Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Referenceopen allclose all

Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
August 1, 2005 to January 23, 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted according to GLP and international test guidelines.
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Species:
rat
Strain:
other: Crl:CD (SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Japan, Inc.
- Age at study initiation: (P) 9 wks; (F1) 4 days
- Weight at study initiation: (P) Males: 307-370 g; Females: 197-239 g
- Diet: ad libitum
- Water: ad libitum, except during fresh urine collection
- Acclimation period: From time of arrival 5 days quarantine, 8 days acclimation


ENVIRONMENTAL CONDITIONS-QUARANTINE ROOM
- Temperature (°C): 21.6-22.4
- Humidity (%): 52.9-64.9
- Air changes (per hr): 6 to 20
- Photoperiod (hrs dark / hrs light): 12/12

ENVIRONMENTAL CONDITIONS-TEST ROOM
- Temperature (°C): 21.8-23.7
- Humidity (%): 45.4-59.1
- Air changes (per hr): 6 to 20
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Test material weighed and disolved in vehicle (purified water), measured by volume.

VEHICLE
- Concentration in vehicle: 2 - 60 mg/kg
- Amount of vehicle (if gavage): Dosed at 5 mL/kg.
Details on mating procedure:
- M/F ratio per cage:Male and female rats were cohabited day and night on a one-to-one basis
- Length of cohabitation: From Day 15 (initial day of mating) for the maximum of 14 days.
- Proof of pregnancy: Vaginal plug and/or sperm in vaginal smear referred to as day 0 of pregnancy: From the day after the initial day of mating, vaginal smears were collected every day in the morning, and examined for the presence of sperm. Copulation was confirmed by the presence of a vaginal plug or sperm in the vaginal smears.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Confirmation of stability and homogeneity:
The dosing solutions of 2 and 200 mg/mL were confirmed to be stable and homogeneous for 8 days in a refrigerator protected from light in a validation study by GC method (study No. B050280) conducted at the test facility.

Confirmation of concentration:
The concentration of the test substance in each dosing solution was measured at the first and final preparation by GC method. In the results, the actual mean concentration of each dosing solution ranged from 92.0 to 108.5% of the nominal concentration (permissible range: 90 to 110%). Since the concentrations of 2 and 12 mg/mL dosing solutions deviated from the criterion, the dosing solutions were reprepared and analyzed. The results of the reprepared solutions met the criterion.

Analytical method of dosing solutions
Each dosing solution was analyzed according to the analytical method validated in the study entitled “Validation of Analytical Method of 1,3-BAC Concentration in Preparation” (study No. B050280) conducted at the test facility.
Duration of treatment / exposure:
MALES: Administration was conducted from 14 days before mating to the day before necropsy (including the mating period; 42 days in total).

PREGNANT AND MATERNAL ANIMALS: Administration was conducted from 14 days before mating to Day 4 of lactation (including the mating period, gestation period, and delivery; the date of delivery was designated as Day 0 of lactation). In the non-delivered females, administration was conducted until the day before necropsy.

RECOVERY (satellite) FEMALES: Administration was conducted for 42 days without mating.


Frequency of treatment:
Once a day between 8:23 and 11:34
Details on study schedule:
Male and female rats were cohabited day and night on a one-to-one basis from Day 15 (initial day of mating) for the maximum of 14 days. From the day after the initial day of mating, vaginal smears were collected every day in the morning, and examined for the presence of sperm. Copulation was confirmed by the presence of a vaginal plug or sperm in the vaginal smears. The day on which evidence of copulation was found was designated as Day 0 of gestation. Based on the results, the following parameters were calculated.
(1) Days until copulation: Number of days from the start of mating to the detection of copulation
(2) Number of estrus stages without copulation
(3) Copulation index (%): (Number of animals with successful copulation/Number of animals cohabited) × 100
(4) Fertility index (%): (Number of pregnant animals/Number of animals with successful copulation) × 100

Remarks:
Doses / Concentrations:
10, 60, 300 mg/kg
Basis:
nominal conc.
No. of animals per sex per dose:
Dosing Period:
Control 7 males, 12 females;
10 mg/kg 12 males, 12 females;
60 mg/kg 12 males, 12 females;
300 mg/kg 7 males, 12 females,

Recovery Period:
Control and 300 mg/kg 5 males, 5 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dosing conditions are recommended in the OECD guidelines.10, 60, and 300 mg/kg
In the dose-range finding study (No. B050279), the test substance was dosed orally by gavage to 3 SD rats/sex/group at 0, 30, 100, 300, and 1000 mg/kg for 14 days. As a result, all animals died or sacrificed moribund at 1000 mg/kg. In the 300 mg/kg group, mucosal edema in the forestomach was noted in both sexes as a major toxic and treatment related change. There were no test substance related changes in any animals at 30 or 100 mg/kg.
In consideration of these results and duration of the study period, the high dose for this study was set at 300 mg/kg, at which obvious toxicity was expected. The middle and low doses were set at 60 and 10 mg/kg, respectively, with a common ratio of about 5. The control group receiving the vehicle (purified water) alone was also set.


- Rationale for animal assignment (if not random):Animals were assigned to the groups by stratified-by-weight randomization method based on the body weight on the day before the first administration to make the group mean body weights even. Forty-eight males and 58 females were used in this study.
Parental animals: Observations and examinations:
Observation during delivery and lactation:
All copulated females were allowed natural delivery. The observation of delivery was conducted twice daily (9:00 and 16:00) from Days 21 to 25 of gestation. The delivery completed by 9:00 was judged as a delivery on the corresponding day (the delivery day was regarded as Day 0 of lactation). The animals that completed delivery by 16:00 were observed for lactation the next day (the day after delivery was regarded as Day 0 of lactation). The females that did not deliver by 25 days after copulation were regarded as non-delivered females. The delivered animals (dams) were allowed to nurse new born infants until day 4 postpartum (Day 4 of lactation), and postpartum behavior such as lactation, nesting, and presence or absence of cannibalism was observed every day.

Observation after lactation :
At necropsy, the ovary and uterus of dams were removed and examined for the numbers of corpora lutea and implantations. Non-delivered females were examined in the same manner to detect implantation sites. Uterus without visible implantation sites was immersed in a 10 vol% ammonium sulfide solution to detect implantation sites. Non-delivered female with no implantation site was regarded as non-pregnant female. Based on the results, the following parameters were calculated.
(1) Gestation length: Days until completion of delivery from Day 0 of gestation
(2) Delivery index (%): (Number of pregnant animals delivered live offspring/Number of pregnant animals) × 100
(3) Implantation index (%): (Number of implantations/Number of corpora lutea) × 100
(4) Gestation index (%): (Total number of delivered offspring/Number of implantations) × 100
Oestrous cyclicity (parental animals):
From the first day of administration to the first day of mating, vaginal smears of all test females of each group were sampled in the morning and examined for estrus cycle. The mean estrus cycle and incidence of females with irregular estrus cycle (with the period of other than 4 to 6 days) were calculated.
Litter observations:
PARAMETERS EXAMINED
The number of delivered offspring (numbers of live offspring and stillborns), sex, and the presence or absence of external anomalies was examined on Day 0 of postpartum.
The body weight gain between days 0 and 4 postpartum was calculated from the litter mean weight.

GROSS EXAMINATION OF DEAD PUPS:
Yes, for all dead offspring other than those found cannibalised.
Postmortem examinations (parental animals):
Refer to repeated dose toxicity study summary (IUCLID section 7.5.1) for full details regarding results seen in the parental animals.
Postmortem examinations (offspring):
External anomaly including that in the oral cavity was examined, and the offspring were euthanized in the same manner as the parental animals, and necropsied. Dead offspring except for those cannibalized were immersed and fixed in 10 vol% neutral phosphate-buffered formalin, and necropsied under a stereomicroscope.
Statistics:
Multiple comparison test:
Body weights, body weight gain, food consumption, hematology, blood chemistry, organ weights, behavioral test (grip strength and motor activity), number of corpora lutea, number of implantations, number of delivered offspring (numbers of live offspring and stillborns)

Kruskal-Wallis test and Dunnett-type multiple comparison test:
Urinalysis (pH, protein, glucose, ketones, bilirubin, occult blood, urobilinogen), days until copulation, number of estrus stages without copulation, mean estrus cycle, gestation length, implantation index, gestation index, live birth index, incidence of offspring with external anomaly, viability index on Day 4

Wilcoxon’s rank sum test:
Histopathological findings

Fisher’s exact probability test:
Incidence of females with irregular estrus cycle, copulation index, fertility index, delivery index, sex ratio (male/female), incidence of dams with externally anormal offspring
Offspring viability indices:
The number of delivered offspring (numbers of live offspring and stillborns), sex, and the presence or absence of external anomalies was examined on Day 0 of postpartum. Thereafter, clinical signs and mortality were observed daily. Based on the results, the following parameters were calculated.

(1) Live birth index (%): (Number of live offspring at birth/Total number of delivered offspring at birth) × 100
(2) Viability index on Day 4 (%): (Number of live offspring on Day 4/Number of live offspring at birth) × 100
All live offspring were weighed individually on days 0 and 4 postpartum. An electronic balance (PB3002-S: Mettler Toledo K.K.) was used for weighing. Moreover, the body weight gain between days 0 and 4 postpartum was calculated from the litter mean weight.
Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Bodyweight gain was supressed in males dosed at 300 mg/kg.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Bodyweight gain was supressed in males dosed at 300 mg/kg.
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
In the 300 mg/kg group, 1 female (animal No. 50405) showed irregular estrus cycle; however, copulation was confirmed by cohabitation
Reproductive function: sperm measures:
effects observed, treatment-related
Reproductive performance:
no effects observed
Refer to repeated dose toxicity study summary (IUCLID section 7.5.1) for full details regarding results seen in the parental animals

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
One male (animal No. 00405) in the 300 mg/kg group was found dead. The animal showed irregular respiration and rale from 3 days before the death, decreases in locomotor activity, ptosis, and abnormal fur on the day before death, and died on Day 25 of dosing.
Salivation was sporadically observed in the 300 mg/kg group (11 males and 16 females) during the dosing period.
Rale and a decrease in locomotor activity were noted in 1 female at 300 mg/kg (animal No. 50409) during the gestation period, and a decrease in locomotor activity was noted continuously in this animal during the lactation period.
Loss of teeth was observed in 1 satellite female of the control group; however, it recovered 3 days later. No test substance-related abnormalities were noted in either sex at 10 or 60 mg/kg.
Salivation disappeared in the recovery males and satellite females at 300 mg/kg by termination of administration.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weight gain suppression was noted in males at 300 mg/kg from Days 8 to 42. During the recovery period, it was comparable to that of the control group, indicating reversibility.
There were no significant differences from the control group in males receiving 10 or 60 mg/kg or test substance-treated females.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Increased absolute and relative adrenal weights were noted in males, and increased relative weights of kidney and adrenal were noted in females at 300mg/kg at the end of the dosing period. Increasing tendency in absolute adrenal weight and increased relative adrenal weight were noted in males at 300 mg/kg at the end of the recovery period.
Increased absolute liver weight was noted in females at 300 mg/kg at the end of the recovery period; however, since there were no changes in males at 300 mg/kg at the end of the dosing period, it was considered to be an incidental change.
There were no significant differences from the control group in either sex at 10 or 60 mg/kg.

GROSS PATHOLOGY (PARENTAL ANIMALS)
The following findings were noted at the scheduled necropsy: thickening of the forestomach wall in both sexes, ulcer of forestomach mucosa, or adhesion of the liver and forestomach in 1 female each in the 300 mg/kg group. The changes in the reproductive system were small testes and epididymides in 2 males (animal No. 00403: bilateral, No. 00406: unilateral) receiving 300 mg/kg. These changes were not noted in the animals necropsied at the end of the recovery period.
In the dead animal, dark reddish change of glandular stomach mucosa, tarry abnormal contents in the duodenum, jejunum, and ileum, and dilatation of the ileum and cecum were noted. In addition, dark reddish changes of the thymus and lung, and small spleen were noted in the dead animal.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Changes considered attributed to the test substance were noted in the stomach of both sexes and in the testes and epididymis of males at 300 mg/kg.In the stomach, focal squamous cell hyperplasia and hyperkeratosis and focal inflammatory cell infiltration were noted in the forestomach in all males and females at 300 mg/kg. The changes in 5 males and 9 females were accompanied by ulcer of the forestomach. Minimal focal squamous cell hyperplasia of the forestomach was noted in all males and females, and minimal focal inflammatory cell infiltration of the forestomach was noted in 3 males and 2 females in the 300 mg/kg group at the necropsy after the recovery period. However, their grades were less than those in the animals necropsied after the dosing period.

In the testis, atrophy of the seminiferous tubule was noted in 4 males at 300 mg/kg and in 2 males at 10 mg/kg. The change noted in 1 of the 4 males at 300 mg/kg was severe, and was accompanied by diffuse hyperplasia of interstitial cells. The atrophy of the seminiferous tubule noted at 10 mg/kg was minimal, and the change was not noted at 60 mg/kg. Accordingly, atrophy of the seminiferous tubule noted at 10 mg/kg was judged to be spontaneous. In 1 of the 5 animals receiving 300 mg/kg, atrophy of the seminiferous tubule was noted at the end of the recovery period.
In the duct of epididymis, cell debris was noted in 2 males, decreased sperm in 2 males, and atrophy in 1 male at 300 mg/kg. The changes in the epididymis were related to the pathological lesions of the testis, and were not noted at the end of the recovery period.
Besides, a variety of pathological changes were noted in each group including the control at the end of the dosing and recovery periods. However, these changes were not considered to be test substance-related, since there were no significant differences in their incidence among the groups and they develop nonspecifically in rats.

In the dead animal, the following changes were noted: focal squamous cell hyperplasia of the forestomach, focal inflammatory cell infiltration of the forestomach, hemorrhage in the glandular stomach, atrophy of the spleen, congestion and edema of the lung, atrophy of the thymus, and incidental cyst of the kidney.

The test substance had no effects on the reproductive parameters such as estrus cycle, copulation index, fertility index, gestation index, gestation length, numbers of corpora lutea or implantations, implantation index, delivery index, parturition, or maternal behavior.
Dose descriptor:
NOEL
Effect level:
300 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
In the observation of lactation, 1 dam receiving 300 mg/kg (animal No. 50405) showed lack of licking pups and retrieving on the day of delivery, and 4 males and 3 females out of 8 males and 9 females delivered by the dam died on the same day. In addition, the rest of delivered offspring except for 1 male died on Day 1 of lactation. There were no abnormalities after Day 1 of lactation.
There were no abnormalities in delivery or lactation in the other dams.
Dose descriptor:
NOEL
Generation:
F1
Effect level:
300 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
gross pathology
Reproductive effects observed:
not specified

In the examination of offspring, no changes attributed to the test substance were noted in the number of offspring, live birth index, sex ratio, viability index on Day 4, external features, clinical signs, body weights, or necropsy findings.

Conclusions:
The no observed effect level of 1,3-Cyclohexanedimethanamine for reproductive/developmental toxicity was considered to be 300 mg/kg/day in parental animals and offspring.
Executive summary:

A combined repeated dose toxicity and reproductive toxicity screening study was conducted on the test substance 1,3-Bis(aminomethyl)cyclohexane (1,3-BAC). The study was performed according to OECD test guideline 422, and in compliance with GLP.

Male and female rats were dosed at levels 0 (vehicle control), 10, 60, and 300 mg/kg bw/day of 1,3-BAC as a solution in water; males were dosed for 42 days in total including the mating period, females were dosed from 14 days before mating until day 4 of lactation. The clinical and pathological effects on reproduction in both the parental generation and the offspring were recorded.

The test substance had no effects on the reproductive parameters of the parental animals, nor on the developmental parameters of the offspring. The no observed effect level of 1,3-Bis(aminomethyl)cyclohexane for reproductive / developmental toxicity was considered to be 300 mg/kg/day in parental animals and offspring.

Endpoint:
extended one-generation reproductive toxicity - basic test design (Cohorts 1A, and 1B without extension)
Type of information:
other: Expert statement
Adequacy of study:
key study
Study period:
2017
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Expert statement based on toxicity data available
Principles of method if other than guideline:
Expert statement based on toxicity data available.
GLP compliance:
no
Justification for study design:
Expert statement based on toxicity data available.
Remarks on result:
not measured/tested
Remarks on result:
not measured/tested
Reproductive effects observed:
not specified
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
300 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

A combined repeated dose toxicity and reproductive toxicity screening study was conducted on the test substance 1,3-Bis(aminomethyl)cyclohexane (1,3-BAC). The study was performed according to OECD test guideline 422, and in compliance with GLP.

Male and female rats were dosed at levels 0 (vehicle control), 10, 60, and 300 mg/kg bw/day of 1,3-BAC as a solution in water; males were dosed for 42 days in total including the mating period, females were dosed from 14 days before mating until day 4 of lactation. The clinical and pathological effects on reproduction in both the parental generation and the offspring were recorded.

The test substance had no effects on the reproductive parameters of the parental animals, nor on the developmental parameters of the offspring. The no observed effect level of 1,3-Bis(aminomethyl)cyclohexane for reproductive / developmental toxicity was considered to be 300 mg/kg/day in parental animals and offspring.


Short description of key information:
NOEL, rat, reproductive / developmental toxicity screen = 300 mg/kg bw/day (Parental animals and offspring).

Effects on developmental toxicity

Description of key information

NOAEL, rat, oral developmental toxicity (OECD Guideline 414 (Prenatal Developmental Toxicity Study)) = 100 mg/kg bw/day (maternal  and fetal NOAEL).

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2 July 2014 to 11 September 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
other: Crl:CD(SD)
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK).
- Age at study initiation: Approximately 71 days old
- Weight at study initiation: 233 to 286 g.
- Housing: for mating, the females were housed 1:1 with identified stock males, on the day of positive evidence of mating (Day0) one female per cage
steel mesh lid;
- Diet (e.g. ad libitum):ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: At least five days before commencement of pairing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):19-23ºC
- Humidity (%):40-70%
- Photoperiod (hrs dark / hrs light):12 hours light : 12 hours dark

IN-LIFE DATES: From: 2 July 2014 To:28 July to 31 July 2014
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test material was weighed out to give the required amount in a suitable container. Starting with the lowest concentration, approximately 50% of the final volume of vehicle was added to the test material.The liquid was then stirred using a magnetic stirrer until evenly mixed. The solution was made up to final volume with purified water and mixed using a magnetic stirrer until homogenous. The remaining formulations were made in ascending order of concentration.

VEHICLE

- Concentration in vehicle: 0, 6, 20 and 60 mg/ml
- Amount of vehicle (if gavage):5 mL/kg body weight
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analytical method GC-FID to check achieved concentration of samples of the first and last formulations prepared for administration and stability from the last formulation. Stability was assessed after six hours at ambient storage.

Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Proof of pregnancy: vaginal plug and sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
From Day 6 to Day 19 of gestation
Frequency of treatment:
Once daily
Duration of test:
20 days (from Day 0 to Day 20 of gestation)
No. of animals per sex per dose:
20 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:The doses used in this study (0, 30, 100 and 300 mg/kg/day) were chosen based on the findings from a Repeated toxicity and
Reproductive/Developmental Toxicity test with 1,3-Cyclohexanedimethanamine in Crl:CD(SD) Rats (Sponsor study Number: B041798).
- Rationale for animal assignment (if not random):To group and cage position in the sequence of mating. Females mating on any one day were evenly
distributed amongst the groups.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: start and end of each working day
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily 1 to 2 hours after completion of dosing and as late as possible in the working day.
detailed physical examination was performed on each animal on Days 0, 5, 12, 18 and 20 after mating to monitor general health

BODY WEIGHT: Yes
- Time schedule for examinations:weight of each adult was recorded on Days 0, 3, 6 to 20 after mating

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: Uterus Gravid uterine weight (including cervix and ovaries)
Full macroscopic examination of the tissues was performed. All external features and orifices were examined visually. Any abnormality in the appearance or size of any organ and tissue (external and cut surface) was recorded and the required tissue samples preserved in appropriate fixative.

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
- Other:Fetuses (live and dead).
Fetal examinations:
- External examinations: Yes: all per litter, sex of each fetus was recorded
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
Statistics:
The following sequence of statistical tests was used for body weight, gravid uterus weight, food consumption, corpora lutea, implantations, live young, fetal, placental and litter weight data:
Bartlett's test for variance homogeneity
Williams’ test for parametric monotonic trend test
Dunnett's test for non monotonic trend test
Kruskal-Wallis’ test for non parametric analysis if Bartlett's test was still significant at the 1% level following both logarithmic and square-root transformations.
Wilcoxon rank sum tests for significant (p<0.05) inter group comparisons
H1 approximate test for monotonicity of dose-response
Shirley's test, a non parametric monotonic trend test, for comparison of the means
Steel's test if the H1 approximate test was significant, suggesting that the dose-response was not monotone

Litter size and survival indices and fetal, placental and litter weight and gravid uterine weight data, if 75% of the data (across all groups) were the same value, for example c, Fisher’s Exact tests.

Pre/post implantation loss and sex ratio were analysed by a random test (Lipsitz).

For resorptions, each treated group was compared to control by exact Wilcoxon rank sum test.
Indices:
Prenatal losses are separated into pre- and post-implantation phases.

Pre-implantation loss (%) = (Number of corpora lutea – Number of implantations)/ Number of corpora lutea x 100

Post-implantation loss (%) = (Number of implantations –Number of live fetuses)/Number of implantations x 100
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
The oral administration of 1,3-BAC at 300 mg/kg/day was not tolerated by pregnant females. Three females were sacrificed for welfare reason as they were considered in general poor clinical condition, with signs including underactivity, gasping, piloerection, salivation and rales (noisy breathing), macroscopic examination revealed abnormal contents (gas) in the colon, ileum, jejunum, and stomach, and also revealed partially blocked nasal turbinates. Females receiving 300 mg/kg/day showed slight bodyweight loss between Days 6 and 7 of gestation and rales was also observed in six females given 300 mg/kg/day and in three females receiving 100 mg/kg/day.
For females which received 1,3-BAC at 30 or 100 mg/kg/day, there was no adverse effect of treatment on bodyweight, bodyweight change, adjusted bodyweight or food consumption.
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Placental weight was slightly reduced, when compare with the controls, in females given 300 mg/kg/day, there was, however, no effect on litter weight, or fetal weight.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): Placental weight was slightly reduced, when compare with the controls, in females given 300 mg/kg/day, there was, however, no effect on litter weight, or fetal weight.
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
There was considered to be no effect of treatment on litter parameters.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
There was considered to be no effect of treatment on litter parameters.
Changes in litter size and weights:
no effects observed
Skeletal malformations:
no effects observed
Description (incidence and severity):
The incidence of major and minor abnormalities and skeletal variants showed no relationship to treatment.
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
All females receiving 1,3-BAC at 300 mg/kg/day were found to be pregnant at macroscopic examination on Day 20 after mating. Litter data, as assessed by the mean numbers of corpora lutea, implantations, resorptions, live fetuses, sex ratio, and the extent of pre- and post-implantation loss were considered unaffected by treatment. Although the mean sex ratio at 300mg/kg/d was statistically lower than in Controls, the magnitude of difference from the ideal value of 50% was similar to controls and there was no effect of treatment on embryo-fetal survival or the morphology of the reproductive organs. Placental weight was slightly reduced, however there was no effect on litter or fetal weights and there were no treatment related fetal pathology examinations.
Thus, the data showed that there is no evidence to indicate that 1,3-BAC is a developmental toxicant at any of the doses tested, up to 300mg/kg/day.
Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
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
external malformations
skeletal malformations
Abnormalities:
not specified
Developmental effects observed:
not specified

Formulation analysis: the stability was confirmed for 1,3-BAC in water formulations and the mean concentrations of 1,3-BAC in test formulations analysed for the study were within ±8% of nominal concentrations, confirming accurate formulation.

Conclusions:
Treatment with 1,3-BAC at 30 or 100 mg/kg/day was well tolerated, with no effects of treatment on embryo-fetal growth, survival or development. Treatment with 1,3-BAC at 300 mg/kg/day exceeded the maximum tolerated dose for pregnant females but was not associated with adverse effects on embryo-fetal development. The maternal and fetal no observed adverse effect level (NOAEL) was therefore considered to be 100 mg/kg/day.
Endpoint:
developmental toxicity
Type of information:
other: Expert statement
Adequacy of study:
key study
Study period:
2017
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Expert statement based on toxicity data available
Principles of method if other than guideline:
Expert statement based on toxicity data available.
GLP compliance:
no
Species:
rabbit
Remarks on result:
not measured/tested
Remarks on result:
not measured/tested
Developmental effects observed:
not specified
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
100 mg/kg bw/day
Study duration:
subchronic
Species:
rat
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

The influence of 1.3-BAC on pregnancy, embryo-fetal survival and development, was assessed when administered to female Sprague-Dawley rats at doses up to 300 mg/kg/day during the organogenesis phase of pregnancy (Days 6 to 19 after mating, inclusive) in a Prenatal Developmental Toxicity Study (OECD 414) and from 14 days before mating until day 4 of lactation in a combined repeated dose toxicity and reproductive toxicity screening study.

In the prenatal developmental toxicity study, treatment with 1,3 -BAC at 30,100 and 300 mg/kg/day showed no effects of treatment on embryo-fetal growth, survival or development however the dose of 300 mg/kg/day was not well tolerated by pregnant females.

In the reproductive/developmental toxicity screening study, the test substance had no effects on estrus cycle, copulation index, fertility index, gestation index, gestation length, numbers of corpora lutea or implantations, implantation index, delivery index, parturition, or maternal behaviour at doses of 10, 60, 300 mg/kg/day. As in the prenatal developmental toxicity study, treatment at 300mg/kg/day had effects on the general conditions of some of the animals and macroscopic changes considered attributed to the test substance were noted in the stomach of both sexes and in the testes and epididymis of males at 300 mg/kg/day.

Data from both studies concurs with the conclusion that the maternal and fetal no observed adverse effect level (NOAEL) for developmental toxicity is 100 mg/kg/day. 

Justification for classification or non-classification

No effects on reproductive or developmental parameters were observed at doses up to 100 mg/kg/day. As there was no evidence for reproductive toxicity in the screening study conducted or developmental toxicity in the the prenatal developmental toxicity study, classification for reproductive toxicity does not apply according to the CLP Regulation (EC regulation 1272/2008).