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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well-documented study. The study fulfils the requirements of the current test guideline for oral sub-chronic exposure to a great extent, but the study was not performed under GLP requirements.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
publication
Title:
Short-term toxicity of strontium chloride in rats
Author:
Kroes, R.; et al.
Year:
1977
Bibliographic source:
Toxicology 7, 11-21

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Rats received SrCl2 hexahydrate in a diet at dose levels of 0, 75, 300, 1200, and 4800 ppm for 90 days. The diet contained adequate levels of Ca, Mg, P and vitamin D3. Growth, food intake, behaviour and mortality were measured, extensive haematology and clinical biochemistry carried out, organ weight determined, X-ray photographs of the bones taken and complete histopathological examination was performed. In addition, the Sr content of blood, bone and muscles was determined in additional groups of rats.
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Strontium chloride hexahydrate
- Molecular formula (if other than submission substance): SrCl6*6H2O
- Substance type: technical product
- Physical state: solid
- Impurities (identity and concentrations): barium content was less than 0.02 %
- Other:Test material was obtained from Mallinckrodt, St. Louis, Missouri, USA

No further information given.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: SPF wistar rats from the breeding stock of the National Institute of Public Health
- Weight at study initiation: 40-60 g bw
- Housing: under conventional conditions, littermate-divided in wire cages, two in a cage according to sex
- Diet (ad libitum): semipurified diet (Muracon SSp-tox, Trouw Ltd., Putten, The Netherlands); the diet contained 0.05 % Mg, 0.75 % P, 0.85 % Ca and 1.8 % I.U. Vit. D.3 per gram
- Water (ad libitum): tap water

No further information given.

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
DIET PREPARATION
- SrCl2 in a semi-purified diet at dose levels of 0, 75, 300, 1200, and 4800 ppm
- the diet contained adequate levels of Ca, Mg, P and vitamin D3 (see above, details on test animals and environmental conditions)

No further information given.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
no data
Duration of treatment / exposure:
90 days
Frequency of treatment:
ad libitum in diet (continuous)
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
75 ppm SrCl2*6H2O
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
300 ppm SrCl2*6H2O
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
1200 ppm SrCl2*6H2O
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
4800 ppm SrCl2*6H2O
Basis:
nominal in diet
No. of animals per sex per dose:
- 10 males and 10 females per group
- additional 15 males in the control group for determination of Sr-levels in blood, bone and muscle
- additional 10 males in the other groups for determination of Sr-levels in blood, bone and muscle
Control animals:
yes
Details on study design:
- Dose selection rationale: A range-finding experiment has been performed (for details see "Any other information on materials and methods incl. tables"; for results see "Remarks on results incl. tables and figures")

No further information given.
Positive control:
no data

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked: behaviour, mortality

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, intake was measured in week 2, 5, 9, 12
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY: Yes
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 90 weeks
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10 female and 10 male rats of each group
- Parameters checked: Hb, Ht and the number of erythrocytes and leucocytes; the MCV, MCHC and MCH were calculated; in addition blood picture was studied

CLINICAL CHEMISTRY: Yes (for method description, see below )
- Time schedule for collection of blood: Ca, Mg and P in the beginning of the experiment and after 4, 8, and 12 weeks; SGPT, Alk. PAse and urea content after 6 and 12 weeks
- Animals fasted: No data
- How many animals: Ca, Mg and P of 5 control males (in the beginning of the experiment) and in 5 males per group after 4, 8, and 12 weeks; SGPT, Alk. PAse and urea conten of 5 males per group
- Parameters checked: Ca, Mg and P analysis; activity of SGPT, Alk. PAse and urea content in the serum

URINALYSIS: Yes
- Time schedule for collection of urine: after 12 weeks
- Metabolism cages used for collection of urine:No data
- Animals fasted: No data
- Parameters checked: urinalysis was carried out with Bililabstix (Ames Cy.) in 10 female and 10 male rats; presence of protein, blood bilirubin and ketones were studied, and pH

NEUROBEHAVIOURAL EXAMINATION: No data

OTHER:
- X-ray photographs of the bones were taken in the ninth and twelfth week of the experiment from 2 females and 2 males of group 1 and from 5 females and 5 males of group 5 (for method description, see below )
- the Sr content of blood, bone and muscles was determined in additional groups of rats (10 males each and 15 control males) at weeks 0, 4, 8, and 12 (for method description, see below )
- the activities of the microsomal liver enzymes AH, and APDM were determined in 5 males per group after 4 and 12 weeks
- the glycogen content in liver was determined in 5 males after 8 week s and in 6 females and 6 males after 12 weeks

No further information given.
Sacrifice and pathology:
After 12 weeks the remaining animals were killed.
GROSS PATHOLOGY: Yes; organ weight determined: brain, pituitary, heart, thyroid, liver, kidneys, spleen, adrenals, ovaries or testes, uterus or prostate
HISTOPATHOLOGY: Yes, complete histopathological examination was carried out by preparing paraffin sections (5 µm) stained by haemalum and eosin: brain, pituitary, heart, thyroid, liver, kidneys, spleen, adrenals, ovaries or testes, uterus or prostate, lungs, thymus, pancreas, mesenterial lymph nodes, stomach, duodenum, ileum, jejunum, coecum, colon, rectum, urinary bladder, nervus ischiadicus, musculus quadriceps and femur
Other examinations:
no
Statistics:
For the purposes of objective quantification of observed differences between separate treatment groups and corresponding controls, Student's t-test was used. The results of these testings are indicated as follows: *=P<0.05, **=P<0.01 and ***=P<0.001. According to the general statistical theory it is advisable not to attach too much weight to a formal significance result (i.e. P<0.05). in such cases of marginal statistical significance it is important to consider carefully the test results obtained for corresponding comparisons regarding the other dose levels.

Results and discussion

Results of examinations

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
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
- no relevant effects
- one control female died during bleeding procedure after more than 11 weeks

BODY WEIGHT AND WEIGHT GAIN
- no relevant effects

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
- food intake was not affected by exposure to strontium chloride hexahydrate
- no data on daily food intake are available in order to calculate daily dose levels
- according to the estimation given in the IUCLID Data Set Strontium carbonate, the NOAEL of 300 ppm corresponds to a dose of 22.5 mg SrCl2/kg bw/d

FOOD EFFICIENCY
- no relevant effects

HAEMATOLOGY
- the slightly elevated erythrocyte count noticed in the range-finding experiment was not confirmed in the 90-day study
- a lower leucocyte count noticed in the males of the 300 ppm group is not considered to be caused by the treatment, since at higher dose levels such an effect was not observed

CLINICAL CHEMISTRY
- Analysis of Alk. Pase, SGPT and urea in serum did not reveal any significant changes although in the highest dose group an indication of an increased activity of Alk. Pase was noticed. No clinical differences in clinical chemistry were noted except an indication of increased alkaline phosphatase activity in the highest dose group
- the levels of Ca, Mg and P in blood were similar for all dose levels and the Ca/P ratio was constant
- blood concentrations of Ca, Mg and P were higher at 8 weeks than 12 weeks, which seems a physiological condition

URINALYSIS
- urinalysis showed no differences in the groups

ORGAN WEIGHTS
- in males, relative thyroid weights were significantly increased in the 1200 and 4800 ppm groups (see Table III in attachment)
- in females, relative pituitary weights were significantly decreased in the 300 and 4800 ppm group, but not in the 1200 ppm group (see Table III)
- the relative prostate weights were significantely decreased at 75 and 1200 ppm (see Table III in attachment); this, however, must be considered with care, because proper preparation of the rat prostate is difficult

HISTOPATHOLOGY: NON-NEOPLASTIC
- there were no changes seen on histopathological examination except slight changes in the liver and thyroid after blind examination (see Table IV in attachment)(the changes consisted of a loss of glycogen in the liver at the highest dose level and a slightly increased activity in the thryroid of the males of the highest dose group)
- glycogen depletion may be caused by several factors other than strontium, such as stress , starvation or dirunal rhythm

OTHER FINDINGS
- The glycogen concentration in the liver after 12 weeks showed a dose-related decrease, which was only significant in the female ath the highest doselevel (see Table II in attachment), which is may be caused by stress, starvation or diurnal rhythm
- microsomal liver enzyme activities did not show any changes
- detectable amounts of Sr in blood and muscle were only noticed at the dose of 4800 ppm (Table Vin attachment )
- the Sr content in bone was increased at all dose levels having a constant level from 4 weeks onwards (steady-state level)(Table V in attachment)
- there were no changes seen in the X-ray photographs
- thus, up to the highest dose of 4800 ppm no rachitic changes occurred

No further information given.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Remarks:
Sr
Effect level:
12.4 mg/kg bw/day (nominal)
Sex:
male
Basis for effect level:
other: see 'Remark'
Dose descriptor:
LOAEL
Remarks:
Sr
Effect level:
49.6 mg/kg bw/day (nominal)
Sex:
male
Basis for effect level:
other: If the increased concentrations of strontium in the bone can be considered a non-toxic effect, a LOAEL of 1200 ppm SrCl2 can be derived from this study which is based on the weight changes of thyroids at the doses of 1200 ppm and 4800 ppm.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

RANGE-FINDING EXPERIMENT

Behaviour, growth, food intake and food efficiency were not affected in the range-finding experiment. Haematological investigation revealed only a slight elevation of the total number of erythrocytes in males and females and a slight increase of the white cell count in the males at the highest dose level. No differences were found in liver and kidney weights and histopathological examination revealed no abnormalities. Sr in blood and muscle were only noted at the highest dose level whereas from 300 ppm onwards increased concentrations were found in bone (see Table I in attachment).

Applicant's summary and conclusion

Conclusions:
If the increased concentrations of strontium in the bone can be considered a non-toxic effect, a NOAEL of 300 ppm SrCl2 can be derived form this study which is based on the weight changes of thyroids at the doses of 1200 ppm (LOAEL) and 4800 ppm.