Sodium O,O-diisobutyl dithiophosphate

Endpoint summary

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Effects on fertility

Description of key information

OECD 422: NOAEL (no-observed-adverse-effect level): 600 mg test item/kg b.w./day, p.o.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-08-29 - 2013-04-24

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

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: Charles River Laboratories,Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: Males/females: Both 59 days
- Weight at study initiation: Males: 272.2 g to 299.9 g, Females: 210.4 g to 233.6 g


- Fasting period before study: 16 hours
- Housing: kept individually in MAKROLON cages (type III plus)
- Diet (e.g. ad libitum): Commercial ssniff® R/M-H V1534 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany; served as food ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C
- Humidity (%): 55% ± 15%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on exposure:

VEHICLE
- Amount of vehicle (if gavage): 10 ml/kg b.w./day

Details on mating procedure:

Sexually mature male and female rats were randomly paired for mating. Matingwas monogamous: 1 male and 1 female animal were placed in one cage during the dark period. The female was placed with the same male until pregnancy had occurred or 2 weeks had elapsed. Each morning the females were examined for the presence of sperm or a vaginal plug. If findings were negative, mating was repeated.
The day of conception (day 0 of gestation) was considered to be the day on which sperm was found. In case pairing was unsuccessful, re-mating of females with proven males of the same group was considered. This procedure was repeated until at least 8 pregnant dams were available for each group.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

For the analysis of the test item-vehicle mixtures samples of approx. 2 x 1 mL were taken at the following time points and stored at ≤ minus 20°C until analysis at LPT:

Start of treatment period:
Concentration and stability:
Immediately after preparation of the test item vehicle mixtures as well as 8 and 24 hours after storage of the test item preparations at room temperature:
3 samples/dose level group (groups 2 to 4)
Number of samples: 3 x 3 = 9
Homogeneity:
At start of administration, during (middle) administration and before administration to the last animal of each dose level group:
3 samples/dose level group (groups 2 to 4).
Number of samples: 3 x 3 = 9

End of treatment period:
Concentration:
During treatment with the test item always before administration to the last animal/dose level group:
1 sample/dose level group (groups 2 to 4).
Number of samples: TD 26 (1 x 3) = 3
Number of samples: TD 39 (1 x 3) = 3
Sum of all samples: 24
Sum of all aliquots: 48

The samples were labelled with the study number, test item, test species, type of sample, aliquot number, concentration, test day, sampling time and date.

Duration of treatment / exposure:

Males
The daily administration of the test item started two weeks before mating and lasted until the day before sacrifice (considering a minimum total dosing period of at least 28 days).
Females
The daily administration of the test item started two weeks before mating and lasted up to at least day 3 of lactation.

Frequency of treatment:

once daily

Details on study schedule:

- Age at mating of the mated animals in the study: 10-11 week

Remarks:

Doses / Concentrations:
0 mg/kg bodyweight/day (control)
Basis:
actual ingested

Remarks:

Doses / Concentrations:
60 mg/kg bodyweight/day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
200 mg/kg bodyweight/day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
600 mg/kg bodyweight/day
Basis:
actual ingested

No. of animals per sex per dose:

80 animals (40 male and 40 female rats), 10 animals/sex/group.
A sufficient number to grant at least 8 pregnant females per group for evaluation of the F0 generation

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: See supporting study: 14-DAY DOSE-RANGE-FINDING STUDY OF IBP1-Na (DANAFLOATTM 245) BY ORAL ADMINISTRATION TO RATS

Parental animals: Observations and examinations:

AGE SIDE OBSERVATIONS: Yes
- Time schedule: at least daily, The frequency was increased when signs of toxicity were observed.
Cageside observations included skin/fur, eyes, mucous membranes, respiratory and circulatory systems, somatomotor activity and behaviour patterns. The onset, intensity and duration of any signs observed were recorded.
Individual animals were observed before and after dosing at each time of dosing for any signs of behavioural changes, reaction to treatment or illness.
In addition, animals were checked regularly throughout the working day from 7:00 a.m. to 3:45 p.m. On Saturdays and Sundays animals were checked regularly from 7:00 a.m. to 11:00 a.m. with a final check performed at approximately 3:30 p.m.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least daily, The frequency was increased when signs of toxicity were observed.
Additionally, once before the first exposure (to allow within-subject comparisons) and once a week thereafter, detailed clinical observations were made in all animals outside the home cage in a standard arena and at the same time, each time preferably by observers unaware of the treatment. Signs noted included changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, piloerection, pupil size, and unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. selfmutilation, walking backwards) were also recorded.
Dated and signed records of appearance, change, and disappearance of clinical signs were maintained on clinical history sheets for individual animals.

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of dosing, weekly thereafter and at termination of the study. During gestation, females were weighed on days 0, 7, 14 and 20 and within 24 hours of parturition (day 1 post-partum) and day 4 postpartum.
Body weights were recorded individually for each adult animal.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

At the intermediate and the high dose level (200 and 600 mg IBP1-Na/kg b.w./day) a slight to extreme salivation was noted in nearly all animals, with a higher incidence at the high dose level. Piloerection and an increased water consumption were noted for a few animals of the intermediate and the high dose group, which is most like related to the alkalinity of the substance.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

No test item-related premature death was noted.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

A slight reduction in body weight was noted until the end of the study on test day 38 for the male rats of the high dose group (600 mg IBP1-Na/kg b.w./day). Body weight at autopsy was reduced accordingly

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

A slight reduction in body weight was noted until the end of the study on test day 38 for the male rats of the high dose group (600 mg IBP1-Na/kg b.w./day). Body weight at autopsy was reduced accordingly

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The histopathological examination revealed test item related changes in the forestomach from male and female rats of the intermediate and the high dose group (200 and 600 mg IBP1-Na/kg b.w./day) in form of hyperplasia and hyperkeratosis of the squamous cell epithelium, infiltration with neutrophilic granulocytes, granulation tissue and ulcerations.
Additionally, test item-related changes were noted in the liver in the form of a centrilobullar hepatocellular hypertrophy at 600 mg/kg bw/day.

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Dose descriptor:

NOAEL

Effect level:

200 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: General health parameters

Dose descriptor:

NOAEL

Effect level:

600 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Reproductive parameters

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

In the high dose group the mean litter weight of the male and female pups was decreased by 5.6% on lactation day 1 and by 9.6% on lactation day 4 (without statistical significance). No noticeable differences were noted for the total litter weight

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

600 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: systemic effects

Reproductive effects observed:

not specified

Conclusions:

NOAEL (no-observed-adverse-effect level): 200 mg test item/kg b.w./day, p.o.

Remark: The test solution contains NaOH due to the production method.

Executive summary:

The test was performed according to OECD guideline 422 under GLP compliance. The test substance was administered orally to rats at dose levels of 60, 200 or 600 mg test item/kg b.w./day.

The following no-observed-effect levels were established for systemic effects: NOAEL (no-observed-adverse-effect level): 200 mg test item/kg b.w./day, p.o.

The following no-observed-effect levels were established for reproductive effects: NOAEL (no-observed-adverse-effect level): 600 mg test item/kg b.w./day, p.o.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

600 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

high quality

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Justification for selection of Effect on fertility via oral route:

Effects on developmental toxicity

Description of key information

OECD 414: There were no treatment-related adverse effects on embryo-fetal development. Thus, the NOAEL for the maternal toxicity and embryo-fetal developmental toxicity is 800 mg/ kg body weight/ day.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material:0001021592
- Expiration date of the lot/batch:December 11th, 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature, protected from light
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: Stable

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, 97633 Sulzfeld, Germany
- Age at study initiation: 12-13 weeks
- Weight at study initiation:
Body weight before
the start of pairing:
males: 349 – 381 g (mean: 366.67 g, ± 20% = 293.34 – 440.01 g)
females: 180 - 236 g (mean: 204.17 g, ± 20% = 163.34 – 245.01 g)
- Housing: - Full barrier in an air-conditioned room
- Diet (e.g. ad libitum): - Free access to Altromin 1324 maintenance diet for rats and mice
- Water (e.g. ad libitum): - Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiological controls at regular intervals)
- Acclimation period: minimum 5 days

DETAILS OF FOOD AND WATER QUALITY:
- Diet (e.g. ad libitum): - Free access to Altromin 1324 maintenance diet for rats and mice
- Water (e.g. ad libitum):- Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control
ENVIRONMENTAL CONDITIONS
- Temperature (°C):22 ± 3 °C
- Humidity (%): 55 ± 10 %
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To:
Delivery of Animals: 03 March 2016
Acclimatisation Period: 03 - 08 March 2016
Experimental Starting Date: 08 March 2016
Experimental Completion Date: 08 July 2016

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): water used
- Concentration in vehicle: 0 23,5, 117,6 and 313,7 mg/l
- Amount of vehicle (if gavage): 5 mL/kg body weight
- Lot/batch no. (if required):
Name: aqua ad iniectabilia
Manufacturer: AlleMan Pharma
Batch No.: 503424
Physical State: liquid
Storage Conditions: at room temperature
Expiry Date: 28/02/2018

- Purity: pure

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The assessment of homogeneity as well as a determination of the nominal concentration of the test item in the vehicle was performed at various intervals using a validated analytic method (BSL Munich study no. 156096).
Samples for analysis of concentration analysis of the active ingredient within the vehicle (nominal concentration) were taken in the first and last week of the study for all doses, including controls (8 samples in total). Samples for homogeneity were taken from the top, middle and bottom of the high dose and low dose preparation. Samples were taken in the first and last week of the study (12 samples in total).
Each sample was retained in duplicates (sample A, sample B, each at least 5 mL). The A- sample were analysed at Eurofins Munich. The procedures followed for the study sample analysis were mentioned in a phase plan (BSL Munich study no. 156108) that was amended to the study plan. The B-samples were retained at -15 to -35°C at BSL Munich until the analysis had been performed, and discarded after completion of the final study report. Analytical results are reported in the appendix of the present report (Appendix 3).
Concentration results were considered acceptable if sample concentration results are within or equal to ± 10% of theoretical concentration. For homogeneity, the criteria for acceptability were a relative standard deviation (RSD) of concentrations of ≤10% for each group.

Details on mating procedure:

Mating was performed using a ratio of 1:2 (male to female). Females were paired for cohabitation in batches in order to control the number of animals for terminal sacrifice on a particular day. At the subsequent morning and the next morning onwards, the vaginal smear of each female was checked to confirm the pregnancy. The day on which sperms were observed in the vaginal smear was considered as gestation day ‘0’. Mated females were assigned in an unbiased manner to the control and treatment groups ensuring that group mean body weights were comparable with each other. Each animal was assigned an unique identification number. After obtaining a sufficient number of sperm positive females, the remaining females and males were discarded without any observations.

Duration of treatment / exposure:

The pregnant females were treated with the test item formulation or vehicle on 7 days per week between gestation day 5 until gestation day 19.

Frequency of treatment:

Daily, between gestation day 5 and gestation day 19.

Duration of test:

Until gestation day 20

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Control group, group name C

Dose / conc.:

60 mg/kg bw/day (nominal)

Remarks:

Lowest dosage group, group name LD

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Medium dosage group, group name MD

Dose / conc.:

800 mg/kg bw/day (nominal)

Remarks:

High dosage group, group name HD

No. of animals per sex per dose:

The control group had 23 sperm positive females, the LD group had 29 sperm positive females and the MD and HD groups each had 24 sperm positive females. However, at necropsy, the number of gravid females was 21 of 23 in the control group, 21 of 29 in the LD group, 20 of 24 in the MD group and 20 of 24 in the HD group.

Control animals:

yes, concurrent vehicle

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: General clinical observations were made once a day, approximately at the same time each day at 30 min after dosing. Twice daily all animals were observed for morbidity and mortality except on weekends and public holidays when observations were made once daily.


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: General clinical observations were made once a day, approximately at the same time each day at 30 min after dosing. Twice daily all animals were observed for morbidity and mortality except on weekends and public holidays when observations were made once daily.

BODY WEIGHT: Yes
- Time schedule for examinations: All animals were weighed once before initiation of pairing to ensure that the body weights are within + 20% variation.
The sperm positive females were weighed during gestations days 0, 5, 8, 11, 14, 17 and 20. Males were not weighed in this study except once before initiation of pairing.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
Food consumption of sperm positive females was measured on gestations days 5, 8, 11, 14, 17 and 20.
Food consumption was not measured for males during the entire study or for both males and females during the mating period

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


POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: At the time of termination, the dam (presumed pregnant female) was examined macroscopically for any structural abnormalities or pathological changes, which may have influenced the pregnancy. Immediately after the termination, the uteri were removed and the pregnancy status of the dams was confirmed. Uteri that appear non-gravid were further examined by staining with 10 % ammonium sulphide solution to confirm the non-pregnant status.
Each gravid uterus with the cervix was weighed. The number of corpora lutea was counted for pregnant animals. The uterine contents were examined for embryonic or fetal deaths as well as the number of viable fetuses. The degree of resorption (late and early) was confirmed in order to help estimate the relative time of death of the conceptus. The position and number of fetuses in each uterine horn was also recorded.


OTHER:

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 ]
Craniofacial examination of the heads of the fetuses used for the soft tissue examination were performed for internal structure including the eyes, brain, nasal passage and tongue by razor blade serial sectioning
technique.
Fetuses scheduled for skeletal examination were eviscerated and the entire litter was transferred into separate(?) plastic bottles containing 95% ethanol. These fetuses were processed using the Alizarin red staining technique. After fixation in 95% ethanol, the fetuses were macerated with a 1% aqueous potassium hydroxide solution for 1 day and transferred to an Alizarin red solution (0.0025% in 1% aqueous potassium hydroxide) for 1 day. After that, the fetuses were again transferred to 1 % KOH. Alizarin stained fetuses were then cleared and dehydrated in a solution containing 2 parts of 70% ethanol, 2 parts of glycerin and one part of benzyl alcohol for 1 day and finally preserved in a 1:1 solution of 70 % ethanol and glycerin.
The stained fetuses were examined under the stereomicroscope, the skull was examined for size, shape and degree of ossification of nasal, parietal, interparietal, supraoccipital, exoccipital, lacrimal, zygomatic (malar), squamosal (temporal), premaxillary, maxillary, basisphenoid, hyoid and tympanic ring (annulus). Similarly, the vertebral centres, ribs and sternal centres were also examined for size, shape and counted for the number of ossification centers. The cervical, thoracic, lumbar, sacral, caudal vertebrae were observed for the ossification of centers and arches. Pelvic girdles, fore limbs and hind limbs were examined for the development of the bones. Any deviation from the normal development was recorded for each fetus.

- Head examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data

Statistics:

A statistical assessment of the results of the body weight, food consumption was performed by comparing values of dosed with control animals using a one-way ANOVA and a post-hoc Dunnett Test. Fetal evaluation parameters like external, visceral, craniofacial and skeletal parameters were analysed using a Fisher’s exact test. The statistics will be performed with GraphPad Prism V.6.01 software (p<0.05 is considered as statistically significant).

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

The observation of moving the bedding started with the onset of treatment in MD and HD group animals. Few animals of LD group also showed moving the bedding from GD 12. This behavior was seen immediately post dose administration, was transient and is considered to be a local and non-specific reaction.
Abnormal breathing was observed in 1 of 24 females of MD group and 4 of 24 females of HD group; alopecia was observed in 3 of 29 females of LD group and 1 of 24 females of HD group; crust formation was observed in 1 of 24 females of HD group; piloerection was observed in 4 of 24 females of HD group; salivation was observed in 2 of 29 females of LD group, 1 of 24 females of MD group and 1 of 24 females of HD group. The clinical signs alopecia and salivation in test item treated groups did not show dose dependency and occurred sporadically in single animals. The abnormal breathing was transient, occurred sporadically and was limited to 4 of 24 animals. Therefore, these clinical findings were not considered to be adverse or treatment-related.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

There were no effects on body weight and body weight gain in test-item treated groups compared to the controls. HD animals showed statistically significantly lower weight gain or body weight loss between GD 5-8 however all animals recovered quickly and between GD 8-11 there was a higher weight gain in the HD group compared to the corresponding control. These changes were transient and are not considered to be an adverse effect of test-item treatment.
There were no statistically significant differences in the mean terminal body weights and mean adjusted maternal body weights in test-item treated groups compared to the corresponding control group.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

There was no adverse effects on food consumption during the treatment period in test-item treated groups compared to the control. There was statistically significantly lower food consumption between GD 5-8 in the HD group compared to the corresponding control however all animals recovered quickly and showed food consumption values similar to controls from GD 8 and onwards. This transient change was not considered to be an adverse effect of test-item treatment.

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

There were no adverse macroscopic findings in test-item treated group animals. In the LD group there were observations of a cyst on the ovary (1 of 29 females), extra growth in fat tissue (1 of 29 females), and fluid filled and distended uterus (2 of 29 females). In the MD group there was one observation of a fluid filled ovary (1 of 24 females). These findings occurred in a limited number of females and without dose dependency. Therefore, these findings are not considered to be related to test-item treatment.

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not specified

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

effects observed, non-treatment-related

Description (incidence and severity):

There were no effects of test-item treatment on prenatal parameters including the number of corpora lutea, number of implantation sites, number of live fetuses and number of resorptions (early and late), number of male and female fetuses, sex ratio and percent pre- and post-implantation losses. Statistically significantly higher mean resorptions (early and total) occurred in the MD group (1.45) compared to the control group (0.48). This finding was not dose dependent and therefore, is not assumed to be related to test-item treatment

Early or late resorptions:

effects observed, non-treatment-related

Description (incidence and severity):

There were no effects of test-item treatment on prenatal parameters including the number of corpora lutea, number of implantation sites, number of live fetuses and number of resorptions (early and late), number of male and female fetuses, sex ratio and percent pre- and post-implantation losses. Statistically significantly higher mean resorptions (early and total) occurred in the MD group (1.45) compared to the control group (0.48). This finding was not dose dependent and therefore, is not assumed to be related to test-item treatment

Dead fetuses:

effects observed, non-treatment-related

Description (incidence and severity):

The finding of 8 dead fetuses in female no. 79 of the HD group was an isolated incidence and is not considered to be a treatment-related effect. The adjusted body weight of this animals was significantly lower compared to the other animals in the group, and the food consumption was reduced from gd 17-20 during which period the animal lost weight

Changes in pregnancy duration:

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

Changes in number of pregnant:

no effects observed

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

800 mg/kg bw/day (nominal)

Based on:

act. ingr.

Basis for effect level:

behaviour (functional findings)

body weight and weight gain

changes in number of pregnant

changes in pregnancy duration

clinical signs

dead fetuses

early or late resorptions

effects on pregnancy duration

food consumption and compound intake

gross pathology

mortality

number of abortions

pre and post implantation loss

total litter losses by resorption

Key result

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

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were slightly higher incidences of incomplete ossification of supraoccipital bones in MD (30%) and HD (35%) compared to control (23.81%); and slightly higher incidences of full 14th rib (right sided) in MD (10%) and HD (10%) group compared to control (4.76%). In addition, in the HD group there was a higher incidence of misshapen 6th sternebrum (10% in HD and 0% in control); a higher incidence of extra ossification sites at vertebral lumbar arch(es) (10% in HD group and 0% in control); a higher incidence of unossified forelimb metacarpals (63.16% in HD group (and 28.57% in control); a higher incidence of pelvic caudal bone (Bilateral) shift (25% in HD and 19.05% in control) a higher incidence of cervical 7th rudimentary rib (bilateral) (15% in HD (and 4.76% in control); and a higher incidence of cervical 7th rudimentary rib (left) (15% in HD group and 9.52% in control).
Higher incidences of cervical 7th rudimentary rib (right) were noted in LD (14.29%) and HD (10%) groups compared to control (4.76%).
The incidences of all of these observations in the test-item treated groups were without statistical significance, without dose dependency and/or were within the historical control data range, except for the misshapen 6th sternebrum in HD group, for which the incidence was outside the historical control data range (0% to 4.55%). The misshapened sternebrae is a malformation, but considering that only 3 of 112 fetuses were affected, and in the complete absence of any other indication of treatment-related fetal effects within the group, the finding was not considered to be related to the test item, but rather a coincidental finding.

Visceral malformations:

effects observed, non-treatment-related

Description (incidence and severity):

There were a range of visceral observations in control and test item treated groups. Most of the observations were noted in single animals and none occurred at statistically significantly higher incidences in test- item treated groups when compared to the corresponding control group. There were higher litter incidences of dilated ureter (bilateral) in MD (30%) and HD (30%) groups compared to control (23.81%) without statistical significance and these were within the historical control data range (see Appendix 4). Therefore, this finding was not an adverse effect of test-item treatment

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

800 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

not specified

Basis for effect level:

reduction in number of live offspring

changes in sex ratio

fetal/pup body weight changes

changes in litter size and weights

changes in postnatal survival

external malformations

skeletal malformations

visceral malformations

Key result

Abnormalities:

no effects observed

Localisation:

external: cranium

external: ear

external: eye

external: face

external: limb

external: paw

external: tail

external: trunk

external: anogenital distance

external: anus

external: genital tubercle

external: large intestine

external: thorax

external: umbilicus

external: pelvic region

skeletal: skull

skeletal: skull, fontanelles

skeletal: skull sutures

skeletal: clavicle

skeletal: scapule

skeletal: forelimb

skeletal: sternum

skeletal: rib

skeletal: supernumerary rib

skeletal: vertebra

skeletal: pelvic girdle

skeletal: hindlimb

visceral/soft tissue: integumentary

visceral/soft tissue: gastrointestinal tract

visceral/soft tissue: hepatobiliary

visceral/soft tissue: urinary

visceral/soft tissue: cardiovascular

visceral/soft tissue: heamatopoietic

visceral/soft tissue: immune system

visceral/soft tissue: musculoskeletal system

visceral/soft tissue: nervous system

visceral/soft tissue: central nervous system

visceral/soft tissue: peripheral nervous system

visceral/soft tissue: somatic nervous system

visceral/soft tissue: autonomic nervous system

visceral/soft tissue: endocrine system

visceral/soft tissue: respiratory system

visceral/soft tissue: male reproductive system

visceral/soft tissue: female reproductive system

visceral/soft tissue: eye

visceral/soft tissue: ear

other:

Key result

Developmental effects observed:

no

Conclusions:

There were no observed effects on maternal animals of test-item treated groups compared to the corresponding control group. There were no treatment-related adverse effects on embryo-fetal development. Thus, the NOAEL for the maternal toxicity and embryo-fetal developmental toxicity is 800 mg/ kg body weight/ day.

Executive summary:

The test was performed according to the OECD guideline 414 under GLP compliance. Wistar starin rat was tested with test item at dose levels of 60, 300, and 800 mg/ kg body weight/ day administered on gestation days 5 to 19. There were no observed effects on maternal animals of test-item treated groups compared to the corresponding control group. There were no treatment-related adverse effects on embryo-fetal development. Thus, the NOAEL for the maternal toxicity and embryo-fetal developmental toxicity is 800 mg/ kg body weight/ day.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

800 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

high quality

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

Justification for selection of Effect on developmental toxicity: via oral route:

Justification for classification or non-classification

Based on the OECD Guideline studies with the test item no classification for developmental toxicity or toxicity to reproduction according to Regulation (EC) No 1272/2008 is warranted for the test item.