4,4'-Methylenedianiline, oligomeric reaction products with 1-chloro-2,3-epoxypropane

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

 In a GLP study performed accoring to OECD 414, the NOAEL for embryo-fetal development was considered to be 90 mg/kg/day based on increased litter and fetal incidence of delayed/absent bone ossification and of malformations at 270 mg/kg/day in a context of severe maternal toxicity.

Effect on fertility: via oral route

Endpoint conclusion:

no study available

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Effects on developmental toxicity

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: see 'Remark'

Remarks:

4,4'-methylenebis[N,N-bis(2,3-epoxypropyl)aniline] is methylenebisbenzenamine with four epoxy groups and is considered as a monoconstituent substance under REACH with a purity > 80% and containing the following impurities: triglycidyl-(2-hydroxy-3-chloropropyl)-methylenedianiline in a range of 3.5 to 4%, dimeric TGMDA in a range of 3 to 5%, triglycidyl-(2,3-dihydroxypropyl)-methylenedianiline in a range of 0.3 to 1%, triglycidyl-(2-hydroxypiperidine)-methylenediamine in a range of 0.5 to 1%, triglycidyl methylenedianiline in a range of 0.3 to 0.8%, (2-hydroxy-3-chloropropyl)-dimeric TGMDA in a range of 0.2 to 0.6%, and trimeric TGMDA in a range of 0.2 to 0.5%. The hypothesis is to read-across some data from the monoconstituent substance described above to the corresponding UVCB substance as described under REACH. The UVCB substance being chemically similar and just differ from the monoconstituent substance by the purity of the main constituent which is < 80% but still stay in the same order of magnitude. The impurities from the monoconstituent substance being also present in the same order of magnitude in the UVCB substance containing also two additional substances, 2-oxiranemethanamine, %{N}-[4-[[4-[[3-chloro-2-(oxiranylmethoxy)propyl](oxiranylmethyl)amino]phenyl]methyl]phenyl]-%{N}-(oxiranylmethyl)- in a range of 0.5 to 2 % and less than 1% 1-[(4-{4-[bis(oxiran-2-ylmethyl)amino]benzyl}phenyl)(oxiran-2-ylmethyl)amino]-3-[(4-{4-[(3-chloro-2-hydroxypropyl)(oxiran-2-ylmethyl)amino]benzyl}phenyl)(oxiran-2-ylmethyl)amino]propan-2-ol. The main assumption is that the minor differences in terms of percentage between the mono and UVCB substance are not significant in respect of all properties under consideration. The full report on read-across approach is attached in section 13 of the IUCLID file.

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

GLP compliance:

yes

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

Animals:
-Number: 96 time-mated females rats were received at CiToxLAB France between 28 November and 18 December 2012.
-Strain and sanitary status: Sprague-Dawley, Crl CD® (SD) IGS BR, Caesarian Obtained, Barrier
Sustained-Virus Antibody Free, (COBS-VAF®).Breeder: Charles River Laboratories Italia, Calco, Italy.
-Age/Weight: at the beginning of the treatmen period, the animals were 10-11 weeks old and had a mean body weight of 273 g(223 g to 323 g). The females were sexually mature and primigravid.
-Mating: Females were mated at the breeder's facilities.
Environmental conditions:
-Temperature: 22 +/- 2°C.
-Relative humidity: 50 +/- 20%.
-Light/dark cycle: 12h/12h.
-Ventilation: about 12 cycles/hour of filtered, non-recycled air.
Housing.
-The animals were individually housed in polycarbonate cages (Techniplast 2154, 940 cm2) with stainless steel lids and containing autoclaved sawdust (SICSA, Alfortville, France). Individual housing was chosen because of software limitations and since it is preferable for pregnant animals. Each cages containes enrichment (rat hut).
Food and water.
-All animals had free access to SSNIFF R/M-H pelleted maintenance diet, batch N° 2537604 (SSNIFF Spezialdiäten GmbH, Soest, Germany) which was distributed weekly. The animals had free access to bottles containing tap water (filtered with a 0.22 microm. filter).

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Remarks:

PEG 400 batch N° MKBG6036V

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): The dose formulations were administred daily from day 6 to day 20 p.c., inclusive.


VEHICLE
-Vehicle (if other than water):The vehicle was PEG 400.
- Lot/batch no. (if required):N° MKBG6036V.
- Amount of vehicle (if gavage): 5 ml/kg/day

Analytical verification of doses or concentrations:

yes

Details on mating procedure:

The females were mated at the breeder's facilities. The day of confirmed mating (detection of a vaginal plug) was designated as a day 0 p.c.

Duration of treatment / exposure:

15 days.

Frequency of treatment:

Daily

Duration of test:

Females were sacrified on day 20 p.c.

No. of animals per sex per dose:

Group 1: 24 time-mated females, dose level: 0 mg/kg/day, concentration: 0 mg/ml.
Group 2: 24 time-mated females, dose level: 30 mg/kg/day, concentration: 6 mg/ml.
Group 3: 24 time-mated females, dose level: 90 mg/kg/day, concentration: 18 mg/ml.
Group 4: 24 time-mated females, dose level: 270 mg/kg/day, concentration: 54 mg/ml.

Control animals:

yes

Maternal examinations:

CAGE SIDE OBSERVATIONS:
- Time schedule: Yes


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:Daily

BODY WEIGHT: Yes
- Time schedule for examinations: day 2, 4 ,6, 9, 12, 15, 18 and 21 p.c., and priormto premature sacrifice.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
The quantity of food consumed by each female was recorded for the following intervals: days2-4, 4-6, 6-9, 9-12, 12-15, 15-18 and 18-21 p.c..

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined:
-Thoracic and abdominal organs
-The ovaries and uterus: All organs examined macroscopically
uterus - live and dead fetuses, resorption sites, placentas
ovaries - corpora lutea


OTHER:
- The weight of the gravid uterus was recorded
- placenta weights
- for apparently non-pregnant females the presence of implantation scars on the uterus was checked using the ammonium sulphide staining technique.
- presence of iron in uterus walls with ammonium sulphide to detect invisible implantation sites

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: Number and distribution of dead and live fetuses, number and distribution of uterine scars.

Fetal examinations:

- External examinations: Yes
Each fetus (excluding the autolyzed fetus) was subjected to a detailed external examination, which included the observation of all visible structures, surfaces and orificies.
- Soft tissue examinations: Yes: half per litter.
- Skeletal examinations: Yes: all per litter

Statistics:

- All data are recorded and calculated using computerized validated software (Reprotox version B.1).
- Data of non-pregnant females are not included in group mean calculations such as body weight, body weight change, food consumption and litter data.

Indices:

No data.

Details on maternal toxic effects:

Maternal toxic effects:yes

Dose descriptor:

NOAEL

Effect level:

90 mg/kg bw/day

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

90 mg/kg bw/day

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
At 270 mg/kg/day, mean feta body weight and mean placenta weight were lower than in controls which correlated with the effect seen on mean gravid uterus. when compared with controls, the differences were statistically significant and considered to be adverse.
There were no effects on mean fetal body weight at 30 and 90 mg/kg/day and on mean fetal/placenta weight ratio at any dose-level.

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

Maternal NOAEL: 90mg/kg/day
Fetal NOAEL: 90 mg/kg/day

Executive summary:

The test item, 4,4’-methylenebis[N,N-bis(2,3-epoxypropyl)aniline] (batch No. AAB0290400), was administered by gavage, once daily, from days 6 to 20 p.c., inclusive, to time-mated female Sprague-Dawley rats at dosages of 30, 90 or 270 mg/kg/day. On the basis of the results obtained in this study: . the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 90 mg/kg/day based on the premature death, adverse clinical signs and effects on mean body weight and mean body weight change (including carcass and net body weight) and mean food consumption at 270 mg/kg/day, . the NOAEL for embryo-fetal development was considered to be 90 mg/kg/day based on increased litter and fetal incidence of delayed/absent bone ossification and of malformations at 270 mg/kg/day in a context of severe maternal toxicity.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

90 mg/kg bw/day

Species:

rat

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Toxicity to reproduction: other studies

Additional information

The test item, 4,4’-methylenebis[N,N-bis(2,3-epoxypropyl)aniline] (batch No. AAB0290400), was administered by gavage, once daily, from days 6 to 20 p.c., inclusive, to time-mated female Sprague-Dawley rats at dosages of 30, 90 or 270 mg/kg/day. On the basis of the results obtained in this study: . the No Observed Adverse Effect Level (NOAEL) for maternal parameters was considered to be 90 mg/kg/day based on the premature death, adverse clinical signs and effects on mean body weight and mean body weight change (including carcass and net body weight) and mean food consumption at 270 mg/kg/day, . the NOAEL for embryo-fetal development was considered to be 90 mg/kg/day based on increased litter and fetal incidence of delayed/absent bone ossification and of malformations at 270 mg/kg/day in a context of severe maternal toxicity.

Based on the above mentioned results and combined with the results observed in the repeated oral toxicity study (90 day) where no adverse effects were noted on the reporductive organs, further investigation on the reproductive toxicity potential of the substance is not necessary.

Justification for classification or non-classification

Based on the results observed in the prenatal developmental toxicity study and as no adverse effects were noted during the oral subchronic repeated dose toxicity study, the substance does not need to be classified for reproductive toxicity.