3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-benzopyran-6-ol

Endpoint summary

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.

Qualifier:

equivalent or similar to guideline

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 or test system 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

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 specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: 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

Key result

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:

not determinable due to absence of adverse toxic effects

Key result

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:

not determinable due to absence of adverse toxic effects

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Ophthalmological findings:

not specified

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

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

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not examined

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

> 2 other: %

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

> 800 mg/kg bw/day

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Reproductive effects observed:

no

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

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 D,L-alpha-tocopherol. Therefore, data from the structurally related vitamine E derivative D-alpha-tocopheryl poly(ethylene glycol) 10000 succinate were used. Please see read-across justification section 13.

In a one-generation reproduction toxicity study (Krasavage 1977), rats were fed up to 2.0% D-alpha-tocopheryl poly(ethylene glycol) 10000 succinate (corresponding to 800 mg/kg bw/d (using default values of 20 g food /day and a body weight of 0.5 kg (REACH guidance Chapter R.8: Characterisation of dose [concentration]-response for human health (Table R8 -17)).

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.

The one-generation study revealed no toxicological differences in rats treated with the vitamin E derivative 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:
The results of a one-generation reproduction rat toxicity study (comparable to OECD 415) with the structurally related vitamin E derivative D-alpha-tocopheryl poly(ethylen glycol) 10000 succinate does not indicate that D,L-alpha-tocopherol has adverse effect on the reproductive function. The reproductive indices of the treated groups were unaffected and the offspring developed normally. The NOAEL in this study was > 800 mg/kg bw/d (corresponding to 2% in the feed).

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.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

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 with D,L-alpha-tocopherol. Therefore, data from the structurally related vitamine E ester D,L

-alpha-tocopheryl acetate were used. This compound is rapidly hydrolysed to the vitamin E active form D,L alpha-tocopherol under physiological conditions.

Please see read-across justification section 13.

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 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 EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.