4,4'-Isopropylidenediphenol, ethoxylated

Endpoint summary

Description of key information

A combined short-term toxicity and screening for toxicity to reproduction study was performed on BPA-5EO according to OECD Testing Guideline 422. On the basis of the significant effects observed on body weight at 1000 mg/kg bw/day it is proposed a NOAEL of 500 mg/kg bw/day.

4,4-isopropylidenediphenol, ethoxylated (BPA-4EO) is being assessed for the purpose of the evaluation of the subchronic toxicity (90 days) via the oral route in rats. The study, according to the OECD TG 408, has been planned and commissioned to Covance Laboratories Limited, Woolley Road, Alconbury, Huntingdon, Cambridgeshire, PE28 4HS, UK
The testing is split into 3 phases
1) Validation of the analytical method

2) Dose range finding study Study

3) OECD 408: histopathology report was provided on 05 October 2021, the final study report is expected in March 2022 

The reason for such a delay in reference to ECHA 30 November 2020 deadline (Refs: TPE- D-2114489554-35-01/F), are explained in a separate letter, attached to this dossier.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
Source: Charles River Japan
Age at study initiation: 10 weeks
Weight at study initiation: Male 361 (334-396) g, Female 235 (216-260) g
Housing: 1/cage
Diet (e.g. ad libitum): MR(pellet), Nosan Corporation, Japan, ad libitum
Water (e.g. ad libitum): tap water(filtered and UV sterilized), ad libitum
Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
Temperature (°C): 22.1-25.0
Humidity (%): 46-60
Air changes (per hr): >10/hr
Photoperiod (hrs dark / hrs light): 12 hr/12 hr

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

VEHICLE
Concentration in vehicle: 0, 30, 120, 500, 1000 mg/kg/day
Amount of vehicle (if gavage): 5 mL/kg
Lot/batch No. (if required): BH17

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

42 days (administration period) followed by a 14 day recovery period.

Frequency of treatment:

Daily.

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

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

Dose / conc.:

500 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:

12 males and 12 females per group, with additional 5 per sex at 0 and 1000 mg/kg bw/day recovery group.

Control animals:

yes, concurrent no treatment

Details on study design:

Satellite group doses: 0, 1000 mg/kg bw/day
Post-exposure recovery period in satellite group: 14 days

Positive control:

No

Observations and examinations performed and frequency:

Detailed observations were made to detect clinical signs in the test animals before the administration period, then on week 1, 2, 3, 4, 5 and 6 of the administration period, and then on week 1 and 2 of the recovery period.

Sacrifice and pathology:

The treated animals were killed at the end of the 14 day recovery period.

Other examinations:

The incidence of response in the sensory/reflex function (e.g. eye sight reaction and pupil reflex) of the test animals was examined on week 6 of the administration period. In addition, the grip strength and motor activity was examined on week 6 of the administration period and week 2 of the recovery period.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Slight salivation was observed at 500 mg/kg bw/day and 1000 mg/kg bw/day in female and male rats during the administration period. No other significant clinical signs were detected in any of the test groups.

Mortality:

no mortality observed

Description (incidence):

There was no mortality recorded in either the male or female test groups.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

All animals gained body weight during the testing period, with the lowest body weight gain generally seen in the highest dose groups. The weight gain in the male group treated at 1000 mg/kg/bw/day was significantly (p<0.05) lower than the control group during the administration period. During the recovery period, animals from the 1000 mg/kg bw/day had a significantly (p<0.05) higher body weight gain when compared to the control animals. Their final body weight was similar at the end of the recovery period between treated and control female and male rats.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

On Day 35 of the administration period, food consumption of male rats treated at 500 mg/kg bw/day was significantly (p<0.05) lower than for the control animals. This difference was not significant on Day 41 at the same dose level, and on Days 35 and 41 at 1000 mg/kg bw/day.
On Day 0 of lactation, food consumption of female rats treated at 500 mg/kg bw/day was significantly (p<0.01) higher than for the control animals. This difference was not significant on Day 3 of lactation at the same dose level, and on Days 0 and 3 at 1000 mg/kg bw/day.
Food consumption of the female rats from the satellite group treated at 1000 mg/kg bw/day was significantly (p<0.05) higher than for the control animals on Day 28 of administration. On Day 35 the food consumption was similar between the treated and control animals.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Description (incidence and severity):

Eye sight reaction and pupil reflex recorded as normal in all groups. No eye discharge recorded.

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

At the end of the recovery period, the number of platelets was significantly (p<0.05) lower in male rats treated at 1000 mg/kg bw/day when compared to the control animals. This effects was not observed at the end of the administration period.
At the end of the administration period, in female rats treated at 1000 mg/kg bw/day the red blood cytes count, hemoglobin, and hematocrit were significantly (respectively p<0.05, p<0.01, and p<0.05) lower to the control animals, while the percentage of reticulocytes was significantly (p<0.05) higher. At the end of the recovery period these altered parameters were comparable between the treated and control female rats, while the activated partial thromboplastin time was significantly (p<0.05) higher in the treated female rats.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

At the end of the administration period, a significantly (p<0.05) lower creatinine level was observed in male rats treated at 30 mg/kg bw/day.
At the end of the administration period, a significantly (p<0.01) lower creatinine level was observed in male rats treated at 120 mg/kg bw/day.
At the end of the administration period, a significantly (p<0.01) lower creatinine level and significantly (p<0.05) higher total-cholesterol and calcium levels were observed in male rats treated at 500 mg/kg bw/day.
At the end of the administration period, a significantly (p<0.01) lower creatinine level and significantly (p<0.01) higher total-cholesterol level were observed in male rats treated at 1,000 mg/kg bw/day.
At the end of the recovery period, these altered parameters were comparable between the treated and control male rats, while the glucose level was significantly (p<0.05) lower and the alanine aminotransferase significantly (p<0.05) higher in the treated animals.

At the end of the administration period, levels of alanine aminotransferase, total cholesterol, and calcium were significantly (respectively p<0.05, p<0.01, p<0.01) higher in female rats treated at 1000 mg/kg bw/day when compared to the control animals. At the end of the recovery period, these altered parameters were comparable between the treated and control female rats.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

At the end of the recovery period, grip strength of hindlimbs and motor activity (30 minutes) were signficantly (respectively p<0.01 and p<0.05) reduced in females treated at 1000 mg/kg bw/day when compared to the control animals. Grip strength of forelimbs and motor activity (60 min) were not significantly different between the treated and control animals. These findings were not significantly different between the threated and control animals (for both sexes) during the administration period.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

At the end of the administration period, significantly (p<0.01) higher relative kidney and liver weights were observed in male rats treated at 500 and 1000 mg/kg bw/day. These were not apparent in recovery group animals suggesting transitory effects.

At the end of the administration period, significantly (p<0.05) lower relative adrenal gland weight was observed in female rats treated at 120 mg/kg bw/day.
At the end of the administration period, significantly (p<0.01 and p<0.05 respectively) higher absolute and relative liver and spleen weights were observed in female rats treated at 500 mg/kg bw/day.
At the end of the administration period, a significantly (p<0.01) higher absolute and relative liver weight and a significantly (p<0.05) lower relative thyroid weight were observed in female rats treated at 1000 mg/kg bw/day
These were not apparent in recovery group animals suggesting transitory effects.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

There was a slight dilatation of the left side pelvis at the end of the recovery period in the male control animals. This effect was not observed in the control animals during the end of the administration period. There was a slight reddish area of the female thymus in the control group at the end of administration. This effect was not observed in the control at the end of the recovery period.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Hypertrophy, hepatocyte, centrilobular were observed in liver of female rats treated at 1000 mg/kg bw/day. These were not apparent in recovery group animals suggesting transitory effects.
Slight to moderate changes were observed in kidneys of male rats treated at 1000 mg/kg bw/day. These were not apparent in recovery group animals suggesting transitory effects.

No abnormalities were detected in the brain, thyroid, parathyroid, thymus, trachea, small intestine, large intestine, adrenal, urinary bladder, testis, epididymis, seminal vesicle, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes and mammary gland from animals of control and 1000 mg/kg bw/day groups of male rats. No abnormalities were detected in the brain, pituitary, thyroid, parathyroid, trachea, small intestine, large intestine, heart, adrenal, urinary bladder, ovary, uterus, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes and mammary glands of control and 1000 mg/kg bw/day groups of female rats.

Histopathological findings: neoplastic:

no effects observed

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

Critical effects observed:

no

Lowest effective dose / conc.:

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

System:

hepatobiliary

Organ:

liver

Critical effects observed:

no

Lowest effective dose / conc.:

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

System:

urinary

Organ:

kidney

Conclusions:

On the basis of the significant effects observed on bodyweight at 1000 mg/kg bw/day it is proposed a NOAEL of 500 mg/kg bw.

Executive summary:

A combined short-term toxicity and screening for toxicity to reproduction study was performed on BPA-5EO according to OECD Testing Guideline 422. Groups of 12 rats per sex were exposed for 42 days to the substance by the oral route. The concentrations selected for this study were 0, 30, 120, 500, and 1000 mg/kg bw/day. Two additional groups of five animals per sex were included for a 14-day recovery period and received 0 or 1000 mg/kg bw/day. Mortality, clinical signs, bodyweights, and food consumption were recorded during the study. Before termination samples of blood and urine were collected to investigate the haematology and blood chemistry parameters and perform the urinalysis. Ophthalmological examination and FOB were performed. At termination, pathological and histopathological examinations were undertaken and organs were weighed. No animal died as a result of the exposure to the substance at up to 1000 mg/kg bw/day. Slight salivation was observed at 500 and 1000 mg/kg bw/day but this was not considered as a significant effect and salivation stopped during the recovery period. A significantly reduced body weight gain was observed in male animals treated at 1000 mg/kg bw/day however the bodyweight was comparable at the end of the recovery period between treated and control animals. A momentary reduced food consumption was observed at 500 mg/kg bw/day and 1000 mg/kg bw/day during the administration period. Some changes were observed at 1000 mg/kg bw/day during the FOB at weeks 2 and 6 of the dosing period, but these changes were not significantly different from controls. Some parameters in the haematology, biochemistry, and urinalysis were altered at the end of the administration period but recovered. Reversible variations in organ weights were observed at the end of the administration period. No effects were observed during the ophthalmological examination. Changes were observed in liver of female rats treated at 1000 mg/kg bw/day. Slight to moderate changes were observed in kidneys of male rats treated at 1000 mg/kg bw/d. These were not apparent in recovery group animals suggesting transitory effects. No abnormalities were detected in the brain, thyroid, parathyroid, thymus, trachea, small intestine, large intestine, adrenal, urinary bladder, testis, epididymis, seminal vesicle, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes, and mammary gland from animals of control and 1000 mg/kg bw/day groups of male rats. No abnormalities were detected in the brain, pituitary, thyroid, parathyroid, trachea, small intestine, large intestine, heart, adrenal, urinary bladder, ovary, uterus, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes, and mammary glands of control and 1000 mg/kg/bw groups of female rats. On the basis of the significant effects observed on body weight at 1000 mg/kg bw/d it is proposed a NOAEL of 500 mg/kg bw/day.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

GLP 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

Only an oral Combined repeated dose toxicity study with the reproduction/developmental screening test according to OECD TG 422 on Grade 5 of BPA EO was available. An oral 90-day repeated dose toxicity assay (according to OECD TG 408) is ongoing (refer to ECHA decision number TPE-D-2114489554-35-01/F).

In the combine repeated dose toxicity study with the reproduction/developmental screening test (according to OECD TG 422), the test substance was administered to 12 rats/sex/dose by oral gavage at dose levels of 0, 30, 120, 500 or 1000 mg/kg bw/day.  Two additional groups of five animals per sex were included for a 14-day recovery period and received 0 or 1000 mg/kg bw/day.

No animal died as a result of the exposure to the substance at up to 1000 mg/kg bw/day. Slight salivation was observed at 500 and 1000 mg/kg bw/day but this was not considered as a significant effect and salivation stopped during the recovery period. A significantly reduced body weight gain was observed in male animals treated at 1000 mg/kg bw/day however the bodyweight was comparable at the end of the recovery period between treated and control animals. A momentary reduced food consumption was observed at 500 mg/kg bw/day and 1000 mg/kg bw/day during the administration period. Some changes were observed at 1000 mg/kg bw/day during the FOB at weeks 2 and 6 of the dosing period, but these changes were not significantly different from controls. Some parameters in the haematology, biochemistry, and urinalysis were altered at the end of the administration period but recovered. Reversible variations in organ weights were observed at the end of the administration period. No effects were observed during the ophthalmological examination. Changes were observed in liver of female rats treated at 1000 mg/kg bw/day. Slight to moderate changes were observed in kidneys of male rats treated at 1000 mg/kg bw/day. These were not apparent in recovery group animals suggesting transitory effects. No abnormalities were detected in the brain, thyroid, parathyroid, thymus, trachea, small intestine, large intestine, adrenal, urinary bladder, testis, epididymis, seminal vesicle, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes, and mammary gland from animals of control and 1000 mg/kg bw/day groups of male rats. No abnormalities were detected in the brain, pituitary, thyroid, parathyroid, trachea, small intestine, large intestine, heart, adrenal, urinary bladder, ovary, uterus, spinal cord, sciatic nerve, bone, bone marrow, lymph nodes, and mammary glands of control and 1000 mg/kg/bw groups of female rats. On the basis of the significant effects observed on body weight at 1000 mg/kg bw/day, the NOAEL was considered to be 500 mg/kg bw/day.

Justification for classification or non-classification

Based on the data available, no critical effects on specific target organs were observed in OECD TG 422. Thus, in accordance with CLP Regulation 1272/2008 classification for Specific Target Organ Toxicant (STOT) is not warranted.