Amines, polyethylenepoly-

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

No data are available. An EOGRTS is proposed.

Effect on fertility: via oral route

Endpoint conclusion:

no study available (further information necessary)

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

Description of key information

OECD 414: NOAEL maternal toxicity = 200 mg/kg bw/day; NOAEL developmental = 400 mg/kg bw/day (at the highest dose of 800 mg/kg bw/day all maternal animals died)

Further, developmental toxicity studies from read across substance TETA are available. Oral in rat and dermal in rabbit. In both studies no teratogenic effect was observed.

Link to relevant study records

Reference

Reference 1

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-10-3 to 2015-06-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Version / remarks:

Jan 2001

Deviations:

yes

Remarks:

These deviations are considered not to have affected the outcome or the achievement of the study objectives.

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3700 (Prenatal Developmental Toxicity Study)

Version / remarks:

Aug . 1998

Deviations:

yes

Remarks:

These deviations are considered not to have affected the outcome or the achievement of the study objectives.

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories France, Domaine des Oncins, 69210 Saint-Germain-Nuelles, France.
- Age at study initiation: 10 to 13 weeks.
- Weight at study initiation: 191 to 252 g.
- Housing: Females were individually housed in plastic cages in compliance with European Regulations (Directive 2010/63/EU).
- Diet: Rat pelleted commercial complete rodent diet ad libitum (Diet reference Safe A04C-10) sterilised by irradiation and analysed for a predefined list of chemical and bacteriological contaminants. Each batch of diet is supplied with a certificate of analysis which is verified and authorized for release by a veterinarian.
- Water: Softened and filtered (0.2 μm) mains drinking water was available ad libitum (via bottles). Water is analysed twice a year for chemical and bacterial contaminants by Laboratoire Santé Environnement Hygiène de Lyon, France.
-contaminants: No known contaminants were present in diet, water or bedding at levels which might have interfered with achieving the objective of the study.Certificates of analysis for the diet, drinking water and bedding are maintained in the archives of the Test Facility.

-Bedding: dust-free wood shavings (SDS/Dietex, Argenteuil, France) made from spruce tree wood, analysed at least twice a year for chemical and bacterial contaminants. Shredded paper (SDS/Dietex) was provided as enrichment.
- Acclimation period: 6 days between animal arrival and the start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C (target range).
- Humidity (%): 35 to 70 % (target range).
- Air changes (per hr): ≥ 10 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light (artificial)/12 hours dark (except when required for technical acts).

IN-LIFE DATES: From: 2014-10-14 To: 2014-11-06

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

VEHICLE
- The test item was prepared as a solution in the vehicle at concentrations of 20, 40, 60 and 80 mg/mL. The pH of each dose formulation was adjusted to pH 9 + 0.05 with 1 N hydrochloric acid (Fluka). Corrected for the volume of hydrochloric acid required for pH correction. No other correction for active ingredient was required.

- Concentration in vehicle: 0, 20, 40,60, 80 mg/L
- Amount of vehicle (if gavage): 10 ml

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

4 x 2 mL samples were taken from each formulation, including the vehicle, from the first preparation to ensure that the analytically determined concentrations are in line with the nominal concentrations. The samples were maintained at room temperature (between +15 and +25 °C), protected from light. Duplicate (2 x 2 mL) samples were dispatched to the Test Site for analysis using a validated method (except for samples at concentration 60 mg/mL which were not dispatched to the Test Site due to the premature termination of the group). The validation of the analytical method as well as the stability analysis is the subject of a separate study (AmatsiAvogadro Ref: A144871).
Backup (2 x 2 mL) samples were discarded without any further notice to the Study Monitor/Study Sponsor.

Details on mating procedure:

The females were mated by the supplier and delivered on gestation day G 0 (day of mating).

Duration of treatment / exposure:

From day 6 (G 6) to day 19 (G 19) of gestation inclusive.

Frequency of treatment:

once daily

Duration of test:

14 days

Dose / conc.:

800 mg/kg bw/day (actual dose received)

Dose / conc.:

400 mg/kg bw/day (actual dose received)

Dose / conc.:

200 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

22

Control animals:

yes, concurrent vehicle

Details on study design:

selected in agreement with the Study Sponsor based on the results of a dose range-finding study in the pregnant rat (AB20654) with doses of 250, 500 and 1000 mg/kg/day. For the present study, a high dose of 800 mg/kg/day was selected in order to try and elicit slight maternal toxicity. The lower doses were expected to be no adverse effect levels for maternal toxicity. in the main study, the high dose was not tolerated leading to the death or early sacrifice of all animals after several days of dosing (between gestation days 10 and 14). There was no influence of treatment at 250 or 500 mg/kg/day on maternal or embryo-foetal parameters.

Maternal examinations:

5.1 Morbidity/mortality
All animals were observed at least twice daily. Animals found dead or judged to be in a moribund condition were necropsied.
5.2 Clinical signs
All animals were observed daily for clinical signs.
During the treatment period, the animals were observed at least once before and at least once after dosing to detect any clinical signs or reaction to treatment. A full clinical examination was performed on each weighing day.
5.3 Body weight
All animals were weighed individually on days 0, 6, 9, 12, 15, 18 and 20 of gestation.
5.4 Food consumption
Individual food consumption was measured for the periods (days) 0 to 6, 6 to 9, 9 to 12, 12 to 15, 15 to 18 and 18 to 20 during gestation.

Ovaries and uterine content:

− pregnancy status
− gravid uterus weight
− number of corpora lutea
− number and distribution of intrauterine implantations, classified as follows:
− live foetus
− dead foetus
− early resorptions
− late resorptions.
− individual foetal weights
− foetal sex.
The uterus of all females was placed in ammonium sulphide solution in order to stain any previously undetected implantation sites.

Fetal examinations:

− individual foetal weights
− foetal sex.
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: [all per litter / half per litter / #? per litter ] / No / No data
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes: half per litter

Statistics:

Statistical analysis was performed, where appropriate, by the data acquisition software, as follows:
The best transformation for the data (none, log or rank) was determined depending upon
− the normality of the data distribution tested by the Shapiro-Wilk's test
− the homogeneity of the variances across groups tested by the Bartlett's test.
Non- or log-transformed data were analysed by parametric methods.
Rank transformed data were analysed using non-parametric methods.
Data were then analysed to test for a dose-related trend to detect the lowest dose at which there was a significant effect, based on the Williams test for parametric data or the Shirley's test for non-parametric data.
Homogeneity of means was assessed by analysis of variances (ANOVA) for parametric data or Kruskal-Wallis test for non parametric data.
If no trend was found and means were not homogeneous, the data were analysed by parametric or non-parametric Dunnett's test to look for significant differences from the control group.
The number of resorptions, number of dead foetuses and all litter-based percentages were analysed using non-parametric methods, i.e. Kruskal-Wallis test followed by non-parametric Dunnett’s test if the Kruskal-Wallis was significant.
Selected incidence data were analysed using a chi2 test for all groups followed by Fisher’s two-tailed test with Bonferroni correction for each treated group versus the control if the chi2 was significant.
Microsoft Excel 2010® was employed to present certain results.

Indices:

Pre-implantation loss (in %): (Number of corpora lutea - Number of implantations) x 100 / Number of corpora lutea
Post-implantation loss (in %):( Number of implantations - Number of viable fetuses) x 100 / Number of implantations

Historical control data:

Values compiled from untreated and control Wistar rats in previous studies.(2006-2009)

Details on maternal toxic effects:

Maternal toxic effects:yes. Remark: at 400 and 800 mg/kg bw

Details on maternal toxic effects:
Treatment at 800 mg/kg/day
Treatment at 800 mg/kg/day was associated with the unscheduled death or sacrifice (moribund or for ethical reasons) of all females between gestation days 10 and 17. Other than one animal found recumbent with slow breathing and cyanosis, there were no remarkable clinical changes noted for the females found dead (nos. 68, 69, 75 and 77). However, reduced food consumption led to body weight loss for 3 out of 5 females sacrificed in a moribund condition (nos. 70, 71, 74, 76 and 80).

These females also presented numerous clinical changes such as irregular/laboured breathing, intermittent tremors, closed/partly closed eyes, decreased activity/subdued behaviour, paresis, recumbency, palor, cold to touch and piloerection prior to sacrifice.

Reduced food consumption and negligible body weight gain/body weight loss was also noted for the majority of the remaining 13 females sacrificed for ethical reasons (nos. 67, 72, 73, 78, 79 and 81 to 88). One of these females (no. 72) had intermittent tremors and irregular breathing prior to sacrifice. These females, together with nos. 70 and 80 found moribund, received a lower dose of 600 mg/kg for the last two administrations before sacrifice.
The principal macroscopic changes included 12 females with pale kidneys and 1 with a pale liver. The kidney changes were associated with histopathology findings – see below. Other changes such as alopecia were considered to be incidental.

Treatment at 200 and 400 mg/kg/day
Mortality
Consistent with the findings in the high dose group, one female (no. 65) in the 400 mg/kg/day group was sacrificed in a moribund condition on gestation day 17. The female became cold to touch, pale, subdued and presented other clinical signs including decreased activity, irregular breathing, paresis of the forelimbs, partly closed eyes and piloerection after dosing on gestation day 17. There were no macroscopic findings at necropsy.
Clinical signs
Consistent with the female that was sacrificed moribund and clinical changes noted in the high dose group, two females in the 400 mg/kg/day group (nos. 50 and 51) had irregular breathing after the final dose on gestation day 19.
One female in the 200 mg/kg/day group (no. 29) had hypersalivation and noisy breathing noted both before and after treatment and piloerection after dosing on gestation day 9 only.
The isolated nature of these findings on a single day for one female in the low dose group suggests that there was no association with treatment.
Other occasional clinical signs noted (such as hairloss, sores and scabs) were noted in the control and treated groups at a similar frequency and were considered to be incidental.
Body weight
Mean body weight gain was statistically significantly lower (-27 %) than in the control group during the last two days of the study only (gestation days 18 to 20) in the 400 mg/kg/day group. One female with irregular breathing after the last dose (no. 51) lost weight during this period. The effect was confirmed by a statistically significantly lower (-37 %) mean body weight change after adjustment for gravid uterus weight. There was no impact on absolute terminal mean body weight (with or without adjustment for gravid uterus weight). There was no influence of treatment on mean body weight gain in the 200 mg/kg/day group.
Food consumption
Consistent with the effect on body weight (see 10.3.3), mean food consumption was slightly reduced (-13 % - not statistically significant) during the last two days of the study (gestation days 18 to 20) in the 400 mg/kg/day group compared with the control. There was no influence of treatment on mean food consumption in the 200 mg/kg/day group.
Necropsy findings of adult females
There were no treatment-related macroscopic findings in either group. Incidental findings in one or more females included alopecia and dilated renal
pelvis.
Pathology
All 18 animals that were sacrificed moribund or for ethical reasons and in which the kidneys were examined histologically (one of which had been given 400 mg/kg/day and the others 800 mg/kg/day) had minimal to severe degeneration and regeneration of the cortical tubular epithelium in the kidney. This was clearly due to HEPA and was considered adverse. Minimal villous atrophy in the jejunum (and sometimes the duodenum), slight centrilobular necrotic inflammation in the liver, minimal or slight decreased splenic periarteriolar lymphoid sheaths and minimal increased apoptosis in the mesenteric lymph node occurred in the moribund animals only and may have been secondary to the effects of the renal lesions. Minimal or slight secondary pulmonary and splenic changes and local intestinal changes were also seen at a low incidence in animals that were sacrificed moribund and animals that were sacrificed for ethical reasons

Dose descriptor:

NOEL

Effect level:

ca. 200 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Dose descriptor:

LOAEL

Effect level:

ca. 400 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Abnormalities:

effects observed, treatment-related

Localisation:

other: kidney

Description (incidence and severity):

minimal to severe degeneration and regeneration of the cortical tubular epithelium

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects: no effects. Remark: no treatment related effects were noted

Details on embryotoxic / teratogenic effects:
External observations

One foetus from the 400 mg/kg/day group (litter no. 54) had gross disruption of the head with a proboscis. Similar findings have been observed in concurrent studies and this isolated finding was therefore considered to have a genetic aetiology rather than having any association with treatment.
There were no other foetal external abnormalities.
Soft tissue findings
There were no soft tissue malformations in any group.
There was no difference from the concurrent control and/or historical control data in the incidences of other less severe soft tissue anomalies and variations, principally including a transposed umbilical artery and dilated ureters, to suggest any association with treatment.
Skeletal observations
There was a single foetus with skeletal malformations in each of the control, 200 and 400 mg/kg/day groups (litter nos. 18, 40 and 54, respectively). Each foetus had multiple vertebral defects in the thoraco-luumbar region which in the case of the foetus from the 400 mg/kg/day group led to scoliosis. This foetus also had multiple skull defects associated with the malformed head and proboscis noted externally. These isolated findings in each of the groups, including the control, were considered to be spontaneous in origin.
There was no dose-related increase in the incidence of other less severe skeletal anomalies or variations in comparison with the concurrent control and/or the historical control data to suggest any association with treatment.

Dose descriptor:

NOAEL

Effect level:

>= 400 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects up to the highest dose tested

Abnormalities:

no effects observed

Developmental effects observed:

no

no effects were noted on any of the parameters with respect to uterine contents upon caesarian section. (see attached .pdf)

Conclusions:

Under the experimental conditions of the study, oral (gavage) administration of Heavy polyamine (HEPA) at 200, 400 or 800 mg/kg bw/day in the pregnant Wistar rat was not tolerated at the high dose with maternal mortality leading to the premature sacrifice of all remaining females despite an attempt to lower the dose from 800 to 600 mg/kg bw/day. Major clinical signs such as irregular/laboured breathing, intermittent tremors, closed/partly closed eyes, decreased activity/subdued behaviour, paresis, recumbency, palor, cold to touch and piloerection were noted for several of these females prior to death/sacrifice. This dose was also associated with adverse renal changes noted histologically (minimal to severe degeneration and regeneration of the cortical tubular epithelium in the kidney) that were clearly due to HEPA.
Treatment at 400 mg/kg/day group was associated with the unscheduled sacrifice of a single female in a moribund condition, a low incidence of irregular breathing, and reduced mean body weight gain and food consumption towards the end of gestation. The same microscopic renal changes as seen in the high dose group were also noted for the animal sacrificed moribund. The low dose of 200 mg/kg/day was a no observed effect level (NOEL) for maternal findings.
Despite the maternal findings at 400 mg/kg bw/day, there was no embryo-foetal toxicity or indication of a teratogenic potential of Heavy polyamine (HEPA). The dose of 400 mg/kg bw/day was therefore the NOEL for embryo-fetal toxicity and teratogenicity.
The higher dose of 800 mg/kg bw/day could not be evaluated due to the premature termination of the group due to marked maternal toxicity. However, it is worthy of note that 21 females in the group were pregnant and all had live implantations at the time of sacrifice.

Executive summary:

1. Summary

1.1 Objectives

The objective of the study was to evaluate the effects of the test item, heavy polyamine (HEPA), CAS number: 68131-73-7, on embryonic and foetal development of the rat when administered by the oral route (gavage) from implantation to the day before caesarean section.

1.2 Procedures

The test item, heavy polyamine (HEPA), CAS number: 68131-73-7, was administered by daily oral gavage at dose levels of 200, 400 and 800 mg/kg bw/day to 3 groups of 22 mated female Wistar rats from days 6 to 19 of gestation inclusive (except for high dose group which was prematurely sacrificed for ethical reasons despite an attempt to lower the dose to 600 mg/kg bw/day). A control group received the same volume (10 mL/kg bw) of the vehicle (water for injection). Clinical condition, body weight and food consumption were monitored throughout the study. The surviving females were submitted to a caesarean examination on day 20 of gestation and litter parameters were recorded. At necropsy, the females were examined macroscopically and foetuses were weighed, sexed and examined for external abnormalities. Half of the foetuses were examined internally prior to processing for skeletal examination. The remaining half of the foetuses in each litter was fixed in Harrison’s fluid for subsequent examination of the head by serial sectioning. Selected organ/tissue samples were fixed and preserved in 10% neutral formalin from the majority of high dose females sacrificed moribund or for ethical reasons together with one similar female from the intermediate dose group and 3 control females with a terminal caesarean section for subsequent examination histopathological examination.

1.3 Results

Treatment at 800 mg/kg bw/day was associated with the unscheduled death or sacrifice (moribund or for ethical reasons) of all 22 females between gestation days 10 and 17. Numerous clinical changes such as irregular/laboured breathing, intermittent tremors, closed/partly closed eyes, decreased activity/subdued behaviour, paresis, recumbency, palor, cold to touch and piloerection were noted prior to death/sacrifice. The principal macroscopic finding included 12 females with pale kidneys.

The principal histopathology finding was minimal to severe degeneration and regeneration of the cortical tubular epithelium in the kidney of all 17 animals that were sacrificed moribund or for ethical reasons and in which the kidneys were examined histologically.

The following summary is based on findings in the 200 and 400 mg/kg bw/day groups.

Consistent with the findings at 800 mg/kg/day, one female in the 400 mg/kg bw/day group was sacrificed in a moribund condition on gestation day 17 after presenting similar clinical changes.

Consistent with the female that was sacrificed moribund and clinical changes noted in the high dose group, two females in the 400 mg/kg bw/day group had irregular breathing after the final dose on gestation day 19.

Mean body weight gain and food consumption was reduced in the 400 mg/kg bw/day group only during the last two days of the study (gestation days 18 to 20) compared with the control group.

There were no treatment-related macroscopic findings or influence of treatment on mean gravid uterus weight in 200 or 400 mg/kg bw/day group. However, the female sacrificed moribund in the 400 mg/kg bw/day group also had microscopic changes in the kidneys.

There were 22, 21 and 19 pregnant females at terminal caesarean section in the control, 200 and 400 mg/kg bw/day groups, respectively, all of which had viable foetuses.

Although the females did not reach terminal necropsy, it is worthy of note that 21 females in the high dose group were pregnant and all had live implantations at the time of sacrifice (between gestation days 10 and 17).

There was no influence of treatment on embryo-foetal survival, mean foetal weight or sex ratio in the 200 or 400 mg/kg bw/day group. Mean live litter size was comparable with that in the control.

There was a single malformed foetus in each of the control, 200 and 400 mg/kg bw/day groups. Each foetus had multiple vertebral defects in the thoraco-lumbar region. The foetus in the 400 mg/kg bw/day group also had a malformed head with a proboscis. Neither this incidence nor the type of malformations detected suggested any association with treatment. There was no dose-related increase in the incidence of other less severe foetal morphological changes in comparison with the concurrent control and/or the historical control data to suggest any association with treatment.

1.4 Conclusion

Under the experimental conditions of the study, oral (gavage) administration of Heavy polyamine (HEPA) at 200, 400 or 800 mg/kg bw/day in the pregnant Wistar rat was not tolerated at the high dose with maternal mortality leading to the premature sacrifice of all remaining females despite an attempt to lower the dose from 800 to 600 mg/kg bw/day. Major clinical signs such as irregular/laboured breathing, intermittent tremors, closed/partly closed eyes, decreased activity/subdued behaviour, paresis, recumbency, palor, cold to touch and piloerection were noted for several of these females prior to death/sacrifice. This dose was also associated with adverse renal changes noted histologically (minimal to severe degeneration and regeneration of the cortical tubular epithelium in the kidney) that were clearly due to HEPA.

Treatment at 400 mg/kg bw/day group was associated with the unscheduled sacrifice of a single female in a moribund condition, a low incidence of irregular breathing, and reduced mean body weight gain and food consumption towards the end of gestation. The same microscopic renal changes as seen in the high dose group were also noted for the animal sacrificed moribund. The low dose of 200 mg/kg bw/day was a no observed effect level (NOEL) for maternal findings.

Despite the maternal findings at 400 mg/kg bw/day, there was no embryo-foetal toxicity or indication of a teratogenic potential of Heavy polyamine (HEPA). The dose of 400 mg/kg bw/day was therefore the NOEL for embryo-fetal toxicity and teratogenicity.

The higher dose of 800 mg/kg bw/day could not be evaluated due to the premature termination of the group due to marked maternal toxicity. However, it is worthy of note that 21 females in the group were pregnant and all had live implantations at the time of sacrifice.

Reference 2

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

refer to analogue justification provided in IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Species:

rabbit

Dose descriptor:

NOAEL

Remarks:

dermal, rabbit

Effect level:

50 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

mortality

Remarks on result:

other:

Remarks:

CAS 112-24-3, Tyl, 1988

Abnormalities:

no effects observed

Dose descriptor:

NOAEL

Remarks:

dermal, rabbit

Effect level:

>= 125 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

external malformations

skeletal malformations

visceral malformations

Remarks on result:

other:

Remarks:

CAS 112-24-3, Tyl, 1988

Abnormalities:

no effects observed

Developmental effects observed:

no

Conclusions:

The source substance TETA was devoid of any embryotoxic activity and did not reveal teratogenic potency in the rabbit after dermal treatement. As explained in the analogue justification, it is considered that the target and the source substances are unlikely to lead to differences in reproductive toxicity.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

400 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex IX, 8.7, of Regulation (EC) No 1907/2006.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

125 mg/kg bw/day

Study duration:

subacute

Species:

rabbit

Quality of whole database:

The available information comprises adequate, reliable and consistent studies, from a reference substances with similar structure and intrinsic properties. Read-across is justified based on similarities in PC/ECO/TOX properties. The selected study is thus sufficient to fulfil the standard information requirements set out in Annex X, 8.7, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Additional information

A reliable study according to OECD guideline 414 in rats is available for the registered substance (Ethyleneamines RC, 2015). Under the experimental conditions of the study, oral (gavage) administration of polyethylenepolyamine (CAS 68131-73-7) at 200, 400 or 800 mg/kg bw/day in the pregnant Wistar rat was not tolerated at the high dose with maternal mortality leading to the premature sacrifice of all remaining females despite an attempt to lower the dose from 800 to 600 mg/kg bw/day. Major clinical signs such as irregular/laboured breathing, intermittent tremors, closed/partly closed eyes, decreased activity/subdued behaviour, paresis, recumbency, palor, cold to touch and piloerection were noted for several of these females prior to death/sacrifice. This dose was also associated with adverse renal changes noted histologically (minimal to severe degeneration and regeneration of the cortical tubular epithelium in the kidney) that were clearly due to polyethylenepolyamine.

Treatment at 400 mg/kg bw/day group was associated with the unscheduled sacrifice of a single female in a moribund condition, a low incidence of irregular breathing, and reduced mean body weight gain and food consumption towards the end of gestation. The same microscopic renal changes as seen in the high dose group were also noted for the animal sacrificed moribund. The low dose of 200 mg/kg bw/day was a no observed effect level (NOEL) for maternal findings.

Despite the maternal findings at 400 mg/kg bw/day, there was no embryo-foetal toxicity or indication of a teratogenic potential of polyethylenepolyamine. The dose of 400 mg/kg bw/day was therefore the NOEL for embryo-fetal toxicity and teratogenicity.

The higher dose of 800 mg/kg bw/day could not be evaluated due to the premature termination of the group due to marked maternal toxicity. However, it is worthy of note that 21 females in the group were pregnant and all had live implantations at the time of sacrifice.

 

There are no data available on developmental toxicity in a second species with polyethylenepolyamine (CAS 68131-73-7). A final decsion from ECHA is available (CCH-D-2114482139 -42 -01/F) including an OECD 414 in rabbit. This study is initiated and will be inlcuded as soon as available. In the meanwhile, in order to fulfil the standard information requirements set out in Annex X, 8.7., in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006, read-across from a structurally related substance was conducted. In accordance with Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across). Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006 whereby substances may be predicted as similar provided that their physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity.

Triethylenetetramine linear, cyclic and branched = TETA (CAS 90640 -67 -8 or 112 -24 -3) is considered to be similar to polyethylenepolyamine (CAS 68131 -73 -7) on the basis of the structural similar properties and/or activities. Therefore, the available developmental toxicity data on the source substance TETA has been read-across to the target substance polyethylenepolyamine (CAS 68131 -73 -7). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

For TETA two reliable studies according to OECD guideline 414 in rats and rabbits are available.

Timed-pregnant Sprague Dawley rats were treated with triethylenetetramine by daily oral gavage on gestational days (gd) 6 up to and including day 15 (CIBA-GEIGY, 1984). Groups of 24 females each received triethylenetetramine formulated in distilled water applied at a volume of 10 mL/kg bw at dosages of 0, 75, 325 or 750 mg/kg bw per day. With regard to the vehicle control, food intake was slightly depressed in all dose groups during the second part of the treatment period. Body-weight gain appeared to be somewhat reduced in the experimental groups; however when corrected body weights were evaluated no differences were observed. No other toxic signs were observed in the mothers. A significant increase in the average body weight of the live foetuses was recorded for the high-dose group in the main study. The gross inspection of the live foetuses did not reveal any finding either in the experimental groups or in the vehicle control. The skeletal assessment revealed instances of irregular sternebral ossification in the dose groups and in the vehicle control. Concerning the status of skeletal maturation of the foetuses shortly before term, apart from a slightly raised number of still unossified calcanei in the high-dose group in the main study, no delay of ossification could be found for the experimental groups in comparison with the vehicle control.

Thus, under the conditions of this study, triethylenetetramine produced no maternal toxicity except for slightly reduced food intake and no developmental toxicity at any dosages employed. Therefore, the NOAEL for maternal toxicity and developmental toxicity was considered to be equal or greater than 750 mg/kg bw/day in rats after oral exposure.

Timed-pregnant New Zealand White rabbits were treated with triethylenetetramine by occluded cutaneous application on gestational days (gd) 6 through 18, six hours per day (Tyl, 1988). Groups of 22 does each received triethylenetetramine formulated in deionized water and applied in a volume of 2.0 mL/animal (approximately 0.5 mL/kg bw) at dosages of 0.0, 5.0, 50.0 or 125.0 mg/kg bw/day. Clinical observations of all study animals were undertaken daily and maternal body weights were measured on gd 0, 6, 12, 15, 18 and 29. In addition, daily observations of the dosing site of treated animals were recorded from the initiation of the treatment period and continued until the termination of the study. At scheduled necropsy on gd 29, does were evaluated for body weight, liver weight, lung and kidney weight, gravid uterine weight and status of implantation sites (i.e., resorptions, dead fetuses, live fetuses). Maternal blood and urine were evaluated for copper content. Live fetuses were dissected from the uterus, counted, weighed, examined for external, visceral and skeletal abnormalities and sexed internally. Maternal kidneys and liver sections were retained in fixative for possible subsequent histopathologic examination. Two does (9.1%) died at 125.0 mg/kg bw/day. Maternal toxicity was also indicated by a significant decrease in body weight gain (in the exposure period) at 125.0 mg/kg bw/day. Also, clinical observations indicated severe skin irritation at 50.0 and 125.0 mg/kg bw/day with some recovery in the post-dosing period. Slight irritation at the dosing site was observed at 5.0 mg/kg bw/day which was transient, with essentially total recovery after exposures ceased. There were no effects on maternal lung, kidney or liver weights (absolute or relative to body weight) or on gravid uterine weight. There were no differences among groups for maternal serum or urinary copper. There was no effect of treatment on number of ovarian corpora lutea, number of total, live, or non-viable implantations per litter, or percent preimplantation loss, percent live fetuses or sex ratio (% male fetuses). Fetal body weights per litter (males, females or total) were also unchanged across groups. There was no significant increase in the incidence of individual malformations or variations or of external, visceral, skeletal, or total (external, visceral, plus skeletal) malformations or variations by fetus or by litter in any triethylenetetramine exposed group relative to the vehicle control group. One finding, poorly ossified hyoid bone exhibited a decreased incidence at 125.0 mg/kg bw/day relative to that in the vehicle control group. Thus, under the conditions of this study, triethylenetetramine produced maternal toxicity other than at the dosing site only at 125.0 mg/kg bw/day but no developmental toxicity at any dosages employed.

Thus, under the conditions of this study, triethylenetetramine resulted in mortality in the highest dose group. No developmental toxicity at any dosages employed was observed. Therefore, the NOAEL for maternal toxicity and was considered to be 50 mg/kg bw/day and the NOAEL for developmental toxicity equal or greater than 125 mg/kg bw/day in rabbits after dermal exposure.

 

In conclusion, the target substance polyethylenepolyamine (CAS 68131-73-7) resulted in no developmental toxicity even under maternal toxicity. The source substance triethylenetetramine (CAS 112-24-3) shows also no developmental toxicity in rats after oral exposure. Further, it was devoid of developmental toxicity in rabbits after dermal exposure. Therefore, it is concluded that the target substance will also show no developmental toxicity in rabbits.

Justification for classification or non-classification

Reliable data on the test substance itself and from a structural analogue on developmental toxicity indicates that the registered substance does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 in respect to developmental toxicity.