Registration Dossier

Administrative data

Description of key information

Administration of DGEBPA via oral gavage resulted in slight body weight effects at 250 mg/kg/day and higher.  Enlarged cecum was noted at necropsy in male rats receiving 250 mg/kg/day.  Slight histopathologic changes were noted in the adrenal gland, cecum and kidney of male and/or female rats ingesting 250 mg/kg/day.  A 3% decrease in body weight was noted in female rats gavaged with 50 mg/kg/day and a slight increase in cholesterol levels was noted which was considered to be not detrimental.
In a dermal study, the only systemic toxicity was a slight decrease in body weights at 1000 mg/kg/day. Thus the NOAEL for systemic toxicity was considered to be 100 mg/kg/day. The NOEL for dermal effects in female rats was 10 mg/kg/day. In male rats a NOEL for dermal effects was not detected.

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:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000-11-20-2001-02-28
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted according to GLP as well as Guidelines.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Qualifier:
according to
Guideline:
EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)
Qualifier:
according to
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Qualifier:
according to
Guideline:
other: japanese MITI guidelines for toxicity testing of chemicals
Principles of method if other than guideline:
A two-year oral gavage toxicity/oncogenicity study of Bisphenol A Diglycidyl Ether (BADGE) was initiated but study was terminated on test days 99 (males) or 101 (females) due to excessive toxicity noted in rats. Standard toxicologic parameters consistent with OECD guideline #408 were evaluatedon a subset of ten rats/sex/dose level.
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals and environmental conditions:
Male and female Fischer 344 rats (7 weeks of age) were received from a commercial supplier, evaluated for general health by a laboratory veterinarian, and were housed 2-3 per cage (suspended stainless steel cages with feed crock and pressure-activated nipple watering system) in rooms designed to maintain adequate environmental conditions and photocycle for rats. They were allowed to acclimate to the laboratory for approximately 3 weeks prior to the start of the study.

The animals were stratified by body weight by a computer randomization program and were assigned to dose groups. They were assigned unique animal identification numbers via subcutaneously-implanted transponders.

Animals were provided food and water ad libitum throughout the study. Food was analyzed for nutritional content and contaminants by the supplier. The water was obtained from a municipal water source, analyzed periodically for chemical parameters and biological contaminants by the municipal water department.

The laboratory operates under AALAC accreditation standards.
Route of administration:
oral: gavage
Vehicle:
other: 0.5% Methocel A4M methyl cellulose ethers with 0.1% Tween 80
Details on oral exposure:
Four groups of 65 male and 65 female rats were orally gavaged with BADGE 7 days per week for 14 weeks as an aqueous suspension in 0.5% Methocel A4M methyl cellulose ethers with 0.1% Tween 80. Concentrations resulting in doses of 0, 50, 250, and 1000 mg/kg/day were used to evaluate systemic toxicity. Dose volume and concentration was calculated based on the most recent animal body weight.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Homogeneity, stability, and concentration of the dose solutions were verified throughtout the study.
Duration of treatment / exposure:
7 days per week for 14 weeks
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
0, 50, 250, and 1000 mg/kg/day
Basis:
other: nominal in dose solution
No. of animals per sex per dose:
65/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
The study was intended to be a 2-year chronic/oncogenicity study. The study was terminated at 99-101 days due to excessive toxicity at 250 and 1000 mg/kg/day dose levels and the study was reported as a subchronic investigation.

Four groups of 65 male and 65 female rats were orally gavaged with BADGE 7 days per week for 14 weeks as an aqueous suspension in 0.5% Methocel A4M methyl cellulose ethers with 0.1% Tween 80. Concentrations resulting in doses of 0, 50, 250, and 1000 mg/kg/day were used to evaluate systemic toxicity. Dose volume and concentration was calculated based on the most recent animal body weight. Homogeneity, stability, and concentration of the dose solutions were verified throughtout the study.

The first 10 rats/sex/dose were selected for urinalysis, hematology, clinical chemistry, necropsy, organ weights, and histopathology. Hematology, clinical chemistry, and urinalysis data were collected following 3 months on study.

Twice daily cageside clinical observations (mortality, morbundity, and feed & water check) monthly detailed clinical observations (replacement animals or first 15 animals/sex/dose only), weekly categorical (clinical) observations on all animals, eye exams (indirect ophthalmology pre-exposure and prior to termination), weekly body weights, weekly food consumption were measured. Hematology, clinical chemistry, urinalysis, and organ weights were evaluated at study termination.

Organs evaluated: brain, liver, kidneys, heart, adrenals, testes, epididymis, uterus, ovaries, and spleen.
Positive control:
no
Observations and examinations performed and frequency:
Twice daily cageside clinical observations (mortality, morbundity, and feed & water check) monthly detailed clinical observations (replacement animals or first 15 animals/sex/dose only), weekly categorical (clinical) observations on all animals, eye exams (indirect ophthalmology pre-exposure and prior to termination), weekly body weights, weekly food consumption were measured. Hematology, clinical chemistry, urinalysis, and organ weights were evaluated at study termination.
Sacrifice and pathology:
The first 10 rats/sex/dose were selected for urinalysis, hematology, clinical chemistry, necropsy, organ weights, and histopathology. Hematology, clinical chemistry, and urinalysis data were collected following 3 months on study.

Organs evaluated: brain, liver, kidneys, heart, adrenals, testes, epididymis, uterus, ovaries, and spleen.
Other examinations:
no additional examinations
Statistics:
Means and standard deviations were calculated for all continuous data. Body weights, food consumption, organ weights, urine volume, urine specific gravity, clinical chemistry data, coagulation, and hematologic data were evaluated by a Bartlett's test for equality of variances. Based on the outcome of the Bartlett's test, exploratory data analysis was performed by a parametric or nonparametric ANOVA. If alpha = 0.05, the ANOVA were followed by a Dunnett's test with a Bonferroni correction for multiple comparisons to the control. Detailed Clinical Observations were statistically analyzed by a z-test of proportions comparing each treated group to the control. Statistical outliers were identified by a sequential test. Outliers may have been excluded for sound scientific reasons. Since the overall false positive rate (Type I errors) was greater than the nominal alpha levels, the interpretation of the data considered other factors such as the dose-response relationship and whether the results were consistent with other biological or pathological findings and historical control values.
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, treatment-related
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
effects observed, treatment-related
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
Analytical
Homogeneity was confirmed pre-study and before last dosing at high and low doses, and the concentration was confirmed pre-study, mid-way through the study, and prior to the last dose delivery to be 104.7-109.3% of the targeted doses at each dose level. Stability analyses demonstrated stability for 13-20 days in two tests.

Mortality
There was no significant treatment-related mortality through the 99-101 day treatment period. Six animals died or were euthanized in moribund condition before study termination at 14 weeks. Three died incidentally during dosing or blood collection, and three high-dose rats had moderate to severe acute tubular necrosis of the kidneys.

Observations
There were no treatment-related observations noted in cageside or clinical examinations, nor were there any noted during detailed clinical observations.

Ophthalmology
No treatment-related ophthalmology was noted. All observations were of variable frequency and did not occur in a dose-related manner.

Body Weights / Weight Gains
Treatment-related statistically-significant decreases in body weights and gains were noted in male and female rats given 250 and 1000 mg/kg/day (19.2% and 17.2% for males and females at 1000 mg/kg/day at day 98), and additionally in females given 50 mg/kg/day. The decrements in body weight were slowly progressive throughout the dosing period. The minimal decrement in female body weight decrease at 50 mg/kg/day was interpreted to be a non-adverse effect.

Food Consumption
Treatment-related statistically-significant decreases in food consumption were noted in male and female rats at 50, 250, and 1000 mg/kg/day.

Clinical Pathology
Minimal, but statistically-significant, decreases in prothrombin time were observed in males given 250 or 1000 mg/kg/day, and in females at 250 mg/kg/day. The alterations were interpreted to be non-adverse, and reflective of normal biologic variation. A significant dose-related decrease in red blood cell count, hematocrit, and hemoglobin were noted in males and females at 250 or 1000 mg/kg/day.

Clinical Chemistry
Statistically-significant increases in urea nitrogen (males and females at 250 and 1000 mg/kg/day), aspartate aminotransferase (males at 1000 mg/kg/day), cholesterol (males and females at all dose levels), total bilirubin (females at 1000 mg/kg/day), and calcium (females at 1000 mg/kg/day). The increased urea nitrogen was consistent with treatment-related renal toxicity. The other clinical chemistry effects were unaccompanied by histopathologic alterations, and were considered to be non-adverse.

Urinalysis
Increases in timed urine volume were noted in males and females at the high dose. Decreases in urine specific gravity were noted in males at the high dose, and females at the high and mid doses. The effects were consistent with treatment-related renal toxicity. In addition, some males and females at 250 and 1000 mg/kg/day had slight, treatment-related decreases in urine pH relative to controls.

Organ Weights
All alterations in organ weights were considered to be reflective of treatment-related decreases in body weights for animals given 250 or 1000 mg/kg/day. In males, decreases in absolute adrenal, heart, spleen weights, and increased relative brain and testes weights were identified. In females, decreases in absolute adrenal, heart, spleen, ovaries, and uterus weights, and increased relative brain weight were recorded.

Gross Pathology
The only treatment-related observation noted was increased cecum size in all males at mid and high doses, and in all females at the high dose.

Histopathology
Effects were noted in adrenal glands, cecum, ileum, kidneys, liver, testes, and uterus of animals at the mid and high dose. Adrenal glands of males at these dose levels had slightly decreased vacuolization of the cortex, relative to controls. The cecal effect was a very slight, diffuse hyperplasia of the mucosal epithelium of males at the mid and high dose, and in females at the high dose. Some males given 1000 mg/kg/day also had slight, diffuse hyperplasia of the ileum. Treatment-related kidney effects were slight, decreased hyalin droplet formation in proximal convoluted tubules of males at the mid and high dose, and slight vacuolization of of females given 250 and 1000 mg/kg/day. In addition, 2 males and 1 female died prior to study termination of moderate or severe acute necrosis of renal proximal tubules. There were 2 treatment-related liver effects: single eosiniphilic focus of altered hepatocytes in females given 1000 mg/kg/day, and altered tinctorial properties of centrilobular hepatocytes of males at the mid and high dose and females at the high dose. The testicular effect was characterized by a slight to moderate degeneration of the seminiferous tubules in all males given 1000 mg/kg/day. Uterine changes included a slight atrophy of the endometrium and myometrium of females given 1000 mg/kg/day, corresponding to a decrease in the mean uterus weight of animals in this group. There were no treatment-related histopathologic effects in any organs or tissues of males or females given 50 mg/kg/day.
Dose descriptor:
NOAEL
Effect level:
50 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Critical effects observed:
not specified

There were no treatment-related histopathologic effects in any organs or tissues of males or females given 50 mg/kg/day.

Conclusions:
Based on alterations in body weights and serum cholesterol in rats given 50 mg/kg/day, a no-observed-effect (NOEL) was not determined. However, since these alterations were not associated with detrimental effects, the dose of 50 mg/kg/day was interpreted to be the no-observed-adverse-effect level (NOAEL).
Executive summary:

A two-year oral gavage toxicity/oncogenicity study of Bisphenol A Diglycidyl Ether (BADGE) was initiated in Fischer 344 rats. Groups of 65 Fischer 344 rats/dose level were dosed with BADGE suspended in a Tween** 80 and methylcellulose vehicle at dose levels of 0, 50, 250 or 1000 mg/kg/day. The study was terminated on test days 99 (males) or 101 (females) due to excessive toxicity noted in rats given 250 or 1000 mg/kg/day. Standard toxicologic parameters consistent with OECD guideline #408 were evaluated on a subset of ten rats/sex/dose level.

Progressive decreases in body weights and feed consumption, relative to controls, occurred throughout the study in males and females given 250 or 1000 mg/kg/day. By the end of the study, body weights of males and females given 1000 mg/kg/day were 19.2 and 10.9% lower than controls, respectively, and body weights of males and females given 250 mg/kg/day were 10.8 and 5.1% lower than controls, respectively. Body weights of females given 50 mg/kg/day were 3.2% lower than controls, while body weights of males given 50 mg/kg/day were comparable to controls throughout the study.

Treatment-related alterations in hematology (decreases in red blood cell count, hemoglobin concentration, and hematocrit), clinical chemistries, urinalysis, and organ weights occurred in rats given 250 or 1000 mg/kg/day. Some clinical chemistry and urinalysis alterations were indicative of renal toxicity, while organ weight alterations were reflective of lower feed consumption and body weights. The only treatment-related clinical

pathology alteration in male and female rats given 50 mg/kg/day was increased cholesterol. The only treatment-related gross pathologic observation was increased size of the cecum in males given 250 or 1000 mg/kg/day, and in females given 1000 mg/kg/day. Two males and one female given 1000 mg/kg/day that died prior to study termination had moderate to severe acute necrosis of renal proximal tubules. Treatment related histopathologic effects in surviving animals given 250 and/or 1000 mg/kg/day were noted in the adrenal glands, cecum, ileum, kidneys, liver, testes, and uterus. Males and females given 50 mg/kg/day had no treatment-related histopathologic effects.

Based on alterations in body weights and serum cholesterol in rats given 50 mg/kg/day, a no-observed-effect level (NOEL) was not determined. However, since these alterations were not associated with detrimental effects, the dose of 50 mg/kg/day was interpreted to be the no-observed-adverse-effect level (NOAEL).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
50 mg/kg bw/day
Study duration:
chronic
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

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1995-02-20-1995-05-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted according to GLP as well as Guidelines.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Qualifier:
according to
Guideline:
EPA OPP 82-3 (Subchronic Dermal Toxicity 90 Days)
Qualifier:
according to
Guideline:
other: EEC Directive 87/302 Sub-Chronic dermal toxicity study
GLP compliance:
yes
Limit test:
no
Species:
mouse
Strain:
B6C3F1
Sex:
male
Details on test animals and environmental conditions:
B6C3F1 male mice were obtained from a commercial supplier, examined by a laboratory veterinarian for general health status, and allowed to acclimate to the laboratory for 12 days prior to study start. Animals were randomized into dose groups based on body weights by a computer randomization program. Animals were identified via a subcutaneously-implanted transponder correlated to a unique animal identificatiom number.

Animal room temperature, humidity, air changes, and photocycle were maintained appropriate to the species. Animals were provided water and food ad libitum, and housed one per cage. Water was analyzed for contaminants at the municipal water source, and feed was analyzed for nutritional content and contaminants by the supplier.
Type of coverage:
open
Vehicle:
acetone
Details on exposure:
Dose solution concentrations were based on the expected average body weights over the duration of the 13 week study. The dose solutions were prepared by dissolving test material in acetone. Mice were dosed dermally with solutions of 0%, 0.05%, 0.5%, or 5% BADGE (w/v). Dose solutions were administered as a fixed volume (50ug/mouse/application). The dose was delivered by pipette to the clipped area of the back (approximately 10% surface area) of each mouse. The treated area was not covered, nor was the skin cleansed following treatment.

Groups of 10 male mice per dose level were dosed dermally with BADGE in acetone. Each animal was dosed 3 times per week
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The stability of the test material in acetone was determined concurrent with the start of the study. Samples of the 0.05% solutions were analyzed periodically to verify stability in the vehicle. Homogeneity of the low and high dose solutions was evaluated concurrently with the conduct of the study. Concentration verification of all dose solutions was confirmed pre-study and after week 1 (as a result of a procedural change in dose solution preparation).
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
3 times/week
Remarks:
Doses / Concentrations:
0, 1, 10, 100 mg/kg/day
Basis:
other: nominal
No. of animals per sex per dose:
10 male mice/dose
Control animals:
yes, concurrent vehicle
Details on study design:
Groups of 10 male mice per dose level were dosed dermally with BADGE in acetone. Each animal was dosed 3 times per week.

Body weights and food consumption were measured weekly, and a daily observation was conducted to evaluate general animal disposition and the availability of feed and water. The dermal application site was evaluated for signs of irritation according to the dermal irritation scoring system recommended by the OECD (1981) on a daily basis the first week of the study, and weekly thereafter for the duration of the study. Clinical examinations were conducted prior to study start and weekly throughout the study and included evaluations of the skin, fur, mucous membranes, respiration, central nervous system function, swelling, masses, and general behavior.

Ophthalmological examinations were conducted on all mice prior to study start and at necropsy.

Clinical Pathology / Chemistry / hematology parameters were evaluated: hematocrit, hemoglobin, erythrocyte count, total leukocyte count, platelet count, differential counts of luekocytes, morphology of erythrocytes, leukocytes, and platelets, enzyme activity (alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase), urea nitrogen, creatinine, total protein, albumin, globulin, glucose, total bilirubin, electrolytes (Na, K, P, Cl, Ca), cholesterol, and triglycerides.

Absolute and relative weights were collected for the brain, liver, heart, kidneys, and testes. All tissues from the control and high dose groups were evaluated, including all reproductive organs and tissues. In addition, tissues in lower dose levels including liver, kidneys, lungs, and skin from the dermal test site and a control site were evaluated.
Positive control:
No
Observations and examinations performed and frequency:
Body weights and food consumption were measured weekly, and a daily observation was conducted to evaluate general animal disposition and the availability of feed and water. The dermal application site was evaluated for signs of irritation according to the dermal irritation scoring system recommended by the OECD (1981) on a daily basis the first week of the study, and weekly thereafter for the duration of the study. Clinical examinations were conducted prior to study start and weekly throughout the study and included evaluations of the skin, fur, mucous membranes, respiration, central nervous system function, swelling, masses, and general behavior.

Ophthalmological examinations were conducted on all mice prior to study start and at necropsy.
Sacrifice and pathology:
Clinical Pathology / Chemistry / hematology parameters were evaluated: hematocrit, hemoglobin, erythrocyte count, total leukocyte count, platelet count, differential counts of luekocytes, morphology of erythrocytes, leukocytes, and platelets, enzyme activity (alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase), urea nitrogen, creatinine, total protein, albumin, globulin, glucose, total bilirubin, electrolytes (Na, K, P, Cl, Ca), cholesterol, and triglycerides.

Absolute and relative weights were collected for the brain, liver, heart, kidneys, and testes. All tissues from the control and high dose groups were evaluated, including all reproductive organs and tissues. In addition, tissues in lower dose levels including liver, kidneys, lungs, and skin from the dermal test site and a control site were evaluated.
Other examinations:
no additional examinations conducted
Statistics:
Descriptive statistics only (means and standard deviations) are reported for feed consumption, feed efficiency and leukocyte differential counts. Body weights, organ weights, clinical chemistry data, and appropriate hematologic data are evaluated by Bartlett's test for equality of variances. Based on the outcome of Bartlett's test, exploratory data analysis is performed by a parametric or nonparamet-ric analysis of variance (ANOVA), followed respectively by Dunnett's test or the Wilcoxon Rank- Sum test with a Bonferroni correction for multiple comparisons. Statistical outliers are identified by a sequential test, but routinely excluded only from feed ' consumption statistics. Outliers may be excluded from other analyses only for documented, scientifically sound reasons, unrelated to treatment.

The nominal alpha levels to be used and test references are as follows:

Bartlett's test (Winer, 1971) α = 0.01
Parametric ANOVA (Steel and Torrie, 1960) α = 0.10
Nonparametric ANOVA (Hollander and Wolfe, 1973) α = 0.10
Dunnett's test (Winer, 1971) α = 0.05 two-sided
Wilcoxon Rank-Sum test (Hollander and Wolfe, 1973) α = 0.05
(Bonferroni correction - Miller, 1966) two-sided
Outlier test (Grubbs, 1969) α = 0.02 two-sided

Because numerous measurements are compared statistically on the same group of animals, the frequency of false positive (Type 1) errors is unknown, but is much greater than the nominal alpha.

The final toxicologic interpretation of the data considers other factors, such as dose- response relationships, biological plausibility and consistency, and historical rates.
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
Concentrations of the dose levels were determined ot be 94-97% of the targeted concentrations. Homogeneity of the solutions was verified; a standard of deviation and percent relative standard deviation were considered within acceptable limits.

There were no treatment-related effects in the following parameters: ophthalmology, in-life observations, body weights and weight gains, feed consumption, and clinical chemistry and hematology parameters that were evaluated. Two control mice were observed on test days 1 and 3 with scab formation where the skin was disturbed during clippind and transponder implantation. The scabs were resolved by test day 5, and no other treatment-related irritation was noted.

Mice administered 1 mg/kg/day had statistically-identified increased absolute brain weights, but a lack of dose-response led authors to dismiss it as not related to treatment. Liver weights at the high dose were increased, but were not accompanied by histopathological findings and were still lower than historical control values for the laboratory. The finding is of doubtful toxicological significance.

There were no gross lesions attributed to treatment, and no indication of systemic pathologic effects supported by histopathology. Histopathologic changes in the skin, however, were not confined to the test site, but were less marked in nature than the test site. The changes in skin were characterized as a very slight to moderate epidermal hyperplasia with very slight to moderate subacute inflammation. In some cases, slight follicular spongiosis, characterized by intraepithelial edema in the epidermis or at the neck of the follicle, was noted. Very slight focal or multifocal parakeratosis was observed in a few mid and high dose animals. Micro abscesses in either the epidermis or neck of the follicle, were observed in one mid dose and one high dose animal. Together, the findings suggest chronic active dermatitis.
Dose descriptor:
NOAEL
Effect level:
100 other: mg/kg/application
Sex:
male/female
Basis for effect level:
other: Systemic toxicity was unaffected by dermal application of 100 mg/kg/day administered three times/week.
Dose descriptor:
NOAEL
Effect level:
>= 1 other: mg/kg/application
Sex:
male/female
Basis for effect level:
other: At the application site, mild to moderate chronic active dermatitis with a weak dose-response was observed histopathologically at doses of 1-100 mg/kg/application DGEBPA administered three times/week.
Remarks on result:
not determinable
Remarks:
no NOAEL identified
Critical effects observed:
not specified

DGEBPA applied to the skin of male BCC3F1 mice three times per week for thirteen weeks at dosages of 1,10 or 100 mg/kg/application caused no

systemic toxicity. Mild to moderate chronic active dermatitis with a weak dose-response was observed histopathologically at dosages up to 100 mg/kg/application DGEBPA. Spongiosis and epidermal micro abscess formation indicated that the maximum-tolerated-dose (MTD) was met in

mice administered 100 mg/kg/application DGEBPA. The no-observed-effect level (NOEL) for dermal effects was not determined.

Conclusions:
There was no systemic toxicity noted up to 100 mg/kg/day. Mild to moderate chronic active dermatitis with a weak dose-response was observed histopathologically at doses up to 100 mg/kg/day. Spongiosis and epidermal micro-abscesses indicated that the maximum-tolerated-dose was met at 100 mg/kg/day. NOEL for DGEBPA in mice dosed dermally was not determined.
Executive summary:

Diglycidyl ether of bisphenol A (DGEBPA), CAS# 1675-54-3, was evaluated for dermal irritation and systemic toxicity potential following approximately 13-weeks of repeated dermal administration. Groups of 10 B6C3F1 male mice/dose level were dosed dermally with DGEBPA in acetone solutions at concentrations of 0, 0.05%, 0.5%, or 5.0% (wlv). The dosing volume was 50 pllapplication which corresponded to approximate dosages of 0, 1, 10, or 100 mg DGEBPA/kg body weighvapplication. Each dose group followed a 3 applications/week (Monday, Wednesday, Friday) dosing regimen resulting in each animal receiving a total of 41 applications during the study. Data were collected on the following: clinical appearance and behavior, dermal irritation at the test site, body weights, food consumption, clinical pathology, gross pathology, and histopathologic appearance of the treated and untreated test sites.

DGEBPA applied to the skin of male B6C3F1 mice three times per week for thirteen weeks at dosages of 1, 10 or 100 mg/kg/application caused no apparent systemic toxicity. Mild to moderate chronic active dermatitis with a weak dose-response was observed histopathologically at dosages up to 100 rng/kg/application DGEBPA. Spongiosis and epidermal micro abscess formation indicated that the maximum-tolerated-dose (MTD) was met in mice administered 100 mg/kg/application DGEBPA. A no-observed-effect level (NOEL) for dermal effects was not determined.

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1995-03-20-1995-06-23
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted according to GLP as well as Guidelines.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Qualifier:
according to
Guideline:
EPA OPP 82-3 (Subchronic Dermal Toxicity 90 Days)
Qualifier:
according to
Guideline:
other: EEC Directive 87/302 sub-chronic dermal toxicity study
Qualifier:
according to
Guideline:
other: MAFF, subchronic dermal toxicity study, 1985
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals and environmental conditions:
Male and female F344 rats were obtained from a commercial supplier. They were examined for general health status, and allowed to acclimate to the laboratory for 12 days prior to study start. Animals were randomized into dose groups based on body weights by a computer randomization program. Animals were identified via a subcutaneously-implanted transponder correlated to a unique animal identificatiom number.

Animal room temperature, humidity, air changes, and photocycle were maintained appropriate to the species. Animals were provided water and food ad libitum, and housed one per cage. Water was analyzed for contaminants at the municipal water source, and feed was analyzed for nutritional content and contaminants by the supplier.
Type of coverage:
open
Vehicle:
acetone
Details on exposure:
Dose solutions were prepared by dissolving test material in acetone. Male rats were dosed dermally with BADGE solutions of 0, 0.9, 9, or 90% (w/v) while female rats were dosed with solutions of 0, 0.6, 6, or 60% (w/v). The dosing solution concentration for each dose level remained constant throughout the duration for the 13-week study. Dose solutions were administered as a fixed volume (300 ul/rat/application). Each appropriate dose was delivered by pipette to the clipped area of the back (approximately 10% surface area) of each rat. The treated area was not covered nor was the skin cleansed following treatment.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Stability of the test material in acetone was determined in a previous study. The homogeneity of the material in the low and high dose level solutions were determined concurrently with the conduct of this study. Analysis of dosing solutions was done by HPLC to verify the concentration on all dose levels at the beginning of the dosing period. Additional analyses were conducted at week 6 and 12.
Duration of treatment / exposure:
5 times per week for 13 weeks
An additional high-dose group of females was dosed 3 times per week for 13 weeks.
Frequency of treatment:
daily (3 or 5 times a week)
Remarks:
Doses / Concentrations:
0, 10, 100, 1000 mg/kg/day
Basis:
other: nominal
No. of animals per sex per dose:
10/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
Groups of 10 male and 10 female rats/sex/dose were dosed dermally with BADGE in acetone 5 times per week for 13 weeks. An additional high-dose group of females was dosed 3 times per week for 13 weeks.

Animals were weighed prior to study start, and weekly throughout the study. In-life observations for signs of toxicity were conducted daily. An additional daily check for availability of feed and water was conducted. The dermal application site was evaluated daily for signs of irritation during week 1, and weekly thereafter based on the OECD (1981) scoring system. Food consumption was recorded weekly. Indirect ophthalmology was evaluated indirectly by a veterinarian pre-study. Clinical pathology was performed on fasted animals at 6 weeks and at necropsy. Urine samples were obtained from non-fasted animals (not including the female satellite group) by manual compression of the bladder pre-study, at week 5, and at necropsy. Hematocrit, hemoglobin concentration, erythrocyte count, total leukocyte count, platelet count, and differential counts of leukocytes and platelets were evaluated. Standard clinical chemistry parameters were additionally evaluated.

A gross necropsy examination was conducted. Tissues collected & evaluated included: adrenals, liver, kidneys, lungs, skin from the test site & from an untreated inguinal site, and reproductive organs and tissues.
Positive control:
no
Observations and examinations performed and frequency:
Animals were weighed prior to study start, and weekly throughout the study. In-life observations for signs of toxicity were conducted daily. An additional daily check for availability of feed and water was conducted. The dermal application site was evaluated daily for signs of irritation during week 1, and weekly thereafter based on the OECD (1981) scoring system. Food consumption was recorded weekly. Indirect ophthalmology was evaluated indirectly by a veterinarian pre-study.
Sacrifice and pathology:
Clinical pathology was performed on fasted animals at 6 weeks and at necropsy. Urine samples were obtained from non-fasted animals (not including the female satellite group) by manual compression of the bladder pre-study, at week 5, and at necropsy. Hematocrit, hemoglobin concentration, erythrocyte count, total leukocyte count, platelet count, and differential counts of leukocytes and platelets were evaluated. Standard clinical chemistry parameters were additionally evaluated.

A gross necropsy examination was conducted. Tissues collected & evaluated included: adrenals, liver, kidneys, lungs, skin from the test site & from an untreated inguinal site, and reproductive organs and tissues.
Other examinations:
no additional examinations conducted
Statistics:
See below
Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
no effects observed
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
Analytical
Homogeneity was confirmed for the dose solutions, with an RSD of 2.36 and 7.44% for two samples of 1000 mg/kg/application male and 0.34% for10 mg/kg/application . Concentration was 97-103% for the first 1000 mg/k/application sample, 92.3-107% for the second 1000 mg/kg/application sample and 0.590-0.594% for the 10 mg/kg/application of the targeted dose levels.

In-life Observations
There were no treatment-related effects noted in ophthalmology or clinical observations. Body weights at the high dose was decreased for males and females of the main study group, but unchanged for the low and mid-dose groups, as well as the satellite female 1000 mg/kg/day group. Irritation at the dermal test site was noted in 1000 mg/kg/day males only. There was a slight decrease in food consumption at the high dose in males and females.

Clinical Pathology
Based on a lack of dose-response relationship (male rats at 10 and 1000 mg/kg/day) and analyses of historical control data, no toxicological significance was assigned to the following findings: decrease in hematocrit, RBC, and hemoglobin. There was no treatment-related finding upon urinalyses. Clinical chemistry findings included an increase in serum cholesterol values for mid and high-dose males and females. No correlated histopathological changes were observed in the kidneys or thyroid glands, and the toxicological significance is questionable. Variations in other parameters (ALT, triglycerides, AST, albumin, total bilirubin, and glucose) did not occur in a dose-response pattern, and had no histopathological correlate.

Anatomic Pathology
There were no gross lesions attributed to treatment. Slight to moderate epidermal hyperplasia with slight chronic inflammation, slight follicular or epidermal spongiosis with intraepithelial edema in the epidermis or at the neck of the follicle was noted at the dermal test site. The findings are characterized as chronic dermatitis. Males were more affected due to higher concentrations of BADGE to achieve the same dose in a fixed volume. Inguinal skin over the mammary gland, the control dermal site, showed an increase in slight chronic inflammation at 100 mg/kg/day in both sexes. The lack of high-dose effect was likely due to migration of the test material based on viscosity. Chronic inflammation of the tongue was noted in both sexes at the high dose, likely the result of licking the application site during grooming.

Lower absolute adrenal, heart, and kidney weights were noted in high-dose males, as well as higher relative testes weigts. Female rats at the high dose had lower absolute and relative adrenal weights, and increased relative brain and liver weights. There were no histological correlates.
Dose descriptor:
NOAEL
Effect level:
>= 10 other: mg/kg/application
Sex:
male
Basis for effect level:
other: Epidermal hyperplasia with chronic inflammation, characterized as chronic dermatitis, was observed histopathologically at all dose levels (10, 100 and 1000 mg/kg/application) in male rats.
Remarks on result:
not determinable
Remarks:
no NOAEL identified
Dose descriptor:
NOEL
Effect level:
10 other: mg/kg/application
Sex:
female
Basis for effect level:
other: Epidermal hyperplasia with chronic inflammation, characterized as chronic dermatitis, was observed histopathologically at dosages of 100 and 1000 mg/kg/application DGEBPA in female rats
Dose descriptor:
LOAEL
Effect level:
>= 100 other: mg/kg/application
Sex:
female
Basis for effect level:
other: Epidermal hyperplasia with chronic inflammation, characterized as chronic dermatitis, was observed histopathologically in female rats at 100 mg/kg/application.
Critical effects observed:
not specified

DGEBPA applied to the skin of male and female rats five times per week for approximately thirteen weeks at dosages of 10,100 or 1000 mg/kg/application caused no apparent systemic toxicity with the exception of decreased body weight and body weight gain in males and females at 1000 mg/kg/application. Food consumption was also slightly lower for both males and females throughout the duration of the study. Serum cholesterol values were increased in a dose-related manner in mid- and high dose level rats of both sexes, but considered of questionable toxicological significance since no correlated histopathological changes were observed. Female rats dosed with 1000 mg/kg/application three times per week (high-dose satellite group) for the same duration showed no signs of systemic toxicity. Histopathologic examination of tissues conducted on all test animals resulted in treatment related effects only at the site of dermal application of DGEBPA. Epidermal hyperplasia with chronic inflammation, characterized as chronic dermatitis, was observed histopathologically at all dose levels in male rats and at dosages of 100 and 1000 mg/kg/application DGEBPA in female rats (including the high-dose satellite group). The no-observed-effect level (NOEL) for dermal effects for female rats was 10 mg/kg/application. A NOEL for dermal effects for male rats was not determined.

Conclusions:
The NOEL for dermal effects for female rats was 10 mg/kg/application. A NOEL for dermal effects for male rats was not determined.
Executive summary:

Diglycidyl ether of bisphenol A (DGEBPA), CAS# 167554-3, was evaluated for dermal irritation and systemic toxicity potential following approximately 13-weeks of repeated dermal administration. Groups of 10 Fischer 344 rats/sex/dose level were dosed dermally with DGEBPA in acetone solutions at concentrations of 0, 0.9%, 9%, or 90% (w/v) for male rats or 0,0.6%, 6%, or 60% (w/v) for female rats, respectively. The dosing volume was 300 ul application which corresponded to approximate dosages of 0, 10, 100, or 1000 mg DGEBPA/kg body weight/application. Each dose group followed a 5 applications/veek (Monday through Friday) dosing regimen. An additional high-dose group of 10 females (satellite group) followed a 3 application/week (Monday, Wednesday, Friday) dosing regimen. Rats dosed 5 dayslweek received a total of 68 (males) or 69 (females) applications, respectively, during the study. The satellite group rats that were dosed 3 days/week received a total of 41 applications during the study. Data were collected on the following: clinical appearance and behavior, dermal irritation at the test site, body weights, food

consumption, clinical pathology, gross pathology and histopathology including the appearance of skin at the treated and untreated test sites and a specialized histopathological evaluation of the testis following principle of Russell et al., (1990). These methods allowed for a thorough evaluation of the process of spermatogenesis and that all stages of spermatogenesis were intact as described by Clermont and Perry (1957).

DGEBPA applied to the skin of male and female rats five times per week for approximately thirteen weeks at dosages of 10, 100 or 1000 mg/kg/application caused no apparent systemic toxicity with the exception of decreased body weight and body weight gain in males and females at 1000 mg/kg/application. Food consumption was also slightly lower for both males and females throughout the duration of the study. Serum cholesterol values were increased in a dose-related manner in mid- and high dose level rats of both sexes, but considered of questionable toxicological significance since no correlated histopathological changes were observed. Female rats dosed with 1000 mg/kg/application DGEBPA three times per week (high-dose satellite group) for the same duration showed no signs of systemic toxicity. Histopathologic examination of tissues conducted on all test animals resulted in treatment-related effects only at the site of dermal application of DGEBPA. Epidermal hyperpiasia with chronic inflammation, characterized as chronic dermatitis, was observed histopathologically at all dose levels in male rats and at dosages of 100 and 1000 mg/kg/application DGEBPA in female rats (including the high-dose satellite group). The no-observed-effect level (NOEL) for dermal effects for female rats was 10 mg/kg/application. A NOEL for dermal effects for male rats was not determined.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
100 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Additional information

A two-year oral gavage toxicity/oncogenicity study of Bisphenol A Diglycidyl Ether (BADGE) was initiated in Fischer 344 rats. Groups of 65 Fischer 344 rats/dose level were dosed with BADGE suspended in a Tween** 80 and methylcellulose vehicle at dose levels of 0, 50, 250 or 1000 mg/kg/day. The study was terminated on test days 99 (males) or 101 (females) due to excessive toxicity noted in rats given 250 or 1000 mg/kg/day. Standard toxicologic parameters consistent with OECD guideline #408 were evaluated on a subset of ten rats/sex/dose level.

Progressive decreases in body weights and feed consumption, relative to controls, occurred throughout the study in males and females given 250 or 1000 mg/kg/day. By the end of the study, body weights of males and females given 1000 mg/kg/day were 19.2 and 10.9% lower than controls, respectively, and body weights of males and females given 250 mg/kg/day were 10.8 and 5.1% lower than controls, respectively. Body weights of females given 50 mg/kg/day were 3.2% lower than controls, while body weights of males given 50 mg/kg/day were comparable to controls throughout the study.

Treatment-related alterations in hematology (decreases in red blood cell count, hemoglobin concentration, and hematocrit), clinical chemistries, urinalysis, and organ weights occurred in rats given 250 or 1000 mg/kg/day. Some clinical chemistry and urinalysis alterations were indicative of renal toxicity, while organ weight alterations were reflective of lower feed consumption and body weights. The only treatment-related clinical pathology alteration in male and female rats given 50 mg/kg/day was increased cholesterol. The only treatment-related gross pathologic observation was increased size of the cecum in males given 250 or 1000 mg/kg/day, and in females given 1000 mg/kg/day. Two males and one female given 1000 mg/kg/day that died prior to study termination had moderate to severe acute necrosis of renal proximal tubules. Treatment related histopathologic effects in surviving animals given 250 and/or 1000 mg/kg/day were noted in the adrenal glands, cecum, ileum, kidneys, liver, testes, and uterus. Males and females given 50 mg/kg/day had no treatment-related histopathologic effects.

Based on alterations in body weights and serum cholesterol in rats given 50 mg/kg/day, a no-observed-effect level (NOEL) was not determined. However, since these alterations were not associated with detrimental effects, the dose of 50 mg/kg/day was interpreted to be the no-observed-adverse-effect level (NOAEL).

Dermal

Diglycidyl ether of bisphenol A (DGEBPA), CAS# 167554-3, was evaluated for dermal irritation and systemic toxicity potential following approximately 13-weeks of repeated dermal administration. Groups of 10 Fischer 344 rats/sex/dose level were dosed dermally with DGEBPA in acetone solutions at concentrations of 0, 0.9%, 9%, or 90% (w/v) for male rats or 0,0.6%, 6%, or 60% (w/v) for female rats, respectively. The dosing volume was 300 ul application which corresponded to approximate dosages of 0, 10, 100, or 1000 mg DGEBPA/kg body weight/application. Each dose group followed a 5 applications/veek (Monday through Friday) dosing regimen. An additional high-dose group of 10 females (satellite group) followed a 3 application/week (Monday, Wednesday, Friday) dosing regimen. Rats dosed 5 dayslweek received a total of 68 (males) or 69 (females) applications, respectively, during the study. The satellite group rats that were dosed 3 days/week received a total of 41 applications during the study. Data were collected on the following: clinical appearance and behavior, dermal irritation at the test site, body weights, food consumption, clinical pathology, gross pathology and histopathology including the appearance of skin at the treated and untreated test sites and a specialized histopathological evaluation of the testis following principle of Russell et al., (1990). These methods allowed for a thorough evaluation of the process of spermatogenesis and that all stages of spermatogenesis were intact as described by Clermont and Perry (1957).

DGEBPA applied to the skin of male and female rats five times per week for approximately thirteen weeks at dosages of 10, 100 or 1000 mg/kg/application caused no apparent systemic toxicity with the exception of decreased body weight and body weight gain in males and females at 1000 mg/kg/application. Food consumption was also slightly lower for both males and females throughout the duration of the study. Serum cholesterol values were increased in a dose-related manner in mid- and high dose level rats of both sexes, but considered of questionable toxicological significance since no correlated histopathological changes were observed. Female rats dosed with 1000 mg/kg/application DGEBPA three times per week (high-dose satellite group) for the same duration showed no signs of systemic toxicity. Histopathologic examination of tissues conducted on all test animals resulted in treatment-related effects only at the site of dermal application of DGEBPA. Epidermal hyperpiasia with chronic inflammation, characterized as chronic dermatitis, was observed histopathologically at all dose levels in male rats and at dosages of 100 and 1000 mg/kg/application DGEBPA in female rats (including the high-dose satellite group). The no-observed-effect level (NOEL) for dermal effects for female rats was 10 mg/kg/application. A NOEL for dermal effects for male rats was not determined.


Justification for classification or non-classification

Changes in organs due to treatment were either consistent with irritation due to a portal of entry effect or were judged conclusive but insufficient for classification.