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Description of key information

An 28-day repeated dose oral toxicity study in rats performed according to OECD 407 guideline and GLP principles is available, the NOAEL was determined to be 1000 mg/kg bw/day.

A reproduction/developmental toxicity screening study in rats performed according to OECD 422 guideline and GLP principles is available. In this study parental toxicity was observed in all dose groeps, and therefore the LOAEL was determined to be 100 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
03 April, 1995 - 01 May, 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to other study
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Version / remarks:
(1981)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))
Version / remarks:
(1992)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Wistar Crl:(WI)BR
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, Germany
- Age at study initiation: Approximately 6 weeks
- Weight at study initiation: males: 182 - 209 g; females: 153 - 170 g
- Fasting period before study: Overnight prior to dosing.
- Housing: Group housing of 5 animals per sex per cage in stainless steel suspended cages with wire mesh floors.
- Diet: Free access to standard pelleted laboratory animal diet (from Carfil Quality BVBA, Oud-Turnhout, Belgium)
- Water: Free access to tap-water
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): Approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 03 April, 1995 To: 30 April, 1995
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: Formulations (w/w) were prepared daily immediately prior to dosing. Adjustment was made for specific gravity of vehicle (1.036).

DOSE VOLUME: 5 mL/kg body weight. Actual dose volumes were calculated weekly according to the latest body weight.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples of formulations, prepared after completion of the in-life phase, were analysed to check homogeneity (highest and middle concentration) and accuracy of preparations (all concentrations).
Duration of treatment / exposure:
28 days
Frequency of treatment:
Once daily, 7 d/w.
Dose / conc.:
50 mg/kg bw/day (actual dose received)
Dose / conc.:
200 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were selected on the basis of a 5-day dose range finding study. No mortality was observed in the 50, 200 and 1000 mg/kg bw. No clinical signs were noted except for 3 animals at the 200 mg/kg bw dose level which showed alopecia and scabs. Based on the results, dose levels for the main study were selected to be 50, 200 and 1000 mg/kg bw.
Positive control:
No.
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least once daily from day 1 onwards. The time of onset, degree and duration were recorded.

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly and on the day preceding termination, prior to overnight fasting.

FOOD CONSUMPTION
- Weekly

WATER CONSUMPTION
- Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination, between 7.30 and 9.30 a.m.
- Anaesthetic used for blood collection: Yes (light ether)
- Animals fasted: Yes, overnight
- How many animals: all rats/sex/group
- Parameters checked: According to test guidelines

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination, between 7.30 and 9.30 a.m.
- Animals fasted: Yes, overnight
- How many animals: all rats/sex/group
- Parameters checked: According to test guidelines

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
NECROPSY:
All animals surv1v1ng to the end of the observation period (day 29) were deeply anaesthetised using ether vapour and subsequently exsanguinated. All animals assigned to the study were necropsied and descriptions of all macroscopic abnormalities recorded.

ORGAN WEIGHTS
The following organ weights (and terminal body weight) were recorded from the surviving animals on the scheduled day of necropsy:
Adrenal glands, Heart, Kidneys, Liver, Spleen and Testes.

HISTOPATHOLOGY: Yes
HISTOTECHNOLOGY
All organ and tissue samples, as defined under Histopathology (following), were processed, embedded and cut at a thickness of 2-4 micrometers and stained with haematoxylin and eosin.

HISTOPATHOLOGY
Slides of adrenals, heart, kidneys, liver, spleen, stomach and testes, collected at the scheduled sacrifice from all animals of the control and the highest dose group, and all gross lesions of all animals were examined by a pathologist.
Based on the treatment related morphologic changes, kidneys were also examined from all rats of the intermediate dose groups.
All abnormalities were described and included in the report.
Statistics:
Univariate one-way analysis of variance was used to assess the significance of intergroup differences.
If the variables could be assumed to follow a normal distribution, the Dunnetttest (many to one t-test) based on a pooled variance estimate was applied for
the comparison of the treated groups and the control groups for each sex.
The Steel-test (many-one rank test) was applied when the data could not be assumed to follow a normal distribution.
All tests were two-sided and in all cases p < 0.05 was accepted as the lowest level of significance.
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):
not examined
Food efficiency:
no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
effects observed, treatment-related
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
No mortality occurred during the study period .
There were no clinical signs of toxicity or behavioural changes over the 28-day observation period that were considered to be related to treatment.

BODY WEIGHT AND WEIGHT GAIN
Body weights and body weight gain of treated animals remained in the same range as controls over the 4-week study period.

FOOD CONSUMPTION
There were no differences in food consumption before or after allowance for body weight between treated and control animals.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study)

HAEMATOLOGY
Haematological parameters of treated rats were considered not to have been affected by treatment.

CLINICAL CHEMISTRY
There were no differences noted between control and treated rats that were considered to be related to treatment with ALCAMIZER 5.

ORGAN WEIGHTS
Spleen weights and spleen:body weight ratios were increased in males receiving 1000 mg/kg when compared to control weights. The spleen weights of the control animals, however, were considered to be slightly low when compared to values in other 28-day toxicity studies with similar rats (where control group mean spleen weights varied from 0.585 to 0.689 gram). The group mean value of 1000 mg/kg treated males remained within this range of historical data. In addition, there were no findings noted in the spleen microscopically and no corroborative findings were noted in the opposite sex. Therefore, it is unlikely that this change represents a toxic effect of the test substance.
Other organ weights and organ:body weight ratios of treated animals were considered to be similar to those of control animals.

GROSS PATHOLOGY
Macroscopic observations at necropsy did not reveal any alterations that were considered to have arisen as a result of treatment.

HISTOPATHOLOGY: NON-NEOPLASTIC
The kidneys of some male and female rats receiving 200 and 1000 mg/kg/day showed degenerative changes which were characterised by cystic dilated tubules containing proteinaceous casts. Regenerative changes, characterised as basophilic tubules, were seen in the kidneys of almost all male and female animals receiving 200 and 1000 mg/kg/day. The severity of both findings was very slight to slight.
One male receiving 1000 mg/kg/day showed a slight inflammatory change diagnosed as pyelonephritis. This finding was considered not to be a clear sign of toxicity as it can incidentally be noted in untreated rats of this age and strain. The small number of other findings recorded are within the normal range of background alterations.

Analysis of dose preparations:
Test substance formulations in propylene glycol formed a homogeneous suspension at the concentrations tested. Analysis of the accuracy of dose preparations revealed values within the range of 90% to 106% of nominal. One analysis resulted in 122% when compared to the nominal concentration based on Mg.
However, this sample revealed 95% based on the analysis of Al. Therefore, the results were considered to represent an acceptable level of accuracy for formulations of this type.
The stability of the test substance in the vehicle was not determined.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effects on kidneys were observed in the OECD421 study, up to and including 1000 mg/kg bw/day, the highest dose tested.
Critical effects observed:
no
Conclusions:
In a 28-day repeated dose oral toxicity study in rats performed according to OECD 407 guideline and GLP principles, the NOAEL was determined to be 1000 mg/kg bw/day.
Executive summary:

A 28-day repeated dose oral toxicity study in rats was performed with Alcamizer5 according to OECD 407 guideline and GLP principles. Based on the results of a 5-day dose range finding study, dose levels for the main study were selected to be 50, 200 and 1000 mg/kg bw/day. No mortality occurred during the study period. There were no clinical signs of toxicity or behavioural changes over the 28-day observation period that were considered to be related to treatment. Body weights and body weight gain of treated animals remained in the same range as controls over the 4-week study period.

Spleen weights and spleen:body weight ratios were increased in males receiving 1000 mg/kg bw/day when compared to control weights. The spleen weights of the control animals, however, were considered to be slightly low when compared to values in other 28-day toxicity studies with similar rats (where control group mean spleen weights varied from 0.585 to 0.689 gram). The group mean value of 1000 mg/kg bw/day treated males remained within this range of historical data. In addition, there were no findings noted in the spleen microscopically and no corroborative findings were noted in the opposite sex. Therefore, it is unlikely that this change represents a toxic effect of the test substance. Other organ weights and organ:body weight ratios of treated animals were considered to be similar to those of control animals.

The kidneys of some male and female rats receiving 200 and 1000 mg/kg bw/day showed degenerative changes which were characterised by cystic dilated tubules containing proteinaceous casts. Regenerative changes, characterised as basophilic tubules, were seen in the kidneys of almost all male and female animals receiving 200 and 1000 mg/kg bw/day. The severity of both findings was very slight to slight.

One male receiving 1000 mg/kg bw/day showed a slight inflammatory change diagnosed as pyelonephritis. This finding was considered not to be a clear sign of toxicity as it can incidentally be noted in untreated rats of this age and strain. The small number of other findings recorded are within the normal range of background alterations.

In a recently performed OECD 421 study in the same rat strain (Wistar), additional parental parameters were included in order to study the reported minimal effects on the kidney observed in the 28-day repeated dose toxicity study. No effects on kidneys were observed in the study, up to and including 1000 mg/kg bw/day, the highest dose tested. Based on the absence of any effects on the kidneys in the same rat strain in the recently performed study, it is concluded that the previously observed minimal effects on the kidneys should not be considered adverse.

Based on the results of the study, the NOAEL was determined to be 1000 mg/kg bw/day.

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Reason / purpose for cross-reference:
reference to same study
GLP compliance:
yes
Limit test:
no
Details on results:
For detailed description of results see sections 7.8.1 and 7.8.2.
Key result
Dose descriptor:
LOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
histopathology: non-neoplastic
organ weights and organ / body weight ratios
other: increased thyroid weight and higher incidence of follicular hypertrophy of the thyroid
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
100 mg/kg bw/day (actual dose received)
System:
endocrine system
Organ:
thyroid gland
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
not specified
Conclusions:
In an oral screening reproduction/developmental toxicity study in rats performed according to OECD 421 and GLP, the parental LOAEL was determined to be 100 mg/kg bw/day based on thyroid effects.
Executive summary:

In an oral screening reproduction/developmental toxicity study performed according to OECD 421 and GLP, rats were administered 0, 100, 300 or 1000 mg/kg bw/day of ALCAMIZER5 daily by gavage. Males were treated for 29 days, i.e. 2 weeks prior to mating, during mating, and up to the day prior to scheduled necropsy. Females were treated for 42-53 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during 4-5 days of lactation (i.e. up to the day prior to scheduled necropsy. Females which failed to deliver healthy offspring were treated for 42 or 53 days.

The following observations and examinations were evaluated: mortality / viability, clinical signs (daily), body weight and food consumption (at least at weekly intervals), clinical biochemistry, macroscopy at termination, organ weights and histopathology on a selection of tissues. In addition, the following reproduction/developmental parameters were determined: mating, precoital time, fertility index, numbers of corpora lutea and implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights, sex and macroscopy).

In males and females of the 1000 mg/kg bw/day dose group, alkaline phosphatase (ALP) was significantly increased. When compared to the concurrent control group, mean values were 51% (males) and 62% (females) higher. However, in the absence of any corroborative findings in the remaining clinical biochemistry parameters and macroscopic findings at necropsy, this observation was not considered to be adverse.

Microscopic examination revealed an increase in incidence and/or severity (up to marked degree) of follicular cell hypertrophy in males and females starting at 100 mg/kg bw/day. This change was accompanied by a substantial increase in thyroid organ weight in males at 100, 300 and 1000 mg/kg bw/day(increase in relative weight of 67%, 83% and 83%, respectively), and in females at 300 and 1000 mg/kg bw/day (increase in relative weight of 25% and 28%, respectively). At the macroscopic level, enlarged thyroid glands were observed in 10/10 males treated at 100, 300 and 1000 mg/kg bw/day and in 6/10 females treated at 1000 mg/kg bw/day.

There were no other test item-related histologic changes. No indication of any adverse effects on kidney was observed (included specifically to investigate the observed effects on kidney in the previously performed 28 day repeated dose toxicity study in the same rat strain). No treatment-related changes were noted in any of the remaining parental parameters investigated in this study (i.e. mortality, clinical appearance, body weight and food consumption).

Conclusion

Treatment with ALCAMIZER5 by oral gavage in male and female Wistar Han rats at dose levels of 100, 300 and 1000 mg/kg bw/day resulted in follicular cell hypertrophy in the thyroid glands, with correlating higher organ weights and enlargement observed at necropsy, in males and females starting at 100 mg/kg bw/day.

The increase in alkaline phosphatase (ALP) noted in males and females at 1000 mg/kg bw/day was regarded treatment-related, but not adverse. No indication of any adverse effects on kidney was observed (included specifically to investigate the observed effects on kidney in the previously performed 28 day repeated dose toxicity study in the same rat strain).

Based on the histologic findings in the thyroid glands, no parental NOAEL could be established and the level of 100 mg/kg bw/day was considered to be a LOAEL.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
100 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
An OECD 407 study is the required study for Annex VIII for the endpoint repeated dose toxicity. In addition, information on repaeted dose toxicity is available from the reproduction/developmental toxicity screening study.
System:
endocrine system
Organ:
thyroid gland

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Two repeated dose toxicity studies with relevant information are available: a 28 day repeated doe toxicity study and a reproduction/developmental toxicity screening study.

An 28-day repeated dose oral toxicity study in rats was performed with Aluminium-magnesium-carbonatehydroxide-perchlorate-hydrate according to OECD 407 guideline and GLP principles. Based on the

results of a 5-day dose range finding study, dose levels for the main study were selected to be 50, 200 and 1000 mg/kg bw/day. No mortality occurred during the study period . There were no clinical signs of toxicity or behavioural changes over the 28-day observation period that were considered to be related to treatment. Body weights and body weight gain of treated animals remained in the same range as controls over the 4-week study period.

Spleen weights and spleen:body weight ratios were increased in males receiving 1000 mg/kg bw/day when compared to control weights. The spleen weights of the control animals, however, were considered to be slightly low when compared to values in other 28-day toxicity studies with similar rats (where control group mean spleen weights varied from 0.585 to 0.689 gram). The group mean value of 1000 mg/kg bw/day treated males remained within this range of historical data. In addition, there were no findings noted in the spleen microscopically and no corroborative findings were noted in the opposite sex. Therefore, it is unlikely that this change represents a toxic effect of the test substance. Other organ weights and organ:body weight ratios of treated animals were considered to be similar to those of control animals.

The kidneys of some male and female rats receiving 200 and 1000 mg/kg bw/day showed degenerative changes which were characterised by cystic dilated tubules containing proteinaceous casts. Regenerative changes, characterised as basophilic tubules, were seen in the kidneys of almost all male and female animals receiving 200 and 1000 mg/kg bw/day. The severity of both findings was very slight to slight.

One male receiving 1000 mg/kg bw/day showed a slight inflammatory change diagnosed as pyelonephritis. This finding was considered not to be a clear sign of toxicity as it can incidentally be noted in untreated rats of this age and strain. The small number of other findings recorded are within the normal range of background alterations.

In a recently performed OECD 421 study in the same rat strain (Wistar), additional parental parameters were included in order to study the reported minimal effects on the kidney observed in the 28-day repeated dose toxicity study. No effects on kidneys were observed in the study, up to and including 1000 mg/kg bw/day, the highest dose tested. Based on the absence of any effects on the kidneys in the same rat strain in the recently performed study, it is concluded that the previously observed minimal effects on the kidneys should not be considered adverse.

Based on the results of the study, the NOAEL was determined to be 1000 mg/kg bw/day.

In an oral screening reproduction/developmental toxicity study performed according to OECD 421 and GLP, rats were administered 0, 100, 300 or 1000 mg/kg bw/day of ALCAMIZER5 daily by gavage. Males were treated for 29 days, i.e. 2 weeks prior to mating, during mating, and up to the day prior to scheduled necropsy. Females were treated for 42-53 days, i.e. during 2 weeks prior to mating, during mating, during post-coitum, and during 4-5 days of lactation (i.e. up to the day prior to scheduled necropsy. Females which failed to deliver healthy offspring were treated for 42 or 53 days.

The following observations and examinations were evaluated: mortality / viability, clinical signs (daily), body weight and food consumption (at least at weekly intervals), clinical biochemistry, macroscopy at termination, organ weights and histopathology on a selection of tissues. In addition, the following reproduction/developmental parameters were determined: mating, precoital time, fertility index, numbers of corpora lutea and implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development (mortality, clinical signs, body weights, sex and macroscopy).

In males and females of the 1000 mg/kg bw/day dose group, alkaline phosphatase (ALP) was significantly increased. When compared to the concurrent control group, mean values were 51% (males) and 62% (females) higher. However, in the absence of any corroborative findings in the remaining clinical biochemistry parameters and macroscopic findings at necropsy, this observation was not considered to be adverse.

Microscopic examination revealed an increase in incidence and/or severity (up to marked degree) of follicular cell hypertrophy in males and females starting at 100 mg/kg bw/day. This change was accompanied by a substantial increase in thyroid organ weight in males at 100, 300 and 1000 mg/kg bw/day(increase in relative weight of 67%, 83% and 83%, respectively), and in females at 300 and 1000 mg/kg bw/day (increase in relative weight of 25% and 28%, respectively). At the macroscopic level, enlarged thyroid glands were observed in 10/10 males treated at 100, 300 and 1000 mg/kg bw/day and in 6/10 females treated at 1000 mg/kg bw/day.

There were no other test item-related histologic changes. No indication of any adverse effects on kidney was observed (included specifically to investigate the observed effects on kidney in the previously performed 28 day repeated dose toxicity study in the same rat strain). No treatment-related changes were noted in any of the remaining parental parameters investigated in this study (i.e. mortality, clinical appearance, body weight and food consumption).

Conclusion

Treatment with ALCAMIZER5 by oral gavage in male and female Wistar Han rats at dose levels of 100, 300 and 1000 mg/kg bw/day resulted in follicular cell hypertrophy in the thyroid glands, with correlating higher organ weights and enlargement observed at necropsy, in males and females starting at 100 mg/kg bw/day.

The increase in alkaline phosphatase (ALP) noted in males and females at 1000 mg/kg bw/day was regarded treatment-related, but not adverse. No indication of any adverse effects on kidney was observed (included specifically to investigate the observed effects on kidney in the previously performed 28 day repeated dose toxicity study in the same rat strain).

Based on the histologic findings in the thyroid glands, no parental NOAEL could be established and the level of 100 mg/kg bw/day was considered to be a LOAEL.

For the endpoint repeated dose toxicity an overall LOAEL of 100 mg/kg bw/day id considered for risk assessment purposes.

Justification for classification or non-classification

Based on the available information on the substance Aluminium-magnesium-carbonatehydroxide-perchlorate-hydrate, the substance does not have to be classified for oral repeated dose toxicity according to Regulation (EC) no. 1272/2008 and its amendments.