct correlation to food consumption. Reduced food consumption was observed in mid- and high-dose animals at the following time points:
P mid-dose males - during the first week of dosing, high-dose males: during weeks 1, 8 and 13-14;
P mid-dose females: during week 3 of lactation, high-dose females: 1 week of dosing and during week 3 of lactation.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
No significant alterations in the estrous cycle were observed in P and F1 females during the premating period. However, persistent diestrus was determined in a few control and test animals.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
A significantly decreased number of cauda epididymal sperm (253.8 ± 61.3 × 1E+6/cauda compared to 286.3 ± 40.3 ×1E+6/cauda in test and control animals, respectively) was determined in high-dose males. However, after normalization to epididymal weight, the number of relative cauda epididymal sperm was comparable among the groups and hence, the effect is considered as non-adverse.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
No significant differences on reproductive function or performance were observed including copulation, fertility, gestation index, precoital interval, gestation length, delivery index, number of implantations and number of litters or pups.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Reduced liver (absolute and relative weight) and spleen (absolute weight) weights were determined in high-dose males. No alterations were observed in females.

GROSS PATHOLOGY (PARENTAL ANIMALS)
No dose-related gross lesions were found in any animal and dose group.

HISTOPATHOLOGY (PARENTAL ANIMALS)
No compound-related alterations were observed.
Dose descriptor:
other: LOAEL (systemic toxicity)
Effect level:
3 000 ppm (nominal)
Based on:
test mat.
Remarks:
(equivalent to 31.2 (males) and 52.0 (females) mg Al/kg bw/day)
Sex:
male/female
Basis for effect level:
other: decreased body weight gain; decreased food consumption
Dose descriptor:
other: NOAEL (systemic toxicity)
Effect level:
600 ppm (nominal)
Based on:
test mat.
Remarks:
(equivalent to 8.6 (males) and 13.5 (females) mg Al/kg bw/day)
Sex:
male/female
Basis for effect level:
other: no adverse effects observed
Dose descriptor:
NOAEL
Remarks:
Fertility
Effect level:
>= 3 000 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 31.2 (males) and 52.0 (females) mg Al/kg bw/day, P generation
Sex:
male/female
Basis for effect level:
other: fertility - no adverse effects observed
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
F1 generation: trauma in the perianal region and tail was observed in one control pup, hemimelia and oligodactyly were observed in one low-dose pup (non-adverse)
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
F1 generation: decreased body weight in high-dose males and females on PND 21 and at scheduled sacrifice; F2 generation: decreased body weight in high-dose females on PND 21 and at sacrifice in both genders
Sexual maturation:
effects observed, treatment-related
Description (incidence and severity):
F1 females, high-dose group: delayed vaginal opening
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
F1 and F2 generation: liver, spleen, thymus, kidney, testes, epididymides, uterus, ovary and brain weights were affected by treatment
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
VIABILITY (OFFSPRING)
Viability was comparable on PND 0, 4 and 21 amomg the groups in any generation (F1 and F2).

CLINICAL SIGNS (OFFSPRING)
F1 generation
No signs indicative for clinical toxicity were observed in test animals. The observed trauma in the perianal region and tail or hemimelia and oligodactyly is not considered as adverse due to the low incidence (observed in 1 control and 1 low-dose pup, respectively).

F2 generation
No signs indicative for clinical signs were observed in any dose group.

BODY WEIGHT (OFFSPRING)
F1 generation
Significantly reduced body weights of high-dose animals (males and females) compared to the control were observed on PND 21 and at sacrifice (for details, please refer to Tables 1 and 2).
F2 generation
Reduced body weight was determined in high-dose females on PND 21 and at sacrifice in both genders (for details, please refer to Tables 1 and 2).


SEXUAL MATURATION (OFFSPRING)
F1 generation
Significantly delayed vaginal opening was observed in high-dose females (31.4 ± 1.7 compared to 29.5 ± 2.1 days in control) correlating to a slightly increased body weight compared to controls (119.0 ± 13.3 versus 109.6 ± 11.6 g).
No significant differences were noted regarding the age of preputial separation. No changes in body weights at the time of preputial completion were noted.


ORGAN WEIGHTS (OFFSPRING)
F1 generation
Significant decreases in liver (absolute and relative organ weight), spleen, thymus, kidney, testes and epididymides weights (absolute weights) and increased relative brain weights were observed in high-dose animals. The absolute uterus weight was decreased in high-dose animals.

F2 generation
Weights of the thymus, spleen (absoulte and relative weights), liver and epididymides (absolute) were significantly reduced. Relative brain weight was increased in high-dose males. Reduced absoulte brain weight was observed in females of the mid-dose group (please refer to Tables 1 and 2).

GROSS PATHOLOGY (OFFSPRING)
No alterations were found in either the F1 or F2 generation during the preweaning period (data not shown).

HISTOPATHOLOGY (OFFSPRING)
No dose-related histopathological changes were determined in the liver or spleen of male and female F1 and F2 weanlings.

OTHER FINDINGS (OFFSPRING)
SEX RATIO
The sex ratio was comparable among all test groups in the F1 or F2 generation.

PHYSICAL DEVELOPMENT
In the F1 generation and F2 males of all test groups, completion rate of pinna unfolding, age at completion of incisor eruption and eye opening were comparable among the groups. F2 females revealed a significantly lower completion rate of pinna unfolding on PND 2 compared to controls in the mid-dose group (17.0 ± 35.4%, compared with 45.8 ± 46.9% in mid-dose females and controls, respectively). Otherwise, females of the F2 generation did not show significant differences in regard to the measured developmental landmarks.

AGD and AGD per cube root of body weight ratio were comparable among the groups in the F1 and F2 generation.

NEUROMOTOR DEVELOPMENT
The development of reflexes (surface righting reflex on PND5, negative geotaxis reflex on PND8 and midair righting reflex on PND 18) was comparable among the groups in any generation. Further, the response times of surface righting and negative geotaxis reflexes were not affected. However, 1 mid-dose F2 female did not achieve the mid-air righting reflex on PND 18 in 1/3 trials without affecting the mean success rate between the control and mid-dose group (100 ± 0.0% versus 98.4 ± 7.3%).

RESULTS ON PARENTAL ANIMALS OF THE F1 GENERATION
CLINICAL SIGNS AND MORTALITY
No clinical signs of toxicity were observed in the parents selected from the F1 generation.
Incidental death occured in the low- and high-dose group: 1 low-dose male died at 9 weeks of dosing. Prior to death, soiling of periocular and perinasal fur and decreased locomotor activity were determined. Necropsy revealed ascitic, accumulation of pleural fluid and dark purple discoloration of the liver and kidneys. Furthermore, 1 high-dose male died at week 12 of dosing without showing clinical signs of toxicity.

WATER CONSUMPTION
Statistically reduced water consumption was determined in the F1 generation in all dose groups: Low-dose males showed significantly decreased water consumption (evident during 3-6, 8, 10 weeks of dosing). Water consumption of mid-and high-dose animals was decreased throughout the dosing period. Decreased water consumption was determined in F1 females of the low-dose group (evident during 9 - 10 weeks of dosing), the mid-dose group (evident during week 10 of dosing and week 3 of lactation) and the high-dose group (evident throughout the entire dosing period).

FOOD CONSUMPTION
Significantly reduced food consumption was determined in the mid- and high-dose group either in week 10 of dosing (males) or week 3 of lactation (females). Moreover, reduced food consumption was observed in low-dose females during week 6 of dosing.

BODY WEIGHT
Body weights were comparable among the groups except for F1 low-dose females which revealed an increased body weight during week 6 - 8 compared to controls.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
Estrous cycles were comparable in F1 females among the groups although some control and test animals had persistent diestrus.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
No effects on the evaluated sperm parameters were determined (the number of cauda epididymal sperm, number of testis sperm, percentage of motile sperm and progressively motile sperm, swimming speed and pattern and percentage of morphologically abnormal sperm).

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Reproductive performance was comparable among the groups: copulation (males, females), fertility (males, females), gestation index, the precoital interval, gestation length, delivery index, the number of implantations, number of litters or pups delivered were not affected by treatment.

ORGAN WEIGHTS
Absolute organ weight of adrenals was significantly decreased in adult F1 males of the high-dose group compared to control animals. Mid-dose males showed significantly reduced absolute testis weight.
Organ weights of females of the F1 generation did not differ from controls.

PATHOLOGY
No dose-related gross lesions were observed in the F1 generation. Reproductive organs revealed no compound-related alterations in adults of the F1 generation.
Dose descriptor:
NOAEL
Remarks:
fertility
Generation:
F1
Effect level:
>= 3 000 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 38.5 (males) and 55.6 (females) mg Al/kg bw/day, F1 generation
Sex:
male/female
Basis for effect level:
viability
Dose descriptor:
LOAEL
Generation:
F1
Effect level:
3 000 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 38.5 (males) and 55.6 (females) mg Al/kg bw/day
Sex:
male/female
Basis for effect level:
sexual maturation
body weight and weight gain
organ weights and organ / body weight ratios
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
600 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 9.78 (males) and 14.0 (females) mg Al/kg bw/day
Sex:
male/female
Basis for effect level:
other: developmental toxicity
Clinical signs:
no effects observed
Mortality / viability:
mortality observed, non-treatment-related
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The bodyweight of female F2 pups were lower than controls on PND 21 (3,000 ppm). No sinificant differences in bodyweight of male F2 pups between control and AS treated groups were noted.
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
liver, spleen, thymus, kidney, testes, epididymides, uterus, ovary and brain weights were affected by treatment.

In males, the absolute and relative weights of the thymus and spleen were significantly decreased in the 3000 ppm group. Significant decreases were also found in the absolute weight of the liver and epididymides at 3000 ppm. The relative brain weight was significantly increased at this dose. At 120 ppm, the only significant change was a non-dose-related decrease in the relative thymus weight.

In females, there were significant decreases in the absolute and relative weights of the liver, and the absolute weight of the spleen, ovary and uterus, and a significant increase in the relative brain weight at 3000 ppm. In addition, a significant decrease in the absolute brain weight was observed only in the 600 ppm group.

Gross pathological findings:
no effects observed
Dose descriptor:
LOAEL
Generation:
F2
Effect level:
3 000 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 38.5 (males) and 55.6 (females) mg Al/kg bw/day
Sex:
male/female
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
Dose descriptor:
NOAEL
Generation:
F2
Effect level:
600 ppm (nominal)
Based on:
test mat.
Remarks:
equivalent to 9.78 (males) and 14.0 (females) mg Al/kg bw/day
Sex:
male/female
Basis for effect level:
other: no adverse effects noted
Reproductive effects observed:
not specified

Table 1.Organ weights of F1 and F2 male weanlings

 

Control

120 ppm

600 ppm

3000 ppm

F1

F2

F1

F2

F1

F2

F1

F2

Number of animals

22

21

20

18

22

22

22

21

Body weight (%)

100%

100%

NS

NS

NS

87.44**

90.31**

Brain

Absolute weight (%)

100%

100%

NS

NS

NS

NS

NS

NS

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

113.22**

112.11**

Thymus

Absolute weight (%)

100%

100%

NS

NS

NS

81.33**

79.84**

Relative weight (%) (g/100 g bw)

100%

100%

NS

89.29*

NS

NS

NS

87.92**

Liver

Absolute weight (%)

100%

100%

NS

NS

NS

NS

80.60**

87.78**

Relative weight (%)(g/100 g bw)

100%

100%

NS

NS

NS

NS

91.61**

NS

Kidneya

Absolute weight (%)

100%

100%

NS

NS

NS

NS

89.62**

NS

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

Spleen

Absolute weight (%)

100%

100%

NS

NS

NS

NS

76.40**

80.43**

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

86.93**

88.36**

Testisa

Absolute weight (%)

100%

100%

NS

NS

NS

NS

90.44*

NS

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

Epididymisa

Absolute weight (%)

100%

100%

NS

NS

NS

NS

88.02**

93.62*

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

NS: no statistically significant differences were observed

**: significantly different from the control, p<0.01;

*: significantly different from the control, p<0.05;

a: total weights of the organs on both sides.

Table 2. Organ weights of F1 and F2 female weanlings

 

 

Control

120

600

3000

F1

F2

F1

F2

F1

F2

F1

F2

Number of animals

22

22

20

18

22

21

21

21

Body weight (%)

100%

100%

NS

NS

89.91**

91.34**

Brain

Absolute weight (%)

100%

100%

NS

NS

NS

102.5*

NS

NS

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

110.20**

110.05**

Thymus

Absolute weight (%)

100%

100%

NS

NS

NS

NS

81.72**

NS

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

Liver

Absolute weight (%)

100%

100%

NS

NS

NS

NS

84.80**

86.24**

Relative weight %) (g/100 g bw)

100%

100%

NS

NS

NS

NS

94.26*

94.56**

Kidneya

Absolute weight (%)

100%

100%

NS

NS

NS

NS

NS

NS

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

Spleen

Absolute weight (%)

100%

100%

NS

NS

NS

NS

86.65*

84.11**

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

Ovarya

Absolute weight (%)

100%

100%

NS

NS

NS

NS

NS

84.52**

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

Uterus

Absolute weight (%)

100%

100%

NS

NS

83.85*

NS

78.47**

81.49*

Relative weight (%) (g/100 g bw)

100%

100%

NS

NS

NS

NS

NS

NS

NS: no statistically significant differences were observed

**: significantly different from the control, p<0.01;

*: significantly different from the control, p<0.05;

a: total weights of the organs on both sides.

Conclusions:
Interpretation of the results of this study is complicated by the effect of aluminium sulphate treatment on fluid consumption. It is noted that high concentrations of aluminium sulphate led to a reduction in the pH of the drinking water which may have reduced the palatability. In addition food consumption of F0 and F1 females was also reduced relative to the controls during week 3 of lactation.

Therefore reductions in food and water consumption during lactation mean that observed effects in the F1 and F2 generations (such as reduced organ weights) may be secondary effects rather than direct effects of aluminium sulphate consumption. Therefore, the utility of this data as stand-alone data is limited.
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
25 Sep 2001 - 24 Jun 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

The source substance and the target substance are considered to be similar enough to facilitate read-across for the following reasons:
(1) Both substances are inorganic salts containing a trivalent aluminium cation and phosphoric acid. Thus, they all share the Al3+ cation and the PO43- anion as common functional groups.
(2) Both substances will ultimately dissociate into the common breakdown products of the Al3+ cations and the PO43- anion.
(3) In general, independently of the cation under consideration, the water solubility of phosphates decreases with increasing degree of phosphate condensation (orthophosphate > diphosphate > triphosphate > polyphosphate).
In accordance with the provisions set out in Annex XI, Section 1.5, the results of the studies used for assessment and read-across are adequate for the purpose of classification and labelling and/or risk assessment; have adequate and reliable coverage of the key parameters addressed in the corresponding test method; cover an exposure duration comparable to or longer than the corresponding test method; and adequate and reliable documentation of the applied method is provided in the technical dossier.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Crj:CD(SD)IGS
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Atsugi Breeding Centre of Charles River Japan
- Age at study initiation: 8 weeks on arrival, 10 weeks at study initiation
- Weight at study initiation: 359-409 g (males), 210-252 g (females), after acclimation period
- Housing: Animals were housed in metal cages (260W x 380D x 180H, mm) at 2 males per cage and 2 females per cage in the quarantine and acclimation periods, 1 animal per cage after group allocation, 1 male and 1 female per cage in the copulation period, 1 dam per cage in the gestation period and 1 litter per cage in the nursing period.
- Diet: γ-Irradiated solid fed CRF-1 from Oriental Yeast Co. Ltd., ad libitum
- Water: Sapporo city water, ad libitum
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3
- Humidity (%): 50±20
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
other: 0.5% carmellose sodium in purified water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Each dose of triphosphoric acid aluminium salt was accurately weighed, finely ground in a mortar then suspended using the control substance to the prescribed concentration and dispersed using a stirrer. A mask and rubber gloves were worn at the time of preparation and procedures were carried out on a clean bench. The prepared solution was put into an airtight container protected from light, stored in a cool dark place (actual range 2-8°C) and used in administration within 1 week after returning it to room temperature. Remaining dosing solution was disposed of by incineration.

VEHICLE
- Concentration in vehicle: 10, 30 and 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 14 days
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after unsuccessful attempts: no
- After successful mating each pregnant female was caged (how): individually (1 dam per cage during gestation and 1 litter per cage during nursing)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Preparation of dosing solution and chemical analysis

(1) Preparation of dosing solution
Each dose of triphosphoric acid aluminium salt was accurately weighed, finely ground in a mortar then suspended using the control substance to the prescribed concentration and dispersed using a stirrer. A mask and rubber gloves were worn at the time of preparation and procedures were carried out on a clean bench. The prepared solution was put into an airtight container protected from light, stored in a cool dark place (actual range 2-8°C) and used in administration within 1 week after returning it to room temperature. Remaining dosing solution was disposed of by incineration.

(2) Chemical analysis of dosing solution
The uniformity of 1 and 200mg/mL dosing solutions of triphosphoric acid aluminium salt prior to administration and the stability of 7-day storage in a cool dark place were consequently analysed using concentration analysis method as below. As a result, the uniformity of prepared solutions of 1 and 200mg/mL triphosphoric acid aluminium salt and the stability of storage for 7 days in a cool dark place were confirmed.
The results of analysis of the concentration of triphosphoric acid aluminium salt in prepared solutions at each concentration at the initial and final times of preparation and used for dosing showed a content of 100.0-105.7% which was within the judgement criterion (85-115%).

(3) Concentration analysis method
Phosphorus standard solution (P: 100mg/L, for use in water quality tests, Wako Pure Chemical Industries) was dissolved in distilled water (Wako Pure Chemical Industries), a standard solution was prepared by adding a colouring agent and absorbance (wavelength 405nm) was measured using an ultraviolet-visible spectrophotometer (UV-160A, Shimadzu Corp.).
A fixed volume of the prepared test substance solution is accurately collected, 35% sodium hydroxide solution, distilled water and nitric acid (1/2) solution were added and this formed the decomposition solution.
After centrifugation of this decomposition solution at 3500rpm, the supernatant was collected then distilled water, nitric acid (1+1) solution and a colouring agent were added and absorbance was measured.
In addition, nitric acid (1+1) solution was added to distilled water to form a blank test solution and the absorbance of this blank test solution was measured. In stability and concentration confirmation tests, the middle layer of the prepared test substance solution was sampled 3 times and the absorbance was measured once after each preparation. In the uniformity test, the prepared test substance solution upper layer and lower layer were each sampled 3 times, and the absorbance of each was measured once after each preparation.
The concentrations of AlH2P3O10x2H20 in the sample solution and in the prepared test substance solution were determined from the absorbance of the standard solution and test sample solution and the content and coefficient of variation were calculated. The content was 85-115% and the coefficient of variation was within 5% and was accepted.
Concentration of AlH2P3O10x2H20 in sample solution (mg/mL) = (absorbance of sample solution - BL)/(mean absorbance of a standard solution - BL) x
standard solution concentration (0.01mg/mL) x conversion factor

BL: mean absorbance of blank test solution
conversion factor (P → AlH2P3O10x2H20): 317.95/ (30.97x3) = 3.422

Concentration of prepared test substance solution (mg/mL) = concentration of AlH2P3O10x2H20 in sample solution x dilution factor
Dilution factor: Total volume of sample solution (mL) x total volume of decomposition solution (mL) / amount of decomposition solution collected (mL) / amount of prepared test substance solution collected (mL)

Content (%) = (mean concentration of prepared test substance solution) / (displayed concentration of prepared test substance solution) x 100
Coefficient of variation (%) = standard deviation of concentration of prepared test substance solution) / (mean concentration of prepared test substance solution) x 100
Duration of treatment / exposure:
males: 46 days (from 14 days prior to mating)
females: from 14 days prior to mating, through mating, gestation and delivery up to Day 4 of lactation
Frequency of treatment:
daily
Details on study schedule:
not appropriate for screening study
Remarks:
Doses / Concentrations:
100, 300 and 1000 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: From the results of a preliminary 14-day experiment in which oral administration at a volume of 10mL/kg of suspensions of triphosphoric acid aluminium salt in 0.5% CMC at doses of 0, 100, 300 and 1000mg/kg were administered to 3 male and 3 female SD rats (Crj:CD(SD)IGS) per group, effects of test substance administration were not observed in any treatment group. Therefore, in both males and females, 1000mg/kg was established as the high dose, 300 and 100mg/kg were established at the intermediate and low doses in the same way as in the preliminary experiment and a group given 0.5% CMC was included as a control making a total of 4 groups.
- Rationale for animal assignment (if not random): After the completion of quarantine and acclimation, 40 healthy males and 40 healthy females (showing no abnormalities in the oestrus cycle) were selected and were used in experiments at 10 weeks of age. Group allocation was carried out using a stratified random sampling method based on body weight at the end of the quarantine and acclimation periods (the day before dosing) so that the mean body weight was uniform. The body weight range at the time of group allocation was 359-409g for males and 210-252g for females and these was within ±20% of the mean body weights (384.1g for males, 228.1g for females). Animals excluded from selection were excluded from the study.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least 4 times a day from Day 1 of administration until the autopsy on the day following Day 46 of administration.
- Cage side observations included: viability, external appearance and behaviour

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: on Days 1, 2, 5, 7, 10 and 14 of administration, every 7 days thereafter before administration, at the end of dosing and on the day of autopsy

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

WATER CONSUMPTION: Yes
- Time schedule for examinations: Days 43-44 of administration

For further information on parental examinations, please refer to 7.5.1 "Repeated dose toxicity"

Oestrous cyclicity (parental animals):
For evaluation of estrous cycle, vaginal smears were prepared in all females daily from day 10 before the start of administration until copulation. Determination of the stage of the estrous cycle (pro-oestrus, the period before estrus, metestrus and diestrus) was determined using a light microscope. Each stage of estrous which was repeated twice or more in the interval from day 4 to day 6 was judged to be normal and the normal estrous period was calculated. When some stages were not observed and when the stage of the same estrous cycle continues for in excess of 3 days, then an anomaly, estrous or diestrous continuing for 7 days or more were taken as constant estrous or constant diestrous which was judged to be abnormal.
Sperm parameters (parental animals):
Parameters examined in male parental generations:
The following reproductive tissues and organs were weight and evaluated at autopsy:
testes (left and right), epididymides (left and right), prostate gland, seminal vesicles and coagulating gland (left and right)
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in offspring: number of surviving offspring and number of dead neonates (determined once a day for each dam on day 0 to day 4), sex of pups and external appearance, weight gain (on day 0, 1 and 4), mean body weight (determined for each litter separately for males and females)


GROSS EXAMINATION OF DEAD PUPS:
yes, for external abnormalities
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals were sacrificed on day 46 of administration.
- Maternal animals: All surviving animals were sacrificed on day 46 of administration.

GROSS NECROPSY
- Gross necropsy consisted of macroscopic evaluation of the following organs and tissues: brain (cerebrum/cerebellum), pituitary gland, thymus, thyroid (including the parathyroid gland, left and right), adrenal glands (left and right), spleen, heart, thoracic aorta, tongue, oesophagus, stomach (forestomach and glandular stomach), liver, pancreas, duodenum, jejunum, ileum (including Peyer's patches), caecum, colon, rectum, larynx, trachea, lung (including bronchus), kidneys (left and right), urinary bladder, testes (left and right), epididymides (left and right), prostate gland, seminal vesicles and coagulating gland (left and right), ovaries (left and right), uterus (horn and neck), vagina, eyeballs and Harder's glands (left and right), skin (taken from the right abdomen), sternal and femoral bone (including bone marrow), spinal cord (neck), mesenteric lymph nodes, mandibular lymph nodes (left and right), submaxillary salivary glands (left and right), sublingual glands (left and right), parotid glands (left and right), skeletal muscle (gastrocnemius muscle), sciatic nerve (right).
Additionally (males): abdominal muscle seen in an umbilical hernia (No. 404).
Additionally (females): sites of macroscopic abnormalities (including normal tissue and boundary area) as well as the spleen adhesion tissue and pancreaticosplenic lymph nodes tissue in No. 351 and left axilla skin in No. 352.

HISTOPATHOLOGY / ORGAN WEIGHTS
Males
Slices from all animal were prepared after embedding in paraffin and staining with haematoxylin-eosin then specimens from all animals in groups 1 and 4 were microscopically examined. All cases where the testes and epididymides showed changes thought to be effects of test substance administration and animals in other treatment groups were microscopically examined. The liver of 1 animal (No. 208) in the 100mg/kg group which showed abnormal finding at autopsy was also microscopically examined.
Females
The same method as used in males was carried out. A microscopic examination was carried out on the pancreas, liver, spleen, pancreaticosplenic lymph nodes and the retroperitoneum in 1 animal (No. 351) in the 300mg/kg treatment group which did not exhibit pregnancy up to day 25 of pregnancy, on the glandular stomach of 1 animal (No. 359) among all dead offspring in the 300mg/kg treatment group, and on the brain of 1 animal (No. 256) showing abnormalities in the 100mg/kg treatment group at autopsy and on the spleen and mammary gland of 1 animal (No. 352) in the 300mg/kg treatment group.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring was sacrificed at 4 days of age following observation of the bodily appearance (including the oral cavity) .

GROSS NECROPSY
- Gross necropsy of sacrificed offspring consisted of systemic organs and tissues.
Dead animals were autopsied immediately on discovery.

Statistics:
Mean values and standard deviations were calculated. For statistical evaluation, Bartlett’s test was used for body weight/weight gain, growth rate, feed consumption, urinalysis, haematology (excluding leucocyte percentage), biochemistry, absolute and relative organ weights, estrous cycle, number of corpora lutea, number of implantation sites and implantation index, number of offspring, number of surviving offspring and dead neonates, delivery index, live birth index, sex ratio, gestation period, number of surviving offspring and viability index.
In the case of equal variance, analysis was carried out using One-Way analysis of variance and, in the case of unequal variance, the Kruskal-Wallis test.
If the results of One-Way analysis of variance showed no significant difference, a Dunnett’s test was used for further analysis. If the results of the Kruskal-Wallis method showed no significant difference, a Mann-Whitney U-test was applied.
The live birth index, sex ratio, viability index and body weights were dealt with taking 1 litter as a sample unit.
For the rate of occurrence of abnormalities in the estrous cycle, the copulation index, fertility index, gestation index, nursing index and histopathological findings showing a grade of 1 or higher, the multiple sample χ2 test was carried out followed by a 2-sample χ2 test. However, if there were mismatches in the 2-sample χ2 test, Fisher's exact test was used.
The level of significance was taken as 5% in comparison test with the control group.
Reproductive indices:
The following reproductive indices were determined:
copulation index (%): (number of paired males and females which copulated/number of males and females housed together) x 100
gestation index: (number of females with live births/number of pregnant females) x 100
fertility index (%): number of females which conceived/number of males and females which copulated) x 100
delivery index: (number of offspring/number of implantation sites) x 100
The following reproductive parameters were determined:
number of corpora lutea
number of implantation sites
gestation period (calculated as the number of days from day 0 of pregnancy (the day copulation was established) to day 0 of nursing (the day delivery was completed)
gestation index (%): number of females with live births/number of pregnant females) x 100
delivery index (%): number of offspring/number of implantation sites) x 100
implantation index (%): number of implantation sites/number of corpora lutea) x 100

Offspring viability indices:
The following offspring indices were determined:
viability index (%): (number of surviving offspring on day 4 of nursing/the number of live offspring at birth) x 100; cannibalism or unexplained disappearance of neonates was treated as a death
live birth index (%): (number of live offspring at birth/number of offspring) x 100
nursing index (%): (number of females with nursing pups on day 4 of nursing/the number of females with live births) x 100
sex ratio: number of male offspring/(the number of male offspring + the number of female offspring)
Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
see Details on results
Other effects:
no effects observed
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
effects observed, treatment-related
Description (incidence and severity):
see Details on results
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
see Details on results
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
Males
Although changes were not observed in the 100 and 300mg/kg treatment groups, central abdominal swelling and soiling of the perioral fur were observed in 1 animal (No. 404) in the 1000mg/kg treatment group on the day after the final dose (day of autopsy).
Females
Although changes were not observed in the 100 and 1000mg/kg treatment groups, 1 animal in the 300mg/kg treatment group showed a subcutaneous mass on day 13 of pregnancy and thereafter.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Males
Significant differences in changes in body weight, amount of body weight gain and rate of body weight gain were not seen in any group during the study period compared to the control group.
Females
Significant differences compared to the control group were not seen in body weight change, amount of body weight gain and rate of body weight gain in each treatment group in the administration period before pregnancy, gestation period and nursing period.

TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)
Not applicable (the test substance was aministered via gavage).

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
No effects on estrous cycle were noted in females of any dose group.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Copulation index, gestation index and nursing index were comparable among the groups. In the 300mg/kg treatment group, 1 animal did not give birth up to day 25 of pregnancy. 2 dead foetuses and 5 surviving foetuses were found within the uterus of this dam at autopsy on day 26 of pregnancy. As surviving fetuses were present in the uterus and no comparable effect was observed in the higher dose group, this effect is not considered as adverse but rather incidental. Further, 1 animal delivered 7 dead foetuses on day 25 of pregnancy thereby reducing the gestation index to 90% compared to 100% in the control and remaining treatment groups, including the high-dose group. Due to the incidental occurrence in the mid-dose group, the decrease in gestation index is not considered as adverse.
Fertility index in the control, low- and mid-dose group reached 100% although 2 control and 4 mid-dose males revealed testis atrophy in histopathology. A decrease in fertility index was determined in the high-dose group, which might be related to infertility of 1 male (severe testis atrophy) and vaginal closure/inflammation in uterus horn in 1 female. Thus, due to the low incidence of infertility (1/10 males and 1/10 females), spontaneous occurrence rather than a treatment-related effect on fertility is considered as reason for infertility. Moreover, based on a comparison with historical control data (Ema et al., 2014; please refer to Table 4), the decreased fertility index observed in the high-dose group is not considered to be of biological significance as pregnancy rates ranging from 80 - 100% were evaluated for the respective tester strain and study period (depending on the feed source).
Number of corpora lutea, number of implantations, implantation index, delivery index and total number of offspring were not affected by treatment. The mid-dose group revealed slightly reduced results in regard to several parameters (incl. number of corpora lutea, number of implantations, number of dams with live offspring, total number of offspring, delivery index, surviving number and delivery index). However, due to a missing dose-response, the observed alterations are considered as incidental and not-treatment related, especially as most parameters fall into the range of historical control data (Ema et al., 2014, please refer to Table 4).

ORGAN WEIGHTS (PARENTAL ANIMALS)
Males
Significant differences were not observed in any organ in any treatment group compared to the control group.
Females
Although the absolute and relative weights of the spleen showed a significantly lower value in the 100 and 1000mg/kg treatment groups compared to the control group, significant differences were not seen in the 300mg/kg treatment group. Thus, due to a missing correlation to a dose-response, the alterations in spleen weights are not considered as adverse.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Males
Deformity of the liver and multifocal fine yellowish-white spots were seen in 1 animal (no. 208) in the 100mg/kg group.
1 Animal (No. 303) in the 300mg/kg group showed atrophy of the bilateral testes and epididymides.
In the 1000mg/kg treatment group, 1 animal (No. 401) showed atrophy of the bilateral testes and epididymides and 1 animal (No. 404) showed umbilical hernia and a dark red protrusion in part of the ileum.
Pregnancy was established in a female (No. 353) paired with No. 303 who showed atrophy of the testes and epididymides and pregnancy in a female (No. 451) paired with No. 401 was not established.
Females
In the 100mg/kg treatment group, 1 animal (No. 256) showed dilation of both cerebral ventricles.
In the 300mg/kg treatment group, 1 animal (No. 352) showed swelling of the spleen and a subcutaneous white mass. Dark reddish macules were seen in the glandular stomach in 1 animal (No. 359) which was among 7 dead offspring delivered, in 1 animal (No. 351) in which delivery was not seen on day 25 of pregnancy, swelling of the liver, deformity of the spleen, adhesion of the intraperitoneal organs (spleen, pancreas, panniculus and retroperitoneum) and swelling of the pancreaticosplenic lymph nodes were observed.
In the 1000mg/kg treatment group, 1 animal (No. 455) showed an ileal diverticulum. In one infertile animal (No. 458), closure of the vagina and dilation of the uterus were seen and retention of a yellowish-white fluid was seen within the uterus.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Males
1 Animal (No. 401) in the 1000mg/kg treatment group showed severe atrophy and interstitial oedema in the seminiferous tubules in the testes, a severe decrease in sperm in the epididymides and intermediate intraluminal cell debris, and this animal did not establish pregnancy with a paired female. Moreover, atrophy of testes of milder severity was observed in 2 control and 4 mid-dose males. However, as all affected control- and mid-dose males were fertile, including the mid-dose male with intermediate atrophy of the seminiferous tubules in the testes and an intermediate decrease in sperm in the epididymides, atrophy of testes observed in the control and mid-dose group is not considered as adverse. Due to missing dose-response and occurence in the control group, atrophy of testes observed in the high-dose group is considered rather incidental than treatment-related.
Mild myocardial degeneration was seen in 1 other animal (No. 408) in which pregnancy with a paired female was not established.
In the low dose group, no histopathological alterations were observed.
Females
In the 1000mg/kg treatment group, 1 animal (No. 256) showed dilation of the cerebral ventricles.
In the 300mg/kg group, 1 animal (No. 352) showed mild extramedullary haemopoiesis in the spleen and adenoma in the mammary gland. In 1 animal (No. 351) in which delivery was not seen, mild extramedullary haemopoiesis in the liver and spleen and intermediate necrosis in the spleen were seen, and 1 animal (No. 359) among all nursing infant deaths showed mild ulceration of the glandular stomach.
Localised necrosis of the liver and atrophy of the thymus were sporadic in the 1000mg/kg treatment group. In 2 animals (Nos. 451 and 458) for which pregnancy was not established, No. 451 showed mild vitreous casts in the kidneys and No. 458 showed mild inflammation of the uterine horn and neck of the uterus and vaginal closure. As inflammatory changes were not seen for the vaginal closure, this was judged to be congenital.

For details, please refer to Tables 1 -4 in the attachment.
Dose descriptor:
NOEL
Remarks:
fertility
Effect level:
>= 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no adverse effects observed on fertility (for details, please refer to "Details on results")
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
VIABILITY (OFFSPRING)
The number of surviving offspring on day 0 of nursing, live birth index, number of surviving offspring on day 4 of nursing and viability index were comparable among the groups. Neonatal deaths at the end of delivery were observed in the control group (2 males and 1 female) and in the 300 mg/kg treatment group (3 males, 4 females and 1 cannibalised neonate of unknown sex).
Deaths or unclear cases in the period up to day 4 of nursing was observed in the control group (4 males and 2 females), in the 100mg/kg treatment group (1 male) and in the 1000mg/kg treatment group (1 female) .

CLINICAL SIGNS (OFFSPRING)
The general condition of offspring was not affected by treatment.

BODY WEIGHT (OFFSPRING)
No effect on body weight was noted in any dose group.

PATHOLOGY (OFFSPRING)
No effects were noted in the autopsy of neonates despite, injury to the lower jaw and a skin crust observed in 1 female in the 100mg/kg treatment group and loss of the tail was seen in 1 female in the 1000mg/kg treatment group.
Dose descriptor:
NOEL
Generation:
F1
Effect level:
>= 1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects noted
Reproductive effects observed:
no

A summary of experimental results is attached (Tables 1- 3). For further details on results, please refer to 7.5.1 "Repeated dose toxicity".

Historical control data are shown in Table 4 (please refer to the attachment).

Conclusions:
Effects of test substance administration were not seen on the reproductive ability of parents and birth of neonates in any treatment group. Consequently, the no observed effect level (NOEL) of repeated dose triphosphoric acid aluminium salt in reproduction in parents and the no observed effect level (NOEL) in the birth of neonates was considered to be 1000mg/kg/day under these experimental conditions.
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 1 - 2) and consistent studies from the target substance and a reference substances with similar structure and intrinsic properties. Read-across is justified based on common functional group(s), common precursors/breakdown products, similarities in PC/ECO/TOX properties (refer to endpoint discussion for further details). The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No. 1907/2006.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Read-across is justified in the endpoint records and in the read-across report attached in Section 13 of this technical dossier.

Effects on developmental toxicity

Description of key information

The dataset submitted is a weight of evidence approach using data on the following source substances; aluminium hydroxide, aluminium citrate and sodium dihydrogenorthophosphate. All studies performed were similar to the OECD 414 guideline or were conducted to older teratogenicity testing and have therefore been assigned a Klimisch reliability 2. None of the studies resulted in developmental toxicity and the NOAELs in all studies were based on the top dose level.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
06/10/1974 - 02/11/1974
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Deficiencies: Food consumption not reported Uterine weights not determined One third used for visceral examination; should be 50% Test substance identification (Batch etc) missing No details on housing conditions/source of animals Administration only during periods of organogenesis, not until day before pregnancy
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or

The source substance and the target substance are considered to be similar enough to facilitate read-across for the following reasons:
(1) Both substances are inorganic salts containing the PO43- anion as a common functional group.
(2) Both substances will ultimately dissociate into the common breakdown products of the PO43- anion. The addition of the sodium ion in the source substance is not considered to have an impact on the toxicological profile since Na+ is a common physiological ion.

In accordance with the provisions set out in Annex XI, Section 1.5, the results of the studies used for assessment and read-across are adequate for the purpose of classification and labelling and/or risk assessment; have adequate and reliable coverage of the key parameters addressed in the corresponding test method; cover an exposure duration comparable to or longer than the corresponding test method; and adequate and reliable documentation of the applied method is provided in the technical dossier.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: no data
Deviations:
not specified
Principles of method if other than guideline:
Adult female albino CD-1 mice were mated with young adult males. Observation of a vaginal sperm plug was considered as Day 0 of gestation. Dosing by oral intubation with a control (Vehicle at level equivalent to group receiving the highest dose or aspirin at 150 mg/kg) or test article in a water suspension (10 mL/kg bw) at 3.7, 17.2, 79.7 or 370.0 mg/kg was carried out daily on Days 6 to 15 of gestation. Observations of body weight, appearence, behaviour, and food consumption were performed. Daily room temperature and humidity were recorded. On Day 17 of gestation all dams underwent Caesarean section. Sex, number of corpora lutea, implantation sites, resorption sites and live/dead foetuses recorded. Body weights of live pups recorded. Urogenital tract of each dam examined for anatomical normality. All foetuses examined grossly for presence of external congenital abnormalities. One third foetuses of each litter underwent detailed visceral examination and the remaining two thirds were cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.
GLP compliance:
no
Remarks:
Study predates GLP
Limit test:
no
Species:
mouse
Strain:
other: albino CD-1
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Outbred
- Age at study initiation: No data
- Weight at study initiation: 29 - 31 g
- Fasting period before study: No data
- Housing: Gang housing in disposable plastic cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: No data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): ave 21 - 26
- Humidity (%): 42 - 74%

IN-LIFE DATES: From: 06/10/1974 To: 02/11/1974
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
VEHICLE: Water
- Amount of vehicle (if gavage): 10 mL/kg bodyweight
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: cohoused
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
Duration of treatment / exposure:
10 days (Day 6 to Day 15 of gestation)
Frequency of treatment:
Daily
Duration of test:
17 days
No. of animals per sex per dose:
Test material and vehicle control: 25 females / dose level
Positive control: 27 females
Table 1 Number of animals dosed
Material Dose (mg/kg) Total
Mated Pregnant
Sham 0.0 25 21
Aspirin 150.0 27 20
FDA 73-2 3.7 25 22
17.2 25 19
79.7 25 20
370.0 25 22
Control animals:
yes, sham-exposed
other: positive control: 150 mg/kg aspirin
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Appearence, behaviour, food consumption and weight observed daily.
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
- Time schedule for examinations: Body weights recorded on days 0, 6, 11, 15 and 17.
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 17
- Organs examined: uterus and urogenital tract
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes
Fetal examinations:
- External examinations: Yes: one third per litter
- Soft tissue examinations: Yes: one third per litter
- Skeletal examinations: Yes: two thirds per litter
- Head examinations: Yes: two thirds per litter
Statistics:
No data
Indices:
No data
Historical control data:
No
Details on maternal toxic effects:
Maternal toxic effects:no effects
Dose descriptor:
NOAEL
Effect level:
> 370 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no effects observed
Abnormalities:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Dose descriptor:
NOAEL
Effect level:
> 370 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Abnormalities:
no effects observed
Developmental effects observed:
no

Table 2 Reproduction data

Dose (mg/kg)

Sham

Aspirin

3.7

17.2

79.7

370.0

Pregnancies

 

 

 

 

 

 

Total No.

21

20

22

19

20

22

Died or aborted (before Day 17)

0

0

0

0

0

0

To term (on Day 17)

21

20

22

19

20

22

Corpora Lutea

 

 

 

 

 

 

Total no.

275

233

255

233

251

279

Average/dam mated

11.0

8.63

10.2

9.32

10.0

11.2

Live litters

 

 

 

 

 

 

Total No.*

21

20

22

19

20

22

Implant Sites

 

 

 

 

 

 

Total No.

259

218

240

224

236

255

Average/dam*

12.3

10.9

10.9

11.8

11.8

11.6

Resorptions

 

 

 

 

 

 

Total No*

8

8

5

9

10

13

Dams with 1 or more sites resorbed

8

6

5

4

9

11

Dams with all sites resorbed

--

--

--

--

--

--

Per cent partial resorptions

38.1

30.0

22.7

21.1

45.0

50.0

Per cent complete resorptions

--

--

--

--

--

--

Live foetuses

 

 

 

215

 

 

Total No

248

208

233

11.3

224

240

Average/dam*

11.8

10.4

10.6

1.01

11.2

10.9

Sex ratio (M/F)

1.04

0.84

0.82

 

0.84

1.07

Dead Foetuses

 

 

 

--

 

 

Total No.*

3

2

2

--

2

2

Dams with 1 or more dead

3

1

2

--

2

2

Dams with all dead

--

--

--

--

--

--

Per cent partial dead

14.3

5.00

9.09

--

10.0

9.09

Per cent all dead

--

--

--

--

--

--

Average foetus weight (g)

0.8

0.85

0.85

0.92

0.93

0.86

* Includes only those dams examined at term

** Positive control: 150 mg/kg

 

Table 3 Summary of skeletal findings

Findings

Dose (mg/kg)

Sham

Aspirin

3.7

17.2

79.7

370.0

Live foetuses examined (at term)

173/21

144/20

165/22

148/149

156/20

167/22

Sternebrae

 

 

 

 

 

 

Incomplete oss.

60/19

65/19

32/14

51/16

13/5

81/19

Scrambled

 

 

 

 

 

 

Bipartite

 

1/1

 

3/2

 

3/2

Fused

 

 

 

 

 

 

Extra

 

 

 

 

 

 

Missing

39/12

36/11

19/9

13/7

2/2

20/12

Other

 

 

 

 

 

 

Ribs

 

 

 

 

 

 

Incomplete oss.

 

 

 

 

 

 

Fused/split

 

 

 

1/1

 

 

Wavy

 

 

 

 

 

 

Less than 12

 

 

 

 

 

 

More than 13

23/10

32/14

23/12

25/10

30/16

25/15

Other

 

 

 

 

 

 

Vertebrae

 

 

 

 

 

 

Incomplete oss.

4/2

11/5

4/3

1/1

 

 

Scrambled

 

 

 

 

 

 

Fused

 

 

 

 

 

 

Extra ctrs. oss.

 

 

 

 

 

 

Scoliosis

 

 

 

 

 

 

Tail defects

 

 

 

 

 

 

Other

 

 

 

 

 

 

Skull

 

 

 

 

 

 

Incomplete closure

2/2

 

 

 

 

 

Missing

 

 

 

 

 

 

Craniostosis

 

 

 

 

 

 

Other

 

 

 

 

 

 

Extremities

 

 

 

 

 

 

Incomplete oss.

6/4

15/5

2/2

1/1

 

1/1

Missing

 

 

 

 

 

 

Extra

 

 

 

 

 

 

Miscellaneous

 

 

 

 

 

 

Hyoid; missing

30/12

60/16

24/12

26/11

27/11

43/15

Hyoid; reduced

26/12

18/13

22/13

26/16

19/10

22/14

* Numerator = Number of foetuses affected; Denominator = Number of litters affected

** Positive control: 150 mg/kg

Summary of soft tissue abnormalities: 1 pup from a litter where the dam was dosed with 3.7 mg/kg test material showed exophthalmos and encephalmeningocele.

Conclusions:
Under the conditions of the study, the test material administered to pregnant mice for 10 days up to a dose level of 370 mg/kg bw showed no maternal or developmental toxicity. The NOAEL for both maternal and foetal toxicity is > 370 mg/kg bw.

This study is considered to be of adewuate reliability for use as part of a weight of evidence for this endpoint.
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
27/09/1974 - 29/10/1974
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
Deficiencies: Food consumption not reported Uterine weights not determined One third used for visceral examination; should be 50% Test substance identification (Batch etc) missing No details on housing conditions/source of animals Administration only during periods of organogenesis, not until day before pregnancy
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or

The source substance and the target substance are considered to be similar enough to facilitate read-across for the following reasons:
(1) Both substances are inorganic salts containing the PO43- anion as a common functional group.
(2) Both substances will ultimately dissociate into the common breakdown products of the PO43- anion. The addition of the sodium ion in the source substance is not considered to have an impact on the toxicological profile since Na+ is a common physiological ion.

In accordance with the provisions set out in Annex XI, Section 1.5, the results of the studies used for assessment and read-across are adequate for the purpose of classification and labelling and/or risk assessment; have adequate and reliable coverage of the key parameters addressed in the corresponding test method; cover an exposure duration comparable to or longer than the corresponding test method; and adequate and reliable documentation of the applied method is provided in the technical dossier.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: no data
Deviations:
not specified
Principles of method if other than guideline:
Adult female albino (Wistar derived stock) rats were mated with young adult males. Observation of a vaginal sperm plug was considered as Day 0 of gestation. Dosing by oral intubation with a control (Vehicle at level equivalent to group receiving the highest dose or aspirin at 250 mg/kg) or test article in a water suspension at 4.1, 19.0, 88.3 or 410.0 mg/kg was carried out daily on Days 6 to 15 of gestation. Observations of body weight, appearence, behaviour, and food consumption were performed. Daily room temperature and humidity were recorded. On Day 20 of gestation all dams underwent Caesarean section. Sex, number of corpora lutea, implantation sites, resorption sites and live/dead foetuses recorded. Body weights of live pups recorded. Urogenital tract of each dam examined for anatomical normality. All foetuses examined grossly for presence of external congenital abnormalities. One third foetuses of each litter underwent detailed visceral examination and the remaining two thirds were cleared in potassium hydroxide, stained with alizarin red S dye and examined for skeletal defects.
GLP compliance:
no
Remarks:
Study predates GLP
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Outbred
- Age at study initiation: No data
- Weight at study initiation: 223 - 237 g
- Fasting period before study: No data
- Housing: Individual housing in mesh bottom cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: No data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 28
- Humidity (%): 42 - 74%

IN-LIFE DATES: From: 27/09/1974 To: 29/10/1974
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
VEHICLE: Water
- Amount of vehicle (if gavage): 1 mL/kg bw at doses equal to or below 250 mg/kg bw and 2 mL/kg at doses up to 500 mg/kg bw
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- Impregnation procedure: No data
- Proof of pregnancy: vaginal plug referred to as day 0 of pregnancy
Duration of treatment / exposure:
10 days (Day 6 to Day 15 of gestation)
Frequency of treatment:
Daily
Duration of test:
20 days
No. of animals per sex per dose:
Test material and vehicle control: 20 females / dose level
Control animals:
yes, sham-exposed
other: positive control: 250 mg/kg aspirin
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Appearence, behaviour, food consumption and weight observed daily.
DETAILED CLINICAL OBSERVATIONS: No
BODY WEIGHT: Yes
- Time schedule for examinations: Body weights recorded on days 0, 6, 11, 15 and 20.
POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus and urogenital tract
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of resorptions: Yes
Fetal examinations:
- External examinations: Yes: one third per litter
- Soft tissue examinations: Yes: one third per litter
- Skeletal examinations: Yes: two thirds per litter
- Head examinations: Yes: two thirds per litter
Statistics:
No data
Indices:
No data
Historical control data:
No
Details on maternal toxic effects:
Maternal toxic effects:no effects
Dose descriptor:
NOAEL
Effect level:
> 410 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no effects observed
Abnormalities:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects
Dose descriptor:
NOAEL
Effect level:
> 410 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects observed
Abnormalities:
no effects observed
Developmental effects observed:
no

Table 1 Reproduction data

Dose (mg/kg)

Sham

Aspirin

4.1

19.0

88.3

410.0

Pregnancies

 

 

 

 

 

 

Total No.

20

20

20

20

20

20

Died or aborted (before Day 20)

0

0

0

0

0

0

To term (on Day 20)

20

20

20

20

20

20

Corpora Lutea

 

 

 

 

 

 

Total no.

267

263

263

252

264

267

Average/dam mated

12.7

12.5

12.5

12.0

12.6

13.4

Live litters

 

 

 

 

 

 

Total No.*

20

19

20

20

20

20

Implant Sites

 

 

 

 

 

 

Total No.

262

238

240

251

239

244

Average/dam*

13.1

11.9

12.0

12.6

12.0

12.2

Resorptions

 

 

 

 

 

 

Total No*

--

35

1

5

1

2

Dams with 1 or more sites resorbed

--

7

1

3

1

1

Dams with all sites resorbed

--

1

--

--

--

--

Per cent partial resorptions

--

35.0

5.00

15.0

5.00

5.00

Per cent complete resorptions

--

5.00

--

--

--

--

Live foetuses

 

 

 

 

 

 

Total No

262

203

239

246

238

242

Average/dam*

13.1

10.2

12.0

12.3

11.9

12.1

Sex ratio (M/F)

1.06

0.92

1.03

0.95

1.00

1.12

Dead Foetuses

 

 

 

 

 

 

Total No.*

--

--

--

--

--

--

Dams with 1 or more dead

--

--

--

--

--

--

Dams with all dead

--

--

--

--

--

--

Per cent partial dead

--

--

--

--

--

--

Per cent all dead

--

--

--

--

--

--

Average foetus weight (g)

3.93

2.92

3.84

3.91

4.00

3.85

* Includes only those dams examined at term

** Positive control: 250 mg/kg

 

Table 2 Summary of skeletal findings

Findings

Dose (mg/kg)

Sham

Aspirin

4.1

19.0

88.3

410.0

Live foetuses examined (at term)

179/20

143/19

166/20

171/20

168/20

170/20

Sternebrae

 

 

 

 

 

 

Incomplete oss.

37/16

89/19

42/17

82/18

49/15

45/11

Scrambled

 

8/6

1/1

 

 

1/1

Bipartite

 

 

 

 

 

 

Fused

 

 

 

 

 

 

Extra

 

 

 

 

 

 

Missing

3/3

70/15

4/3

10/7

 

7/6

Other

 

 

 

 

 

 

Ribs

 

 

 

 

 

 

Incomplete oss.

2/2

15/8

7/2

 

 

1/1

Fused/split

 

5/2

 

 

 

 

Wavy

23/8

60/17

19/8

23/11

11/6

13/5

Less than 12

 

 

 

 

 

 

More than 13

2/2

59/15

 

4/4

 

1/1

Other

 

 

 

 

 

 

Vertebrae

 

 

 

 

 

 

Incomplete oss.

4/4

68/18

2/1

1/1

9/6

10/4

Scrambled

 

 

 

 

 

 

Fused

 

 

 

 

 

 

Extra ctrs. oss.

 

1/1

 

 

 

 

Scoliosis

 

 

 

 

 

 

Tail defects

 

 

 

 

 

 

Other

 

 

 

 

 

 

Skull

 

 

 

 

 

 

Incomplete closure

38/13

88/19

33/14

30/12

37/11

40/11

Missing

 

 

 

 

 

 

Craniostosis

 

 

 

 

 

 

Other

 

 

 

 

 

 

Extremities

 

 

 

 

 

 

Incomplete oss.

 

3/3

 

 

 

 

Missing

 

 

 

 

 

 

Extra

 

 

 

 

 

 

Miscellaneous

 

 

 

 

 

 

Hyoid; missing

21/10

54/16

21/12

19/10

12/8

20/11

Hyoid; reduced

26/13

21/10

33/14

28/11

41/14

47/14

* Numerator = Number of foetuses affected; Denominator = Number of litters affected

** Positive control: 250 mg/kg

 

Table 3 Summary of soft tissue abnormalities

Material

Dose level (mg/kg)

Dam

Number of pups

Description

Aspirin

250.0

44235

1

Hydrocephalus

 

 

44240

1

Hydrocephalus; Encephalomeningocele

 

 

44246

2

Hydrocephalus

 

 

44252

1

Spina bifida; Encephalomeningocele

 

 

44255

1

Hydrocephalus

FDA 73-2

88.3

44337

1

Umbilical hernia

 

Conclusions:
Under the conditions of the study, the test material administered to pregnant rats for 10 days up to a dose level of 410 mg/kg bw showed no maternal or developmental toxicity. The NOAEL for both maternal and foetal toxicity is > 410 mg/kg bw.

This study is considered to be of adequate reliability for use as part of a weight of evidence for this endpoint.
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

The source substance and the target substance are considered to be similar enough to facilitate read-across for the following reasons:
(1) Both substances are inorganic salts containing a trivalent aluminium cation.
(2) Both substances will ultimately dissociate into the common breakdown products of the Al3+ following chemical or biological hydrolysis and dissolution of the ionic bonds.
In accordance with the provisions set out in Annex XI, Section 1.5, the results of the studies used for assessment and read-across are adequate for the purpose of classification and labelling and/or risk assessment; have adequate and reliable coverage of the key parameters addressed in the corresponding test method; cover an exposure duration comparable to or longer than the corresponding test method; and adequate and reliable documentation of the applied method is provided in the technical dossier.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.

Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
Animals were only dose during the period of organogenesis.
GLP compliance:
not specified
Limit test:
no
Species:
mouse
Strain:
Swiss
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Panlab, Barcelona, Spain
- Age at study initiation: specific information not provided, animals were declared to be 'sexually mature'.
- Weight at study initiation: 28-32 g
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±2°C
- Humidity (%):50±10%
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
No information on preparation of dosing solutions provided.
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 5 females to 2 males
- Length of cohabitation: until copulation was detected
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
10 days - from day 6 to day 15 of gestation
Frequency of treatment:
daily
Duration of test:
Dams were killed on gestation day 18
Dose / conc.:
66.5 mg/kg bw/day
Dose / conc.:
133 mg/kg bw/day
Dose / conc.:
266 mg/kg bw/day
No. of animals per sex per dose:
20 dams per group
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: daily

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 18
- Organs examined: heart, lungs, spleen, liver, kidneys, brain
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 data
- Number of implantations: Yes

- Other: Live, dead and resorbed foetuses were recorded.
Fetal examinations:
- External examinations: Yes:
- Skeletal examinations: Yes [> half the viable foetuses were examined]
- Soft tissue examinations: Yes - remaining viable foetuses were evaluated
Statistics:
Maternal body weight gain and food consumption were evaluated by analysis of variance with differences among groups determined by Student's t test or Mann-Whitney U test. Nonparametric data were analysed by the Kruskal-Wallis test.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Dead fetuses:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Dose descriptor:
NOAEL
Effect level:
266 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no effects noted
Abnormalities:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Dose descriptor:
NOAEL
Effect level:
266 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects noted
Abnormalities:
no effects observed
Developmental effects observed:
no

Effects of aluminium hydroxide administered to mice on days 6 to 15 of gestation

Values are expressed as mean values ± SE

Parameter observed

Dose (mg/kg/day)

0

66.5

133

266

No. of litters

20

18

19

19

No of total implants/litter

11.5±0.7

12.4±0.7

11.8±0.7

11.1±1.0

No. of resorptions / litter

 

Early

0.4±0.2

3.0±1.7

2.5±1.1

1.3±1.1

Late

0.0±0.0

0.1±0.2

0.1±0.3

0.1±0.3

No. of live foetuses / litter

11.1±0.7

9.4±1.1

9.2±1.1

9.8±1.2

No. dead foetuses / litter

0.0±0.0

0.0±0.0

0.0±0.0

0.0±0.0

Sex ratio (M:F) / litter

1.34±0.25

0.96±0.07

1.02±0.17

1.13±0.25

Foetal body weight (g)

1.40±0.02

1.41±0.03

1.43±0.02

1.42±0.03

Foetal body length (cm)

3.11±0.02

3.11±0.03

3.25±0.03

3.10±0.03

Conclusions:
In the present study, aluminium hydroxide was administered at 3 dose levels up to 266 mg/kg bw day to pregnant mice during the period of organogenesis. No maternal toxicity nor developmental toxicity (embryotoxicity/teratogencity) were observed. Aluminium absorption from aluminium hydroxide is considered to be low due to the low water solubility.
Endpoint:
developmental toxicity
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
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

The source substance and the target substance are considered to be similar enough to facilitate read-across for the following reasons:
(1) Both substances are inorganic salts containing a trivalent aluminium cation.
(2) Both substances will ultimately dissociate into the common breakdown products of the Al3+ following chemical or biological hydrolysis and dissolution of the ionic bonds.
In accordance with the provisions set out in Annex XI, Section 1.5, the results of the studies used for assessment and read-across are adequate for the purpose of classification and labelling and/or risk assessment; have adequate and reliable coverage of the key parameters addressed in the corresponding test method; cover an exposure duration comparable to or longer than the corresponding test method; and adequate and reliable documentation of the applied method is provided in the technical dossier.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.

Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
yes
Remarks:
lack of test item characterisation. Animals were only dosed during organogenesis.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Interfauna Iberica (Barcelona, Spain)
- Weight at study initiation: 225-240 g
- Diet: Food (commercial chow, Panlab, Barcelona, Spain), ad libitum
- Water: tap water: ad libitum
- Acclimation period: 10 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23 °C
- Humidity (%): 45±5%
- Photoperiod (hrs dark / hrs light): 12 h light and 12 h dark
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: not reported.

Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
Not reported - rats were already pregnant.
Duration of treatment / exposure:
Pregnant rats were dosed from day 6 to day 15 of gestation
Frequency of treatment:
Daily
Duration of test:
Up tp gestation day 20.
Dose / conc.:
192 mg/kg bw/day
Remarks:
equivalent to 66.5 mg Al/kg bw/day
Dose / conc.:
384 mg/kg bw/day
Remarks:
equivalent to 133 mg Al/kg bw/day
Dose / conc.:
768 mg/kg bw/day
Remarks:
266 mg Al/kg bw/day
No. of animals per sex per dose:
20 females
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: 3 day intervals during pre-treatment, treatment and post-treatment period


BODY WEIGHT: Yes
- Time schedule for examinations:

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


POST-MORTEM EXAMINATIONS: Yes / No / No data
- Sacrifice on gestation day #
- Organs examined: absolute/relative organ weight (kidney, liver, gravid uterine weight).

OTHER: Maternal-placental-fetal Al contents following gestational exposure have been examined. Al concentrations (µg/g wet weight) in maternal liver, brain, bone, placenta; and whole fetus (detection limit 0.05 µg/g).
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes
- Soft tissue examinations: Yes
- Skeletal examinations: Yes (approximately 1/3 of foetuses)
- Head examinations: No data

Three foetuses from each dam were used for whole body analyses of Al by AAS.
Statistics:
Results of the quantitative continuous variables (e.g., maternal body weights, organ weights, fetal weights, etc.) were compared using analysis of variance (ANOVA) with significant F values further analyzed using Student’s t-test or Mann-Whitney U-test.
Nonparametric data were statistically evaluated using the Kruskal-Wallis test when appropriate. The incidence of developmental abnormalities was not analysed statistically because no differences were evident between treated and control groups.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Statistically significant decreases in maternal food consumption during the treatment periods (6-15) and during the whole study period. Differences were sporadic and not dose-related. Maternal stress due to gavage is most likely to be the cause.
Food consumption recovered during the post-treatment period on GD 15-20.
Food efficiency:
not examined
Haematological findings:
no effects observed
Description (incidence and severity):
Tabulated data not reported.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Tabulated data not reported.
Organ weight findings including organ / body weight ratios:
no effects observed
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
High variability was evident in all Al treated and control groups.
Description (incidence and severity):
Group I animals an increased number of early resorption (statistically non-significant) was reported.
Group III animals an increased number of early resorption (statistically non-significant) was reported.
Details on maternal toxic effects:
No Al related treatment effects were observed on maternal hematology and biochemical parameters at termination (GD 20).
No increased Al levels in maternal liver, bone, brain and placenta in Al treated animals compared to the control group was detected.
The maternal/placental Al concentrations were not statistically different between control and treated rats (high variability is evident in all groups).
Dose descriptor:
NOAEL
Effect level:
768 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: no effects noted
Remarks on result:
other:
Remarks:
dose equivalent to 266.0 mg Al /kg bw /day
Abnormalities:
no effects observed
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
External malformations:
no effects observed
Skeletal malformations:
no effects observed
Visceral malformations:
no effects observed
Dose descriptor:
NOAEL
Effect level:
768 mg/kg bw/day
Based on:
test mat.
Remarks:
Al content
Sex:
male/female
Basis for effect level:
other: No effects noted
Remarks on result:
other:
Remarks:
dose equivalent to 266.0 mg Al /kg bw /day
Abnormalities:
no effects observed
Developmental effects observed:
no
Conclusions:
The goal of study is to assess the developmental toxicity and embryotoxic/teratogenic potential of high doses of target compound (aluminium hydroxide) administered orally to rats during the period of active organogenesis.

No significant general/maternal toxicity was observed in any Al treated groups that were orally exposed to Al hydroxide at doses 66.5, 133 and 266 mg Al/kg bw/day.



Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
no examination of food consumption performed.
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
See read-across justification report under Section 13 ‘Assessment Reports’.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

The source substance and the target substance are considered to be similar enough to facilitate read-across for the following reasons:
(1) Both substances are inorganic salts containing a trivalent aluminium cation.
(2) Both substances will ultimately dissociate into the common breakdown products of the Al3+ following chemical or biological hydrolysis and dissolution of the ionic bonds.
In accordance with the provisions set out in Annex XI, Section 1.5, the results of the studies used for assessment and read-across are adequate for the purpose of classification and labelling and/or risk assessment; have adequate and reliable coverage of the key parameters addressed in the corresponding test method; cover an exposure duration comparable to or longer than the corresponding test method; and adequate and reliable documentation of the applied method is provided in the technical dossier.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report under Section 13 ‘Assessment Reports’.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report under Section 13 ‘Assessment Reports’.

4. DATA MATRIX
See read-across justification report under Section 13 ‘Assessment Reports’.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
equivalent or similar to guideline
Guideline:
other: OCED guideline 426 and 452
Deviations:
yes
Remarks:
(no examination of food consumption)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Housing: Animals were housed individually in shoebox or ventilated cages except during breeding (wire bottom cages) and until weaning (grouped with rest of litter) for the pups
- Diet: AIN-93G growth food (Purina TestDiet) until postnatal day (PND) 95-99, followed by AIN-93M (Purina TestDiet) maintenance food
- Acclimation period: at least 9 days
Route of administration:
oral: drinking water
Vehicle:
water
Remarks:
fresh filtered, de-ionized water adjusted to pH 6 - 7
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Fresh filtered solutions were prepared weekly using de-ionized water and the pH was adjusted between 6 and 7.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The final Al concentrations have been independently assessed using inductively coupled plasma mass spectrometric analyses by Maxxam Analytics Inc. (Burnaby, Canada). Specifically, aluminum (Al), iron (Fe), manganese (Mn), copper (Cu) and zinc (Zn) were quantified using a Perkin Elmer ELAN 6000 ICP-MS apparatus according to the manufacturer's standardized GLP protocols.
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: up to 5 consecutive nights
- Proof of pregnancy: vaginal plug
Duration of treatment / exposure:
Dams
GD 6 to PND 21

Pups
Cohort 1 – GD 6-21, PND 1-22
Cohort 2 – GD 6-21, PND 1-64
Cohort 3 – GD 6-21, PND 1-120
Cohort 4 – GD 6-21, PND 1-364
Frequency of treatment:
daily, 7 days/week
Duration of test:
1 year
No. of animals per sex per dose:
20 (dams)
80 (offspring, main study)
Control animals:
other: placebo control solution (contained of sodium citrate at concentration of 27.2 g/L, which mimics the citrate ionic strength of the highest aluminum citrate solution).
Details on study design:
- Dose selection rationale: doses were selected based on the results of a previous 90-day pilot toxicity study and the maximum solubility of aluminum citrate in water (high-dose).
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: all dams underwent daily morbidity and mortality checks and Functional Observational Battery (FOB) examinations on gestational Day (GD) 7 and 13 and PND 3 and 10.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: day of delivery

BODY WEIGHT: Yes
- Time schedule for examinations: GD 6, 13, 20 and PND 8, 15 and 22

WATER CONSUMPTION AND COMPOUND INTAKE: Yes
Fetal examinations:
PUPS
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: all pups underwent daily morbidity and mortality checks on a daily basis from PND 1 to PND 366.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: pups from the citrate and high-dose group at least weekly from PND 33 to PND 153

BODY WEIGHT: Yes
- Time schedule for examinations: on days of age 4, 11, 17, 29, 56, 84, 112, 140, 168, 196, 224, 252, 280, 308, 336 and 364

WATER CONSUMPTION AND COMPOUND INTAKE : Yes

FUNCTIONAL OBSERVATIONAL BATTERY (FOB)
- Time schedule: one male and female pup from each litter on PNDs 5, 11, 22, 36, 45 and 56 and biweekly thereafter until the week of PND 350
- Examinations/Tests: T-maze, Morris water maze and motor activity

OTHERS
Developmental landmarks:
Female pups were monitored for vaginal opening starting on PND 26. Male pups were monitored for preputial separation starting on PND 35.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: on the day of schedulded sacrifice
- Parameters checked: alanine aminotransferase, albumin, albumin/globulin ratio, alkaline phosphatase, aspartate aminotransferase, calcium, chloride, cholesterol, creatine kinase, creatinine, globulin, glucose, sorbitol dehydrogenase, phosphorus, potassium, sodium, total bilirubin, total protein, triglycerides, urea nitrogen were measured.
Statistics:
STATISTICS
Statistical analyses were conducted using SAS® Release 9.1 for Windows. Data collected on dams and pups were analyzed separately and all statistical analyses on the pups were performed separately for each sex. Analyses for pups were done separately for each test day group unless specified otherwise Statistical significance was declared when P ≤ 0.05. All statistical analyses were done based on the principle of intention-to-treat.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
MORTALITY
No mortality was observed in the dams during the gestation and postnatal period in all groups.

CLINCAL OBSERVATION
4 animals in the control group, 8 in the Na-citrate group, 4 in the low-dose group, 6 in the mid-dose group, and 12 in the high dose group exhibited clinical signs. Most signs were considered mild, for example alopecia and porphyrin staining. One dam from the Na-citrate group stopped nursing and was euthanized early. Diarrhoea was observed in 8 dams in the high-dose group.

BODY WEIGHT
There were no significant differences in mean body weights in dams between the treatment groups and controls during the gestational and postnatal period.

FLUID CONSUMPTION
During gestation and lactation, animals of the low- and mid-dose group consumed more fluid in comparison to the controls in week 1 and 2. The dose-dependent increases in fluid consumption in the Al-citrate groups was also seen in the Na-citrate group; consistent with a citrate ion-specific effect.

DOSAGE LEVEL
Actual dosage, although not at 100% of target followed fluid consumption which varied similarly in all groups (according to the physiological needs of the dams), and group differences in dosage were maintained.
Dose descriptor:
NOAEL
Effect level:
3 225 mg/kg bw/day
Based on:
test mat.
Remarks:
Aluminum citrate (equivalent to 300 mg Al/kg bw)
Basis for effect level:
other: maternal toxicity
Abnormalities:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:not examined
Dose descriptor:
NOAEL
Effect level:
3 225 mg/kg bw/day (nominal)
Based on:
test mat.
Remarks:
Al citrate (equivalent to 300 mg Al / kg bw/day
Sex:
male/female
Basis for effect level:
other: no effects noted
Abnormalities:
no effects observed
Developmental effects observed:
no

Details on toxic effects on adult offspring

MORTALITY

A number of animals from each treatment group died or were euthanized prior to their planned sacrifice dates. The main finding was the presence of urinary tract lesions of hydronephrosis, ureteral dilation, obstruction and/or presence of calculi. Obstruction of the urinary tract by calculi was found especially in the high-dose males (see Table 1).

 

CLINICAL OBSERVATION

In the Day 23 group, a few animals in the high-dose group were observed to have abdominal distention or were small and cold.

In the Day 64 group, 3 animals of the Na-citrate group were thin and with poor hair coats. One control group female was bloated, had diarrhea and was euthanized. In the high-dose group, one female and seven males were thin, had diarrhea, mild dehydration and poor hair coat. The postmortem signs of urinary tract pathology (see Table 1) lead to the decision to euthanize the remaining high-dose males.

 

BODY WEIGHT

Weights of male and female pups (Day 364 group) during the pre-weaning phase of the Na-citrate and high-dose groups were significantly lower. During the post-weaning, there were no significant differences in male mean body weights of Na-citrate, low-dose or mid-dose groups compared with controls in Day 364 group. High-dose males were sacrificed from study Day 98 onwards because of excessive clinical signs and body weight loss (see Table 1). At the time of weaning, male pups of the Na-citrate group had 17% lower body weights relative to controls but recovered. High-dose males had 12% lower weights in comparison to controls and continued to lose weight. By PND 84 the mean weight of male high-dose pups was 30% below controls (animals were euthanized).

 

FLUID CONSUMPTION

Mid-dose males in the Day 64 group had higher fluid consumption than low-dose or control groups and fluid consumption of the Na-citrate group was higher than control group consumption. In the Day 120 group males, fluid consumption of the mid-dose group exceeded that of the controls, low and Na-citrate groups and the Na-citrate group fluid consumption exceeded that of the controls. Day 364 group males, higher fluid consumption of mid-dose and Na-citrate group males was observed in comparison to controls and the low-dose group (high-dose males were euthanized, see above). For the Day 364 group mid-dose group, and other groups followed a similar pattern, mean fluid consumption during the first post-weaning week was 16.0 mL/day (171 mL/kg bw/day) compared with 13.7 mL/day (129 mL/kg bw/day) for controls, resulting in a difference of 17% (33% on a per body weight basis). From about Day 70 to the end of the study, mean fluid consumption of all groups declined gradually. Day 64 group females of the high-dose group showed higher fluid consumption when compared to controls. For day 364 group females, the mean fluid consumption of the high-dose group exceeded that of the rest. The low-dose group had lower fluid consumption than the NA-citrate, mid- and high-dose groups.

The results indicate that dosing with either aluminum citrate or sodium citrate resulted in a significant increase in fluid consumption compared to controls. The differences between the males and females in the high-dose group fluid consumption pattern may be related to the disturbed fluid balance caused by the more serious urinary tract lesions in male animals.

DOSAGE LEVELS

In post-weaned pups significant differences between groups in regard to dosage levels during the study period, with the low, medium and high-dose groups following the expected pattern of receiving the lowest to highest dosages, respectively. Unexpectedly, for the majority of the study period, dosage was significantly below target for all dose groups due to reduced fluid consumption of animals. The anticipated benchmark was 120 mL/kg bw/day. The achieved doses were highest at the beginning of the post-weaning period, declining linearly for the first 9-13 weeks, then leveling off and falling gradually for the remainder of the study period (see Table 2). The authors concluded that the decline in dosage was due to the advancing age of the rats, as dosage followed fluid consumption. The impact of this on the study was deemed to be low and the study design and results remained valid for the reasons that: First, the dosages received by the groups were significantly different from one another. Second, signs of toxicity were observed at the highest dose. Third, the concentration of Al-citrate in the high-dose solutions was near saturation; therefore, a higher intake level was not possible. Fourth, dosage was calculated based on total body weight. If Al mostly distributes into the blood compartment, lean body weight, which is lower, may be a better measure of body mass to use with respect to aluminum exposure, and the dosage on a lean body weight basis would be higher.

 

DEVELOPMENTAL LANDMARKS

High dose and Na-citrate females and males took longer to reach landmarks as compared with the other groups (see Table 6).

 

FOB

No significant group differences were recorded for the neonatal rat pups of either sex. However, significant differences emerged overtime in a subset of adult animals.

In females, no abnormal observations were detected for tonic convulsions (home cage), clonic convulsions (home cage), tremors (home cage and open field), posture (home cage and open field), conjunctivitis (handling observations) or total gait (open field).

Non-normal observations with significant group differences include wasting, fur appearance, mouth and nose deposits, eye opacity, salivation, arousal, defecation, defecation characteristics, pupil response, rearing, pupil size, tail pinch, urination, and urine characteristics.

In males, no non-normal observations were found for tonic convulsions (home cage and open field), clonic convulsions (home cage and open field), tremors (home cage and open field), posture (home cage and open field), palpebral closure (home cage), conjunctivitis (handling observations), ocular exudate (handling observations), writhing (handling observations). Non-normal observations with significant group differences include fur appearance, mouth and nose deposits, eye opacity, red crusty deposits, exophthalmus, piloerection, defecation, defecation characteristics, tail pinch, rearing, urination, and urine characteristics.

In summary, high-dosage Al exposure was not associated with autonomic or sensimotor dysfunction. There was, however, a weak association between high dosage Al exposure and reduced home cage activity and excitability.

 

MOTOR ACTIVITY

There were no consistent patterns of significant group differences over the course of the study and only mild differences occurred.

The high-dose group of Day 64 group males had low ambulatory counts indicative for less active animals in later intervals, and this may be a mild treatment-related effect.

T-maze

No statistically significant differences were noted among the groups.

Morris water maze

There were no significant differences among groups in either sex in any of the sacrifice day groups.

There were no significant group-wise differences in the platform-removed probe tests.

There were no group-wise differences in the latencies or the search strategies in the visible platform test.

There were no significant group-wise differences on the visible platform probe tests.

 

HEMATOLOGY

In Day 23 group females, mean cell volume (MCV) in the control group was significantly higher in comparison to the low-dose group. Nucleated red blood cells were different in the low-dose group than in the control, mid- or high-dose groups and platelet counts were less in the low-dose group than in the high-dose group. In Day 23 group males, MCV in the high-dose group was significantly decreased. In Day 64 group females, the high-dose group hematocrit (HCT), mean cell hemoglobin (MCH) and MCV were lower in comparison to the other groups. Hemoglobin (HGB) was lower in the high-dose group than in the mid-dose, low-dose or control groups. White blood cells (WBC) were higher in the high-dose group than in the control or low-dose groups; however, absolute agranulocyte counts were less in the high- dose group than in the controls. In Day 64 group males, MCH and MCV were lower in the high-dose group. HCT was decreased in the high-dose group in comparison to the control. Red blood cell counts were slightly higher in the high-dose group than in the mid-dose or sodium citrate groups. In Day 120 group females, MCV in the high-dose group is less than in the other groups. MCH in the high-dose group is slightly less than in the mid-dose, Na-citrate and control groups. WBC, absolute granulocytes and absolute agranulocytes in the high-dose group are higher than in the other Al citrate groups and controls.

In the Day 64 and 120 group females and Day 64 group males, the high-dose group exhibited a low grade, slightly microcytic anemia that was more apparent during the fast growth phase at the Day 64 resolving by the Day 120 and completely resolved in comparison to the other groups by Day 364.

 

CLINICAL CHEMISTRY

Differences were observed between the high-dose group and some or all of the other groups (see Table 3 and 4 under “Any other information on results incl. tables”). In males of the Day 64 group, the following parameters were slightly affected: A_G, ALP, CA, CL, CRE, GLOB, GLU, NA, PHOS, TG, TP and UREA (for abbreviations, please refer to Table 3). Serum chemistry features of aluminum toxicity include elevated ALP and CA levels; both observed especially in the Day 64 group. The high-dose animals of both sexes were less robust and parameters such as total protein, albumin and globulin were slightly lower especially in the Day 64 group. Creatinine and urea elevations in the Day 64 group males may be due to renal calculi.

ORGAN WEIGHTS

Brain weights did not differ among the groups except in high-dose males of the Day 64 group which had significantly lower mean brain weight. Furthermore, in the Day 120 high-dose group females significantly lower mean brain weight were observed.

Total body weights tended to differ more among the groups than brain weight and brain to body weight ratios tended to follow body weight.

 

MACROSCOPIC EXAMINATION

The main gross postmortem finding was the presence of white precipitates in the urinary tracts of males and females. Secondary pathologies such as hydronephrosis and ureteral dilation occurred in most affected animals, most common in high-dose group animals, especially males.

A second finding in some high-dose group animals, especially males (5/20 from Day 64 group, 2/20 from Day 120 group and 2/20 from Day 364 group), but also one female from each of Day 23 and 364 groups and one Day 364 group male from the Na-citrate group, was fluid colonic content.

 

HISTOPATHOLOGY

In Day 23, 64 and 120 groups, there were no abnormal microscopic findings except for one female in the Day 23 group, low-dose group, that had both a necrotic neuron and a neuron with satellitosis in the section of basal ganglia, and in Day 64 group, control group female with very mild inflammation of the connective tissue around the sciatic nerve. In Day 364 group, a proportion of the animals had neuronal cytoplasmic vacuolation in mainly the thoracic, but also some lumbar dorsal root ganglia (see Table 5).

Cortical and hippocampal areas were unremarkable.

METAL LEVELS IN RAT TISSUE

In general, metal tissue concentrations were associated with Al exposure, with the exception of blood, where the intergroup differences became much less distinct in the Day 64 and Day 364 group. Tissue concentrations in Day 23 group were generally ranked in order of highest to lowest: blood, 10- to 100-fold > brainstem > femur = spinal cord > cerebellum > liver > cerebral cortex. Blood, femur and liver concentrations generally remained similar or increased from one sacrifice day group to the next, while CNS tissue concentrations remained similar or decreased from Day 23 group to Day 64 group, then stabilized from Day 64 group to Day 364 group. The concentrations of Cu, Fe, Mn and Zn in blood and tissues remained relatively stable and were similar in all groups throughout the study.

Table 1. Male rats with urinary tract lesions.

Group

Collection time

Day 23 group

Day 64 group

Day 120 group

Day 364 group

Na citrate

0

1

0

0

Control

0

0

0

0

Low-dose

0

0

0

1

Mid-dose

0

3

1

0

High-dose

0

11

7

5

 

Table 2. Dosage levels in offspring.

Dose Group

Post weaning

Week 9

Week 15-49

Target dose

[mg Al per kg bw per day]

Low-dose

Males

40.2

15.4

10.7 to 2.0*

30

Females

43.5

17.4

13.5 to 10.0

30

Mid-

Males

174.3

69.8

37.6 to 22.9

100

Females

199.2

70.4

49.8 to 40.9

100

High-dose

Males

496.2

379.4

-

300

Females

614.9

338.5

243.3 to 124.6

300

*: between Week 15-49 the decline in dosage levels from maximum to minimum is given.

-: animals were euthanized after Week 9

Table 3. Summary of clinical chemistry parameters in females showing significant group differences.

Parameter

Day 23

Day 64

Day 120

Day 364

ALB (albumin)

High < Low, Control and Mid

High < Low and Mid

 

ALP (alkaline

phosphatase)

 

High > Low, Na citrate, Control, Mid

High > Low, Control and Mid

 

CA (calcium)

Na < Mid, High

Control < High

High > Low, Na citrate, Control, Mid

High > Na citrate, Control

 

CL (chloride)

 

 

Na-citrate < Control

Na-citrate < Control Low

GLOB (globulin)

 

High < Low, Na citrate, Control, Mid

High < Mid

 

GLU (glucose)

Control > Low, Na citrate, High

 

 

K (potassium)

Control > Low, Na citrate

 

 

Na (sodium)

Na-citrate > Low, Control, Mid, High

Mid > Na citrate, High

 

 

Phos (phosphorus)

Control > High

 

 

 

TG (triglycerides)

 

High dose < Low, Control

 

 

TP (total protein)

 

High < Low, Na citrate, Control, Mid

High < Low, Control, Mid

 

Urea

Na-citrate > Mid, High

 

High > Low, Control

High > Low, Control, Mid

 

Table 4. Summary of clinical chemistry parameters in males showing significant group differences.

Parameter

Day 23

Day 64

Day 120*

Day 364*

A_G (albumin/ globulin ratio)

 

High > Low, Na citrate, Control, Mid

 

ALP (alkaline phosphatase)

High > Control

High > Low, Na citrate, Control, Mid

 

 

AST (aspartate aminotrans-ferase)

Na-citrate > Low

 

 

 

CA (calcium)

 

High > Low, Na citrate, Control, Mid

 

 

CL (chloride)

 

High < Low, Na citrate, Control, Mid

 

 

CRE (creatinine)

 

High > Low, Na citrate, Control, Mid

 

 

GLOB (globulin)

 

High < Low, Na citrate, Control, Mid

Na citrate < Low, Control, Mid

 

GLU (glucose)

 

High > Low, Control, Mid

 

 

Na (sodium)

Na-citrate> Low, Control, Mid, High

High < Low, Control, Mid

 

 

Phos (phosphorus)

 

High > Low, Na citrate, Control, Mid

 

 

TBIL (total bilirubin)

 

Na-citrate < Low, Mid, Migh

 

 

TG (triglycerides)

 

High < Low, Control

 

 

TP (total protein)

 

High < Low, Control, Mid

Na-citrate < Low, Control

 

Urea

 

High > Low, Na-citrate, Control, Mid

 

 

*: no high-dose group.

Table 5. Animals with vacuolated neurons in thoracic ganglia on Day 364 per group and sex. 

Dose Group

Sex

Number

of animals

Low-dose

Males

2

 

Females

4

Na-citrate

Males

1

 

Females

3

Control

Males

2

 

Females

3

Mid-dose

Males

1

 

Females

3

High-dose

Males

0*

 

Females

3

*: all animals were pre-terminally sacrificed, no high-dose group.

 

Table 6. Summary statistics for developmental landmarks (females: vaginal opening; males: preputial separation).

Parameter

Na-citrate

Control

Low-dose

Mid-dose

High-dose

Number of days to landmark

Males

41.1 ± 2.4

39.6 ± 2.1

39.3 ± 1.4

39.4 ± 1.9

42.5 ± 3.2

Females

35.3 ± 2.9

31.3 ± 2.1

32.1 ± 2.5

32.4 ± 2.1

39.7 ± 5.6

Conclusions:
No developmental toxicity observed in this study.
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises adequate, reliable (Klimisch score 1 - 2) and consistent studies from the target substance and a reference substances with similar structure and intrinsic properties. Read-across is justified based on common functional group(s), common precursors/breakdown products, similarities in PC/ECO/TOX properties (refer to endpoint discussion for further details). The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.6, in accordance with Annex XI, 1.5, of Regulation (EC) No. 1907/2006.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Read-across is justified in the endpoint records and in the read-across report attached in Section 13 of this technical dossier.

Justification for classification or non-classification

Based on a weight of evidence approach consisting of data on analogous substances, aluminium orthophosphate is not considered to be a reproductive or developmental toxicant in accordance with Regulation (EC) No. 1272/2008 (EU CLP).

Additional information