Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
one-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented publication which meets basic scientific principles, please refer to IUCLID section 13 for read across justification.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
other: COBS, CD, albino
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River
- Age at study initiation: no data
- Weight at study initiation: no data
- Fasting period before study: no data
- Housing: five per cage in suspended wire-bottom cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on mating procedure:
- M/F ratio per cage: 1/1
- Length of cohabitation: 2 weeks
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: no data
- After successful mating each pregnant female was caged: the male was removed from the cage, a nesting pan was introduced and the female was allowed to litter
- Any other deviations from standard protocol: no data
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
The parent generation was treated for 264-268 days.
All offspring were killed after 5 weeks of weaning.
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:
0.002, 0.2, 2.0% (8, 80 and 800 mg/kg bw/day)
Basis:
actual ingested
No. of animals per sex per dose:
15
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: no data
- Rationale for animal assignment: random
Positive control:
None
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: no data


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: no data


BODY WEIGHT: Yes
- Time schedule for examinations: prior to treatment, twice during the first week of feeding and weekly thereafter


FOOD CONSUMPTION AND COMPOUND INTAKE :
- Food consumption for each animal determined and mean daily diet consumption calculated as mg food/rat/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data
Oestrous cyclicity (parental animals):
No data
Sperm parameters (parental animals):
No data
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no


PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
insemination, fertility, gestation, viability, lactation, average gestation (days), average litter size, sex (M/F), mean mortality at birth, mean mortality 0-8 weeks


GROSS EXAMINATION OF DEAD PUPS:
No data
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals after 265-268 days of treatment when it was certain no additional matings were necessary.
- Maternal animals: All surviving animals after 265-268 days of treatment when it was certain no additional matings were necessary.


GROSS NECROPSY
- Gross necropsy consisted of: no data


HISTOPATHOLOGY / ORGAN WEIGHTS
organ weights, hematocrit, hemoglobin concentration, total and differential white cell counts, serum glutamic oxalacetic transaminase, serum alkaline phosphatase, urea nitrogen, lactic acid dehydrogenase, serum glucose, serum total protein, triglyceride and cholesterol.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring were sacrificed at 5 weeks after weaning.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:


GROSS NECROPSY
- Gross necropsy consisted of: no data


HISTOPATHOLOGY / ORGAN WEIGTHS
No data
Statistics:
Data in all studies were analyzed statistically by analysis of variance, Duncan's multiple range ste, or Student's T-test p<0.05.
Reproductive indices:
Yes
Offspring viability indices:
Yes
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:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Dose descriptor:
NOAEL
Effect level:
> 2 other: %
Sex:
male/female
Basis for effect level:
other: Dose was converted to 800 mg/kg bw/day using default values. No treatment related effects were observed.
Remarks on result:
other: Generation: P and F1 (migrated information)
Dose descriptor:
NOAEL
Effect level:
800 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: No treatment-related effects.
Remarks on result:
other: Generation: P and F1 (migrated information)
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
Reproductive effects observed:
not specified

No apparent differences in reproductive indices were seen between controls and treated groups of parents. Mean gestation period, litter size, sex ratio, and mortality of pups and parents were unaffected by the test substance.

Hematology and clinical chemistry done after 255 days of treatment revealed no toxicologic differences between control and high dose group. None of the organ weights (neither absolute nor relative) of the treated groups were significantly different from control. Microscopic examination of tissues of high dose and control F0 and F1b animals revealed no morphological changes that were attributable to ingestion of the test substance.

Ingestion of the test substance had no effect on body weight gain of the pups.

Based on default values in the REACH guidance Chapter R.8: Characterisation of dose [concentration]-response for human health (Table R8 -17); using the daily intake of 20 gram/day: and the average body weight of the rats of 0.5 kg (data of the 90 day study), the NOAEL of 2% is converted to 800 mg/kg bw/day.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
800 mg/kg bw/day
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No studies have specifically conducted with RRR-(alpha-, beta-, gamma-, delta)-tocopherol. Therefore, the substance is evaluated on the basis of data from the structurally related substances, e.g. vitamine E derivative D-alpha-tocopheryl-poly(ethylene glycol) 10000 succinate and D-alpha-tocopheryl acetate. For further information please refer to read-across hypothesis and justification.

The key study is done with the vitamine E derivative D-alpha-tocopheryl-poly(ethylene glycol) 10000 succinate.

In an one-generation reproduction toxicity study (Krasavage 1977), rats were fed with up to 2.0% D-alpha-tocopheryl poly(ethylene glycol) 10000 succinate (corresponding to 800 mg/kg bw/d).

No apparent differences in reproductive indices were seen between controls and treated groups of parents. Mean gestation period, litter size, sex ratio, and mortality of pups and parents were unaffected by the test substance.

Hematology and clinical chemistry done after 255 days of treatment revealed no toxicologic differences between control and high dose group. None of the organ weights (neither absolute nor relative) of the treated groups were significantly different from control. Microscopic examination of tissues of high dose and control F0 and F1b animals revealed no morphological changes that were attributable to ingestion of the test substance. Ingestion of the test substance had no effect on body weight gain of the pups.

No two-generation reproduction toxicity study is available for vitamin E. However, after the evaluation of the reproduction toxicity data and other repeated dose toxicity data it is concluded that a two-generation study is not needed (as discussed below).

 

In the repeated dose toxicity studies (Morrissey, 1988) no effects were noted for the reproductive organs (ovary, uterus, prostate, epididymis and testes), only the absolute testis weights were decresed.There was no significant effect of treatment with the test substance on body weight, absolute and relative right cauda weight, absolute and relative right epididymis weight, relative right testis weight, sperm motility, sperm density, or incidence of abnormal sperm (Morrissey, 1988).

 

In another study female rats were fed DL-alpha-tocopherol for 3 months prior to mating with untreated males (Yang and Desai. 1977). This study described effects in the highest group tested at 10,000 ppm (~ 555 mg/kg bw): only 1/5 rat of the high-dose group was pregnant, while pregnancy was 100% in all other groups (treated at 25, 250 and 2500 ppm). The study is not sufficiently reported. The health status and behaviour of the females is not properly described. Thus, it cannot be excluded that the lower fertility index which was observed in the highest dose group is due to maternal toxicity. Furthermore, 5 females per dose group are not sufficient for a statistical evaluation.

 

The one-generation study as the most reliable guideline study, revealed no toxicological differences in rats of d-alpha-tocopheryl poly(ethylene glycol) 10000 succinate treated group compared to the control group and no effects were observed in rats of the treated group after microscopic examination or in body weight gain of the pups.

 

Taken together, these data are considered sufficient to cover the endpoint effects on reproduction.


Short description of key information:
Key: RA:59-02-9, OECD 415, NOAEL = 800 mg/kg

Justification for selection of Effect on fertility via oral route:
Most reliable study which is used for classification.

Effects on developmental toxicity

Description of key information
In teratogenicity studies (OECD 414, pre-GLP) with rats and rabbits with the Vitamin E ester D,L-alpha tocopheryl acetate, no significant difference in malformations between the control and the treated groups. The NOAEL is >= 1600 mg/kg bw/d for both species.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented study report which meets basic scientific principles; according to OECD Guideline 414 and pre-GLP study, please refer to IUCLID section 13 for read across justification.
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
no
Remarks:
pre-GLP study
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: virgin adult; no further data
- Weight at study initiation: no data
- Fasting period before study: no data
- Housing: individually in mesh bottom cages
- Diet (e.g. ad libitum): no data
- Water (e.g. ad libitum): no data
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS: no data
Route of administration:
oral: gavage
Vehicle:
corn oil
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
days 6 through 15 of gestation
Frequency of treatment:
daily
Duration of test:
until day 20 of gestation
Remarks:
Doses / Concentrations:
16, 74.3, 345, 1600 mg/kg bw/d
Basis:
no data
No. of animals per sex per dose:
24-25
Control animals:
yes, sham-exposed
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: day 0, 6, 11, 15, 20 of gestation

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day # 20
- Organs examined: urogenital tract
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No data
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: one third per litter
- Skeletal examinations: Yes: two thirds per litter
- Head examinations: No data
- Other: number of dead and live fetuses, weight of live fetuses
Statistics:
no data
Historical control data:
no data
Dose descriptor:
NOAEL
Effect level:
> 1 600 mg/kg bw/day
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEL
Effect level:
> 1 600 mg/kg bw/day
Basis for effect level:
other: embryotoxicity
Dose descriptor:
NOAEL
Effect level:
> 1 600 mg/kg bw/day
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
not specified

No deaths occurred. Treatment with the test substance had no clearly discernible effect on nidation or on maternal or fetal survival. The number of abnormalities of soft or skeletal tissues of the test group was not significantly different from sham-treated controls.

Survival of dams at gd 20:

25/25 (16 mg/kg tocopheryl acetate)

25/25 (74.3 mg/kg tocopheryl acetate)

25/25 (345 mg/kg tocopheryl acetate)

24/24 (1600 mg/kg tocopheryl acetate)

25/25 (sham treated control group)

25/25 (positive control group)

Maternal body weight at gd 0:

211 - 228 g (tocopheryl acetate groups)

204 g (sham treated control group)

214 g (positive control group)

Maternal body weight at gd 20:

333 - 348 g (tocopheryl acetate groups)

316 g (sham treated control group)

298 g (positive control group)

Pregnancies:

21 - 23 (tocopheryl acetate groups)

25 (sham treated control group)

23 (positive control group)

Abortions until gd 20:

0 (tocopheryl acetate groups)

0 (sham treated control group)

0 (positive control group)

Total number of live litters:

21 - 22 (tocopheryl acetate groups)

25 (sham treated control group)

19 (positive control group)

Total number of implantation sites:

246 - 266 (tocopheryl acetate groups)

241 (sham treated control group)

262 (positive control group)

Total numbers of resorptions:

1 - 7 (tocopheryl acetate groups); no complete resorption was observed

5 (sham treated control group); no complete resorption was observed

79 (positive control group); complete resorption was observed in 17.4% of the dams

Total number of live fetuses:

239 - 257 (tocopheryl acetate groups)

236 (sham treated control group)

180 (positive control group)

Average number of live fetuses per dam:

10.9 - 11.5 (tocopheryl acetate groups)

9.44 (sham treated control group)

7.83 (positive control group)

Sex ratio (m/f):

0.80 - 1.05 (tocopheryl acetate groups)

0.79 (sham treated control group)

0.87 (positive control group)

Total number of dead fetuses:

8 (74.3 mg/kg bw tocopheryl acetate group), 0 (other tocopheryl acetate groups)

0 (sham treated control group)

3 (positive control group)

Average fetus weight:

3.83 - 3.95 g (tocopheryl acetate groups)

3.82 g (sham treated control group)

2.49 g (positive control group)

There were no significant differences in any parameter between the groups treated with the test substance. No dose-response relationship was noted. Malformations of the sternebrae, ribs, vertebrae, skull, and extremities were recorded; however, the incidence of these malformations in vitamin E-treated groups were not significantly different from those observed in sham-treated control. Soft tissue variations in the tocopheryl acetate groups were distributed over all groups without indicating any dose-response. These variations included cardiomegaly (3 fetuses), anophthalmia (1 fetus), hydrocephalus (3 fetuses), gastroschisis (2 fetuses), apulmonism (1 fetus), and petechiae (1 fetus). No soft tissues are reported for the sham-treated control fetuses. In the positive control group treated with Aspirin, the expected statistically significant increase in the incidence of skeletal malformations and soft tissue variations was observed.

Executive summary:

The administration of up to 1600 mg/kg bw of the test material to pregnant rats for 10 consecutive days had no clearly discernible effect on nidation or on maternal or fetal survival . The number of-abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham-treated controls.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 600 mg/kg bw/day
Species:
rat
Quality of whole database:
The whole database is reliable.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

No studies have specifically been conducted withRRR-(alpha-, beta-, gamma-, delta)-tocopherol. Therefore, data from the structurally related vitamine E esterDL--alpha-tocopheryl acetate were used. This compound is rapidly hydrolysed to the vitamin E active form D,L alpha-tocopherol under physiological conditions. For lease see read-across hypothesis and justification.

In a teratogenicity study, rats were fed up to 1600 mg/kg bw/day D,L-alpha-tocopheryl acetate (Food&Drug Research Laboratory, 1973).

There were no significant differences in any parameter between the groups treated with the test substance. No dose-response relationship was noted. Malformations of the sternebrae, ribs, vertebrae, skull, and extremities were recorded.However, the incidences of these malformations in vitamin E-treated groups were not significantly different from those observed in sham-treated control.Soft tissue findings in the tocopheryl acetate groups were distributed over all treated groups without indicating any dose-response. These included cardiomegaly (3 fetuses), anophthalmia (1 fetus), hydrocephalus (3 fetuses), gastroschisis (2 fetuses), apulmonism (1 fetus), and petechiae (1 fetus). No soft tissues findings are reported for the sham-treated control fetuses. In the positive control group treated with Aspirin, the expected statistically significant increase in the incidence of skeletal malformations and soft tissue variations was observed.

Additionally, the teratogenic activity of D,L-alpha-tocopheryl acetate was investigated in rabbits (Food&Drug Research Laboratory, 1973). The test substance was administered by gavage at dose levels of 0 (sham-treated control group), 16, 74.3, 345, and 1600 mg/kg bw/d. The administration of up to 1600 mg/kg bw/d of the test material to pregnant rabbits for 13 consecutive days had no clearly discernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham-treated controls.


Justification for selection of Effect on developmental toxicity: via oral route:
Most reliable study was chosen for classification.

Justification for classification or non-classification

Based on the available data, vitamin E does not need to be classified for effects on fertility and developmental toxicity according to Annex I of Directive 67/548/EEC and according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.