Reaction products of Soya and Linseed Oil Fatty Acids with Bisphenol A diglycidylether

Endpoint summary

Description of key information

subchronic NOAEL = 50 mg/kg bw/d (rat, similar to OECD TG 408) (read-across: BADGE)

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:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
This read-across hypothesis corresponds to scenario 2 of the Read-Across Assessment Framework (RAAF), ECHA, March 2017 - different compounds have qualitatively similar properties - of the read-across assessment framework i.e. properties of the target substance are predicted to be quantitatively equal to those of the source substance. Namely, the source substance BADGE predicts the toxicological and ecotoxicological properties of the target substance Soya/Linseed Oil Fatty Acid-BADGE reaction product.

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
for details see Justification for read-across attached to iuclid section 13

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
for details see Justification for read-across attached to iuclid section 13

3. ANALOGUE APPROACH JUSTIFICATION
for details see Justification for read-across attached to iuclid section 13

4. DATA MATRIX
for details see Justification for read-across attached to iuclid section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

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 or test system 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:

no

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:

subchronic

Species:

rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

No experimental data on repeated dose toxicity are available for Soya/Linseed Oil Fatty Acid-BADGE reaction product. However, a two-generation study as well as a one-generation study with the source substance BADGE are available. Further data on repeated dose toxicity are available for fatty acids. A justification for read-across is attached to iuclid section 13.

 

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).

 

Additional data are also available for fatty acids:

As a result of repeated oral administration of 100, 300 and 1000 mg/ kg bw/d of docosanoic acid to male and female rats, no deaths were observed in any of the administration groups, and no effects of docosanic acid administration on general condition, body weight transition and food intake were observed. In males, the mean corpuscular hemoglobin concentration was significantly decreased in the mid and high dose groups, ALP was significantly reduced in all test groups, and the glucose concentration was significantly low in the high dose group. However, since all the changes were mild and no concomitant changes were observed in other parameters including histopathology tests, these are considered to be incidental changes and have no toxicological relevance. Histopathological examination showed very slight atrophy of the seminiferous tubules in 2 animals in the 1000 mg/kg bw/d dose group, both of which were spontaneous histological images due to localized changes withou effects on reproductive performance. No effects of docosanoic acid administration were observed on maternal gestation length, fertility, parturition and lactation status, as well as the viability, weight, sex ratio and morphology of the offspring.

Based on the above results, the no-effect level for repeated dose toxicity and reproductive and developmental toxicity of docosanoic acid is considered to be 1000 mg/kg bw/day under the conditions of this study.

 

Weanling male Fischer 344 rats were fed either control or diet containing 1% CLA (conjugated linoleic acid) for 18 months. Weight gain and survival rate were not different between treatments, but CLA-fed animals ate slightly less food. CLA feeding did not significantly reduce body fat compared to that of control. Clinical chemistry and hematology analyses were performed on blood samples at week 69–72. CLA had no effects except on blood glucose, which was reduced in CLA-fed animals compared to control. All animals had chronic renal failure at the end of the study; however, CLA decreased the amount of protein in urine at week 70 of feeding. Necropsy and histopathology results indicated that there was no difference between treatment groups. Although this study used a limited number of animals and a single dose of CLA, our results suggest that long term CLA feeding did not cause any adverse effects in rats.

 

The no-effect level for repeated dose toxicity of the fatty acids was far above the NOAEL of BADGE; therefore, the fatty acid moiety is not expected to contribute to a relevant extent to the overall toxicity of Soya/Linseed Oil Fatty Acid-BADGE reaction product. The main determinant is BADGE. The NOAEL of 50 mg/kg bw/d obtained in the subchronic toxicity study with BADGE is used as key value for chemical safety assessment for the target substance Soya/Linseed Oil Fatty Acid-BADGE reaction product.

Justification for classification or non-classification

Based on reliable, adequate and relevant data, Soya/Linseed Oil Fatty Acid-BADGE reaction product does not require classification for specific organ toxicity after repeated exposure according to regulation (EC) 1272/2008.