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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-03-28 to 2018-04-24
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Version / remarks:
22 January 2001
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Version / remarks:
August 1998
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Version / remarks:
30 May 2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Constituent 1
Test material form:
solid: crystalline

Test animals

Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Toxi-Coop Zrt., 1103 Budapest, Cserkesz u. 90., Hungary
- Age at study initiation: males: 35-52 weeks of age; females: 10 weeks
- Weight at study initiation: 157-236 g (females)
- Housing:
Before mating: 1-3 females per cage 1-2 males per cage
During mating: 1 male and 1-3 females / cage
During gestation: 2 sperm positive females per cage, if not possible 1 sperm positive female per cage

- Diet: ad libitum, ssniff® SM R/M-Z+H "Autoclavable complete feed (ssniff Spezialdiäten GmbH, D-59494 Soest, Germany)
- Water: ad libitum
- Acclimation period: 16 days for females, ca. 20 weeks for males

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 22
- Humidity (%): 33 - 68
- Air changes (per hr): above 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test item was formulated in the vehicle (water, Aqua purificata) in concentrations of 20 mg/mL, 6 mg/mL and 2 mg/mL. Formulations were prepared in the formulation laboratory of the Test Facility with a frequency of daily to every three days and stored at room temperature in accordance with the results of stability measurements in the frame of the analytical method validation.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The suitability of the chosen vehicle for the test item was analytically proven. Recovery was 102 and 101 % of nominal concentrations at 0.1 mg/mL and 200 mg/mL in distilled water, respectively. Diammonium peroxodisulphate (APS) was proved to be stable in the formulations for three days at room temperature and in the refrigerator (5 ± 3°C). A separate analytical report provided these results (Toxi-Coop study no. 552-100-3004). Analytical control of dosing solutions (control of test item concentration and homogeneity) was performed in the Analytical Laboratory of Test Facility two times during the study.
Five samples from different places were taken from each concentration for analysis of concentration and homogeneity on two occasions. Similarly, five samples were taken from the vehicle (Control) and analyzed. The formulations proved to be homogeneous. Test item concentrations in the dosing formulation samples varied in the range from 95% to 104% of the nominal values. Results of analysis are added to the Study Report
Details on mating procedure:
- Impregnation procedure: cohoused
- M/F ratio per cage: one male: one to three females
- Length of cohabitation: 2 - 4 hours
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
from gestational day 5 to 19
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Dose / conc.:
10 mg/kg bw/day
Remarks:
2 mg/mL
Dose / conc.:
30 mg/kg bw/day
Remarks:
6 mg/mL
Dose / conc.:
100 mg/kg bw/day
Remarks:
20 mg/mL
No. of animals per sex per dose:
26
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose setting was based on findings obtained in previous studies (Acute Toxicity of Potassium Persulfate - 28 day Subacute Feeding Study in Male Albino Rats, ICG/T-79-024; Acute Toxicity of Ammonium Persulfate - 28 day Subacute Feeding Study in Male Albino Rats, ICG/T-79-025; Oral Reproductive/Developmental Toxicity Screening Test in Rats with Ammonium Persulfate in Feed, I2003-2337; Safety Evaluation of: Sodium persulfate - A 90-Day Dietary Feeding Study In Rats, I1990-1151) provided by the Sponsor. The 300 mg/kg bw/day dose in the Preliminary Prenatal Developmental Toxicity Study with Diammonium peroxodisulphate (APS) in the Rat by Oral (Gavage) Administration (Toxi-Coop Zrt. study number: 552-410-3000) caused death of 3/ 9 animals. Two animals of 7 had temporary clinical signs in the mid dose (100 mg/kg bw/day) and slight reductions in the body weight gain and food consumption. The high dose 100 mg/kg bw/day in this main study was selected accordingly with the aim to induce some maternal/developmental toxicity but not death or severe suffering. The low dose 10 mg/kg bw/day was chosen to induce no toxic effect. The mid dose 30 mg/kg bw/day was interpolated geometrically.

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily (morbidity and mortality were checked twice daily)
- Cage side observations included check of behavior and general condition, duration and severity of the clinical signs and observations for signs of morbidity and mortality.

BODY WEIGHT: Yes
- Time schedule for examinations: gestation days 0, 3, 5, 8, 11, 14, 17 and 20

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: Uterus with cervix and ovaries

OTHER:
Examination of placental signs: All sperm positive animals were examined for vaginal bleeding (placental sign of gestation) on the 13th gestational day. If negative on day 13, the examination was repeated on day 14 of gestation.
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: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter
Statistics:
The statistical evaluation of data was performed with the program package SPSS PC+4.0.
The homogeneity of variance between groups was checked by Bartlett’s homogeneity of variance test.Where no significant heterogeneity is detected a one-way analysis of variance (ANOVA) is carried out. If the obtained result is significant Duncan’s Multiple Range test was used to assess the significance of inter-group differences. If significance is the result of the Bartlett’s test, the Kruskal-Wallis analysis of variance was used and the inter-group comparisons were performed using Mann-Whitney U-test.
Dams or litters were excluded from the data evaluation in cases of:
- Disease or death of the dam unrelated to the treatment (total exclusion)
- Non pregnant females i.e. females with no implantation and no corpora lutea (total exclusion)
- Body weight, body weight gain, food consumption, clinical signs and necropsy findings of females with no implantation but corpora lutea i.e. total preimplantation loss (only the intrauterine parameters were evaluated, partial exclusion)
- Body weight, body weight gain, food consumption of females with total post-implantation loss
- Circumstances unrelated to the test item which are considered to be reason for exclusion, at the discretion of the Study Director
Although these animals were excluded from the data evaluation the Study Report contains all data of these animals, too. A male/female fetus was considered as retarded in body weight, when its weight was below the average minus twofold standard deviation of the control male/female fetuses.
Historical control data:
The results were compared to the laboratory's historical control data.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
There was no statistical significance indicated in the body weight values. The body weight gain was slightly lower in the 100 mg/kg bw/day dose group between GD 5 and 8, 17 and 20 as well 0 to 20. The corrected body weight and body weight gain was slightly lower in the 100 mg/kg bw/day group without a statistical significance. These differences were in association with the results of the DRF study (Dose Range Finding Prenatal Developmental Toxicity Study with Diammonium Peroxodisulphate (APS) in the Rat by Oral (Gavage) Administration Toxi-Coop study number: 552-410-3000) and were attributed to the test item.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Slight, but statistically significant decreases of the food consumption were observed in the 100 mg/kg bw/day group on GD 5 to 8 (-16%) and 11 to 14 (-9%) if compared to the control. These differences were in association with the results of the DRF study (Dose Range Finding Prenatal Developmental Toxicity Study with Diammonium Peroxodisulphate (APS) in the Rat by Oral (Gavage) Administration Toxi-Coop study number: 552-410-3000) and were attributed to the test item. A slight, statistically significant increase in the 10 mg/kg bw/day dose group between GD 14 and 17 without a dose response was not attributed to the treatment.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Reddish mottled lungs or pinhead sized haemorrhages in the lungs were recorded without a dose response. Gas filled uterine horn was observed in the uterus for a dam in the 30 mg/kg bw/day group. These observations were not attributed to the test item.

Maternal developmental toxicity

Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
There was no statistically significant increase indicated in the mean percent of pre- and post-implantation loss (early embryonic, late- and fetal death). The CH square test indicated a statistically significant increase (p<0.01) in the pre-implantation loss in all test item treated groups and in the total intrauterine mortality (pre- and post-implantation loss summarized) in the 100 mg/kg bw/day dose group. Considering that the mean percentage pre-implantation loss was between the historical control range (2.4-21.4% and 21.0-22.82% respectively) or slightly above (30 mg/kg bw/day), this effect was not attributed to the test item.

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
food consumption and compound intake

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Fetal body weight changes:
no effects observed
Reduction in number of live offspring:
no effects observed
Changes in sex ratio:
no effects observed
Changes in litter size and weights:
no effects observed
Changes in postnatal survival:
not examined
External malformations:
no effects observed
Description (incidence and severity):
Malformations
One fetus in the 10 mg/kg bw/day dose group was found with umbilical hernia which was not attributed to the test item since one fetus of the control group showed the same malformation.

Variations
Body weight retardation (below 2.56 g for males and 2.51 g for females) was found without a dose response. One fetus had a slightly shorter mandible in the 100 mg/kg bw/day doss group and one was judged to be edematous in the 30 mg/kg bw/day group. These variations were not attributed to an effect of the test item considering the low incidence and that similar alteration may occur in control fetuses.

Placentas, amnion
Seven edematous placentas were observed in the 100 mg/kg bw/day dose group. Considering that it was found in one single litter and that edematous placentas can occur in control animals, this placental change was not attributed to the treatment of the dams with the test item.
Skeletal malformations:
no effects observed
Description (incidence and severity):
Malformations
The number of the affected litters was 0, 1, 1 and 3 in in the control, 10, 30 and 100 mg/kg bw/day group, respectively.
A misshapen xiphoid cartilage in one fetus each in the 10 and 30 mg/kg bw/day groups and one split sternum in the 30 mg/kg bw/day group was observed. One fetus in the 30 mg/kg bw/day group had bipartite cartilage of a thoracic centrum as well as two fetuses in the 100 mg/kg bw/day had dumb-bell shaped cartilage of thoracic centrum. One fetus was found with bent scapula and slightly bent radius in the 100 mg/kg bw/day dose group.
The overall incidence of malformations in the 100 mg/kg bw/day dose group showed a statistical significance (3 fetuses with malformations, within the historical control range. The different type of malformations occurred with low incidences or/and without a dose response. Furthermore most of these changes occur in control fetuses without a relationship to the treatment according to the historical control data. Therefore these malformations were judged to be unrelated to an effect of the test item.

Variations
There was no dose related increase observed in the incidence of variations. Moreover, it was lowest in the 100 mg/kg bw/day dose group.
Incompletely, not ossified or retarded skull bones, bipartite supra occipital, unossified hyoid bone, incompletely ossified sternebra, interrupted or wavy ribs, dumb-bell shaped, bipartite, asymmetrically ossified, incompletely ossified or unossified vertebral centra, unossified pubic bone, and asymmetric or incomplete ossification of metacarpal and metatarsal were evaluated as variations during the skeletal examination. There was no dose related statistical significance indicated in the different type of variations.
There was a statistical significant increase (p<0.05) in the occurrence of litters with markedly incomplete ossification of skull bones at 10 mg/kg bw/day and fetuses with wavy ribs at 30 mg/kg bw/day but no dose response was indicated. There were no significant differences in the incidence of the other types of variations.
Visceral malformations:
no effects observed
Description (incidence and severity):
Malformations
The number of the affected litters was 1, 2, 1 and 0 in the control, 10, 30 and 100 mg/kg bw/day groups, respectively.
The fetuses found with umbilical hernia at external examination in the control (No.: 1030187/9) and 10 mg/kg bw/day (No.: 1451169/7) dose groups (discussed at external examination) were recoded also at visceral examination. Fetus No.: 1451169/7 in the 10 mg/kg bw/day had rudimentary brain tissue in addition and was cathegorized as multiply malformed. These findings were not attributed to the treatment considering the low incidence and that absent brain tissue may occur also in control fetuses according to the Background Pregnancy and Fetal data.
Situs inversus cordis and an absent lung lobe in one fetus in the 10 mg/kg bw/day as well as situs inversus totalis in one fetud in the 30 mg/kg bw/day were observed. These alterations may occur with low incidence in control fetuses according to the experience of this laboratory which is in line with the historical control data. These malformations were judged as incidental.

Variations
Hydro- or dilated ureter was found with a slight increase in the 100 mg/kg bw/day group (statistical significance p<0.05). This was not proved to be an effect due to the treatment since the incidence was within the background control data. Hydro ureter with dilated renal pelvis was observed in one fetus each in the 30 and 100 mg/kg bw/day groups. Considering the low incidence this was not attributed to the treatment.

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No treatment related effects were observed.

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Applicant's summary and conclusion

Conclusions:
Oral treatment of pregnant Han: Wistar rats from gestation day 5 up to day 19 (the day before Caesarean section) with Diammonium peroxodisulphate (APS) at the dose levels of 10, 30 and 100 mg/kg bw/day did not cause death, or revealed clinical signs and treatment related macroscopic changes at gross pathology. Slightly reduced food intake on GD 5 to 8 and 11 to 14 and slightly lower body weight gain on GD 5 to 8, 17 to 20 and 0 to 20, as well as slightly lower corrected body weight/gain were attributed to the test item at the dose level of 100 mg/kg bw/day. Diammonium peroxodisulphate (APS) did not reveal any adverse effect on the pre- and post-implantation loss, number of implantation, sex distribution, body and placental weight, external, visceral and skeletal development of the fetuses. Based on these observations the No Observed Adverse Effect Level (NOAEL) was determined as follows:
NOAEL (maternal toxicity): 30 mg/kg bw/day
NOAEL (developmental toxicity including teratogenicity): 100 mg/kg bw/day
Executive summary:

Diammonium peroxodisulphate (APS) was examined for its possible prenatal developmental toxicity in accordance with OECD guideline 414. Groups of 26 sperm-positive female Han: Wistar rats were treated with Diammonium peroxodisulphate (APS) by oral administration daily at three dose levels of 10, 30 and 100 mg/kg bw/day respectively from day 5 up to and including day 19 post coitum. A control group of 26 sperm positive females was included and the animals were given the vehicle water (aqua purificata). The treatment volume was 5 mL/kg bw.

A sufficient stability and homogeneity in the chosen vehicle was verified over the range of relevant concentrations at the appropriate frequency of preparation. Diammonium peroxodisulphate (APS) in water was stable at room temperature and in the refrigerator (5 ± 3 oC) for three days at the concentrations of ca. 0.1 and 200 mg/mL. Analytical control of dosing solutions was performed during the first and last week of treatment. Concentrations of the test item in the dosing formulations varied in the acceptable range between 95 and 104 % of nominal concentrations at both analytical occasions confirming proper dosing.

During the study, mortality was checked and clinical observations were performed. Body weight and food consumption of the dams were also recorded. The day, when sperm was detected in the vaginal smear, was regarded as day 0 of gestation. Caesarean section and gross pathology were performed on gestational day (GD) 20. The number of implantations, early and late resorptions, live and dead fetuses in each uterine horn and the number of corpora lutea were recorded. Each fetus was weighed and examined for sex and gross external abnormalities. The placentas were weighed and examined externally. About half of each litter was preserved for visceral examination and the other half of the litters were preserved for skeletal evaluation. At visceral examination the bodies were micro dissected by means of a dissecting microscope. The heads were examined by Wilson's free-hand razor blade method. After cartilage-bone staining the skeletons were examined by means of a dissecting microscope. All abnormalities found during the fetal examinations were recorded.

In total, on gestation day 20 there were dams with 23, 20, 16 and 16 implantation sites in the control, 10, 30 and 100 mg/kg bw/day group, respectively. None of the females died before scheduled necropsy and there were no test item related clinical signs recorded. No treatment related necropsy findings were observed. Slight, statistically significant decreases of food consumption were indicated in the 100 mg/kg bw/day dose group on GD 5 to 8 and 11 to 14. Slightly lower body weight gain was observed between GD 5 and 8 and 17 and 20. A slightly lower corrected body weight/gain was observed between GD 0 to 20 without a statistical significance.

These differences were in association with the results of the DRF study (Dose Range Finding Prenatal Developmental Toxicity Study with Diammonium Peroxodisulphate (APS) in the Rat by Oral (Gavage) Administration Toxi-Coop study number: 552-410-3000) and were therefore attributed to the test item.

The CH square test indicated a statistical significance in the increase of pre-implantation loss in all test item treated groups. A statistical significant increase of the total intrauterine mortality was observed in the 100 mg/kg bw/day dose group. The mean percentage pre-implantation loss was between the historical control range or slightly above (30 mg/kg bw/day). The mean percentage intrauterine mortality was also between the historical background values. Thus, this was not attributed to treatment.Moreover the mean number of implantations was highest in the 100 mg/kg bw/day dose. There were no dose related increases seen in the post-implantation loss and sex distribution of the fetuses was not influenced by the treatment.

There were no test item related adverse effects on the fetal- and placental weight. There were no test item related external malformations and variations found. The visceral malformations were not attributed to the treatment. The

Skeletal malformations were found in three fetuses with a statistical significance. However the incidence was low and the type of alterations less severe and partially different. There was no dose related increase seen in external and skeletal variations. Based on these observations the No Observed Adverse Effect Level (NOAELs) were determined as follows: NOAEL (maternal toxicity): 30 mg/kg bw/day, NOAEL (developmental toxicity): 100 mg/kg bw/day, NOAEL (teratogenicity): 100 mg/kg bw/day (high dose).