Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Screening study; oral (gavage); rat (Sprague Dawley), m/f (OECD guideline 422, GLP): NOAEL(general toxicity) = 300 mg/kg bw/d; NOAEL(reproductive / developmental toxicity) = 120 mg/kg bw/d

Link to relevant study records
Reference
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:
2013-08-14 to 2013-10-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study, GLP
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
adopted on 22 March 1996
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Hsd: Sprague Dawley SD
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Italia SpA, Italy
- Age at study initiation: 6 to 7 weeks
- Weight at study initiation: 176 to 200 g (males), 151 to 175 g (females)
- Housing: up to 5 of one sex/cage, in polisulphone solid bottomed cages measuring 59.5x38x20 cm; nesting material was provided inside suitable bedding bags and changed at least twice a week
- During mating, animals were housed one male to one female in clear polycarbonate cages measuring approximately 43x27x19 cm with a stainless steel
mesh lid and floor, each cage tray held absorbent material which was inspected and changed daily
- After mating, the males were re-caged as they were before mating, the females were transferred to individual solid bottomed cages for the gestation period, birth and lactation; suitable nesting material was provided and changed as necessary
- Diet (e.g. ad libitum): laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI), Italy), ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C±2°C
- Humidity (%): 55%±15%
- Air changes (per hr): 15 to 20
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The formulations were prepared daily.

VEHICLE
- Concentration in vehicle: 10, 24 and 60 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg body weight
Details on mating procedure:
- M/F ratio per cage: 1:1
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 1 of pregnancy
- The female was paired with the same male until positive identification occurred.
- After successful mating each pregnant female was transferred to individual solid bottomed cage for the gestation period, birth and lactation; suitable nesting material was provided and changed as necessary
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentration and homogeneity of the low and high dose level were assessed by taking six analytical aliquots (approximately 1 g) in different positions. For the intermediate levels, only concentration was assessed by taking two different analytical aliquots (approximately 1 g).
Duration of treatment / exposure:
males: 29 d (2 consecutive weeks prior to pairing and thereafter through the day before necropsy)
females: 29 d (2 consecutive weeks prior to pairing and thereafter during pairing, post coitum and post partum periods until Day 3 post partum or the day before sacrifice)
Frequency of treatment:
daily, 7 d / week
Details on study schedule:
F1 animals were not mated (screening study)
Remarks:
Doses / Concentrations:
0, 50, 120, 300 mg/kg bw/d
Basis:
actual ingested
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for animal assignment: computerised stratified randomisation to give approximately equal initial group mean body weights
Parental animals: Observations and examinations:
CLINICAL SINGS AND MORTALITY: Yes
- Time schedule: at least once daily, approximately 0.5-1 hour after dosing

DETAILED CLINICAL OBSERVATIONS: Yes (Functional Observation Battery Tests)
- Time schedule: once before commencement of treatment and at least once a week thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: weekly; females additionally on Days 0, 7, 14 and 20 post coitum, dams with live pups were also weighed on Days 1 and 4 post partum

FOOD CONSUMPTION AND COMPOUND INTAKE: yes
-Time scedule: weekly during the pre-mating period, Individual food consumption for the females was measured on Days 7, 14 and 20 post coitum starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: day of necropsy
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: 5 males and 5 females randomly selected from each group
- Parameters: Haematocrit, Haemoglobin, Red blood cell count, Reticulocyte count, Mean red blood cell volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration, White blood cell count, Differential leucocyte count (Neutrophils, Lymphocites, Eosinophils, Basophils, Monocytes, Large unstained cells), Platelets, Prothrombin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: day of necropsy
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: 5 males and 5 females randomly selected from each group
- Parameters: Alkaline phosphatase, Alanine aminotransferase, Aspartate aminotransferase, Gamma-glutamyltransferase, Urea, Creatinine, Glucose, Triglycerides, Bile acids, Inorganic phosphorus, Total bilirubin, Total cholesterol, Total protein, Albumin, Globulin, A/G Ratio, Sodium, Potassium, Calcium, Chloride

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: once during the study, towards the end of treatment,
- Dose groups that were examined: all groups, 5 males and 5 females
- Battery of functions tested: sensory activity / grip strength / motor activity
Oestrous cyclicity (parental animals):
Vaginal smears were taken daily in the morning starting two weeks before pairing until a positive identification of copulation was made. The vaginal
smear data were examined to determine the following:
1. anomalies of the oestrous cycle
2. the pre-coital interval (i.e., the number of nights paired prior to the detection of mating)
Sperm parameters (parental animals):
Parameters examined in P males:
testis weight, epididymis weight, staging of spermatogenic cycle
Litter observations:
PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, body weight Days 1 and 4 post partum

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals. The males were killed after the mating of all females on Day 30 of study.
- Maternal animals: All surviving animals . The females with live pups were killed on Day 4 post partum. The females with total litter loss were killed on the day the total litter loss occurred or shortly after.
The females which did not give birth 25 days after positive identification of mating were sacrificed on Days 26, 27 or 28 post coitum.

All females were examined also for the following:
– number of visible implantation sites (pregnant animals)
– number of corpora lutea (pregnant animals)

GROSS PATHOLOGY: Yes (see table)

HISTOPATHOLOGY: Yes (see table)
Postmortem examinations (offspring):
SACRIFICE / GROSS NECROPSY
All pups found dead in the cage were examined for external and internal abnormalities. All live pups sacrificed at termination were killed and examined for external abnormalities and sex confirmation by gonadal inspection.
Statistics:
Standard deviations were calculated as appropriate. For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data.
Statistical analysis of histopathological findings was carried out by means of the non-parametric Kolmogorov-Smirnov test if n was more than 5. The non-parametric Kruskal-Wallis analysis of variance was used for the other parameters. Intergroup differences between the control and treated groups were assessed by the non-parametric version of the Williams test. The criterion for statistical significance was p<0.05.
Reproductive indices:
Copulation Index, Fertility Index
Offspring viability indices:
Pre-birth loss, Pup loss at birth, Cumulative pup loss on Day 4 post partum
Clinical signs:
no effects observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
effects observed, treatment-related
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No mortality occurred during the study. No significant clinical signs were observed.
Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
No relevant differences were found in body weight and body weight gain between male groups throughout the study and between female groups up to Day 14 of post coitum period. On Day 20 post coitum, a decrease in body weight and body weight gain (statistically significant) was evident in females dosed at 300 mg/kg bw/day respect to controls.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
Oestrous cycle and reproductive parameters were unaffected by treatment. A slight increase in mean pre-coital interval was observed in the mid- (120 mg/kg bw/day) and high dose (300 mg/kg/day) animals compared to controls. The increase in the mid-dose group was related to 1 female which mated 14 days after pairing.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages; regular layering in the germinal epithelium was noted.

IMPLANTATION; PRE-BIRTH LOSS DATA AND GESTATION LENGTH (FEMALES)
Gestation length of the low (50 mg/kg bw/day) and mid-dose (120 mg/kg bw/day) groups was slightly higher than control group in which the majority of dams gave birth on Day 22 of gestation. Most of low and mid-dose females gave birth on Day 23 post coitum. High dose females (300 mg/kg bw/day) had more prolonged gestation length, statistically significant, compared to controls. In particular, 4 females gave birth on Day 25 post coitum, 2 gave birth on Day 24 post coitum and only 1 on Day 22 post coitum. The pre-birth loss was significantly increased at statistical analysis in high dose females, compared to controls. This increase could be attributable to the prolonged gestation period which caused most probably pup suffering and the death during or shortly after the birth.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Terminal body weight was unaffected by treatment. Absolute and relative thymus weights were slightly reduced in mid- and high dose males compared to controls. High dose females (300 mg/kg bw/day) showed a slight reduction in absolute and relative adrenals weights.

GROSS PATHOLOGY (PARENTAL ANIMALS)
No remarkable changes were noted at post mortem examination in treated animals, when compared with controls.

HISTOPATHOLOGY (PARENTAL ANIMALS)
No treatment-related changes were noted. Periportal hepatocytic vacuolation, consistent with fatty change like associated in most instances with inflammatory cell foci and in few instances with single cell apoptosis/necrosis, was seen in a single control male and female and with an increased incidence in treated males and females. However, considering that the liver pathologyshowed a different incidence between treated males and females, without any dose-relation and that the pathological picture was similar to that observed in male and female controls, such changes could be attributed to spontaneous or incidental pathology rather than to the treatment. Lymphoid depletion in the thymus was only noted in 2 out of 10 high dose females (300 mg/kg bw/day) and was considered unrelated to treatment. The sporadic lesions reported in control and treated animals were considered to be an expression of spontaneous and/or incidental pathology, commonly seen in this species and age under our experimental conditions.

OTHER FINDINGS (PARENTAL ANIMALS)
More details are given in IUCLID section "Repeated dose toxicity".
Dose descriptor:
NOAEL
Remarks:
general toxicity
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: no treatment-related adverse effects
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
120 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
female
Basis for effect level:
other: significantly increased pre-birth loss, prolonged gestation period in high dose females
Dose descriptor:
NOAEL
Remarks:
reproductive toxicity
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male
Basis for effect level:
other: no treatment-related adverse effects
Critical effects observed:
no
Clinical signs:
no effects observed
Mortality / viability:
mortality observed, treatment-related
Body weight and weight changes:
not examined
Sexual maturation:
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
Considering that no high dose females (300 mg/kg bw/day) had live pups from Day 1 post partum, the group litter data and sex ratios were evaluated between control, low (50 mg/kg bw/day) and mid-dose (120 mg/kg bw/day) females only.
No relevant differences in litter data were noted between the control, the low and the mid-dose groups. Decreases in litter weights, not statistically significant, were seen in low and mid-dose groups. These differences were due to the lower number of pups in treated groups respect to control (more evident in mid-dose group). The increased pup loss observed in mid-dose group was attributed to single females. Sex ratios at birth and on Day 4 post partum did not show any differences.

CLINICAL SIGNS (OFFSPRING)
No treatment-related findings were noted in pups. An increased presence of missing or dead pups was noted in the high dose females (300 mg/kg bw/day).

GROSS PATHOLOGY (OFFSPRING)
No treatment-related findings were noted in pups which died or in pups sacrificed on Day 4 post partum.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
120 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: significantly increased pre-birth loss; no high dose females had live pups from Day 1 post partum
Reproductive effects observed:
yes
Lowest effective dose / conc.:
300 mg/kg bw/day (actual dose received)
Treatment related:
yes
Relation to other toxic effects:
reproductive effects in the absence of other toxic effects
Dose response relationship:
not specified
Relevant for humans:
yes
Conclusions:
A combined repeated toxicity study with a reproduction and developmental toxicity study was conducted in Sprague Dawley rats in order to provide information on systemic toxicity (body weight and body weight gain, clinical signs including neurotoxicity assessment, motor activity and sensory reaction to stimuli, food consumption, clinical pathology parameters, organ weights, macroscopic and microscopic examination) as well as any possible effects of the test item on male and female reproductive performance (gonadal function, mating behaviour, conception, development of the conceptus, parturition and early lactation).The dosages were 50, 120 and 300 mg/kg bw/day.
On the basis of the results obtained in the study, the NOAEL (No Observed Adverse Effect Level) for systemic toxicity was considered 300 mg/kg bw/day for males and females.
The NOAEL for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day for males and 120 mg/kg bw/day for females and their litters.
Executive summary:

In a combined repeated toxicity study with a reproduction and developmental toxicity study according to OECD Guideline 422 (adopted on 22 March 1996) Tetrahydrofurfuryl methacrylate was administered to 10 Sprague Dawley rats / sex / dose by gavage at dose levels of 0 (control), 50, 120 and 300 mg/kg bw/day in corn oil at a constant volume of 5 mL/kg bw.

 

Males were treated for a total of 29 days including 2 weeks prior to pairing and continuously thereafter, up to the day before necropsy. Females were dosed throughout the study including 2 weeks before pairing, thereafter during pairing, gestation and lactation periods until the day of necropsy or Day 3 post partum for females with live pups at Day 4 post partum.

The following parameters were evaluated in parental animals: body weight, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, oestrous cycle, mating performance, clinical pathology investigations (haematology and clinical chemistry), litter data, macroscopic observations, organ weights and histopathological examination. Pup weight, pup clinical signs and pup macroscopic observations were also performed.

 

No mortality occurred in the study. Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item. No significant clinical signs were observed.

 

Males

No relevant changes were recorded during the study and at the post mortem examinations in males at any dose level investigated. In particular no effects were seen on body weight and body weight gain,

clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, clinical pathology investigations (haematology and clinical chemistry), macroscopic observations, organ weights and histopathological examination, including identification of the stages of

the spermatogenic cycle. The mating performance including the pre-coital interval (i.e. the number of days paired to sperm positive day) and the copulatory evidence (i.e. the presence of sperm and/or copulation plug in situ or in the cage) did not show differences between groups.

 

Females

A total of 5 females were found not pregnant at necropsy: 4 in the control group and 1 in the mid-dose (120 mg/kg bw/day) group. Unilateral implantation was observed in one low dose (50 mg/kg bw/day) female. In the high dose group (300 mg/kg bw/day), 3 females had total resorption and 7 females had total litter loss within 1 day of parturition.

Therefore, the number of females with live pups on Day 4 post partum was: 6 in the control, 10 in the low dose (50 mg/kg bw/day), 9 in the mid-dose (120 mg/kg bw/day) group and none in the high dose (300 mg/kg bw/day) group.

On Day 20 post coitum, a decrease in body weight and body weight gain (statistically significant) was evident in females dosed at 300 mg/kg bw/day respect to controls. Decreases in food consumption were seen in high dose females (300 mg/kg bw/day) when compared with controls during the post coitum and post partum periods with statistically significance on Days 7 and 14 post coitum and 4 post partum. Gestation length of all treated groups was higher than controls and significantly increased at statistical analysis, in the high dose group. The pre-birth loss was significantly increased at statistical analysis, in high dose females. This increase could be attributable to the prolonged gestation period which caused most probably pup suffering and the death during or shortly after the birth.

 

Offspring

An increased presence of missing or dead pups was noted in females receiving 300 mg/kg bw/day. No other treatment-related findings were noted in pups.

At necropsy no treatment-related findings were noted in pups which died or in pups sacrificed on Day 4 post partum.

No difference in sex ratios was noted between the control and treated groups.

No relevant differences in litter data were seen. Decreases in litter weights, seen in low and mid-dose groups were due to the lower number of pups in treated groups respect to control, more evident in mid-dose group in which the increased pup loss was attributed to single females.

 

On the basis of the results obtained in the study, the NOAEL (No Observed Adverse Effect Level) for systemic toxicity was considered 300 mg/kg bw/day for males and females.

The NOAEL for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day for males and 120 mg/kg bw/day for females and their litters.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Effect on fertility: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
120 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
OECD guideline 422 study, no deviations, GLP
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

For THFMA, a reliable (RL=1), relevant and adequate study is available on toxicity to reproduction:

 

In a combined repeated toxicity study with a reproduction and developmental toxicity study according to OECD Guideline 422 (adopted on 22 March 1996) Tetrahydrofurfuryl methacrylate was administered to 10 Sprague Dawley rats / sex / dose by gavage at dose levels of 0 (control), 50, 120 and 300 mg/kg bw/day in corn oil at a constant volume of 5 mL/kg bw.

 

Males were treated for a total of 29 days including 2 weeks prior to pairing and continuously thereafter, up to the day before necropsy. Females were dosed throughout the study including 2 weeks before pairing, thereafter during pairing, gestation and lactation periods until the day of necropsy or Day 3 post partum for females with live pups at Day 4 post partum.

The following parameters were evaluated in parental animals: body weight, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, oestrous cycle, mating performance, clinical pathology investigations (haematology and clinical chemistry), litter data, macroscopic observations, organ weights and histopathological examination. Pup weight, pup clinical signs and pup macroscopic observations were also performed.

 

No mortality occurred in the study. Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item. No significant clinical signs were observed.

 

Males

No relevant changes were recorded during the study and at the post mortem examinations in males at any dose level investigated. In particular no effects were seen on body weight and body weight gain,

clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, clinical pathology investigations (haematology and clinical chemistry), macroscopic observations, organ weights and histopathological examination, including identification of the stages of

the spermatogenic cycle. The mating performance including the pre-coital interval (i.e. the number of days paired to sperm positive day) and the copulatory evidence (i.e. the presence of sperm and/or copulation plug in situ or in the cage) did not show differences between groups.

 

Females

A total of 5 females were found not pregnant at necropsy: 4 in the control group and 1 in the mid-dose (120 mg/kg bw/day) group. Unilateral implantation was observed in one low dose (50 mg/kg bw/day) female. In the high dose group (300 mg/kg bw/day), 3 females had total resorption and 7 females had total litter loss within 1 day of parturition.

Therefore, the number of females with live pups on Day 4 post partum was: 6 in the control, 10 in the low dose (50 mg/kg bw/day), 9 in the mid-dose (120 mg/kg bw/day) group and none in the high dose (300 mg/kg bw/day) group.

On Day 20 post coitum, a decrease in body weight and body weight gain (statistically significant) was evident in females dosed at 300 mg/kg bw/day respect to controls. Decreases in food consumption were seen in high dose females (300 mg/kg bw/day) when compared with controls during the post coitum and post partum periods with statistically significance on Days 7 and 14 post coitum and 4 post partum. Gestation length of all treated groups was higher than controls and significantly increased at statistical analysis, in the high dose group. The pre-birth loss was significantly increased at statistical analysis, in high dose females. This increase could be attributable to the prolonged gestation period which caused most probably pup suffering and the death during or shortly after the birth.

 

Offspring

An increased presence of missing or dead pups was noted in females receiving 300 mg/kg bw/day. No other treatment-related findings were noted in pups.

At necropsy no treatment-related findings were noted in pups which died or in pups sacrificed on Day 4 post partum.

No difference in sex ratios was noted between the control and treated groups.

No relevant differences in litter data were seen. Decreases in litter weights, seen in low and mid-dose groups were due to the lower number of pups in treated groups respect to control, more evident in mid-dose group in which the increased pup loss was attributed to single females.

 

On the basis of the results obtained in the study, the NOAEL (No Observed Adverse Effect Level) for systemic toxicity was considered 300 mg/kg bw/day for males and females.

The NOAEL for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day for males and 120 mg/kg bw/day for females and their litters.


Effects on developmental toxicity

Description of key information
Screening study; oral (gavage); rat (Sprague Dawley), m/f (OECD guideline 422, GLP): NOAEL(general toxicity) = 300 mg/kg bw/d; NOAEL(reproductive / developmental toxicity) = 120 mg/kg bw/d
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2013-08-14 to 2013-10-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study, GLP
Qualifier:
according to
Guideline:
other: OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) (adopted on 22 March 1996)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
other: Hsd: Sprague Dawley SD
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Italia SpA, Italy
- Age at study initiation: 6 to 7 weeks
- Weight at study initiation: 176 to 200 g (males), 151 to 175 g (females)
- Housing: up to 5 of one sex/cage, in polisulphone solid bottomed cages measuring 59.5x38x20 cm; nesting material was provided inside suitable bedding bags and changed at least twice a week
- During mating, animals were housed one male to one female in clear polycarbonate cages measuring approximately 43x27x19 cm with a stainless steel
mesh lid and floor, each cage tray held absorbent material which was inspected and changed daily
- After mating, the males were re-caged as they were before mating, the females were transferred to individual solid bottomed cages for the gestation period, birth and lactation; suitable nesting material was provided and changed as necessary
- Diet (e.g. ad libitum): laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI), Italy), ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C±2°C
- Humidity (%): 55%±15%
- Air changes (per hr): 15 to 20
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The formulations were prepared daily.

VEHICLE
- Concentration in vehicle: 10, 24 and 60 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg body weight
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
- M/F ratio per cage: 1:1
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 1 of pregnancy
- The female was paired with the same male until positive identification occurred.
- After successful mating each pregnant female was transferred to individual solid bottomed cage for the gestation period, birth and lactation; suitable nesting material was provided and changed as necessary
Duration of treatment / exposure:
males: 29 d (2 consecutive weeks prior to pairing and thereafter through the day before necropsy)
females: 29 d (2 consecutive weeks prior to pairing and thereafter during pairing, post coitum and post partum periods until Day 3 post partum or the day before sacrifice)
Frequency of treatment:
daily, 7 d / week
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Maternal examinations:
CLINICAL SINGS AND MORTALITY: Yes
- Time schedule: at least once daily, approximately 0.5-1 hour after dosing

DETAILED CLINICAL OBSERVATIONS: Yes (Functional Observation Battery Tests)
- Time schedule: once before commencement of treatment and at least once a week thereafter

BODY WEIGHT: Yes
- Time schedule for examinations: weekly; females additionally on Days 0, 7, 14 and 20 post coitum, dams with live pups were also weighed on Days 1 and 4 post partum

SACRIFICE
- Male animals: All surviving animals. The males were killed after the mating of all females on Day 30 of study.
- Maternal animals: All surviving animals . The females with live pups were killed on Day 4 post partum. The females with total litter loss were killed on the day the total litter loss occurred or shortly after.
The females which did not give birth 25 days after positive identification of mating were sacrificed on Days 26, 27 or 28 post coitum.

All females were examined also for the following:
– number of visible implantation sites (pregnant animals)
– number of corpora lutea (pregnant animals)

more details are given in IUCLID section "repeated dose toxicity" and "toxicity to reproduction"
Fetal examinations:
SACRIFICE / GROSS NECROPSY
All pups found dead in the cage were examined for external and internal abnormalities. All live pups sacrificed at termination were killed and examined for external abnormalities and sex confirmation by gonadal inspection.
Dose descriptor:
NOAEL
Effect level:
120 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: developmental toxicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
A combined repeated toxicity study with a reproduction and developmental toxicity study was conducted in Sprague Dawley rats in order to provide information on systemic toxicity (body weight and body weight gain, clinical signs including neurotoxicity assessment, motor activity and sensory reaction to stimuli, food consumption, clinical pathology parameters, organ weights, macroscopic and microscopic examination) as well as any possible effects of the test item on male and female reproductive performance (gonadal function, mating behaviour, conception, development of the conceptus, parturition and early lactation).The dosages were 50, 120 and 300 mg/kg bw/day.
On the basis of the results obtained in the study, the NOAEL (No Observed Adverse Effect Level) for systemic toxicity was considered 300 mg/kg bw/day for males and females.
The NOAEL for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day for males and 120 mg/kg bw/day for females and their litters.
Executive summary:

In a combined repeated toxicity study with a reproduction and developmental toxicity study according to OECD Guideline 422 (adopted on 22 March 1996) Tetrahydrofurfuryl methacrylate was administered to 10 Sprague Dawley rats / sex / dose by gavage at dose levels of 0 (control), 50, 120 and 300 mg/kg bw/day in corn oil at a constant volume of 5 mL/kg bw.

 

Males were treated for a total of 29 days including 2 weeks prior to pairing and continuously thereafter, up to the day before necropsy. Females were dosed throughout the study including 2 weeks before pairing, thereafter during pairing, gestation and lactation periods until the day of necropsy or Day 3 post partum for females with live pups at Day 4 post partum.

The following parameters were evaluated in parental animals: body weight, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, oestrous cycle, mating performance, clinical pathology investigations (haematology and clinical chemistry), litter data, macroscopic observations, organ weights and histopathological examination. Pup weight, pup clinical signs and pup macroscopic observations were also performed.

 

No mortality occurred in the study. Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item. No significant clinical signs were observed.

 

A total of 5 females were found not pregnant at necropsy: 4 in the control group and 1 in the mid-dose (120 mg/kg bw/day) group. Unilateral implantation was observed in one low dose (50 mg/kg bw/day) female. In the high dose group (300 mg/kg bw/day), 3 females had total resorption and 7 females had total litter loss within 1 day of parturition.

Therefore, the number of females with live pups on Day 4 post partum was: 6 in the control, 10 in the low dose (50 mg/kg bw/day), 9 in the mid-dose (120 mg/kg bw/day) group and none in the high dose (300 mg/kg bw/day) group.

On Day 20 post coitum, a decrease in body weight and body weight gain (statistically significant) was evident in females dosed at 300 mg/kg bw/day respect to controls. Decreases in food consumption were seen in high dose females (300 mg/kg bw/day) when compared with controls during the post coitum and post partum periods with statistically significance on Days 7 and 14 post coitum and 4 post partum. Gestation length of all treated groups was higher than controls and significantly increased at statistical analysis, in the high dose group. The pre-birth loss was significantly increased at statistical analysis, in high dose females. This increase could be attributable to the prolonged gestation period which caused most probably pup suffering and the death during or shortly after the birth.

 

An increased presence of missing or dead pups was noted in females receiving 300 mg/kg bw/day. No other treatment-related findings were noted in pups. At necropsy no treatment-related findings were noted in pups which died or in pups sacrificed on Day 4 post partum. No difference in sex ratios was noted between the control and treated groups. No relevant differences in litter data were seen. Decreases in litter weights, seen in low and mid-dose groups were due to the lower number of pups in treated groups respect to control, more evident in mid-dose group in which the increased pup loss was attributed to single females.

 

On the basis of the results obtained in the study, the NOAEL (No Observed Adverse Effect Level) for systemic toxicity was considered 300 mg/kg bw/day for males and females.

The NOAEL for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day for males and 120 mg/kg bw/day for females and their litters.

NOTE: Any of data in this dataset are disseminated by the European Union on a right-to-know basis and this is not a publication in the same sense as a book or an article in a journal. The right of ownership in any part of this information is reserved by the data owner(s). The use of this information for any other, e.g. commercial purpose is strictly reserved to the data owners and those persons or legal entities having paid the respective access fee for the intended purpose.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
120 mg/kg bw/day
Study duration:
subacute
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

For THFMA, a reliable (RL=1), relevant and adequate study is available on developmental toxicity:

 

In a combined repeated toxicity study with a reproduction and developmental toxicity study according to OECD Guideline 422 (adopted on 22 March 1996) Tetrahydrofurfuryl methacrylate was administered to 10 Sprague Dawley rats / sex / dose by gavage at dose levels of 0 (control), 50, 120 and 300 mg/kg bw/day in corn oil at a constant volume of 5 mL/kg bw.

More details on general toxicity are presented in section “Repeated dose toxicity”.

Females were dosed throughout the study including 2 weeks before pairing, thereafter during pairing, gestation and lactation periods until the day of necropsy or Day 3 post partum for females with live pups at Day 4 post partum.

The following parameters were evaluated in parental animals: body weight, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction to stimuli), food consumption, oestrous cycle, mating performance, clinical pathology investigations (haematology and clinical chemistry), litter data, macroscopic observations, organ weights and histopathological examination. Pup weight, pup clinical signs and pup macroscopic observations were also performed.

 

No mortality occurred in the study. Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item. No significant clinical signs were observed.

 

A total of 5 females were found not pregnant at necropsy: 4 in the control group and 1 in the mid-dose (120 mg/kg bw/day) group. Unilateral implantation was observed in one low dose (50 mg/kg bw/day) female. In the high dose group (300 mg/kg bw/day), 3 females had total resorption and 7 females had total litter loss within 1 day of parturition.

Therefore, the number of females with live pups on Day 4 post partum was: 6 in the control, 10 in the low dose (50 mg/kg bw/day), 9 in the mid-dose (120 mg/kg bw/day) group and none in the high dose (300 mg/kg bw/day) group.

 

On Day 20 post coitum, a decrease in body weight and body weight gain (statistically significant) was evident in females dosed at 300 mg/kg bw/day respect to controls. Decreases in food consumption were seen in high dose females (300 mg/kg bw/day) when compared with controls during the post coitum and post partum periods with statistically significance on Days 7 and 14 post coitum and 4 post partum. Gestation length of all treated groups was higher than controls and significantly increased at statistical analysis, in the high dose group. The pre-birth loss was significantly increased at statistical analysis, in high dose females. This increase could be attributable to the prolonged gestation period which caused most probably pup suffering and the death during or shortly after the birth.

 

An increased presence of missing or dead pups was noted in females receiving 300 mg/kg bw/day. No other treatment-related findings were noted in pups.

At necropsy no treatment-related findings were noted in pups which died or in pups sacrificed on Day 4 post partum.

No difference in sex ratios was noted between the control and treated groups.

No relevant differences in litter data were seen. Decreases in litter weights, seen in low and mid-dose groups were due to the lower number of pups in treated groups respect to control, more evident in mid-dose group in which the increased pup loss was attributed to single females.

 

On the basis of the results obtained in the study, the NOAEL (No Observed Adverse Effect Level) for systemic toxicity was considered 300 mg/kg bw/day for males and females.

The NOAEL for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day for males and 120 mg/kg bw/day for females and their litters.


Justification for selection of Effect on developmental toxicity: via oral route:
OECD guideline 422 study, no deviations, GLP

Justification for classification or non-classification

Based on the available data, THFMA is classified for toxicity to reproduction and developmental toxicity Category 1B and assigned the hazard phrase H360D according to the criteria given in regulation (EC) 1272/2008.