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Administrative data

Description of key information

Both 28-day and 90-day repeated toxicity studies were performed with 2 moles ethoxylated bisphenol A dimethacrylate by oral gavage in male and female rats. No adverse effects were observed up to the maximal dose of 1000 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 May 2017 - 12 September 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
21 September 1998
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: breeder: Janvier, le Genest-Saint-Isle, France
- Age: on the first day of treatment, the animals were approximately 6 weeks old
- Mean body weight: on the first day of treatment, the males had a mean body weight of 241 g (range: 198 g to 268 g) and the females had a mean body weight of 186 g (range: 163 g to 199 g)
- Fasting period before study: no
- Housing: the animals were housed in twos, by sex and group, in polycarbonate cages with stainless steel lids (Tecniplast 2000P, 2065 cm²) containing autoclaved sawdust
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: for a period of 11 days before the beginning of the treatment period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 8 to 15 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 12 June 2017 to 12 September 2017.
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
PREPARATION OF DOSING FORMULATIONS: Emulsion in the vehicle
- Concentration in vehicle: 20, 60 and 200 mg/mL
- Amount of vehicle: 5 mL/kg/day.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Type of method: High Performance Liquid Chromatography with UV detection (HPLC/UV)
Test item concentrations: remained within an acceptable range of -4.8% to +11.7% when compared to the nominal values ( ± 15% of the nominal concentrations)
Homogeneity: the dose formulations containing the test item and prepared at 2 mg/mL and 200 mg/mL in PEG 400 were found to be homogeneous after preparation. Dose formulations ranging from 2 mg/mL to 200 mg/mL are therefore considered to be suitable for routine administration in GLP Toxicological studies, and have to be maintained under magnetic agitation after preparation and during the administration procedure to animals in follow-up studies.
Stability: not assessed, dose formulation prepared daily
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for dose selection:
The dose levels were selected by the Sponsor Representative based on the results of a previous OECD 422 study performed in Wistar Han rats.
In this study, rats received the test item daily by gavage at dose levels of 62, 250 or 1000 mg/kg/day for 29 days (males) or 43 to 46 days (females).
There were no relevant changes in any of the parental parameters (i.e. clinical signs, functional observation battery and motor activity, body weight, body weight change, food consumption, clinical laboratory investigations and histopathology examination).
Thus, based on these available data, the dose levels selected for the present study were: 100, 300 and 1000 mg/kg/day.

- Rationale for animal assignment: computerized stratification procedure.
Positive control:
no (not required)
Observations and examinations performed and frequency:
MORBIDITY/MORTALITY:
- Time schedule: each animal was checked for mortality and morbidity once a day during the acclimation period and at least twice a day during the treatment period, including weekends and public holidays.

CLINICAL SIGNS:
- Time schedule: each animal was observed once a day, at approximately the same time on the days of treatment, for the recording of clinical signs.

DETAILED CLINICAL OBSERVATIONS:
- Time schedule: detailed clinical examinations were performed on all the animals once before the beginning of the treatment period and then once a week until the end of the study.

BODY WEIGHT:
- Time schedule: the body weight of each animal was recorded once before the beginning of the treatment period, on the first day of treatment and at least once a week until the end of the study.

FOOD CONSUMPTION:
- Time schedule: the quantity of food consumed by the animals in each cage was recorded at least once a week, over a 7-day period, during the study.

OPHTHALMOSCOPIC EXAMINATION:
- Time schedule:
Ophthalmological examinations were performed:
- on all animals, before the beginning of the treatment period,
- on control and high-dose animals on one occasion at the end of the treatment period (i.e. in Week 13).

HAEMATOLOGY:
- Time schedule for collection of blood: the animals were deprived of food for an overnight period of at least 14 hours.
- Time schedule for collection of peripheral blood: at the end of the treatment period
- Time schedule for collection of bone marrow: at the end of the treatment period
- Anaesthetic used for blood collection: Yes, light isoflurane anesthesia
- Animals fasted: Yes
- How many animals: all
- Parameters checked in table [1] were examined.

CLINICAL CHEMISTRY:
- Time schedule for collection of blood: at the end of the treatment period
- Animals fasted: Yes
- How many animals: all
- Parameters checked in table [2] were examined.

THYROID HORMONES:
- Time schedule: An additional blood sample (approximately 0.5 mL) was taken from each animal euthanized at the end of the treatment period into tubes containing K3-EDTA as anticoagulant.
- Animals fasted: Yes
Sacrifice and pathology:
ORGAN WEIGHTS: see table 3.
The body weight of each animal was recorded before euthanasia at the end of the treatment period. The organs specified in the Tissue Procedure Table were weighed wet as soon as possible after dissection.
The ratio of organ weight to body weight (recorded immediately before euthanasia) was calculated.

GROSS PATHOLOGY:
A complete macroscopic post-mortem examination was performed on all animals.

PRESERVATION OF TISSUES:
For all study animals, the tissues specified in the Tissue Procedures Table were preserved in 10% buffered formalin (except for the eyes and optic nerves and Harderian glands, and the testes and epididymides which were fixed in Modified Davidson's Fixative).
Two bone marrow smears for potential determination of the bone marrow differential cell count (see § Bone marrow) were prepared from the femur of each animal euthanized on completion of the treatment period.

PREPARATION OF HISTOLOGICAL SLIDES:
All tissues required for microscopic examination were trimmed according to the RITA guidelines, when applicable, embedded in paraffin wax, sectioned at a thickness of approximately four microns and stained with hematoxylin-eosin.
The tissue processing was performed at CiToxLAB France.

HISTOPATHOLOGY:
A microscopic examination was performed on all tissues listed in the Tissue Procedure Table:
- for the control and high-dose animals (groups 1 and 4) euthanized at the end of the treatment period and for the group 2 female that was euthanized prematurely,
- for all macroscopic lesions from all low- and intermediate-dose animals (groups 2 and 3) euthanized on completion of the treatment period.
Based upon the organ weights and microscopic results of the high-dose group, liver and spleen of females from the low- and intermediate-dose groups (groups 2 and 3) were examined.
Statistics:
Citox software was used to perform the statistical analyses of body weight, food consumption, hematology, and blood biochemistry.
PathData software was used to perform the statistical analysis of organ weight data.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
See Table 1.
Test item-related clinical signs consisted in hunched posture in 1/10 males given 1000 mg/kg/day for 2 days, half-closed eyes in 1/10 males given 300 mg/kg/day and 1/10 males given 1000 mg/kg/day up to for 16 days, and/or chromorhynorrhea in one to two males and/or females at all dose-levels up to for 2 days. These findings were considered as non-adverse as they were minimal and/or transiently noted.

Ptyalism was transiently observed in control females and in all test item-treated groups in a dose-related manner, from the second week of treatment in group 4 animals. This non-adverse finding is commonly reported when formulations are administered by gavage.
Slight soft feces were observed in all control and test item-treated animals from Week 10. This non-adverse finding is commonly reported when PEG 400 is administered, resulting from its effects on the intestinal transit.

The other clinical signs recorded during the study (thinning of hair, alopecia, scabs, abnormal growth of teeth, reflux at administration, chromodacryorrhea and/or exophthalmos), were of isolated occurrence, observed both in control and test item-treated animals, observed with no dose-relationship, commonly observed when a test item is administered by gavage, and/or common in this strain of rats under laboratory conditions. They were therefore considered to be unrelated to the test item.
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No test item-related deaths occurred during the study.

One female given 100 mg/kg/day was euthanized prematurely for humane reason on Day 10 (Week 2). Prior to sacrifice (from Day 9), this female showed signs of poor clinical condition (i.e. thin appearance with a slight body weight gain of 4 g over the first week of treatment, hunched posture, abdominal and loud breathing, piloerection and half-closed eyes). Ptyalism was also noted on Day 9.
At necropsy, this female had no gross findings. The cause of moribundity could not be established but this premature sacrifice was considered to be probably unrelated to the test item administration in view of the isolated occurrence at the low dose level. This female had minimal centrilobular hepatocellular hypertrophy that was considered to be related to the test item.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
See table 2.
At 1000 mg/kg/day in males, when compared with controls, lower mean body weight gain was recorded during the Day 1-36 interval (-8%), mainly due to a statistically significant, lower mean body weight gain over the second week of treatment (-18%, p<0.05). This was followed by a statistically significant, lower mean body weight gain (-25% vs. controls) until Day 64, and then by a mean body weight gain similar to controls until the end of the study. These transient differences, which were attributed to the test item, did not affect the final mean body weight.

At 100 and 300 mg/kg/day in males, when compared with controls, the same tendency was noted during the Day 36-64 interval, but to a lesser extent (-19% and -15%, respectively) and not dose-related. Although they were not dose-related, a test item relationship could not be excluded.

In females, no relevant effects were noted on body weight or body weight gain.
When compared with controls, slight but statistically significant, lower mean body weight gain was recorded in the mid part of the treatment period at 100 mg/kg/day. This transient variation was considered to be fortuitous as it was not dose-related.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
See table 3.
At 1000 mg/kg/day in males, when compared with controls, slightly lower mean food consumption was recorded during the Day 43-71 interval (down to -14%, p<0.05). This mainly correlated with the minimally lower mean body weight gain recorded over the Day 36-64 interval. These differences, which were attributed to the test item, were transient and of slight to moderate magnitude.

There were no test item-related effects at 100 and 300 mg/kg/day in males, or in any test item-treated females.
Food efficiency:
not examined
Ophthalmological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No test item-related findings were observed at the end of the treatment period.

Chorioretinopathy was observed in control and test item-treated animals with similar incidences.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
See table 4.
At 1000 mg/kg/day in males and when compared with controls, statistically significant, lower mean white blood cell count (-23%) was noted. This correlated with a statistically significant, lower mean lymphocyte and basophil counts (-28% and -29% vs. controls, respectively).
In both sexes and when compared with controls, statistically significant lower reticulocyte counts (-24%) were noted. In females, they were accompanied by statistically significant, lower mean hemoglobin concentration (-4% vs. controls) and mean packed cell volume (-4% vs. controls). These variations correlated with the minimal decreased severity of extramedullary splenic hematopoiesis seen in females.
In view of the slight magnitude and/or as mean values were within or close to the range of the HCD, the findings detailed above were not considered as adverse.

At 100 and 300 mg/kg/day in females and when compared with controls, statistically significant lower reticulocyte count (-19% and -24%, respectively) were recorded. These variations correlated also with the minimal decreased severity of extramedullary splenic hematopoiesis noted in females treated at 300 mg/kg/day. As these differences were of minor magnitude, they were not considered as adverse.

The other differences between control and test item-treated animals, namely in white blood cell, lymphocyte count (males given 100 mg/kg/day), basophil count (males given 100 mg/kg/day), and prothrombin time (males given 1000 mg/kg/day) were considered to be of no toxicological importance as they were of low magnitude and/or noted with no dose-relationship.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
See table 5.
When compared with mean control values, statistically significant blood biochemistry changes were noted:
- lower mean potassium level in females at 300 mg/kg/day (-8%) and in males and females at 1000 mg/kg/day (-7% and -8%, respectively),
- higher mean protein level in females at 100, 300 and 1000 mg/kg/day (+9%, +6% and +7%, respectively),
- lower mean albumin level in males at 300 and 1000 mg/kg/day (-5%),
- lower mean albumin to globulin ratio in females at 1000 mg/kg/day (-9%),
- higher mean cholesterol level in males and females at 100 mg/kg/day (+33% and +56%, respectively), at 300 mg/kg/day (+50% and +60%, respectively) and at 1000 mg/kg/day (+42% and +90%, respectively),
- higher mean triglyceride level in males at 100, 300 and 1000 mg/kg/day (x 1.8, x 2.6 and x 2.1, respectively),
- higher mean urea level in females at 1000 mg/kg/day (+11%),
- lower mean alkaline phosphatase activity in males at 1000 mg/kg/day (-29%).

Variations in potassium, albumin, protein and urea levels and in the albumin to globulin ratio were considered to be of minor toxicological importance as they were poorly dose-related, of minimal magnitude and/or within the range of the HCD.
Increases in cholesterol levels in all test item-treated males (poorly dose-related) and females and in triglyceride levels in all test item treated males (poorly dose-related) were considered as "minimal to moderate" in view of the magnitude of the differences from controls and as most of these findings were above the upper limit of the HCD. As no histopathological findings correlated with these changes, they were considered as non-adverse.

The other statistically significant differences between control and test item-treated animals, namely in sodium (males at 100 mg/kg/day), calcium (females at 100 and 1000 mg/kg/day), albumin (females at 100 mg/kg/day), albumin to globulin ratio (males at 300 mg/kg/day), total bilirubin (females at 100 and 300 mg/kg/day) levels, and alkaline phosphatase (males at 1000 mg/kg/day and females at 300 mg/kg/day), aspartate aminotransferase (males at 300 mg/kg/day) activity were considered to be incidental, of no toxicological importance and/or not test item-related as they were marginal, without a dose relationship and without any correlating microscopic findings.
Endocrine findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
See table 6.
Slightly higher absolute and relative-to-body liver weights were noted in females treated at 100, 300 or 1000 mg/kg/day (up to +20% in females treated at 300 or 1000 mg/kg/day in relative-to-body weights; p<0.01). These differences were considered to be related to the test item administration and correlated with microscopic hepatocellular hypertrophy.

Slightly lower absolute and relative-to-body spleen weights were noted in females treated at 1000 mg/kg/day (up to -17% in absolute weights; p<0.05). These differences were considered to be related to the test item administration and correlated with microscopic lower severity of extramedullary hematopoiesis.

All other weight differences were minimal, did not correlate with macroscopic or microscopic findings and were considered not to be test item treatment-related. They included the lower adrenal gland weights in males treated at 300 or 1000 mg/kg/day with no microscopic correlates, the lower absolute and relative to-body thymus weights in males treated at 1000 mg/kg/day with no microscopic correlates, the higher relative-to-body weights of kidneys in males treated at 1000 mg/kg/day (probably due to the contribution of lower body weights), and the lower uterus absolute and relative-to-body weights in females treated at 100, 300 or 1000 mg/kg/day related to higher incidence of proestrus/estrus in controls than in test item-treated females.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Unscheduled death
One female treated at 100 mg/kg/day and sacrificed on Day 10 had no gross findings. The cause of moribundity of this female could not be established.

Terminal sacrifice
There were no test item-related findings.
The few macroscopic changes were seen with similar incidence in both control and test item-treated groups and were consistent with spontaneous findings commonly described in the rats of these strain and age.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
See table 7.
Unscheduled death
One female sacrificed on Day 10 had no noteworthy microscopic findings except minimal centrilobular hepatocellular hypertrophy that was considered to be related to the test item administration.

Terminal sacrifice
Minimal to slight, dose-related hepatocellular hypertrophy was recorded in the liver from females treated at 100, 300 or 1000 mg/kg/day and correlated with the increased liver weights.
In the absence of degenerative process and in view of the low magnitude of this change, it was considered to be non-adverse.
Minimally decreased severity of extramedullary splenic hematopoiesis was recorded in females treated at 300 or 1000 mg/kg/day and correlated with the lower spleen weights at 1000 mg/kg/day.
In view of the very low magnitude of this variation, it was considered to be non-adverse.

The other findings were considered not to be related to test item administration since there were seen with similar incidence and severity in controls and/or were consistent with the spontaneous findings described in the literature. This included the increased severity/incidence of plasma cell numbers in the mandibular lymph node of males treated at 1000 mg/kg/day and the mineralization seen in the pelvis or tubules of kidneys from occasional females treated at 1000 mg/kg/day (incidence up to 2/10) that are commonly seen in the rats of these strain and age.
Histopathological findings: neoplastic:
not examined
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Critical effects observed:
no

Table 1: Clinical signs

 

Sex

Male

Female

Dose level (mg/kg/day)

0

100

300

1000

0

100

300

1000

Hunched posture

-

-

-

1

-

-

-

-

Half-closed eyes

-

-

1

1

-

-

-

-

Chromorhynorrhea

-

1

1

2

-

1

-

1

Total affected animals

0/10

1/10

2/10

3/10

0/10

1/9

0/10

1/10

Ptyalism

-

3

8

9

2

5

9

10

Soft feces

10

10

10

10

10

9

10

10

Total affected animals

10/10

10/10

10/10

10/10

10/10

9/9

10/10

10/10

-: no clinical signs.

Table 2: Body weight

 

 

Sex

Male

Female

Dose level (mg/kg/day)

0

100

300

1000

0

100

300

1000

 

Mean BW gain Days 1/36

+221

+224

+225

+203

+93

+92

+88

+92

 

Mean BW gain Days 36/64

+75

+61

+64

+56*

+28

+21*

+25

+28

 

Mean BW gain Days 64/91

+20

+25

+27

+22

+9

+9

+6

+7

 

Mean BW gain Days 1/91

+316

+310

+315

+282

+130

+122

+118

+127

 

% from controls

-

-2

0

-11

-

-6

-9

-2

 

Mean body weight on Day 1

244

243

240

239

190

184

186

186

 

Mean body weight on Day 64

540

528

529

498

310

297

298

306

 

% from controls

-

-2

-2

-8

-

-4

-4

-1

 

Mean body weight on Day 91

559

553

555

520

319

305

304

313

 

% from controls

-

-1

-1

-7

-

-4

-5

-2

 

Statistically significant from controls: *: p<0.05; -: not applicable.

Table 3: Food consumption

 

 

Sex

Male

Female

Dose level (mg/kg/day)

0

100

300

1000

0

100

300

1000

 

Days 43 to 50

34.7

32.8

33.2

31.4

23.1

22.8

21.7

23.4

 

Days 50 to 57

33.3

31.1

32.0

30.0

22.8

21.2

20.7

20.9

 

Days 57 to 64

32.4

30.9

31.1

28.0*

22.0

21.0

21.0

21.2

 

Days 64 to 71

33.0

31.5

31.7

29.2

21.4

21.1

20.8

21.1

 

Statistically significant from controls: *: p<0.05.

Table 4: Haematology

 

Sex

Male

Female

Dose level (mg/kg/day)

0

100

300

1000

0

100

300

1000

White blood cell count (G/L)

20.42

(5.51-25.13)

17.29

18.40

15.63**

11.48

10.69

10.28

10.43

Hemoglobin (g/dL)

16.1

15.9

15.7

15.9

15.3

(13.2-17.0)

15.2

14.8

14.7*

Packed Cell Volume (L/L)

0.49

0.49

0.48

0.48

0.45

(0.39-0.51)

0.45

0.44

0.43*

Lymphocytes (G/L)

16.98

(4.80-21.91)

14.13

15.12

12.25**

9.83

8.48

8.53

8.60

Reticulocytes (%)

2.64

(1.38-4.49)

2.19

2.28

2.02*

2.39

(0.95-7.12)

1.95**

1.92**

1.87**

Reticulocytes (T/L)

0.25

0.20

0.21

0.19*

0.21

0.17**

0.16**

0.16**

Statistically significant from controls: *: p<0.05; **: p<0.01.

( ): minimum-maximum values from Historical Control Data (HCD).

Bold: test item-related.

Table 5: Blood biochemistry

 

Sex

Male

Female

Dose level (mg/kg/day)

0

100

300

1000

0

100

300

1000

Potassium (mmol/L)

3.86
(3.19-5.49)

3.77

3.71

3.59**

3.79
(3.17-5.43)

3.62

3.48**

3.47**

Proteins (g/L)

64.7
(56.8-69.2)

63.3

63.6

62.8

61.8
(55.3-82.4)

67.1**

65.2*

65.9**

Albumin (g/L)

38
(35-42)

37

36*

36*

37
(33-55)

39*

38

38

A/G ratio

1.43
(1.25-1.82)

1.37

1.33*

1.38

1.47
(1.32-2.05)

1.42

1.38*

1.34**

Cholesterol (mmol/L)

2.06
(0.85-3.04)

2.75**

3.10**

2.92**

2.33
(1.22-3.19)

3.64**

3.72**

4.43**

Triglycerides (mmol/L)

0.77
 (0.24-1.28)

1.40**

1.97**

1.62**

0.63
(0.16-0.85)

0.72

0.59

0.69

Urea (mmol/L)

6.4
(3.3-7.6)

6.6

6.9

6.5

6.4
(4.1-7.0)

6.7

6.6

7.1*

Statistically significant from controls: *: p<0.05; **: p<0.01.

( ): minimum-maximum values from Historical Control Data (HCD).

A/G ratio: albumin to globulin ratio

Bold: test item-related.

Table 6: Organ weights

 

Sex

Male

Female

Group

2

3

4

2

3

4

Dose level (mg/kg/day)

100

300

1000

100

300

1000

Number of animals

10

10

10

9

10

10

- Final body weight

-1

0

-7

-5

-6

-3

- Liver  

. absolute

+6

+10

0

+14*

+13

+16*

. relative

+7

+10

+9

+19**

+20**

+20**

- Spleen  

. absolute

-2

-1

-13

-8

-16*

-17*

. relative

0

0

-5

-3

-11

-14*

Statistically significant from controls: *: p<0.05, **: p<0.01

The significance concerned the organ weights values and not the percentages.

Table 7: Microscopic findings

 

Sex

Male

Female

Group

1

2

3

4

1

2

3

4

Dose-level (mg/kg/day)

0

100

300

1000

0

100

300

1000

n

10

10

10

10

10

9

10

10

Liver; hepatocellular hypertrophy

 

 

 

 

 

 

 

 

. grade 1

-

na

na

-

-

3

6

5

. grade 2

-

na

na

-

-

-

-

5

Spleen; hematopoiesis

 

 

 

 

. grade 1

7

na

na

4

2

1

3

5

. grade 2

3

na

na

6

7

6

7

5

. grade 3

-

na

na

-

1

2

-

-

-: not recorded.

*: Out of five grades (minimal; slight; moderate; marked; severe).

na: not applicable.

Conclusions:
The test item was administered daily for 13 weeks by gavage to male and female Sprague-Dawley rats at dose levels of 100, 300 or 1000 mg/kg/day in PEG 400.
There were no test item-related or adverse effects on mortality, clinical signs, body weight, food consumption, clinical pathology and histopatholocical parameters.
Under the experimental conditions and according to the results of this study, the No Observed Adverse Effect Level (NOAEL) was set at 1000 mg/kg/day.
Executive summary:

The objective of this GLP study was to evaluate the potential toxicity of the test item following daily oral administration (gavage) to rats for 13 weeks.


Methods


Three groups of ten male and ten female Sprague-Dawley rats received the test item daily by the oral route (gavage) for 13 weeks. The test item was administered at dose levels of 100, 300 or 1000 mg/kg/day as an emulsion in the vehicle (PEG 400) under a constant dosage volume of 5 mL/kg/day. A control group of ten males and ten females per sex received the vehicle, alone, under the same experimental conditions.


The actual test item concentrations in the dose formulations prepared for use in Weeks 1, 5, 9 and 13 were determined using a High Performance Liquid Chromatography with UV detection method (HPLC/UV).


The animals were checked daily for mortality and morbidity and clinical signs. Detailed clinical observations were performed once a week. Body weight was recorded pre-test, on the first day of treatment and then once a week. Food consumption was recorded weekly.


Ophthalmological examinations were performed on all animals before the beginning of the study and on control and high-dose animals at the end of the treatment period (Week 13).


Hematology and blood biochemistry investigations were performed in all animals at the end of the treatment period (Week 13).


On completion of the treatment period, all animals were euthanized and a full macroscopic post-mortem examination was performed. Designated organs were weighed and selected tissues were preserved. A microscopic examination was performed on designated tissues from animals in the control and high-dose groups sacrificed at the end of the treatment period, from the low-dose female that was euthanized prematurely, and on all macroscopic lesions from all low- and intermediate-dose animals (100 and 300 mg/kg/day) euthanized on completion of the treatment period. The liver and spleen of females from the low- and intermediate-dose groups(groups 2 and 3) were also microscopically examined.


 


Results


The test item concentrations in the administered dose formulations remained within an acceptable range of -4.8% to +11.7% when compared to the nominal values (± 15% of the nominal concentrations) and no test item was detected in control formulation.


 


No test item-related deaths occurred during the study.


Ptyalism was observed with a dose-related incidence at all dose levels. Other non-adverse clinical signs consisted of hunched posture in 1/10 males given 1000 mg/kg/day,half-closed eyes in 1/10 males given 300 mg/kg/day and 1/10 males given 1000 mg/kg/day, and/or chromorhynorrhea in one to two males and/or females at all dose levels.


 


Lower body weight gain was recorded in males given 1000 mg/kg/day until Day 64. This was more marked over the second week of treatment (-18%) and then over the Day 36-64 interval (-25% vs. controls). It was accompanied by slightly lower food consumption (down to -14% vs. controls) during the Day 43-71 interval. At 100 and 300 mg/kg/day in males, a similar trend was noted during the Day 36-64 interval, but to a lesser extent and not dose-related (-19% and -15%, respectively). As these transient differences did not affect the final body weight, they were considered to be non-adverse.


There were no effects on body weight or body weight gain in females.


There were no significant effects on mean food consumption in males at 100 and 300 mg/kg/day and in all test item-treated females.


 


No test item-related ophthalmogical findings were observed at the end of the treatment period.


 


At hematology investigations, slightly lower white blood cell and lymphocyte counts, together with slightly shortened prothrombin time were noted in males given 1000 mg/kg/day. Slightly lower reticulocyte counts were noted in females from 100 mg/kg/day and in males given 1000 mg/kg/day. This was accompanied by slight decreases in hemoglobin concentration and packed cell volume in females given 1000 mg/kg/day. Red blood mass changes correlated with non-adverse decreased splenic hematopoiesis in females from 300 mg/kg/day. Hematological variations were considered to be of minor toxicological importance.


 


At blood biochemistry investigations, decreases in potassium level in females from 300 mg/kg/day and in males at 1000 mg/kg/day, decreases in albumin level in males from 300 mg/kg/day and in albumin/globulin ratio in females at 1000 mg/kg/day, increases in protein levels in females from 100 mg/kg/day and an increase in urea level in females at 1000 mg/kg/day were recorded. All these changes were considered to be of minor importance. From 100 mg/kg/day, males and females showed a non-adverse increase in cholesterol levels and males showed a non-adverse increase in triglyceride levels.


 


At histopathology,dose-related slightly higher liver weights were noted in females treated at 100, 300 and 1000 mg/kg/day. These differences correlated with microscopic non-adverse hepatocellular hypertrophy in test item-treated females at all dose levels.


Slightly lower spleen weights were noted in females treated at 1000 mg/kg/day and correlated with microscopic non-adverse lower severity of extramedullary hematopoiesis. This microscopic finding was also seen in females treated at 300 mg/kg/day.


There were no gross test item-related findings.


 


Conclusion


The test item was administered daily for 13 weeks by gavage to male and female Sprague-Dawley rats at dose levels of 100, 300 or 1000 mg/kg/day in PEG 400.


 


There were no test item-related or adverse effects on mortality, clinical signs, body weight, food consumption, clinical pathology and histopatholocical parameters.


 


Under the experimental conditions and according to the results of this study, the No Observed Adverse Effect Level (NOAEL) was set at 1000 mg/kg/day. The objective of this GLP study was to evaluate the potential toxicity of the test item following daily oral administration (gavage) to rats for 13 weeks.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
the 90-day study is a reliable study, performed according to an OECD guideline study.

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

Combined 28-day repeated dose toxicity study with the reproduction/developmental toxicity screening test (WIL 2013):


This study was performed in rats with 2 moles ethoxylated bisphenol A dimethacrylate by oral gavage (OECD guideline 422).


Based on the results of a 14-day dose range finding study, the dose levels for this study were 62, 250 and 1000 mg/kg. After acclimatization, four groups of ten male and ten female Wistar Han rats were exposed by oral gavage to the test substance. Males were exposed for 29 days, i.e. 2 weeks prior to mating, during mating, and up to termination. Females were exposed for 43-46 days, i.e. during 2 weeks prior to mating, during mating, during postcoitum, and during at least 4 days of lactation.


The following observations and examinations on parental animals were evaluated: mortality / viability, clinical signs (daily), functional observations and locomotor activity (end of treatment), body weight and food consumption (at least at weekly intervals), clinical pathology (end of treatment), macroscopy at termination, organ weights and histopathology on a selection of tissues.


 


Accuracy, homogeneity and stability of formulations were demonstrated by analyses.


No parental toxicity was observed up to the highest dose level tested (1000 mg/kg).


No toxicologically relevant changes were noted in any of the parental parameters investigated in this study (i.e. clinical appearance, functional observations, body weight, food consumption, clinical laboratory investigations, macroscopic examination, organ weights, and microscopic examination).


 


Treatment with 2 moles ethoxylated bisphenol A dimethacrylate by oral gavage in male and female Wistar Han rats at dose levels of 62, 250 and 1000 mg/kg revealed no parental toxicity up to 1000 mg/kg. Based on these results, a parental No Observed Adverse Effect Level (NOAEL) of at least 1000 mg/kg was derived.


 


 


90-day repeated dose toxicity study (CIToxLAB 2017):


The objective of this GLP study was to evaluate the potential toxicity of the test item following daily oral administration (gavage) to rats for 13 weeks.


Three groups of ten male and ten female Sprague-Dawley rats received the test item daily by the oral route (gavage) for 13 weeks. The test item was administered at dose levels of 100, 300 or 1000 mg/kg/day as an emulsion in the vehicle (PEG 400) under a constant dosage volume of 5 mL/kg/day. A control group of ten males and ten females per sex received the vehicle, alone, under the same experimental conditions.


 


No test item-related deaths occurred during the study.


Ptyalism was observed with a dose-related incidence at all dose levels. Other non-adverse clinical signs consisted of hunched posture in 1/10 males given 1000 mg/kg/day,half-closed eyes in 1/10 males given 300 mg/kg/day and 1/10 males given 1000 mg/kg/day, and/or chromorhynorrhea in one to two males and/or females at all dose levels.


Lower body weight gain was recorded in males given 1000 mg/kg/day until Day 64. This was more marked over the second week of treatment (-18%) and then over the Day 36-64 interval (-25%vs.controls). It was accompanied by slightly lower food consumption (down to -14%vs.controls) during the Day 43-71 interval. At 100 and 300 mg/kg/day in males, a similar trend was noted during the Day 36-64 interval, but to a lesser extent and not dose-related (-19% and -15%, respectively). As these transient differences did not affect the final body weight, they were considered to be non-adverse.


There were no effects on body weight or body weight gain in females.


There were no significant effects on mean food consumption in males at 100 and 300 mg/kg/day and in all test item-treated females.


No test item-related ophthalmogical findings were observed at the end of the treatment period.


At hematology investigations, slightly lower white blood cell and lymphocyte counts, together with slightly shortened prothrombin time were noted in males given 1000 mg/kg/day. Slightly lower reticulocyte counts were noted in females from 100 mg/kg/day and in males given 1000 mg/kg/day. This was accompanied by slight decreases in hemoglobin concentration and packed cell volume in females given 1000 mg/kg/day. Red blood mass changes correlated with non-adverse decreased splenic hematopoiesis in females from 300 mg/kg/day. Hematological variations were considered to be of minor toxicological importance.


At blood biochemistry investigations, decreases in potassium level in females from 300 mg/kg/day and in males at 1000 mg/kg/day, decreases in albumin level in males from 300 mg/kg/day and in albumin/globulin ratio in females at 1000 mg/kg/day, increases in protein levels in females from 100 mg/kg/day and an increase in urea level in females at 1000 mg/kg/day were recorded. All these changes were considered to be of minor importance. From 100 mg/kg/day, males and females showed a non-adverse increase in cholesterol levels and males showed a non-adverse increase in triglyceride levels.


At histopathology,dose-related slightly higher liver weights were noted in females treated at 100, 300 and 1000 mg/kg/day. These differences correlated with microscopic non-adverse hepatocellular hypertrophy in test item-treated females at all dose levels.


Slightly lower spleen weights were noted in females treated at 1000 mg/kg/day and correlated with microscopic non-adverse lower severity of extramedullary hematopoiesis. This microscopic finding was also seen in females treated at 300 mg/kg/day.


There were no gross test item-related findings.


 Under the experimental conditions and according to the results of this study, the No Observed Adverse Effect Level (NOAEL) was set at 1000 mg/kg/day.


 


 

Justification for classification or non-classification

Based on the 28 -day and 90 -day repeated toxicity studies, no adverse organ toxicity was showed in rats treated with 2 moles ethoxylated bisphenol A dimethacrylate. Therefore no classification is required for the repeated toxicity according to the Regulation EC n°1272/2008.