Registration Dossier

Administrative data

Endpoint:
three-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not Specified
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study report does not specify a guideline followed, however results are quite deatiled for F2 & F3 generations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1970
Report date:
1970

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: Not specified
Principles of method if other than guideline:
Ionox 330 was fed to rats at dietary concentrations of 0 and 5,000 ppm. Observations were made on the health, body weight and food intake of each animal At 14 weeks of age one male and one female from a treatment group were selected at random and caged together for mating. Three litters were bred from each pair for three successive generations. The final litter from the third generation was examined for any evidence of teratogenic effects.
GLP compliance:
no
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol
EC Number:
216-971-0
EC Name:
3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol
Cas Number:
1709-70-2
Molecular formula:
C54H78O3
IUPAC Name:
3,3',3'',5,5',5''-hexa-tert-butyl-α,α',α''-(mesitylene-2,4,6-triyl)tri-p-cresol
Details on test material:
A 100 kg sample of Ionox 330, batch No, 89/5/6232, purity 98%, was supplied by Shell Chemicals U.K.

Test animals

Species:
rat
Strain:
other: Carworth Farm E
Sex:
male/female
Details on test animals or test system and environmental conditions:
80 rats of the Carworth Farm E strain were used. These animals were bred and maintained under specific-pathogen-free conditions at this laboratory.
Groups of 20 male and 20 female rats, aged three weeks (FO), received diets (Diet 86, Scientific Products Farm).
The animals were caged in groups of two of each sex and given the appropriate diet. At 14 weeks of age, one male and one female from within a treatment group were selected at random and caged together for mating.

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
Not specified
Details on mating procedure:
At 14 weeks of age, one male and one female from within a treatment group were selected at random and caged together for mating. The pair remained together throughout the experiment and three litters were bred (F1a, F1b and F1c).
Litters F1a and F1c were killed at weaning. From each treatment group, one male and one female from each of the weaned second litters (F1b) were selected at random and fed the appropriate treatment diet. The rats were kept as for the FO generation, paired and mated when 100 days old. Three litters were bred (F2a, F2b and F2c), a selection made as before from F2b to produce the third generation litters, F3a, F3b and F3c.
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
Not specified
Frequency of treatment:
Daily
Details on study schedule:
At 14 weeks of age, one male and one female from within a treatment group were selected at random and caged together for mating. The pair remained together throughout the experiment and three litters were bred (F1a, F1b and F1c). Litters F1a and F1c were killed at weaning. From each treatment group, one male and one female from each of the weaned second litters (F1b) were selected at random and fed the appropriate treatment diet. The rats were kept as for the FO generation, paired and mated when 100 days old. Three litters were bred (F2a, F2b and F2c), a selection made as before from F2b to produce the third generation litters, F3a, F3b and F3c.
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 5000 ppm
Basis:
nominal in diet
No. of animals per sex per dose:
FO generation: 20 males & 20 females per dose group
Control animals:
yes, plain diet
Details on study design:
Not specified
Positive control:
Not specified

Examinations

Parental animals: Observations and examinations:
Daily observations were made on general health and behaviour of all animals. Records were kept of dates of mating, intervals between mating and number of pregnancies. The number of young in each litter were counted at birth, after five days and at weaning (21 days). Litters were culled to 12 at birth.
Oestrous cyclicity (parental animals):
Not specified
Sperm parameters (parental animals):
Not specified
Litter observations:
Daily observations were made on general health and behaviour of all animals. Records were kept of dates of mating, intervals between mating and number of pregnancies. The number of young in each litter were counted at birth, after five days and at weaning (21 days). Litters were culled to 12 at birth.
Postmortem examinations (parental animals):
Not specified
Postmortem examinations (offspring):
Not specified
Statistics:
Statistical analyses of body and organ weights were made using the initial body weight as a covariate in covariance analysis.
The data from the litters were examined using analysis of variance.
The significance of any difference between the treatment and control groups of animals was measured using the Student 't' test.
Reproductive indices:
Data produced in the results
Offspring viability indices:
Data produced in the results

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed

Details on results (P0)

Not specified

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
> 5 000 ppm (nominal)
Sex:
male/female
Dose descriptor:
NOAEL
Effect level:
> 5 000 ppm (nominal)
Sex:
male/female
Remarks on result:
other: Generation: F3 (migrated information)

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Details on results (F1)

Fetal findings: F3c Weanling Rats – Findings
Dietary concn (ppm): 0
Pair No 80a: Male and female – peribronchial cuffing; medial hypertrophy of branches of pulmonary artery; foci of alveolar macrophages in lung; few basophilic tubles in kidney; serous cysts in ovary.
Pair No 81: Male – Focal myocardial fibrosis; moderately severe chronic nephrosis. Female – Focal centrilobular fatty change in liver; few basophilic tubles in kidneys.
Pair No 82: Male and female – broken colloid in thyroid; focal myocardial fibrosis; mild to focal peribronchial cuffing and mild to moderately severe chronic nephrosis. Medial hypertrophy of branches of pulmonary artery; focal ductule hyperplasia in liver; a focus of fatty change in liver parenchyma (male only); mild peribronchial cuffing; generalised fatty change in liver with increased mitotic activity (female only).
Pair No 83: Male – Peribronchial cuffing; mild chronic nephrosis. Female – Peribronchial cuffing and aggregates of alveolar macrophages in lung.
Pair No 84: Male and female – some broken colloid in thyroid; peribronchial cuffing and aggregates of alveolar macrophages in lung; mild to chronic nephrosis. Centrilobular fatty change in liver with foci of ductile hyperplasia; focal myocardial fibrosis (male only).
Pair No 85: Male – some broken colloid in thyroid; focal myocardial fibrosis; moderately severe chronic nephrosis. Female – few basophilic tubes in kidneys.
Pair No 86: Male and female – focal myocardial fibrosis; mild and focus of peribronchial cuffing and medial hypertrophy in branches of pulmonary artery; mild chronic and focal nephrosis. Medial hypertrophy of branches of splenic artery (female only).
Pair No 87: Male – focal myocardial fibrosis; focal peribronchial cuffing; mild chronic nephrosis. Female – poorly stained thyroid colloid; peribronchial cuffing; medial hypertrophy of a branch of pulmonary artery; foci of alveolar macrophages; focal fatty change in the liver; mild chronic nephrosis; deciduoma in uterus.
Pair No 88: Male – poorly stained thyroid colloid; medial hypertrophy of branches of coronary artery; focal peribronchial cuffing; a focus of ductile hyperplasia in liver; mild chronic nephrosis; unilateral atrophy of semiferous epithelium of testes. Female – Focal myocardial fibrosis; peribronchial cuffing; raised mitotic rate in liver; mild chronic nephrosis.
Pair No 89: Male and female –peribronchial cuffing and medial hypertrophy of branch of pulmonary artery; mild chronic nephrosis. Poorly stained thyroid colloid (male only); few aggregates of alveolar macrophages in lung (female only).
Pair No 91: Male – poorly stained thyroid colloid; focal myocardial fibrosis; centrilobular fatty change in liver. Female – peribronchial cuffing and aggregates of alveolar macrophages in lung; mild chronic nephrosis.
Pair No 92: Male and female – focal myocardial fibrosis; mild peribronchial cuffing and medial hypertrophy of branches of pulmonary artery. Poorly stained thyroid colloid; minute foci of myocarditis in papillary muscle of heart; mild ductile proliferation in liver; mild chronic nephrosis (male only); dilated kidney tubles, a few lined by basophilic epithelium (female only).
Pair No 93: Male and female –mild peribronchial cuffing; medial hypertrophy of a branch of pulmonary artery. Poorly stained thyroid colloid; aggregates of alveolar macrophages with lymphocytes; isolated nephritic tubles in kidneys (male only); foci of alveolar macrophages; mild chronic nephrosis; medial hypertrophy of a branch of renal artery (female only).
Pair No 94: Male and female –mild to focal peribronchial cuffing; focal lipid vacuolation in liver. Poorly stained thyroid colloid; foci of myocardial fibrosis; severe chronic nephrosis (male only); foci of alveolar macrophages in lung; few basophilic tubles in kidney; uterine carcinoma (female only).
Pair No 95: Male and female –mild peribronchial cuffing; foci and few aggregates of alveolar macrophages in lung; mild chronic nephrosis. Poorly stained thyroid colloid; focus of ductile hyperplasia in liver (male only); medial hypertrophy of branches of pulmonary artery (female only).
Pair No 96: Male and female –medial hypertrophy of a branch of pulmonary artery. Mild peribronchial cuffing; foci of alveolar macrophages; mild chronic nephrosis with protein droplet formation in tubular epithelium (male only); few basophilic tubles in kidneys (female only).
Pair No 97: Male and female – mild peribronchial cuffing in lungs; few basophilic tubles in kidneys. Minute foci of myocarditis; foci of alveolar macrophages; medial hypertrophy of a branch of pulmonary artery (female only).
Pair No 98: Male – poorly stained thyroid colloid; mild peribronchial cuffing in lung; mild chronic nephrosis. Female – focal myocardial fibrosis; focal fatty change and patchy congestion in liver; mild chronic nephrosis.
Pair No 99: Male – poorly stained thyroid colloid; mild peribronchial cuffing; medial hypertrophy of branch of pulmonary artery; mild chronic nephrosis. Female – mild peribronchial cuffing; few basophilic tubles in kidney; deciduoma in uterus.

Dietary concn (ppm): 5000
Pair No 100: Male and female – poorly stained thyroid colloid; focal myocardial fibrosis; peribronchial cuffing; foci of alveolar macrophages in lung; mild chronic nephrosis. Medial hypertrophy of branches of pulmonary artery; focal centrilobular fatty change in liver (male only).
Pair No 101: Male and female –mild peribronchial cuffing; mild chronic nephrosis. Poorly stained thyroid colloid (male only); focal myocardial fibrosis; medial hypertrophy of branches of pulmonary artery; foci of alveolar macrophages in lungs (female only).
Pair No 102: Male – poorly stained thyroid colloid; focal myocardial fibrosis; mild peribronchial cuffing; mild chronic nephrosis. Female – focal peribronchial cuffing; medial hypertrophy of a branch of pulmonary artery; foci of alveolar macrophages in lung; mild chronic nephrosis; ovarian cyst, probably of follicular origin.
Pair No 103: Male and female – focal myocardial fibrosis; mild peribronchial cuffing; medial hypertrophy of a branch of pulmonary artery; foci of alveolar macrophages in lungs; mild chronic nephrosis. Minute congenital cyst in parathyroid (male only); small hyperplastic nodules in adrenal cortex (female only).
Pair No 104: Male – poorly stained thyroid colloid; peribronchial cuffing; centrilobular fatty change in liver; moderately severe chronic nephrosis; focal ductile proliferation in pancreas. Female – focal myocardial fibrosis; mild chronic nephrosis.
Pair No 105: Male and female –mild to focal peribronchial cuffing; medial hypertrophy of a branch of pulmonary artery; fatty change in liver. Poorly stained thyroid colloid; focal myocardial fibrosis; mild chronic nephrosis (male only); a few basophilic tubles in kidneys with calcification of renal papillae (female only).
Pair No 106: Male and female –focal peribronchial cuffing; medial hypertrophy of branches of pulmonary artery; mild to severe chronic nephrosis. Poorly stained thyroid colloid; focal myocardial fibrosis; foci of alveolar macrophages; an area of vaculoation and degeneration of acinar cells of exocrine pancreas; associated with mild ductile hyperplasia (male only).
Pair No 107: Male – poorly stained thyroid colloid; focal peribronchial cuffing; medial hypertrophy of some branches of pulmonary artery; a focus of ductile hyperplasia in liver. Female – focal peribronchial cuffing; a focus of alveolar macrophages in lung; periportal fatty change and increase in mitotic rate in liver; a focus of ductile hyperplasia in pancreas and vacuolation and degeneration of a few acinar cells.
Pair No 108: Male and female –focal myocardial fibrosis; peribronchial cuffing; medial hypertrophy of some branches of pulmonary artery; mild to severe chronic nephrosis. Poorly stained thyroid colloid; a focus of ductile proliferation in pancreas (male only); focal ductile hyperplasia in a few portal areas in liver (female only).
Pair No 109: Male and female –peribronchial cuffing; medial hypertrophy of some branches of pulmonary artery; mild chronic nephrosis. Poorly stained thyroid colloid; focal myocardial fibrosis (male only); congenital cyst in the thyroid; foci of alveolar macrophages in lung; a decidumoa in uterus (female only).
Pair No 110: Male and female –mild peribronchial cuffing; medial hypertrophy of some branches of pulmonary artery; focal fatty change in liver; mild to moderately severe chronic nephrosis. Poorly stained thyroid colloid; minute foci of ductile hyperplasia in pancreas (male only); focal myocardial fibrosis (female only).
Pair No 111: Male – poorly stained thyroid colloid; advanced fibrosis of papillary muscle in the heart; peribronchial cuffing; arteriosclerotic lesion of part wall of branch of pulmonary artery; foci of alveolar macrophages in lungs; excess haemosiderin in red pulp of the spleen; severe chronic nephrosis. Female – focal myocardial fibrosis; focal peribronchial cuffing; medial hypertrophy of a branch of pulmonary artery; a few basophilic tubles in kidneys.
Pair No 112: Male and female –focal myocardial fibrosis; peribronchial cuffing; medial hypertrophy of branches of pulmonary artery. Poorly stained thyroid colloid; chronic nephrosis (male only); a few basophilic tubles in kidneys (female only).
Pair No 113: Male and female –medial hypertrophy of branches of pulmonary artery; mild chronic nephrosis. Poorly stained thyroid colloid; focal myocardial fibrosis (male only); peribronchial cuffing; foci of alveolar macrophages in lung (female only).
Pair No 114: Male – poorly stained thyroid colloid; peribronchial cuffing; medial hypertrophy of branches of pulmonary artery; foci of alveolar macrophages in lung; moderately severe chronic nephrosis; focal vacuolation of exocrine secretory cells and mild ductile proliferation in pancreas. Female – peribronchial cuffing; minute focus of ductile hyperplasia in liver; moderately severe chronic nephrosis.
Pair No 115: Male – poorly stained thyroid colloid; ultimo-branchial cysts; focal myocardial fibrosis; peribronchial cuffing; mild chronic nephrosis.
Pair No 116: Male – poorly stained thyroid colloid; focal myocardial fibrosis; medial hypertrophy of branches of pulmonary artery; very severe chronic nephrosis. Female – medial hypertrophy of a branch of pulmonary artery; foci of alveolar macrophages in lung; pyelonephritis; a focus of ductile hyperplasia in pancreas; cystitis; focal arteriosclerosis of the aorta.
Pair No 117: Male and female – poorly stained thyroid colloid; focal myocardial fibrosis; mild peribronchial cuffing; medial hypertrophy of branches of pulmonary artery; foci of alveolar macrophages in lung. Mild chronic nephrosis (male only); few basophilic tubles in kidneys; arteriosclerot lesion in wall of major artery in thorax (female only).
Pair No 118: Male – poorly stained thyroid colloid; focal peribronchial cuffing; focus of ductile hyperplasia liver; mild chronic nephrosis. Female – mild peribronchial cuffing; medial hypertrophy of some branches of pulmonary artery; centrilobular fatty change in lover; mild chronic nephrosis; medial hypertrophy of branch of pancreatic artery.
Pair No 119: Male and female – peribronchial cuffing and focal hypertrophy of media branches of pulmonary artery; mild chronic nephrosis.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
> 5 000 ppm (nominal)
Sex:
male/female

Results: F2 generation

Effect levels (F2)

Dose descriptor:
NOAEL
Generation:
F2
Effect level:
> 5 000 ppm (nominal)
Sex:
male/female

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

Reproductive performance of rats fed Ionox 330

Dietary concn (ppm)

No of pregnancies

No of young born

No of young born alive

No of young alive after 21 days

Fertility (a) Index

Gestation (b) Index

Viability (c) Index

Lactation (d) Index

 

GENERATION 1

 

Litter F1a

0

19/20

220

(11.6)+

218

(11.5)+

181

95

100

99.1

83.0

5000

19/20

221

(11.6)

218

(11.5)

158

95

100

98.6

72.5

 

Litter F1b

0

19/20

178

(9.4)

178

(9.4)

140

95

100

100

78.7

5000

19/20

171

(9.0)

171

(9.0)

130

95

100

100

76.0

 

Litter F1c

0

19/20

187

(9.8)

187

(9.8)

160

95

100

100

85.6

5000

19/20

192

(10.1)

192

(10.1)

167

95

100

100

87.0

 

GENERATION 2

 

Litter F2a

0

20/20

236

(11.8)

236

(11.8)

173

100

100

100

73.3

5000

20/20

223

(11.2)

223

(11.2)

207

100

100

100

92.8

 

Litter F2b

0

20/20

190

(9.5)

190

(9.5)

175

100

100

100

92.1

5000

20/20

190

(9.5)

190

(9.5)

183

100

100

100

96.3

 

Litter F2c

0

20/20

209

(10.5)

209

(10.5)

189

100

100

100

90.4

5000

*19/20

169

(8.9)

169

(8.9)

157

95

100

100

92.9

 

GENERATION 3

 

Litter F3a

0

19/20

219

(11.5)

219

(11.5)

204

95

100

100

93.2

5000

20/20

232

(11.6)

232

(11.6)

217

100

100

100

93.5

 

Litter F3b

0

18/20

189

(10.5)

189

(10.5)

181

90

100

100

95.8

5000

20/20

189

(9.5)

189

(9.5)

172

100

100

100

91.0

 

Litter F3c

0

18/20

178

(9.9)

178

(9.9)

165

90

100

100

92.7

5000

20/20

207

(10.4)

207

(10.4)

191

100

100

100

92.3

       * Female sent to pathology

+ Average number of pups/litter

(a)      - % matings resulting in pregnancies

(b)                - % pregnancies resulting in birth of live litters

(c)     - % of young born alive after 2 days

(d)  - % young alive after 2 days surviving to weaning at 21 days

Applicant's summary and conclusion

Conclusions:
In conclusion feeding Ionox 330 to rats at a dietary concentration of 5,000 ppm for three generations produced no effects upon reproduction. No gross or histological evidence was found of congenital abnormalities attributable to Ionox 330.
Executive summary:

Ionox 330 was fed to rats at dietary concentrations of 0 and 5,000 ppm. Observations were made on the health, body weight and food intake of each animal At 14 weeks of age one male and one female from a treatment group were selected at random and caged together for mating. Three litters were bred from each pair for three successive generations. The final litter from the third generation was examined for any evidence of teratogenic effects.

Ionox 330 when fed to rats at a dietary concentration of 5,000 ppm for three generations produced no effects upon reproduction. No gross or histological evidence was found of any congenital abnormalities attributable to Ionox 330.