Registration Dossier

Administrative data

Description of key information

In a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test according to OECD Guideline 422 (28 July 2015) 4-{Methacryloyloxy)benzophenone was administered to 10 Hsd: Sprague Dawley SD rats/sex/dose orally by gavage at dose levels of 0 (control), 100, 300 and 1000 mg/kg bw/d. The treatment schedule included 2 weeks before pairing, during pairing, post coitum and post partum periods up to day 3 post partum. Animals were administered for approximately 5 and 8 weeks for males and females, respectively. 

 

The purpose of this study was to generate information concerning toxic effects on SpragueDawley rats of both sexes after repeated oral dosing with 4-(Methacryloyloxy)benzophenone,as well as any effects of the test item on male and female reproductive performance, such

asgonadal function, conception, parturition and lactation of offspring. The dose levels usedduring the study were 70, 200 and 600 mg/kg/day.

Males

One control male was sacrificed on Day 14 of the study for poor health condition. On thebasis of the macroscopic and microscopic findings, the factor contributory to the death wasmainly attributed to a misdosing.

Males were treated for 2 weeks prior to pairing and during pairing with females until the daybefore necropsy, for a total of 48 days.

During thein-lifephase, mortality check, clinical signs (including neurotoxicity assessment,motor activity and sensory reaction tostimuli), body weight, body weight gain, food consumptionand mating performance were evaluated.

Clinical pathology investigations (haematology, clinical chemistry, urinalysis), organ weights,macroscopic observations and histopathological examination were also performed. In

addition, thyroid hormone levels were determined in all adult males. The histopathologicalexamination was performed only on control and high dose groups (5 animals/sex/grouprandomly selected). It included identification of the stages of the spermatogenic cycle in the

same five control and high dose males.

No relevant signs of toxicological significance were observed in treated males, apart from atransient salivation, observed for a few days in the high dose animals.

Body weights and food consumption did not show any treatment related effects. Fertilityindex and copulatory index were unaffected by treatment. No effects considered treatmentrelated were observed in haematology or coagulation, clinical chemistry and urinalysis.

Hormone analysis did not showany relation to treatment. Some fluctuations in organweights,absolute and/or relative, were noted, but in the absence of histopathological correlated

findings, these changes were considered of no toxicological significance.

Females

Females were treated for 2weeks prior to pairing, during pairing and throughout the gestationand lactation periods until Day 12post partum.

During thein-lifephase, mortality check, clinical signs (including neurotoxicity assessment,motor activity and sensory reaction tostimuli), body weight, body weight gain, food

consumption and mating performance were evaluated. Clinical pathology investigations(haematology and clinical chemistry) and thyroid hormone determination in pups on Day 13post partumwere also evaluated. The histopathological examination was carried out in five

females of control and high dose groups, selected randomly. Clinical signs of pups, as well asnecropsy examination of decedent pups or pups sacrificed on Days 4 and 13post partum(including thyroid weight) were recorded. Litter data, sex ratios and gestation length were

recorded.

No relevant signs of toxicological significance were observed. As noted for males, salivationwas recorded occasionally in treated females receiving 600 mg/kg/day. Body weights andfood consumption did not show changes of toxicological significance.

Concerning the reproductive parameters, no relevant differences were found in terms ofmating performance including the pre-coital interval (number of days paired to spermpositive day) and the copulatory evidence (the positive identification of mating i.e. the

presence of spermand/or copulation plugin situor in the cage), as well as fertility index.

The slight changes noted at clinical chemistry investigation (haematology, clinical chemistryand coagulation) were not considered treatment related. Some fluctuations in organ weights,absolute and/or relative, were noted but in the absence of histopathological correlated

findings, these changes were considered of no toxicological significance.

Conclusion

Based on the results of the present study, the NOAEL (No Observed Adverse Effect Level) forgeneral toxicity and for reproductive and developmental toxicity was considered to be 600mg/kg/day both for males and females.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: oral, other
Remarks:
COMBINED REPEATED DOSE TOXICITY STUDY WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST IN RATS
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19.09.2016 to 02.11.2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
adopted on 28 July 2015.
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Specific details on test material used for the study:
98.408 % 4-(Methacryloyloxy)benzophenone
0.581 % Benzophenone acetate
0.054 % 4-Hydroxy benzophenone

Expiry date 10 November 2016 (extended to 10 February 2017 as communication of the Sponsor of the 17 November 2016)
Storage conditions Room temperature protected from light
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone (UD), Italy.
- Age at study initiation: 8 to 9 weeks
- Weight at study initiation: 199 to 213 g for females, 168 to 197 g for males
- Fasting period before study:
- Housing:
- Diet : commercially available laboratory rodent diet (4 RF 21,Mucedola S.r.l., Via G. Galilei, 4, 20019, SettimoMilanese (MI), Italy) was offered ad libitum throughout the study
- Water (e.g. ad libitum): yes except in the case of urinalysis investigations
- Acclimation period: 19 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 hours

IN-LIFE DATES: from 28 September (day of allocation) to 15. November 2016
Route of administration:
oral: gavage
Details on route of administration:
The test item was administered orally by gavage at a dose volume of 5mL/kg body weight.
Control animals received the vehicle alone at the same dose volume.
The dose was administered to each animal on the basis of the most recently recorded body weight and the volume administered was recorded for each animal.
Vehicle:
corn oil
Details on oral exposure:
Dose level:
Group 1: Control (vehicle)
Group 2: 70 mg/kg/day
Group 3: 200 mg/kg/day
Group 4: 600 mg/kg/day
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis was performed to confirmthat the proposed formulation procedure was acceptable and that the stability of the formulation was satisfactory. In the present study, samples of the formulations prepared during the study were also analysed to check the concentration and homogeneity (the first and the last week of treatment, where possible).
The software used for this activity was the Empower® Pro build No. 2154. Results of formulation analyses were within the acceptability limits
Duration of treatment / exposure:
5mL/kg body weight
Frequency of treatment:
Once daily
Dose / conc.:
70 mg/kg bw/day (actual dose received)
Dose / conc.:
200 mg/kg bw/day (actual dose received)
Dose / conc.:
600 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10 male and 10 female rats
Control animals:
yes, concurrent vehicle
Details on study design:
This study design was in agreement with the procedures described in OECD Guideline no. 422 adopted on 28 July 2015.
The test item was administered orally by gavage to parental animals at 5 mL/kg body weight.
The dosages, selected in consultation with the Sponsor, were 100, 300 and 1000 mg/kg/day.
The oral route was selected as it is a possible route of exposure of the test item in man.
Positive control:
no
Observations and examinations performed and frequency:
In vivo observations
Full records were maintained for all measurements and observations.
Mortality
Throughout the study, all animals were checked early in each working day and again in the afternoon. At weekends and Public Holidays a similar procedure was followed except that the final check was carried out at approximately mid-day.

Clinical signs
Once before commencement of treatment and at least once daily during the study, each animal was observed and any clinical signs were recorded. Observations were performed at the same time interval each day, the interval was selected taking into consideration the presence of post-dose reactions.

Clinical Observations (Functional Observation Battery Tests)
Once before commencement of treatment and at least once aweek from the start of treatment until termination, each animal was given a detailed clinical examination. Each animal was removed from the home cage and observed in an open arena. The tests included observation of changes in gait and posture, reactivity to handling, presence of clonic or tonic movements, stereotypies or bizarre behaviour and effects on the autonomic nervous system (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern). Changes in fur, skin, eyes, mucous membranes, occurrences of secretions and excretions were also recorded.

Grip strength and sensory reactivity to stimuli
Once during the study, towards the end of treatment, 5 males and 5 females were randomly selected from each group for evaluation of sensory reactivity to stimuli of different modalities (e.g. auditory, visual and proprioceptive stimuli) and for assessment of grip strength.
Measurements were performed using a computer generated random order. For the females, the tests were performed on Day 12 post partum.

Motor activity assessment (MA)
Once during the study, towards the end of treatment, 5 males and 5 females were randomly selected from each group and the motor activity was measured (for approximately 5 minutes) by an automated activity recording device. Measurements were performed using a computer generated random order. For the females, the tests were performed on Day 12 post partum.

Body weight
Males were weighed weekly from allocation to termination. Females were weighed weekly from allocation to positive identification of mating and on Days 0, 7, 14 and 20 post coitum.
Dams were also weighed on Days 1, 4, 7 and 13 post partum.

Food consumption
The weight of food consumed by each cage ofmales and females was recorded weekly during the pre-mating period starting from allocation. Individual food consumption for the females was measured on Days 7, 14 and 20 post coitum starting from Day 0 post coitum and on Days 4, 7 and 13 post partum starting from Day 1 post partum.

Vaginal smears
Females were evaluated for oestrous cyclicity 7 days before allocation to groups and animals that exhibited irregular cycle were not included in the study. Before the start of treatment a total of 5 females were replaced with spare animals from the batch initially ordered for the study.
Vaginal smears were taken daily in the morning starting from two weeks before pairing throughout the mating period 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)

Mating
Matings were monogamous (one male to one female). Vaginal smears were taken from the day after the start of pairing until positive identification of copulation (spermidentification, vaginal plug in situ or copulation plug found on the cage tray). The female was paired with the same male until positive identification occurred or 14 days had elapsed.

Parturition check and duration of gestation
A parturition check was performed from Day 20 to Day 25 post coitum. Gestation length was calculated as the time between the day of successful mating (Day 0 post coitum) and the day of birth, when the parturition was defined complete (Day 0 post partum).
Female nos. X0390007 (Group 1), X0390031 (Group 2) and X0390059 (Group 3), which did not give birth after 25 days of post coitum period, were sacrificed on Day 27. These animals were found not pregnant at necropsy. In addition, female no. X0390021 (Group 2) did not mate during the 14 days of mating period. The female was sacrificed thereafter and was found not pregnant.
One female of Group 2 (no. X0390023) lost its litter on Day 0 post partum and was sacrificed on Day 1 post partum.

Pups identification, weight and observation
As soon as possible after parturition was considered complete, all pups (live and dead) were counted, sexed and only live pups were identified. Live pups were individually weighed on Days 1, 4, 7 and 13 post partum. Pups killed or dying during the lactation period were weighed before the despatch to necropsy. Observations were performed once daily for all litters.

Culling
On Day 4 post partum, all pups were weighed and litters in excess of 8 offspring were culled to 8 (4 males and 4 females, where possible) by a random selection. Litters were not culled to less than 8 pups.

Pups selection for blood collection at necropsy – Day 4 post partum On Day 4 post partum, blood samples were collected from two of the non selected pups (males or females). Pooled samples from different pups were applied, if necessary. For litters
with less than 8 pups, two of the pups of the litter were sacrificed for serum assessments and preference was given to remove two female pups in order to retain more male pups for nipple retention on Day 13 post partum. However, retained female pups in each litter were not below 2.
Since no differences in hormone determination were seen in pups on Day 13 post partum, samples collected from pups on Day 4 post partum will not be analysed and will be destroyed after the finalisation of the report (after Sponsor communication).

Anogenital distance (AGD)
The AGD of each pup was measured on Day 1 post partum. The AGD was normalized to the cube root of pup body weight recorded on Day 1 post partum.

Clinical pathology investigations
As a part of the sacrificial procedure, samples of blood were withdrawn under isofluorane anaesthesia from the abdominal vena cava from 5 males and 5 females (females with viable litters) randomly selected from each group, under condition of food deprivation.
In addition, as part of the necropsy procedure, blood samples were taken from the abdominal vena cava of all parental males and females rats from each group, as mentioned above, under isofluorane anaesthesia, to performthe determination of serum T3, T4 and TSH levels

The blood samples collected were divided into tubes as follows:
– EDTA anticoagulant for haematological investigations
– Heparin anticoagulant for biochemical tests
– Citrate anticoagulant for coagulation tests
The measurements performed on blood samples are listed below:

Haematology
– 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
These parameters were analysed by Siemens Advia 120.
Coagulation
– Prothrombin time
– Activated partial thromboplastin time
These parameters were analysed by Instrumentation Laboratory ACL 3000 PLUS.

Clinical chemistry
– Alkaline phosphatase
– Alanine aminotransferase
– Aspartate aminotransferase
– Gamma-glutamyltransferase
– Urea
– Creatinine
– Glucose
– Triglycerides
– Phosphorus
– Total bilirubin
– Total cholesterol
– Bile acids
– Total protein
– Albumin
– Globulin
– A/G Ratio
– Sodium
– Potassium
– Calcium
– Chloride
These parameters were analysed by Siemens Advia 1200.

Urinalysis (Only 5 males randomly selected)
At the same time interval as the clinical pathology investigations, individual overnight urine
samples were also collected from the same males under the same conditions. Before starting
urine collection, water bottles were removed from each cage and each animal received
approximately 10 mL/kg of drinking water by gavage, in order to obtain urine samples
suitable for analysis.
The measurements performed on urine samples are listed below:
– Appearance
– Volume
– Specific gravity
– pH
– Protein
– Glucose
– Ketones
– Bilirubin
– Urobilinogen
– Blood
These parameters were analysed byMenarini AUTION MAX 4280/AUTION ELEVEN AE 4020.
The sediments, obtained from centrifugation at approximately 3000 rpm for 10 minutes, were
examined microscopically for:
– Epithelial cells
– Leucocytes
– Erythrocytes
– Crystals
– Spermatozoa and precursors
– Other abnormal components



Sacrifice and pathology:
Terminal studies
Euthanasia
Parental animals killed for humane reasons were killed under carbon dioxide asphyxiation.
Parental animals that had completed the scheduled test period and selected for blood collection were killed by exsanguination under isofluorane anaesthesia.
Pups
Pups that had completed the scheduled test period (Day 4 post partum orDay 14 post partum) were euthanised by intraperitoneal injection of Sodium Thiopenthal. Pups selected for blood collection for hormone determination were killed on the day of blood sampling.

Parental males
The males were killed after the mating, after 48 days of dosing.
Parental females
The females with live pups were killed on Day 13 post partum. The females which did not give birth 25 days after positive identification of mating were killed shortly after.

Necropsy
The clinical history of adult animals was studied and a detailed post mortem examination was conducted (including examination of the external surface and orifices). Changes were noted, the requisite organs weighed and the required tissue samples preserved in fixative and processed for histopathological examination.

Females
All females were examined also for the following:
– external and internal abnormalities;
– number of visible implantation sites (pregnant animals);
– number of corpora lutea (pregnant animals).
Uteri of females with no visible implantationswere immersed in a 10% solution of ammonium sulphide to reveal evidence of implantation

Pups
All pups found dead in the cage were examined for external and internal abnormalities.
All culled pups sacrificed on Days 4 and 13 post partum were subjected to an external examination.

Sex was determined by internal gonads inspection.
Nipple retention on Day 13 post partum
No nipples were found in pups to be retained at Day 13 post partum. Data were not tabulated, but will be archived with all raw data.

Organ weights
Parental animals
From all animals completing the scheduled test period, the organs indicated in Annex 1 of Study Protocol (section 4.5.6) were dissected free of fat and weighed. The ratios of organ weight to body weight were calculated for each animal.

Pups on Day 13 post partum
Thyroid was weighed from one male and one female pup from each litter (the same pups in which blood collection was performed on Day 13 post partum) and preserved. The thyroid weight was determined after fixation in 10% neutral buffered formalin.

4.5.4 Tissues fixed and preserved
Samples of all the tissues listed in Annex 1 of Study Protocol (section 4.5.6) were fixed and preserved in 10% neutral buffered formalin (except eyes, optic nerves and Harderian glands, testes and epididymides which were fixed in modified Davidson’s fluid and preserved in 70% ethyl alcohol).

4.5.5 Histopathological examination
The tissues required for histopathological examination are listed Annex 1 of Study Protocol (section 4.5.6). After dehydration and embedding in paraffin wax, sections of the tissues were cut at 5 micrometer thickness and stained with haematoxylin and eosin. In addition, the testes and epididymides were cut at 2-3 micrometer thickness and stained with Periodic Acid Schiff (PAS). Themorphological evaluation of the seminiferous epithelium (staging of spermatogenic cycle) was also performed.
Other examinations:
Blood collection for thyroid hormone determination (T3, T4 and TSH)
Blood collection from pups on Days 4 and 13 post partum Blood samples were withdrawn under light ether anaesthesia from the heart (by intracardiac puncture). Pooled samples from different pups per sex were performed when necessary on
Day 4 post partum. On Day 4 post partum, approximately 0.2 mL of blood samples was taken from two pups: 1 pup/sex/ litter. On Day 13 post partum, approximately 0.5 mL of blood samples was taken from two pups: 1 pup/sex/litter.
Parental animals
As a part of the necropsy procedure, approximately 0.8 mL of blood samples was taken from all parental males and females, from the abdominal vena cava.
All samples were transferred into tubes containing no anticoagulant and centrifuged at room temperature. The serum obtained was divided in several aliquots for analysis as indicated in section 4.4.5. The aliquots obtained were stored at -80°C until nalysis.
In the first instance, the determination was restricted as detailed below):
1. All parental males from all groups
2. Samples collected in pups on Day 14 post partum
Since no treatment related effects were seen in the determination performed in parental males and in pups on Day 13 post partum, samples obtained from females and from pups on Day 4 post partum will not be analysed and will be destroyed after the finalisation of the report (after Sponsor communication).

Bioanalysis - Thyroid hormone determination (T3, T4 and TSH)
Samples from all adult males and from 1 pup/sex/litter sacrificed on Day 13 post partum were assayed to determine the serum levels of Total triiodothyronine (total T3), Total thyroxine (total T4) and thyroid stimulating hormone (TSH) by a multiplex assay, using LuminexMagpix system and the MILLIPLEX MAP Rat ThyroidMagnetic Bead Panel kit (MerkMillipore, cat. no. RTHYMAG-30K).
When the serum levels for Total triiodothyronine (total T3) were BLOQ (below the limit of quantification - 625 pg/mL), the values were not included in the mean calculation.
The results of these analyses are presented as individual data, mean and standard deviation.
Statistics:
Standard deviationswere 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. The non-parametric Kruskal-Wallis analysis of variance (non-continuous variables) 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.
Statistical analysis of histopathological findings was carried out by means of the non-parametric Kolmogorov-Smirnov test if ‘n’ was more than 5.
The criterion for statistical significance was p < 0.05. The mean values, standard deviations and statistical analysis were calculated from actual values in the computer without rounding off.
Clinical signs:
no effects observed
Description (incidence and severity):
Salivation was noted for a limited number of days in males and females receiving 600 mg/kg/day before pairing.
Thereafter no clinical signs were noted both in control and treated animals.
Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item.
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No treatment related effects were seen in animals of both sexes compared to the control group, throughout the study.
Haematological findings:
no effects observed
Description (incidence and severity):
The slight changes noted were not considered treatment related.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No adverse findings were observed in urinalysis assessed in males only.
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Terminal body weight was unaffected by treatment in both sexes. Some changes noted in absolute and relative organ weights in treated groups were dose unrelated and/or not accompanied by histopathological findings and therefore they were considered unrelated to treatment.
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no compound-related effects.
Neuropathological findings:
no effects observed
Description (incidence and severity):
No relevant differenceswere noted in all parameters investigated between control and treated groups at the end of the treatment period.
Other effects:
no effects observed
Description (incidence and severity):
Thyroid hormone determination
Thyroid hormones in adult males.
No differences in hormone levels were described between the control and the treated parental males.

Thyroid hormones in male and female pups performed on Day 13 post partum.
Hormone levels in treated pups were similar to controls.
Key result
Dose descriptor:
NOAEL
Effect level:
600 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effect observed
Critical effects observed:
no

TISSUE PROCESSING

Organs / Tissues

Weight

Fixation Preservation

Microscopic Examination

  Abnormalities

 

ü

ü

  Adrenal glands

ü

ü

ü

  Bone marrow (from sternum)

 

ü

ü

Brain(cerebrum, cerebellum, medulla/pons)

ü

ü

ü

  Clitoral gland

 

ü

ü

  Caecum

 

ü

ü

  

 

ü

ü

  Duodenum

 

ü

ü

  Epididymides

ü

ü

ü

  Heart

ü

ü

ü

  Ileum

 

ü

ü

  Jejunum (including Peyer’s patches)

 

ü

ü

  Kidneys

ü

ü

ü

  Liver

ü

ü

ü

  Lungs (including mainstem bronchi)

 

ü

ü

  Lymph nodes - cervical

 

ü

ü

  Lymph nodes - mesenteric

 

ü

ü

  Nasal cavity

 

ü

*

  Oesophagus

 

ü

*

  Ovaries with oviducts

ü

ü

ü

 

  Parathyroid glandsa

ü

ü

ü

  Pituitary gland

 

ü

ü

  Penis

 

ü

ü

  Prostate gland

ü

ü

ü

  Rectum

 

ü

ü

  Sciatic nerve

 

ü

ü

  Seminal vesicles with coagulating glands

ü

ü

ü

  Spinal column

 

ü

*

  Spinal cord (cervical, thoracic, lumbar)

 

ü

ü

  Spleen

ü

ü

ü

  Stomach (forestomach and glandular)

 

ü

ü

  Testes

ü

ü

ü

  Thymus (where present)

ü

ü

ü

  Thyroid

ü

ü

ü

  Trachea

 

ü

ü

  Urinary bladder

 

ü

ü

  Uterus - cervix

ü

ü

ü

  Vagina

 

ü

ü

a= weighed and preserved with thyroid

*: To be examined if indicated by signs of toxicity or target organ involvement.

Conclusions:
Based on the results of the present study, the NOAEL (No Observed Adverse Effect Level) for general toxicity and for reproductive and developmental toxicity was considered to be 600 mg/kg/day both for males and females.
Executive summary:

In a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test according to OECD Guideline 422 (28 July 2015) 4-{Methacryloyloxy)benzophenone was administered to 10 Hsd: Sprague Dawley SD rats/sex/dose orally by gavage at dose levels of 0 (control), 70, 200 and 600 mg/kg bw/d. The treatment schedule included 2 weeks before pairing, during pairing, post coitum and post partum periods up to day 3 post partum. Animals were administered for approximately 5 and 8 weeks for males and females, respectively. 

 

The purpose of this study was to generate information concerning toxic effects on SpragueDawley rats of both sexes after repeated oral dosing with 4-(Methacryloyloxy)benzophenone,as well as any effects of the test item on male and female reproductive performance, such as gonadal function, conception, parturition and lactation of offspring. The dose levels used during the study were 70, 200 and 600 mg/kg/day.

Males

One control male was sacrificed on Day 14 of the study for poor health condition. On thebasis of the macroscopic and microscopic findings, the factor contributory to the death wasmainly attributed to a misdosing.

Males were treated for 2 weeks prior to pairing and during pairing with females until the daybefore necropsy, for a total of 48 days.

During thein-lifephase, mortality check, clinical signs (including neurotoxicity assessment,motor activity and sensory reaction tostimuli), body weight, body weight gain, food consumptionand mating performance were evaluated.

Clinical pathology investigations (haematology, clinical chemistry, urinalysis), organ weights,macroscopic observations and histopathological examination were also performed. In

addition, thyroid hormone levels were determined in all adult males. The histopathologicalexamination was performed only on control and high dose groups (5 animals/sex/grouprandomly selected). It included identification of the stages of the spermatogenic cycle in the

same five control and high dose males.

No relevant signs of toxicological significance were observed in treated males, apart from atransient salivation, observed for a few days in the high dose animals.

Body weights and food consumption did not show any treatment related effects. Fertilityindex and copulatory index were unaffected by treatment. No effects considered treatmentrelated were observed in haematology or coagulation, clinical chemistry and urinalysis.

Hormone analysis did not showany relation to treatment. Some fluctuations in organweights,absolute and/or relative, were noted, but in the absence of histopathological correlated

findings, these changes were considered of no toxicological significance.

Females

Females were treated for 2weeks prior to pairing, during pairing and throughout the gestationand lactation periods until Day 12post partum.

During the in-life phase, mortality check, clinical signs (including neurotoxicity assessment,motor activity and sensory reaction tostimuli), body weight, body weight gain, food

consumption and mating performance were evaluated. Clinical pathology investigations(haematology and clinical chemistry) and thyroid hormone determination in pups on Day 13post partumwere also evaluated. The histopathological examination was carried out in five

females of control and high dose groups, selected randomly. Clinical signs of pups, as well asnecropsy examination of decedent pups or pups sacrificed on Days 4 and 13post partum(including thyroid weight) were recorded. Litter data, sex ratios and gestation length were

recorded.

No relevant signs of toxicological significance were observed. As noted for males, salivation was recorded occasionally in treated females receiving 600 mg/kg/day. Body weights andfood consumption did not show changes of toxicological significance.

Concerning the reproductive parameters, no relevant differences were found in terms ofmating performance including the pre-coital interval (number of days paired to spermpositive day) and the copulatory evidence (the positive identification of mating i.e. the

presence of spermand/or copulation plugin situor in the cage), as well as fertility index.

The slight changes noted at clinical chemistry investigation (haematology, clinical chemistryand coagulation) were not considered treatment related. Some fluctuations in organ weights,absolute and/or relative, were noted but in the absence of histopathological correlated

findings, these changes were considered of no toxicological significance.

Conclusion

Based on the results of the present study, the NOAEL (No Observed Adverse Effect Level) for general toxicity and for reproductive and developmental toxicity was considered to be 600 mg/kg/day both for males and females.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
600 mg/kg bw/day
Study duration:
subacute
Species:
rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification