Reaction product of propargyl chloride and sodium bisulfite

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Feb 2017 - Mar 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

water content of 68.3%

Content: w (C3H3SO3Na): 18.7 g/100 g (main component)
w(C3H4S2O6Na2): 2.0 g/100 g (II component)
w(C3H4S2O6Na2): 0.6 g/100 g (III component)
w(C3H3SO2Na): 0.3 g/100 g (IV component)
W(C3H5S3O9Na3): 1.0 g/100 g (V component)
w (NaCL): 8.74 g/100 g
Homogeneity: Given (visually)
Physical state/Appearance: Liquid/ yellowish, clear
Storage conditions: Room temperature

Specific details on test material used for the study:

A.I. 31.7%
test material contains 68.3% water

Test animals

Species:

rat

Strain:

Wistar

Details on species / strain selection:

strain Crl:WI(Han), supplied by Charles River Laboratories, Research Models and Services, Germany GmbH/ Charles River Laboratories, France, which were free from any clinical signs of disease

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/ Charles River Laboratories, France
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation (when arrived from breeder): males (11 weeks old) and females (10 weeks old)
- Weight at study initiation: see raw data
- Fasting period before study:
- Housing: During the pretreatment period, the rats were housed together (up to 5 animals per sex and cage) in Polysulfonate cages Typ 2000P (H-Temp) supplied by TECHNIPLAST, Hohenpeißenberg, Germany.
During the study period, the rats were housed individually in Polycarbonate cages type III supplied by TECHNIPLAST, Hohenpeißenberg, Germany and Becker & Co., Castrop-Rauxel, Germany, with the following exceptions: During overnight matings, male and female mating partners were housed together in Polycarbonate cages type III and Pregnant animals and their litters were housed together until PND 13 in Polycarbonate cages type III.
- Diet (e.g. ad libitum): ground Kliba maintenance diet mouse/rat “GLP” meal, supplied by Provimi Kliba SA, Kaiseraugst, Switzerland, which was available to the animals ad libitum throughout the study
- Water (e.g. ad libitum): Drinking water was supplied from water bottles (ad libitum).
- Acclimation period: 3 weeks

DETAILS OF FOOD AND WATER QUALITY: see above

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24C
- Humidity (%): 30-70%
- Air changes (per hr): 15 times per hour
- Photoperiod (hrs dark / hrs light): 12 hours light from 6.00 h to 18.00 h and 12 hours darkness

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

Test substance was administered to the parental animals orally by gavage, once daily at approximately the same time in the morning.Females in labor were not treated. The treatment lasted up to one day prior to sacrifice.

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
The test substance solutions in drinking water were prepared in intervals, which took into account the analytical results of the stability verification. For the preparation of the administration solutions the test substance was weighed in a
graduated flask depending on the dose group, topped up with drinking water and intensely mixed with a magnetic stirrer.


VEHICLE
- Justification for use and choice of vehicle (if other than water): water

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analytical investigations of the test substance preparations were carried out as a separate study at the test facility Competence Center Analytics of BASF SE, 67056 Ludwigshafen, Germany. (internal study nr.)

Analytical verifications of the stability of the test substance in drinking water for a period of 7 days at room temperature were carried out before the study was initiated and confirmed.

At the beginning (during premating), twice during gestation and once during lactation of the study each 1 sample was taken from the low, mid and high concentration for a concentration
control analysis. All test samples, plus a duplicate set of reserve samples, were withdrawn by staff of the Reproduction Toxicology.
The samples collected at the beginning of the administration period and during lactation were analyzed in the Analytical Laboratory. The samples of the gestation were analyzed only if any imprecision occurs during the analysis
of the samples from the beginning and lactation of the study. The samples within this study were labeled with serial numbers. The same number was not used for several samplings.

Duration of treatment / exposure:

The duration of treatment covered a 2-week pre-mating and a mating period, approximately 1 day post-mating in males, and the entire gestation period as well as approximately 21 days of the lactation period and 29 days post-mating in females (for
sperm negative female).

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Remarks:

test material contains 68.3% water

Doses / concentrations 2

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Remarks:

test material contains 68.3% water

Doses / concentrations 3

Dose / conc.:

3 000 mg/kg bw/day (actual dose received)

Remarks:

test material contains 68.3% water

Doses / concentrations 4

Dose / conc.:

95.1 other: mg/kg a.i.

Remarks:

corrected to a.i. (31.7% of actual dose)

Doses / concentrations 5

Dose / conc.:

317 other: mg/kg a.i.

Remarks:

corrected to a.i. (31.7% of actual dose)

Doses / concentrations 6

Dose / conc.:

951 other: mg/kg a.i.

Remarks:

corrected to a.i. (31.7% of actual dose)

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on range finder study with 1000 and 3000 mg/kg bw actual received.
- Rationale for animal assignment (if not random): random

Positive control:

no

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
A cageside examination was conducted at least daily for any signs of morbidity, pertinent behavioral changes and signs of overt toxicity before the administration as well as within 2 hours and within 5 hours after the administration.

DETAILED CLINICAL OBSERVATIONS: Yes
A detailed clinical observation (DCO) was performed in all animals once before the start of the administration period and thereafter at weekly intervals.
Mortality: A check for moribund or dead animals was made twice daily on working days or once daily (Saturday, Sunday or on public holidays).
The parturition and lactation behavior of the dams was generally evaluated in the mornings in combination with the daily clinical inspection of the dams.
On weekdays (except Saturday, Sunday and public holidays) the parturition behavior of the dams was inspected in the afternoons in addition to the evaluations in the mornings.

DCO examinations are:
1. Abnormal behavior in “handling”
2. Fur
3. Skin
4. Posture
5. Salivation
6. Respiration
7. Activity/arousal level
8. Tremors
9. Convulsions
10. Abnormal movements
11. Gait abnormalities
12. Lacrimation
13. Palpebral closure
14. Exophthalmos (Protruding eyeball)
15. Assessment of the feces excreted during the examination (appearance/consistency)
16. Assessment of the urine excreted during the examination
17. Pupil size

BODY WEIGHT: Yes
In general, the body weight of the male and female parental animals was determined once a week at the same time of the day (in the morning) until sacrifice.
Females without positive evidence of sperm, without litter or waiting for necropsy, were weighed weekly.

FOOD CONSUMPTION:
Generally, food consumption was determined once a week for male and female parentalanimals, with the following exceptions: Food consumption was not determined after the 2nd premating week (male parental
animals) and during the mating period (male and female F0 animals). Food consumption of the F0 females with evidence of sperm was determined on gestation days (GD) 0 - 7, 7 - 14 and 14 - 20. Food consumption of F0 females which gave birth to a litter was determined on PND 1 - 4, 4 - 7, 7 - 10 and 10 - 13.
Food consumption was not determined in females without positive evidence of sperm during the mating and the gestation period and in females without litter during the lactation period.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
Water consumption of the females with evidence of sperm was determined for GD 7-8, 14-15 and 19-20.
Water consumption of the F0 females, which gave birth to a litter was determined for PND 1-2, 3-4, 6-7, 9-10 and 12-13.
Water consumption was not determined in the females without positive evidence of sperm during mating and gestation periods and in the females without litter during lactation period.

HAEMATOLOGY: Yes
In the morning blood was taken from the retro-bulbar venous plexus from fasted animals. The animals were anaesthetized using isoflurane. The blood sampling procedure and subsequent
analysis of blood and serum samples were carried out in a randomized sequence. The parameters listed below were examined in the first 5 surviving parental males per group
at termination and in the first 5 females with litters (in order of delivery) per group at PND 14.
Parameters checked:
Leukocyte count (WBC)
Erythrocyte count (RBC)
Hemoglobin (HGB)
Hematocrit (HCT)
Mean corpuscular volume (MCV)
Mean corpuscular hemoglobin (MCH)
Mean corpuscular hemoglobin concentration (MCHC)
Platelet count (PLT)
Differential blood count
Reticulocytes (RETA)
Prothrombin time (Hepato Quick’s test) (HQT)


CLINICAL CHEMISTRY: Yes
see Haematology for details of blood sampling.
Parameters measured are:
Enzyme:
Alanine aminotransferase (ALT) (L-alanine: 2-oxoglutarate aminotransferase)
Aspartate aminotransferase (AST) (L-aspartate: 2-oxoglutarate aminotransferase)
Alkaline phosphatase (ALP) (orthophosphoric acid monoester phosphohydrolase)
g-Glutamyltransferase (GGT) (g -glutamyl) peptide: aminoacid-g-glutamyl-transferase)
Blood Chemistry Parameters:
Sodium (NA)
Potassium (K)
Chloride (CL)
Inorganic phosphate (INP)
Calcium (CA)
Urea (UREA)
Creatinine (CREA)
Glucose (GLUC)
Total bilirubin (TBIL)
Total protein (TPROT)
Albumin(ALB)
Globulins (GLOB)
Triglycerides (TRIG)
Cholesterol (CHOL)
Bile acids (TBA)

Thyroid Hormones
Blood samples were taken from all surplus pups or 2 preferably female pups per litter at PND 4 as well as one male and one female pup per litter at PND 13 by decapitation under isoflurane
anesthesia. Additionally, blood samples from all dams at PND 14 and all males at termination were taken by puncturing the retrobulbar venous plexus under isoflurane anesthesia. The adults were fastened before the blood sampling.
Parameters:
Total thyroxine (T4)
Thyroid stimulating hormone (TSH)

URINALYSIS: Not specified

NEUROBEHAVIOURAL EXAMINATION: Yes
A functional observational battery (FOB) was performed in the first 5 male and the first 5 female animals with litter per group (in order of delivery) at the end of the administration period starting at about 10.00 h.The FOB started in a randomized sequence with passive observations without disturbing the animals, followed by removal from the home cage, open field observations in a standard arena and sensorimotor tests as well as reflex tests.

Home cage observations:
The animals were observed in their closed home cages (for a short period: about 10-30 seconds); any disturbing activities (touching the cage or rack, noise) were avoided during these examinations in order not to influence the behavior of the animals.Attention was paid to:
1. Posture
2. Tremors
3. Convulsions
4. Abnormal movements
5. Gait
6. Other findings

Open field observations:
The animals were transferred to a standard arena (50 × 50 × 25 cm) and observed.The following parameters were examined:
1. Behavior on removal from cage
2. Fur
3. Skin
4. Salivation
5. Nasal discharge
6. Lacrimation
7. Eyes/pupil size
8. Posture
9. Palpebral closure
10. Respiration
11. Tremors
12. Convulsions
13. Abnormal movements/stereotypy
14. Gait
15. Activity/arousal level
16. Feces (appearance/ consistency) within 2 minutes
17. Urine (amount/color) within 2 minutes
18. Rearing within 2 minutes
19. Other findings

Sensory motor tests/Reflexes:
The animals were removed from the open field and subjected to following sensory motor or reflex tests:
1. Reaction to an object being moved towards the face (Approach response)
2. Touch sensitivity (Touch response)
3. Vision (Visual placing response)
4. Pupillary reflex
5. Pinna reflex
6. Audition (Startle response)
7. Coordination of movements (Righting response)
8. Behavior during handling
9. Vocalization
10. Pain perception (Tail pinch)
11. Other findings
12. Grip strength of forelimbs
13. Grip strength of hindlimbs
14. Landing foot-splay test

Motor activity measurement
The Measurement of motor activity (MA) was measured at the end of the administration period in the first 5 surviving parental males and the first 5 surviving females with litter (in order of delivery) per group.

Estrous cycle determinations:
For all females in a pool of up to 50 animals, estrous cycle normality was evaluated before the randomization (the estrous cycle data of these individuals were not be reported and can be found in the raw data).
For a minimum of 2 weeks prior to mating estrous cycle length was evaluated by daily analysis of vaginal smear for all F0 female parental rats. Determination was continued throughout the pairing period until the female exhibited evidence of copulation. At necropsy, an additional vaginal smear was examined to determine the stage of estrous cycle for each F0 female with scheduled sacrifice.

Male reproduction data
The pairing partners, the number of mating days until vaginal sperm was detected in the female animals, and the gestational status of the females were recorded for F0 breeding pairs. For the males, mating and fertility indices were calculated

Female reproduction and delivery data
The pairing partners, the number of mating days until vaginal sperm was detected and gestational status was recorded for F0 females. For the females, mating, fertility and gestation indices were calculated.

Litter data:

Pup number and status at delivery:
All pups delivered from the F0 parents (F1 litter) were examined as soon as possible on the day of birth to determine the total number of pups, the sex and the number of liveborn and stillborn pups in each litter.
Pup viability/mortality:
In general, a check was made for any dead or moribund pups twice daily on workdays (once in the morning and once in the afternoon) or as a rule, only in the morning on Saturdays, Sundays or public holidays.
Sex ratio:
On the day of birth (PND 0) the sex of the pups was determined by observing the distance between the anus and the base of the genital tubercle; normally, the anogenital distance is considerably greater in male than in female pups. Later, during the course of lactation, this initial sex determination was followed up by surveying the external appearance of the anogenital region and the mammary line. The sex of the pups was finally confirmed at necropsy.
Pup clinical observations
The live pups were examined daily for clinical symptoms (including gross-morphological findings) during the clinical inspection of the dams and documented for each pup.
Pup body weight data
The pups were weighed on the day after birth (PND 1) and on PND 4 (before standardization), 7, 10 and 13. Pups' body weight change was calculated from these results.

Sacrifice and pathology:

All parental animals were sacrificed by decapitation under isoflurane anesthesia. The exsanguinated animals were necropsied and assessed by gross pathology, special attention being given to the reproductive organs.

GROSS PATHOLOGY: Yes (see table)

Organ weights

The following weights were determined in all animals sacrificed on schedule:
1. Anesthetized animals
2. Epididymides
3. Ovaries
4. Prostate
5. Seminal vesicles with coagulating glands
6. Testes
7. Thyroid glands (fixed)
8. Uterus (with cervix)

The following weights were determined in 5 animals per sex/test group:
1. Adrenal glands
2. Brain
3. Heart
4. Kidneys
5. Liver
6. Spleen
7. Thymus

Organ/tissue fixation
The following organs or tissues of all parental animals were fixed:
1. All gross lesions
2. Adrenal glands
3. Aorta
4. Bone marrow (femur)
5. Brain
6. Cecum
7. Cervix
8. Coagulating glands
9. Colon
10. Duodenum
11. Eyes with optic nerve (males in 4% buffered formaldehyde solution; females in modified
Davidson’s solution)
12. Esophagus
13. Extraorbital lacrimal glands
14. Epididymides (modified Davidson’s solution)
15. Femur with knee joint
16. Heart
17. Ileum
18. Jejunum (with Peyer’s patches)
19. Kidneys
20. Larynx
21. Liver
22. Lungs
23. Lymph nodes (axillary and mesenteric)
24. Mammary gland (male and female)
25. Nose (nasal cavity)
26. Ovaries (modified Davidson’s solution)
27. Oviducts
28. Pancreas
29. Parathyroid glands
30. Pharynx
31. Pituitary gland
32. Prostate gland
33. Rectum
34. Salivary glands (mandibular and sublingual)
35. Sciatic nerve
36. Seminal vesicles
37. Skeletal muscle
38. Spinal cord (cervical, thoracic and lumbar cord)
39. Spleen
40. Sternum with marrow
41. Stomach (forestomach and glandular stomach)
42. Testes (modified Davidson’s solution)
43. Thymus
44. Thyroid glands
45. Trachea
46. Urinary bladder
47. Uterus (uteri of all apparently nonpregnant animals or empty uterus horns were stained
according to Salewski E,1964)
48. Vagina

Pups
On PND 13, one selected male and one female pup per litter was sacrificed under isoflurane
anesthesia by decapitation. Blood was sampled for determination of thyroid hormone
concentrations.

HISTOPATHOLOGY: Yes (see table)
Fixation was followed by histotechnical processing, examination by light microscopy and assessment of findings according to the following table was performed for the highest dose group and the control at minimum:
1. All gross lesions
2. Adrenal glands
3. Brain
4. Cecum
5. Cervix
6. Coagulating glands
7. Colon
8. Duodenum
9. Epididymides
10. Eyes with optic nerve
11. Heart
12. Ileum
13. Jejunum
14. Kidneys
15. Liver
16. Lungs
17. Lymph nodes (axillary and mesenteric)
18. Ovaries
19. Oviducts
20. Prostate gland
21. Peyer’s patches
22. Rectum
23. Sciatic nerve
24. Seminal vesicles
25. Skeletal muscle
26. Spinal cord (cervical, thoracic, lumbar)
27. Spleen
28. Sternum with marrow
29. Stomach (forestomach and glandular stomach)
30. Testes
31. Thymus
32. Thyroid glands
33. Trachea
34. Urinary bladder
35. Uterus
36. Vagina

Other examinations:

Special attention was given to the stages of spermatogenesis and histopathology of interstitial testicular cell structure.

Statistics:

The following statistical analyses were carried out:see any other

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Five high-dose males (Nos.: 31, 35, 37, 38 and 39 - 3000 mg/kg bw/d) showed salivation (slight to moderate) during mating days 11 – 14. One high-dose male (No.: 39) showed salivation
(slight) on post-mating day 0. Four high-dose females (Nos.: 134, 137, 138 and 140) showed salivation (slight to moderate) during several parts of the gestation period.
One high-dose female (No. 137) showed a skin lesion (head region) during GD 14 - 22 and PND 1 - 5 and skin lesion (entire body) during PND 6 - 19.
One control male (No. 2) showed gasping and labored respiration on premating day 9. One high-dose female (No.: 140) had a complete litter loss on PND 8.
These isolated findings were considered as incidental, as they occurred without any relation to the test substance administration.

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

The statistically significantly decreased food consumption of the high-dose F0 females during PND 1 - 4 was assessed as incidental as it was mainly caused by one animal (No. 140) which
delivered only two pups and subsequently no increasing food consumption during lactation was seen.

Food efficiency:

not examined

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

no effects observed

Ophthalmological findings:

not specified

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

In males of test group 3 (3000 mg/kg bw/d), absolute and relative monocyte counts were significantly lower compared to controls. This was also true for absolute monocyte counts in
males of test group 1 (300 mg/kg bw/d). However, the means were within historical control ranges (males, absolute monocytes 0.07-0.14 Giga/L; relative monocytes 1.3-2.7 %), whereas
at least absolute monocyte counts of the controls were above the historical control range. Therefore, these changes were regarded as incidental and not treatment-related.
In females of test group 3 (3000 mg/kg bw/d) hemoglobin values were significantly lower compared to controls, but the values were within the historical control range (females,
hemoglobin 8.4-9.2 mmol/L). Therefore, this alteration was regarded as incidental and not treatment-related.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Clinical chemistry: No treatment-related changes among clinical chemistry parameters were observed.
Thyroid hormones: In parental males (300, 1000 and 3000 mg/kg bw/d) and in male and female pups at PND13
(300, 1000 and 3000 mg/kg bw/d), no treatment-related alterations of T4 and TSH levels were
observed.

Urinalysis findings:

not examined

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

The statistically significantly increased value in the high-dose males during interval 1 was assessed as incidental as the summation of all intervals was not statistically significantly
increased and no similar effect was seen in female animals which were treated for a considerably longer period.

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

Absolute organ weights
When compared to control group 0 (set to 100%), the mean absolute weights of following organs were significantly decreased: Adrenal glands.
All other mean absolute weight parameters did not show significant differences when compared to the control group 0.
Relative organ weights
All mean relative weight parameters did not show statistical significant differences when compared to the control group 0 (set to 0).
The significant decrease of the absolute weight of the adrenal glands in test group 2 and 3 (83.6 and 85.0 mg, respectively) were within the historical control range (69.20 – 93.0 mg).
Furthermore, no histopathological correlate was found, that could explain the weight decrease.

Gross pathological findings:

no effects observed

Description (incidence and severity):

All findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.
Fertility:
The non-pregnant female animal No. 111 showed dilation of the ureters, a mass in the urethra (4 mm in diameter), and enlargement of the iliac lymph nodes. All other not-pregnant females
(Nos. 105, 138, and 139) as well as all male mating partners (Nos. 5, 11, 38, and 39) did not show relevant gross lesions.

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

All findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without
any relation to treatment. The stages of spermatogenesis in the testes of males of the high dose test group were comparable to those of the controls. In high dose females the different stages of functional
bodies in the ovaries were present and comparable to the control animals.
Fertility
The female animals (Nos. 105 and 111), which were not pregnant showed increased mucification of the vagina and cervix with atrophy of the epithelium beneath. This finding represents a spontaneous background lesion which might influence fertility. In animal No. 111, following findings were also noted: inflammation of the urinary bladder and urethra (both accompanied by epithelial hyperplasia), dilation of the ureters and plasmocytosis in the iliac lymph nodes. These findings were consistent with the macroscopic changes. (see above)

Histopathological findings: neoplastic:

no effects observed

Other effects:

no effects observed

Description (incidence and severity):

Pup number and status at delivery
The mean number of delivered F1 pups per dam and the rates of liveborn, stillborn, found dead and cannibalized F1 pups were evenly distributed among the test groups. The respective values reflect the normal range of biological variation inherent in the strain used in this study.
Pup viability
The viability index indicating pup mortality during lactation (PND 0 - 4) varied between 100%/
99.3% /95.6% and 92.9% in test groups 0 - 3, respectively.
The pups surviving index indicating pup mortality during lactation (PND 4 - 13) varied between 100%/ 100% /100% and 87.5% in test groups 0 - 3, respectively. The supposedly decreased values in the high dose group are attributable to one female (No.: 140) which had only two pups (one found dead at day 1 and one cannibalized at day 8). These isolated finding was assessed as incidental and not related to the treatment.
Sex ratio
The sex distribution and sex ratios of live F1 pups on the day of birth and PND 4 did not show substantial differences between the control and the test substance-treated groups; slight
differences were regarded to be spontaneous in nature.
Pup clinical observations
There were no test substance-related adverse clinical signs observed in any of the F1 generation pups of the different test groups.
Pup body weight data
No test substance-related influence on body weights and body weight change values of F1 pups were noted in test groups 1 - 3 (300, 1000 and 3000 mg/kg bw/d). One male runt was seen in test group 2.
Anogenital distance/anogenital index
Neither on anogenital distance nor anogenital index test substance-related effects were noted in all treated F1 offspring (test groups 1 - 3 [300, 1000 and 3000 mg/kg bw/d]).
Nipple/ areola anlagen
The apparent number and percentage of male pups having areolae was not influenced by the test substance when examined on PND 13.
Pup necropsy observations
A few pups showed spontaneous findings at gross necropsy, such as discolored testis (left, brown), small testis, empty stomach and partly cannibalized.
These findings occurred without any relation to dosing and/or can be found in the historical control data at comparable or even higher incidences

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

>= 3 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects up to and including the highest tested dosage

2

Key result

Dose descriptor:

NOAEL

Effect level:

>= 951 other: mg/kg bw/d corrected to active ingredient

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: no adverse effects up to and including the highest tested dosage

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

In conclusion, under the conditions of the present OECD 422 combined repeated dose toxicity study with the reproductive/developmental screening test in Wistar rats the oral administration
of [trade name] by gavage to male and female Wistar rats did not result in signs of systemic toxicity up to a dose level of 3000 mg/kg bw/d (951 mg/kg bw/d a.i. due to 68.3% water content) . Thus, the no observed adverse effect level
(NOAEL) for general systemic toxicity was 3000 mg/kg bw/d (951 mg/kg bw/d a.i.) for male and female Wistar rats. The NOAEL for reproductive performance and fertility was also set to 3000 mg/kg bw/d (951 mg/kg bw/d a.i.) for
male and female Wistar rats. The NOAEL for developmental toxicity was 3000 mg/kg bw/d (951 mg/kg bw/d a.i.).