Aluminium sulphate

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Reliable without restrictions. Well-presented study, with relevant measurement of chemical concentrations

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Remarks:

Tested by LG Life Science/Toxicology Center, Korea,Test No. S506)

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Calcium sulfate, dihydrate

IUPAC Name:

Calcium sulfate, dihydrate

Constituent 2

Reference substance name:

10101-41-4

EC Number:

600-148-1

Cas Number:

10101-41-4

IUPAC Name:

10101-41-4

Test material form:

solid: compact

Details on test material:

- Name of test material: Calcium sulfate, dihydrate
- Molecular formula (if other than submission substance): CaSO4•2H2O
- Molecular weight (if other than submission substance): 172.171
- Smiles notation (if other than submission substance): O.O.[O-]S(=O)(=O)[O-].[Ca+2]
- Structural formula attached as image file (if other than submission substance): see Fig.1
- Substance type:inorganic
- Physical state:solid
- Density -2.32 g/cm3)
- Solubility in water - 2.05 g/L at 20 °C
- Synonyms: Alabaster
Annaline
C.I. Pigment white 25
Gypsum
Gypsum stone
Land and plaster
Light spar
Magnesia white
Mineral white
Native calcium sulfate
Precipitated calcium sulfate
Sainite
Satin spar
Sulfuric acid, calcium(2+) salt, dihydrate
Terra alba
- Calcium sulfate, dihydrate consists of colorless, monoclinic and hygroscopic crystals

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Test organism
- Sex: male/female
- Age of animals at study: 8 weeks old for males and females
- Weight at study repeated dose toxicity: 254.2 - 297.8 g for males and
182.7 - 208.2 g for females
- Number of test animals: 60 animals for each sex
- Group composition: twelve animals (6 male + 6 female) were
allocated as a recovery group for the control (0 mg/kg/day) and T3 (100
mg/kg/day) group.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

not specified

Details on oral exposure:

Dose level: 0, 100, 300 and 1,000 mg/kg/day; pre-treatment (Test No. P705) had conducted with 0, 125, 250, 500 and 1,000 mg/kg/day of the test
substance for 7 days to determine the appropriate starting dose level.
Exposure period : 35 days for male animals and 41 to 45 days for female animals
Frequency of treatment : Daily
Control group : Yes (Concurrent no treatment)

Duration of treatment / exposure:

35 days for male animals and 41 to 45 days for female animals

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300 and 1,000 mg/kg/day; pre-treatment (Test No. P705) had conducted with 0, 125, 250, 500 and 1,000 mg/kg/day of the test substance for 7 days to determine the appropriate starting dose level.
Basis:
nominal in diet

No. of animals per sex per dose:

Number of test animals: 60 animals for each sex
- Group composition: twelve animals (6 male + 6 female) were allocated as a recovery group for the control (0 mg/kg/day) and T3 (100
mg/kg/day) group.

Control animals:

yes, concurrent no treatment

Details on study design:

Post-exposure period: no data

Examinations

Observations and examinations performed and frequency:

- Clinical observations performed and frequency: Clinical symptoms were observed once a day but were observed once a week in detail; a
death rate was observed twice a day; and body weight was observed once a week and just before the necropsy, but in case of pregnant
females, it was measured on the day 0, 7, 14, 20 of gestation period, date of delivery, and 4 days after the delivery; consumption rate of
fodder was observed once a week except mating period.

- Tests for sensory organ and reflex action: 5 animals were randomly selected from each test group. Both preyer reflex test and corneal reflex
test were performed before necropsy and during lactation for males and females, respectively.

- Behaviour test: 5 animals were randomly selected from each test group to do grip strength test in terms of behaviour test. This test was
performed before necropsy and during lactation for males and females, respectively.

- Haematological and biochemical test of blood: randomly selected 5 male and female animals from each test group were fasted a day before
necropsy for both tests. Animals were anesthetized using ether and cut the abdomen open to collect blood. In case of the haematological test, blood
coagulation preventative chemicals for the test of blood coagulation and the calculation of blood-corpuscles were 3.2 % sodium citrate and EDTA-
2K, respectively. On the other hand, blood coagulation preventative chemical was not used for the biochemical test, but gathered blood was
left itself in the room temperature then the sera were separated using a centrifuge. For haematological test, 6 following items were measured;
Haematocrit, hemoglobin concentration, erythrocyte count, total and different leucocyte count, platelet count, prothrombin time, and active
partial thromboplastin time. For biochemical test of blood, eleven following items were measured; sodium, potassium, glucose, total cholesterol,
blood urea nitrogen, creatinine, total protein, albumin, alanine aminotransferase, aspatate aminotransferase, and total bilirubin.

Sacrifice and pathology:

Organs examined at necropsy:
Organ weight: testes, epididymider (all males) liver, kidney, adrenals, thymus, spleen, brain and heart (5 male and female animals from each
test group).
Fixation: 22 kinds of tissues were fixed to do histopathologic tests such as testes, epididymides, ovaries, accessory sex organs for all animals,
brain (including cerebrum, cerebellum and pons), spinal cord, stomach, small and large intestines (including peyer’s patches), liver, kidneys,
adrenals, spleen, heart, thymus, thyroid, trachea, lungs, uterus, urinary bladder, lymph nodes (cervical mesenteric), peripheral nerve (sciatic or
tibial), and bone marrow

Statistics:

Statistical decision tree, but in case of recovery group, either twoside Student’s t-test or two-side Aspin-Welch t-test was used.
In case of categorical data, two-sided Fisher’s exact test was used.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

There was one death at day 8 for male, and each death on the day 7 and 14 for female in the treatment group of 300 mg/kg/day. These were occurred during the administration process, so it did not have relationship with test substance.

Mortality:

no mortality observed

Description (incidence):

There was one death at day 8 for male, and each death on the day 7 and 14 for female in the treatment group of 300 mg/kg/day. These were occurred during the administration process, so it did not have relationship with test substance.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

In male groups, temporarily, a case of diminishment in the amount of body weight change was observed at week 2 within the control group in which some clinical signs were observed such as damage to esophagus. Body weight loss for female animals was observe

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No crucial difference between the treatment group and the control group was observed for both male and female animals during test period. For recovery group, no significant change was observed within themselves.

Food efficiency:

no effects observed

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

In the 100 mg/kg/day BUN (Blood urea nitrogen) was decreased .The values of TP (Total Protein), ALB (Albumin), BUN (Blood Urea Nitrogen), and CREA (Creatinine) were decreased significantly at the 300 mg/kg bw/day and 1,000 mg/kg bw/day treatment for males

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

In the 100 mg/kg/day BUN (Blood urea nitrogen) was decreased .The values of TP (Total Protein), ALB (Albumin), BUN (Blood Urea Nitrogen), and CREA (Creatinine) were decreased significantly at the 300 mg/kg bw/day and 1,000 mg/kg bw/day treatment for males

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

- Mortality: There was one death at day 8 for male, and each death on the day 7 and 14 for female in the treatment group of 300 mg/kg/day.
These were occurred during the administration process, so it did not have relationship with test substance.
- Body weight: In male groups, temporarily, a case of diminishment in the amount of body weight change was observed at week 2 within the
control group in which some clinical signs were observed such as damage to esophagus. Body weight loss for female animals was observed several
times during lactation period in every treatment group including the control group. However, these were occurred temporarily because of the lactation.
- Clinical signs: In male control group, a case of salivation and bloodylike secretion was observed on the day 11 and 12. In the 1,000
mg/kg/day treatment group, a case of depilation, dcab and pus was observed on the left cheek between the day 25 and the closing day.
However, the frequency of occurrence was low and no dose-response correlation. Thus these symptoms were not influenced by test substance.
In female control group, a case of genitalia bloody-like secretion was observed at day 29. In the 100 mg/kg/day treatment
group, each case of hypoactivity and depilation was observed on the day 8 and 9, and between day 44 and the closing day, respectively.
However, these symptoms were disappeared in short, thus these did not have relationship with test substance.
- Amount of fodder consumption: No crucial difference between the treatment group and the control group was observed for both male and
female animals during test period. For recovery group, no significant change was observed within themselves.
- Test of reflex action: Five male and female animals were randomly selected from each test group, in which no specific reaction was observed.
- Grip strength test: For male animals, 6 animals were left out from the treatment group. For female animals, 5 animals were left out from the
control group, and the treatment group. All things being considered, there was no dose-response correlation and was no illness at the related organs
such as the cerebellum and muscle.
- Organ weight: Both absolute weight of the liver and the left kidney were increased at the recovery group with administration of 1,000 mg/kg/day
as compared with that of the control group within the recovery group. There was no histopathological illness at the organs, so increased organ
weight did not have relationship with test substance.
- Necropsy opinions: For male animals, in the control group within the recovery group, a case of left and right caput epididymis cyst was
observed and the 1,000 mg/kg/day recovery group had symptom of right caput epididymis cyst. However, its frequency of occurrence was low
and it was even observed at the control group within the recovery group, so it did not have relationship with test substance. For female animals, in
the 300 mg/kg/day treatment group, each animal was dead on the day 7 and 14 and; each case of lung dark-red discolouration was observed,
but white particle in a lobe of the lung was observed just from one of carcasses. A case of spleen white nodule was observed for an animal
in the 300 mg/kg/day treatment group. There was a case of right adrenal gland white spots at the 1,000 mg/kg/day treatment group. In the control
group within the recovery group, each case of right adrenal gland hemorrhagia and atrophy and liver adhesion with diaphragm was observed.
- Analysis of haematological test of blood: In the 1,000 mg/kg/day male recovery group, segments were increased (p < 0.05) in contrast with that
of the control group within the recovery group. Prothrombin time (PT) was decreased in the 1,000 mg/kg/day female recovery group in
comparison with that of the control group within the recovery group. However, these symptoms were not observed in the definitive test
groups, so these symptoms were not influenced by test substance. In addition, level of WBC (white blood cell) was increased (p < 0.001) in the
100 mg/kg/day female treatment group in contrast with that of the control group. However, its increased value was in the normal range and no
dose-response correlation, so it did not have relationship with test substance.
- Analysis of biochemical test of blood: In the 100 mg/kg/day treatment group for male animals, BUN (Blood urea nitrogen) was decreased as
compared with that of the control group. The male treatment groups with both administration of 300 mg/kg/day and the 1,000 mg/kg/day
decreased in TP (Total protein), ALB (Albumin), AST (Aspatate aminotransferase), ALT (Alanine aminotransferase), BUN (Blood urea nitrogen), CREA (Creatinine), Na (Sodium), TCHO (Total cholesterol), and Cl (Chloride) as compared with those of the control group. In case of the 1,000 mg/kg/day male recovery group, the value of AST was decreased significantly in contrast with that of the control group within the recovery group. No significant difference was found at every test item between female control and treatment group. The female recovery group with administration of 1,000 mg/kg/day decreased in AST in contrast with that of the control group within the recovery group, but TCHO and GLU (Glucose) were increased. In fact, decreased values of AST and ALT could be no toxicological effects. In addition, the changed values of AST, TCHO, and GLU in this test were in the normal range and no histopathological opinion in terms of related organs, so these changed values were not influenced by test substance. However, decreased values of TP, ALB, BUN, and CREA were possibly influenced by excretion process or metabolism of test substance in relation to the kidney, and these symptoms were possibly recovered in 2 weeks from reversible effects.
- Histopathology: For male animals, in the control group, each case of heart focal inflammatory cell infiltration, submadibular lymph node
blood absorption, liver mononuclear cell foci, and adrenal gland cortical vacuolation was observed. In the 300 mg/kg/day treatment
group, two cases of pancreas vacuolation, and a case of liver mononuclear cell foci were observed. In the treatment group with administration of 1,000 mg/kg/day, there were three cases of liver mononuclear cell foci; and a case of heart focal inflammatory cell infiltration was found.
For female animals, in the control group, two cases of liver mononuclear cell foci, a case of kidney cortical scaring, and a case of pancreas
vacuolation were observed. In the treatment group with administration of 100 mg/kg/day, one case of esophagus submucosal gland proliferation
was observed. In the 300mg/kg/day treatment group, a case of trachea submucosal gland proliferation was observed. In the 1,000 mg/kg/day
treatment group, each case of pancreas vacuolation and liver mononuclear cell foci was observed. However, these symptoms for both sexes were just subtle level and were occurred spontaneously, so there was no significant difference between the treatment group and the control group.

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: see 'Remark'

2

Dose descriptor:

LOAEL

Effect level:

300 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

3

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: see 'Remark'

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

RS-Freetext:
Results for F0
- Mortality: There was one death at day 8 for male, and each death on the  day 7 and 14 for female in the treatment group of 300 mg/kg/day. These  were occurred during the administration process, so it did not have  relationship with test substance. 
- Body weight: In male groups, temporarily, a case of diminishment in the  amount of body weight change was observed at week 2 within the control  group in which some clinical signs were observed such as damage to  esophagus. Body weight loss for female animals was observed several times  during lactation period in every treatment group including the control  group. However, these were occurred temporarily because of the lactation. 
- Clinical signs:  In male control group, a case of salivation and  bloody-like secretion was observed on the day 11 and 12. In the 1,000  mg/kg/day treatment group, a case of depilation, dcab and pus was  observed on the left cheek between the day 25 and the closing day.  However, the frequency of occurrence was low and no dose-response  correlation. Thus these symptoms were not influenced by test substance.  In female control group, a case of genitalia bloody-like secretion was  observed at day 29. In the 100 mg/kg/day treatment group, each case of  hypoactivity and depilation was observed on the day 8 and 9, and between  day 44 and the closing day, respectively. However, these symptoms were  disappeared in short, thus these did not have relationship with test  substance.
- Amount of fodder consumption: No crucial difference between the  treatment group and the control group was observed for both male and  female animals during test period. For recovery group, no significant  change was observed within themselves.  
- Test of reflex action: Five male and female animals were randomly  selected from each test group, in which no specific reaction was observed.
- Grip strength test: For male animals, 6 animals were left out from the  treatment group. For female animals, 5 animals were left out from the  control group, and the treatment group. All things being considered,  there was no dose-response correlation and was no illness at the related  organs such as the cerebellum and muscle.
- Organ weight: Both absolute weight of the liver and the left kidney  were increased at the recovery group with administration of 1,000  mg/kg/day as compared with that of the control group within the recovery  group. There was no histopathological illness at the organs, so increased  organ weight did not have relationship with test substance.
- Necropsy opinions: For male animals, in the control group within the  recovery group, a case of left and right caput epididymis cyst was  observed and the 1,000 mg/kg/day recovery group had symptom of right  caput epididymis cyst. However, its frequency of occurrence was low and  it was even observed at the control group within the recovery group, so  it did not have relationship with test substance. For female animals, in  the 300 mg/kg/day treatment group, each animal was dead on the day 7 and  14 and; each case of lung dark-red discolouration was observed, but white  particle in a lobe of the lung was observed just from one of carcasses. A  case of spleen white nodule was observed for an animal in the 300  mg/kg/day treatment group. There was a case of right adrenal gland white  spots at the 1,000 mg/kg/day treatment group. In the control group within  the recovery group, each case of right adrenal gland hemorrhagia and  atrophy and liver adhesion with diaphragm was observed. 
- Analysis of haematological test of blood: In the 1,000 mg/kg/day male  recovery group, segments were increased (p < 0.05) in contrast with that  of the control group within the recovery group. Prothrombin time (PT) was  decreased in the 1,000 mg/kg/day female recovery group in comparison with  that of the control group within the recovery group. However, these  symptoms were not observed in the definitive test groups, so these  symptoms were not influenced by test substance. In addition, level of WBC  (white blood cell) was increased (p < 0.001) in the 100 mg/kg/day female  treatment group in contrast with that of the control group. However, its  increased value was in the normal range and no dose-response correlation,  so it did not have relationship with test substance.
- Analysis of biochemical test of blood: In the 100 mg/kg/day treatment  group for male animals, BUN (Blood urea nitrogen) was decreased as  compared with that of the control group. The male treatment groups with  both administration of 300 mg/kg/day and the 1,000 mg/kg/day decreased in  TP (Total protein), ALB (Albumin), AST (Aspatate aminotransferase), ALT  (Alanine aminotransferase), BUN (Blood urea nitrogen), CREA (Creatinine),  Na (Sodium), TCHO (Total cholesterol), and Cl (Chloride) as compared with  those of the control group. In case of the 1,000 mg/kg/day male recovery  group, the value of AST was decreased significantly in contrast with that  of the control group within the recovery group. 
No significant difference was found at every test item between female  control and treatment group. The female recovery group with  administration of 1,000 mg/kg/day decreased in AST in contrast with that  of the control group within the recovery group, but TCHO and GLU  (Glucose) were increased. 
In fact, decreased values of AST and ALT could be no toxicological  effects. In addition, the changed values of AST, TCHO, and GLU in this  test were in the normal range and no histopathological opinion in terms  of related organs, so these changed values were not influenced by test  substance. However, decreased values of TP, ALB, BUN, and CREA were  possibly influenced by excretion process or metabolism of test substance  in relation to the kidney, and these symptoms were possibly recovered in  2 weeks from reversible effects.
- Histopathology: For male animals, in the control group, each case of  heart focal inflammatory cell infiltration, submadibular lymph node blood  absorption, liver mononuclear cell foci, and adrenal gland   cortical  vacuolation was observed. In the 300 mg/kg/day treatment group, two cases  of pancreas vacuolation, and a case of liver mononuclear cell foci were  observed. In the treatment group with administration of 1,000 mg/kg/day,  there were three cases of liver mononuclear cell foci; and a case of  heart focal inflammatory cell infiltration was found.
For female animals, in the control group, two cases of liver mononuclear  cell foci, a case of kidney cortical scaring, and a case of pancreas  vacuolation were observed. In the treatment group with administration of  100 mg/kg/day, one case of esophagus submucosal gland proliferation was  observed. In the 300mg/kg/day treatment group, a case of trachea  submucosal gland proliferation was observed. In the 1,000 mg/kg/day  treatment group, each case of pancreas vacuolation and liver mononuclear  cell foci was observed. However, these symptoms for both sexes were just  subtle level and were occurred spontaneously, so there was no significant  difference between the treatment group and the control group.

For more details or results, see attached document.

Applicant's summary and conclusion

Conclusions:

There was no specific opinion about test items such as clinical symptoms, body weight change, food consumption, reflex action, and necropsy, etc. under the influence of test substance. However, in case of male animals, the repeated dose toxicity test of more than 35 days affect at the kidney subtle level, so LOAEL and NOAEL were determined as 300 mg/kg/day and 100 mg/kg/day, respectively. In case of female animals, no effects were observed at the top dose tested.

Executive summary:

In a gavage study in Sprague-Dawley rats in accordance with OECD TG 422, Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Screening Test was conducted. The rats were exposed to calcium sulfate, dihydrate at level of 0, 100, 300 and 1,000 mg/kg bw/day for more than 35 days and 41 - 45 days for male and female animals, respectively. For male animals, in the control group within the recovery group, a case of left and right caput epididymis cyst was observed and the 1,000 mg/kg bw/day recovery group had symptom of right caput epididymis cyst. For female animals, in the 300 mg/kg bw/day treatment group, one animal was died on day 7 and another on day 14 and in each case dark-red discolouration of the lung was observed, but white particles in a lobe of the lung were observed just in one dead animal. However, its frequency of occurrence was quite low.

After all, calcium sulfate, dihydrate had no critical influence on test items such as mortality, body weights (organ weight), food consumption, necropsy and behaviour of the animals. However, the values of TP (Total Protein), ALB (Albumin), BUN (Blood Urea Nitrogen), and CREA (Creatinine) were decreased significantly at the 300 mg/kg bw/day and 1,000 mg/kg bw/day treatment for male animals in accordance with analysis of biochemical test of blood, for the test substance might affect the excretion process, distribution or metabolism of the test substance in relation to the kidney. On the other hand, no significant difference was found at every test item between female controls and treatment groups. There were some changed values in the female recovery group for AST, TCHO and GLU, but these were in the normal range and there was no dose-response relationship. Therefore, LOAEL and NOAEL for male animals were determined to be 300 mg/kg bw/day and 100 mg/kg bw/day, respectively. (National Institute of Environmental Research (NIER), 2002, Calcium sulfate dihydrate: Combined Repeated Dose Toxicity with the Reproduction/Developmental Toxicity Screening Test (Test No. S506)). For female rats, no effects were observed at the top dose tested (1,000 mg/kg bw/day).