Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

No full animal studies are available for(Z)-N-9-octadecenylpropane-1,3-diamine (Oleyl-diamine)on effects on fertility. No adverse effects on reproductive organs were identified in the 28-day study on (Z)-N-9-octadecenyl-1,3-diaminopropane (Oleyl diamine) and a 90 day study in rats on N-C12/14 alkyl-1,3-diaminopropane. A developmental toxicity study performed on Oleyl diamine also included endpoints relevant to assessing an effect on fertility. No effects on pre/post implantation rate, late/early resorptions, corpora lutea or number of live fetuses were seen in this study. Although the available data on Oleyl diamine does not include information on sperm parameters and oestrus cycle, there are currently no indications of an effect of alkyl-diamines on fertility.

Further read-across with the category of Polyamines to a full 2-generation study and various developmental toxicity studies available indicate a clear lack for concern for reproduction toxicity.

In addition, there is no consumer exposure Oleyl diamine. Further, manufacture is highly controlled and occupational measurements of exposure to workers are available limiting the possibility of exposures.

Link to relevant study records
Reference
Endpoint:
two-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From 22 November 1993 to 01 August 1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to the OECD Guideline 416 and in compliance with GLP.
Justification for type of information:
Cross-reading to data available on other diamines and polyamines is acceptable on the basis of identical alkyl-diamine structure, resulting to the same functional groups with similar properties leading to common biological activity, and common metabolic degradation. Further information on the applicability of the read-across from various diamines to Oleyl-diamine can be obtained from the document "Category polyamines - 20170518.pdf" added to IUCLID Ch. 13.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent, England
- Age at study initiation: (P) 4 weeks; (F1) 25 d
- Weight at study initiation: (P) ca. 90 g
- Housing: Initially 2 per cage in popypropylene cages with stainless steel grid bottoms, mesh tops and food hoppers (42 x 27 x 20 cm). 3 d prior to mating the males were transferred to individual cages. For mating the females were transferred to the cage of the appropriate co-group male. Mated females were transferred to individual solid bottomed cages of the same dimensions, where they remained with the litters until termination. F1 animals selected as parents for the next generation were housed 2 per grid bottomed cage, sexes separated, and then followed the same caging regime as P animals.
- Diet: Rat and Mouse Breeder Diet No. 3 (Expanded) SQC, ad libitum
- Water: Domestic mains water, ad libitum
- Acclimation period: 13 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 2
- Humidity (%): 50 ± 15
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Formulations were prepared at intervals of up to 1 week. For the high dose concentration, the test substance was weighed into a labelled dose container and the requisite quantity was added and mixed by inversion as required. For the lower dose level, formulation was by serial dilution of the next highest concentration. A quitable aliquot of each formulation was dispensed to the animal room for dosing on that day. Solutions were used within 8 days of preparation.

VEHICLE
- Concentration in vehicle: 0, 1, 3 and 8 mg/mL (for 30.2% aqueous solution of the test substance)
- Amount of vehicle (if gavage): 10 mL/kg
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: Until mating or 7 nights elapsed
- Proof of pregnancy: Vaginal plug or sperm in vaginal smear, referred to as Day 0 of pregnancy
- After 7 d of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: No
- After successful mating each pregnant female was caged (how): Individual solid bottomed cages (42 x 27 x 20 cm)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
On 6 occasions during the study, triplicate samples of dosing solutions were taken and analysed for concentration and homogeneity. The method of analysis was reported the provision of IRI Project 370340.
Duration of treatment / exposure:
Parental: 10 weeks prior to mating to weaning of F1 generation
F1: From Day 25 after birth to weaning of F2 generation.
Frequency of treatment:
Daily
Details on study schedule:
F1 parental animals not mated until 11 weeks after weaning
Remarks:
Doses / Concentrations:
0, 10, 30 and 90 mg/kg bw/day (30.2% aqueous solution)
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
0, 3, 9 and 27 mg/kg bw/day (pure substance)
Basis:
actual ingested
No. of animals per sex per dose:
28/sex for parental generation and 24/sex for F1 generation
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on evaluation of existing data.
- Rationale for animal assignment (if not random): Computer generated randomly sequenced number, but ensuring that siblings were not placed in the same treatment group.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily, plus at the beginning of each day and as late as possible for viability.

BODY WEIGHT: Yes
- Time schedule for examinations: One week prior to commencement of treatment, then on each day of dosing.

FOOD CONSUMPTION AND COMPOUND INTAKE:
Food consumption was recorded weekly for each animal, commencing 1 week prior to treatment. Food consumption monitoring was suspended during the mating period and then, for males, recommenced as before. For females, consumption was measured over Days 0-7, 7-14 and 14-20 of gestation, and Days 0-7 and 7-14 of lactation.

Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: No

PARAMETERS EXAMINED
The following parameters were examined in F1/F2 offspring: Number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical abnormalities

GROSS EXAMINATION OF DEAD PUPS: Yes where appropriate, for external and internal abnormalities.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals, termination after weaning of the litters
- Maternal animals: All surviving animals, termination after weaning of the litters

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cranial, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following organs were weighed: Ovaries, uterus, cervix and vagina, testes (weighed individually), epididymides (weighed individually), seminal vesicle, coagulating gland, prostate gland, pituitary gland. The female reproductive tract was examined for signs of pregnancy and the number of visible implantation sites was recorded.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at 7 weeks of age without necropsy.

GROSS NECROPSY
Offspring found dead or killed before Day 14 of lactation were sexed, examined for the externally visible abnormalities and for the presence of milk in the stomach. Offspring dying on or after Day 14 were subjected to a gross necropsy, in which the cranial, thoracic and abdominal contents were examined macroscopically. From each litter of F1 and F2 pups, 2 male and 2 female pups were necropsied at weaning. The necropsy consisted of an external examination, followed by macroscopic examination of the tissues and organs of the cranial, thoracic and abdominal cavities. The remaining pups (except F1 weanlings selected for rearing to produce the next generation) were killed after external examination and the carcasses discarded without necropsy.
Statistics:
Where considered appropriate to assist with interpretation, statistical analysis was applied to determine the statistical significance of differences from control. Organ weight data were analysed by analysis of variance. Testes and epididymides weights were also analysed by analysis of covariance using the terminal body weight as a single covariate. For prostate and seminal vesicles weights, tests for linearity of relationship to body weight and for homogeneity of slopes indicated that analysis of covariance was inappropriate for these organs. Treatment means were compared using an F-protected Least Significant Difference procedure (Snedecor and Cochran, 1980). For other parameters, interpretation was based on examination of the individual and group values.
Reproductive indices:
The following reproductive indices were calculated for each group: Fertility index and gestation index
Offspring viability indices:
The following offspring viability indices were calculated for each litter and group: Birth index, live birth index, viability index, lactation index and overall survival index.
Clinical signs:
effects observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
not examined
Other effects:
not examined
Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
At 27 mg/kg bw/day, most animals of both generations showed signs of reaction to treatment, consisting of dyspnea, piloerection and hunched posture, and many animals also had episodes of post-dosing salivation. For occasional animals, the outline of the spine was prominent. A total of 8 animals died or were killed after showing marked signs of reaction, and a 9th death may also have been related to treatment.
At 9 mg/kg bw/day occasional animals in both generations showed post dosing salivation; this observation was probably associated with treatment.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
In both generations, at 27 mg/kg/day, mean weight gain was markedly lower than control; this effect was apparent for males, and for females in the premating period and during gestation.

FOOD CONSUMPTION (PARENTAL ANIMALS)
Food consumption in both generations was slightly lower than in controls at 27 mg/kg bw/day.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
Mating performance, fertility, duration of gestation, litter size and pup survival were considered to be similar in all groups.

ORGAN WEIGHTS (PARENTAL ANIMALS)
Mean seminal vesicle weights at 27 mg/kg bw/day in both generations were significantly lower than controls, however, it was considered that this reduction was an indirect effect of the lower body weights, rather than a direct effect on the seminal vesicles.
At 27 mg/kg bw/day, mean epidymides weights in both generations were lower than control, with the value for F0 animals attaining statistical significance. However, after adjustment for bodyweight (covariance analysis) the epididymides weights were similar to control in both generations. Mean epididymides weights at the lower levels were not obviously affected by treatment.
Mean absolute testes weights in both generations at 90 mg/kg/day were slightly lower than control, with the value for F1 males attaining statistical significance. These lower values were considered to reflect the lower body weights, and after adjustment for bodyweight (covariance analysis) mean values were similar to control. Testes weights at 3 and 9 mg/kg/day were similar to control.
Mean prostate weights were essentially similar in all groups of both generations.
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
9 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: Based on observed signs of toxicity and a marked reduction in food consumption and body weight gain of males and of females during the premating and gestation periods at 27 mg/kg bw/day.
Remarks on result:
other: Generation: P and F1 (migrated information)
Dose descriptor:
NOAEL
Remarks:
fertility
Effect level:
27 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: No adverse effects on fertility up to the highest tested dose.
Remarks on result:
other: Generation: P and F1 (migrated information)
Clinical signs:
effects observed, treatment-related
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
CLINICAL SIGNS (OFFSPRING)
Two pups at 27 mg/kg bw/day showed body tremors in late lactation; these tremors may have been associated with treatment.

BODY WEIGHT (OFFSPRING)
At 27 mg/kg bw/day, mean litter and pup weights of the F2 pups were slightly lower than controls. Although these differences were probably incidental, the possibility that they were related to treatment could not be entirely discounted. The litter and pup weights of the F1 pups at 27 mg/kg bw/day and of both generations at 3 and 9 mg/kg bw/day were similar to controls.
Dose descriptor:
NOAEL
Remarks:
developmental toxicity / systemic toxicity
Generation:
F2
Effect level:
9 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: Based on slightly reduced pup and litter weights and observed clinical signs (2 pups with body tremors) at 27 mg/kg bw/day.
Reproductive effects observed:
not specified
Conclusions:
The study was performed in accordance with the guideline specified.
The performance of the 2 generations was very similar.
Under the conditions of the study 30 mg/kg/day produced only minor parental toxicity but no reproductive effects.
At 90 mg/kg/day there was very marked parental toxicity, a possible minor effect on pup and litter weights, and 2 pups with body tremors. There were no obvious adverse effects on mating and littering performance at any of the levels tested.
Executive summary:

An oral two-generation study was conducted to assess the reproductive toxicity potential of N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine in rats according to the OECD Guideline 416 and in compliance with GLP.

 

Sprague Dawley rats were dosed orally by gavage, once daily at dose levels, 0, 10, 30 and 90 mg/kg/day (i.e., equivalent to 0, 3, 9 and 27 mg a.i./kg bw/day). F0 animals were randomised into the 4 treatment groups, each containing 28 males and 28 females. From each treatment group, 24 male and 24 female F1 weanlings were selected for rearing to maturity and mating to produce the F2 generation. The F0 animals were dosed for 10 weeks prior to mating, and then throughout the mating, gestation and lactation periods until sacrifice at the time of weaning of the F1 animals. The F1 animals were exposed to possible effect of the test substance from conception through to weaning. Clinical observations were performed daily. Body weight and food consumption were recorded at various intervals throughout the study. Females were allowed to litter normally and observations on the females and litters were recorded. All F0 and F1 animals were subjected to necropsy, consisting of an external examination, followed by macroscopic examination of the tissues and organs of the cranial, thoracic and abdominal cavities. Gross lesions were monitored and applicable organs were weighed. Only 2 male and 2 female pups that were weaned from each litter underwent necropsy. No histopathology was performed as this was available from a 90-day repeat oral dose study.

 

Toxicity to parent (adult) animals at 27 mg a.i./kg bw/day was indicated in both generations by a marked reduction in body weight gain of males, and of females during the pre-mating and gestation periods. Food consumption at this level was slightly lower than control. Additionally most animals in both generations receiving 27 mga.i./kg bw/day showed clinical signs of reaction to treatment (principally dyspnoea, piloerection and hunched posture, many animals also had episodes of post dose salivation, and for occasional animals the outline of the spine was prominent). A total of 8 animals (2 F0 males, 3 F0 females, 1 F1 male and 2 F1 females) died or were killed after showing marked signs of reaction, and a ninth death (of an F1 female) may also have been related to treatment. At 9 mga.i./kg bw/day, the only finding that was considered to have been probably associated with treatment was occasional animals in both generations with post dosing salivation; this finding might not be indicative of systemic toxicity.

Mean seminal vesicle weights in both generations at 27 mga.i./kg bw/day were significantly lower than control; it was considered that this reduction was an indirect effect of the lower body weights rather than a direct effect on the seminal vesicles.Mating performance, fertility, duration of gestation, litter size and pup survival were considered to be similar in all groups of both generations. At 27 mga.i./kg bw/day, mean litter weight and pup weights of the F2 pups were slightly lower than control; although these differences were probably incidental, the possibility that they indicated a slight effect of treatment could not be entirely discounted. The litter and pup weights of the F1 pups at 27 mga.i./kg bw/day, and of both generations at 3 and 9 mga.i./kg bw/day, were similar to control. Two pups at 27 mga.i./kg bw/day showed body tremors in late lactation; these may have been associated with treatment. At the 3 mga.i./kg bw/day dose level no marked toxicity was noted.

 

Hence, under the conditions of the study, the NOAEL for reproductive toxicity was established at 27 mg a.i./kg bw/day and the NOAEL for parental and developmental toxicity was established at the next tested lower dose level of 9 mg a.i./kg bw/day.

Effect on fertility: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
30 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
The study is GLP compliant and has Klimisch score 1.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

The evaluation for repeat dose toxicity of(Z)-N-9-octadecenyl-1,3-diaminopropane (Oleyl diamine)is based onsome highly reliable GLP studies performed over the category of alkyl-diamines. (Information on the applicability of the read-across from various diamines to Oleyl-diamine can be obtained from the document "Category polyamines - 20170518.pdf" added to IUCLID Ch. 13.).Specifically read-across is applied from N-C12/14 alkyl-1,3-diaminopropane, CAS 90640-43-0, as a worst case representative of the category of alkyl-diamine.

 

No adverse effects on reproductive organs were identified in the 90 day study in rats on C12-14-diamine. This study was performed according to OECD 408 and does not include detailed reproductive parameters as indicated in OECD 416. Nevertheless, the 90 day study did not indicate any effects on weight of testes, prostate, seminal vesicles, ovaries, and uterus. Histological examination of these tissues also confirmed no treatment-related effects. There was a very slight increase in relative weight of the epididymis in the high dose animals but there were no corresponding histological changes. There was also no indication of a dose-response relationship. Accordingly, the effect on epididymis is not believed to be treatment related.

 

A developmental toxicity study performed on Oleyl-diamine also included endpoints relevant to assessing an effect on fertility. No effects on pre/post implantation rate, late/early resorptions, corpora lutea or number of live fetuses were seen in this study. Although the available data on Oleyl-diamine does not include information on sperm parameters and oestrus cycle, it supports the conclusion that there are no indications of an effect of diamines on fertility.

 

Cross-reading to data available on other diamines and polyamines is acceptable on the basis of identical alkyl-diamine structure, resulting to the same functional groups with similar properties leading to common biological activity, and common metabolic degradation. Further information on the applicability of the read-across from various diamines to Oleyl-diamine can be obtained from the document "Category polyamines - 20170518.pdf" added to IUCLID Ch. 13.

The table below further presents available data from reproduction toxicity studies in rat on various polyamines:

Sub-group

Alkyl chain

Study

Results NOAEL (in mg/kg bw/d)

Diamines

Oleyl

Development rat (OECD 414)

No effects on pre/post implantation rate, late/early resorptions, corpora lutea or number of live fetuses were seen in this study.

There was a slight increase in visceral and skeletal variations. No malformations were observed.

NOAEL development: 20 mg

Triamines

Coco

OECD 422

NOAEL reproduction and development: 30 mg (highest dose since all animals were sacrificed at 100 mg/kg/day, and hence no potential effect on reproductive/developmental parameters could be determined at this dose level.)

 

Tallow

Development rat (OECD 414)

NOAEL maternal = 30 mg/kg based on signs of toxicity and lower BW seen from 60 mg/kg.

NOAEL development: 30 mg/kg, based on the observation of pale adrenals in 7 foetuses, and signs of retarded skeletal ossification seen at 60 and 120 mg/kg.

Y-triamines

C12

Development rat (OECD 414)

NOAEL maternal = 7.5 mg

NOAEL development = 22.5 mg, based on secondary effects due to maternal toxicity observed in the foetuses (increase in incidence of early embryonic deaths and slightly decreased mean foetal weight seen at 60 mg).

 

 

Development rabbit (OECD 414)

NOAEL maternal = 9 mg

NOAEL teratology > 20 mg

NOAEL development = 9 mg, based on incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity seen at 20 mg.

 

 

2-Gen reproduction (OECD 416)

NOAEL maternal, F1 and F2: 9 mg. Based on reduced body weight gain, clinical signs of reaction to treatment at higher levels. There were no obvious adverse effects on mating and littering performance at any of the levels tested.

Tetramines

Tallow

OECD 422

NOAEL reproduction and development: 100 mg (highest dose since all animals were sacrificed at 300 mg/kg/day, and hence no potential effect on reproductive/developmental parameters could be determined at this dose level.)

 

Additionally, no adverse effects on reproductive organs were identified in any of the available repeated dose studies on Polyamines.

There are various reproduction screening studies, a full 2-generation study and various developmental toxicity studies available, covering the range of Polyamines, which show a clear lack for concern for reproduction toxicity.

 

(Z)-N-9-octadecenyl-1,3-diaminopropane (Oleyl diamine) is a liquid/paste with a vapour pressure less than 0.0015 Pa at 20°C (value is an overestimation as it is based on read-across from shorter chain C12-14-diamine). Also the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur.

 

In addition, there is no consumer exposure to Oleyl-diamine.Manufacture and use are highly controlled. Its use is limited to industrial and professional users where following its severe corrosive properties sufficient protection measures will be in place to prevent exposure. Furthermore, diamines are not expected to easily pass the skin, and in view of their severe corrosive properties, testing via dermal route for reproduction toxicity is not a first choice.

Effects on developmental toxicity

Description of key information

Prenatal developmental toxicity (OECD 414, GLP) is available on Oleyl-diamine: NOAEL for embryo-foetal toxicity was found to be 20 mg/ kg body weight.

On overall, there are various reproduction screening studies, a full 2-generation study and various developmental toxicity studies available, covering the range of Polyamines, which show a clear lack for concern for reproduction toxicity. This includes a developmental toxicity study in rabbits on N-(3-aminopropyl)-N-

dodecylpropane-1,3-diamine, resulting to a NOAEL maternal of 9 mg/kg bw, a NOAEL teratology > 20 mg/kg bw, and a NOAEL development also of 9 mg, based on incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity seen at 20 mg.

Link to relevant study records

Referenceopen allclose all

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2008-05-14 - 2010-04-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.31 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier S.A.S, Route des Chênes secs-B.P.4105-53941 LE GENEST-ST-ISLE-France
- Age at study initiation: 11 - 12 weeks
- Weight at study initiation: within ± 20% of the mean weight (no more information given)
- Housing: The animals were housed individually in IVC cages (except during mating
period where 2 females were paired with one male), type III H, polysulphone cages on Altromin saw fiber bedding
- Diet: Free access to Altromin 1324 maintenance diet for rats and mice
- Water: Free access to tap water, sulphur acidified to a pH of approximately 2.8 (drinking water, municipal residue control, microbiol. controlled periodically)
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 10%
- Air changes (per hr): 10 x / hour
- Photoperiod (hrs dark / hrs light): Artificial light, sequence being 12 hours light, 12 hours dark

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The test item was dissolved or suspended corn oil. The vehicle was chosen as suggested by sponsor and the test item’s solubility. The test item formulation was prepared freshly on each administration day before the administration procedure.

VEHICLE
- Lot/batch no. (if required): 058K0070 and 128K0040 (Sigma)

- Administered dose volume: 4 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The assessment of homogeneity as well as a determination of the nominal concentration of the test item in the vehicle was performed at specified intervals. Analysis of the dose formulations of the test item in the vehicle (nominal concentration) was performed in the first and last week of the study for all doses. Samples for homogeneity were taken from the top, middle and bottom of the high dose and low dose preparation. The determination was performed in the first and last week of the study. The dose formulation analysis was performed at BSL BIOSERVICE Scientific Laboratories GmbH under the BSL study Nr. 081567.
Details on mating procedure:
- Impregnation procedure: cohoused
- if cohoused:
- M/F ratio per cage: 1/2
- Length of cohabitation: overnight
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
The test item was administered to sperm positive females (presumed pregnant) from respective GD 0 to GD 19.
Frequency of treatment:
daily
Duration of test:
Duration of test: 20 days (animals were killed on day 20)
Remarks:
Doses / Concentrations:
1.25, 5.0, 20.0 mg/kg bw
Basis:
actual ingested
No. of animals per sex per dose:
20
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Each animal was observed twice daily during entire gestation period except during weekends and holidays where clinical observation was made only once. Mortality, morbidity, pertinent behavioural changes and all signs of overt toxicity were recorded.

BODY WEIGHT: Yes
- Time schedule for examinations: The sperm positive females were weighed during GD 0, 3, 5, 8, 11, 14, 17 and 20. Males were not weighed in this study.

FOOD CONSUMPTION: Yes
- Food consumption of sperm positive females was measured on respective GD 3, 5, 8, 11, 14, 17 and 20.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: uterus, ovaries

OTHER:
- Inflammatory Markers:
Serum samples were collected at terminal sacrifice from all females and stored at ≤ -20 °C for the possible analysis of inflammatory markers by ELISA technique.
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
Fetal examinations:
- External examinations: Yes: all per litter
- Soft tissue examinations: Yes: half per litter
- Skeletal examinations: Yes: half per litter
- Head examinations: Yes
Statistics:
For statistical analysis one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison test was carried out to reveal any differences between control and test groups. Statistical significance for fetal anamolies were determined by Chi Square analysis. Statistical analysis was performed with GraphPad Prism (Version V) software (p < 0.05 was considered as statistical significant).
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
Test item related clinical signs were observed in high dose females during the entire treatment period. 4 decedents were found (1 in the MD and 3 in the HD group). But the death of only 2 animals might be considered due to toxicity. Statistically significant decrease for the body weight and for the food consumption in the HD group was observed. Also statistical anlysis of parental data revealed significance in the parental parameters gravid uterus weight and adjusted maternal weight in HD group compared to corresponding controls. Decrease in pregnancy rate was observed in HD group (69.56% compared to LD (96%), MD (96.8%) and, control (95.8%)).
The terminally sacrificed animals belonging to the HD group revealed incidences of few lesions at necropsy, which were as gas filled stomach and intestine, whitish spots on adrenals, discoloured liver, small spleen and thymus, enlarged adrenals, bloody lung, discoloured heart, bloody lung.
Dose descriptor:
NOAEL
Effect level:
1.25 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:yes

Details on embryotoxic / teratogenic effects:
Few gross abnormalities were seen in fetuses among the control and the treatment groups. Typical external finding noted were protruding tongue,
malrotated limbs, micrognathia, edematous neck, small neck and hematoma (localised). But no statistical deviation was observed for these above findings except for hematoma which were localised and did not show dose related pattern.
Internal observation of the viscera by free hand micro discussion technique revealed range of visceral abnormalities in all groups including control.
However, the statistical analysis revealed differences for findings viz., hemorrhagic kidney-bilateral (HD group), convoluted ureter-bilateral (HD
group), dilated renal pelvis-left side (MD group), split thymus (MD), small spleen (MD group). Most of the above findings (except for hemorrhagic kidney and convoluted ureter in HD group) were not attributed to toxicity due to lack of dose dependent effect.
Skeletal examination of the Alizarin red stained fetuses revealed a range of abnormalities which were of a type or which occurred at an incidence in both treatment and control groups. The statistical difference observed for supernumerary 14th rib-right side –bilaterally (MD group) and right (HD group), large naso-frontal suture (MD group), incomplete ossification of 4th sternebrum (LD group), split interparietal (HD group) and small hyoid (HD group) were attributed to toxicity, but the statistical difference observed for other anamalies were not considered of toxicological relevance due to lack of dose related pattern.
Dose descriptor:
NOAEL
Effect level:
20 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: fetotoxicity
Abnormalities:
not specified
Developmental effects observed:
not specified

Summary of clinical observations

Clinical Finding

Group

C (0 mg/kg)

LD (1.25 mg/kg)

MD (5 mg/kg)

HD (20 mg/kg)

Total number of animals examined

24

25

25

26

Regurgitation

0

1

1

0

Dyspnoea

0

1

0

1

Moving the bedding

0

0

2

1

Salivation

0

0

3

15

Vocalization

0

0

4

9

Sneezing

0

0

0

4

Bloody nasal discharge

0

0

0

1

Weight loss

0

0

0

5

Piloerection

0

0

0

3

Half eye lid closure

0

0

0

1

Apnoea

0

0

0

2

Swollen abdomen

0

0

0

1

Swollen snout

0

0

0

1

Cyanosis

0

0

0

1

Diarrhea

0

0

0

1

Summary of prenatal data

Parameters

 

C (0 mg/kg)

LD (1.25 mg/kg)

MD (5 mg/kg)

HD (20 mg/kg)

Terminal Body weight (g)

Mean

± SD

N

420.17

24.94

23

412.33

38.72

24

408.83

24.85

23

369.13*

53.44

16

Uterus weight (g)

Mean

± SD

N

73.16

13.83

23

68.79

26.44

24

74.52

16.57

23

67.5*

24.25

16

Adjusted maternal weight (g)

Mean

± SD

N

347.04

20.66

23

343.54

19.42

24

334.3

18.97

23

301.63*

34.13

16

Corpora lutea

Mean

± SD

N

14.26

1.48

23

14.88

2.76

24

15.0

2.58

23

15.19

1.42

16

Implantation

Mean

± SD

N

12.74

2.14

23

12.63

4.22

24

12.74

3.15

23

13.5

3.2

16

Live Fetuses

Mean

± SD

N

12.22

2.26

23

11.75

4.70

24

12.74

3.15

23

11.88

4.5

16

Early resorptions

Mean

± SD

N

0.48

0.79

23

0.88

1.03

24

0.52

0.9

23

0.75

1.44

16

Late resorptions

Mean

± SD

N

0.04

0.21

23

0.0

0.0

24

0.0

0.0

23

0.88

3.5

16

Total resorptions

Mean

± SD

N

0.52

0.79

23

0.88

1.03

24

0.52

0.9

23

1.63

3.59

16

Dead Fetuses

Mean

± SD

N

0.0

0.0

23

0.04

0.2

24

0.09

0.29

23

0.0

0.0

16

Sex Ratio (M/F)

Mean

± SD

N

2.27

2.37

23

0.96*

0.61

23

1.02*

0.66

22

12.67

3.31

15

Pre-implantation loss

Mean

± SD

N

10.86

10.55

23

16.96

22.51

24

15.17

15.3

23

11.07

19.09

16

Post-implantation loss

Mean

± SD

N

4.24

6.04

23

11.88

22.48

24

0.0

0.0

23

11.38

25.65

16

Summary of Fetal Visceral Examination

Observations

Group

Control

Low Dose

Mid Dose

High Dose

Dose

0 mg/kg bw

1.25 mg/kg bw

5 mg/kg bw

20 mg/kg bw

No. Of litters evaluated

23

24

23

16

No. Of pups evaluated

147

147

152

99

 

A

B

A

B

A

B

A

B

Hemorrhagic Kidney (B)

0

0.0

1*

0.75

3

2.11

5

5.49

Convoluted Ureter (B)

7

5.19

11

8.21

16

11.27

15*

16.48

Dilated Renal Pelvis (L)

0

0.0

3

2.24

5*

3.52

0

0.0

                Thymus

0

0.0

2

1.49

4*

2.82

1

1.1

Small Spleen

2

1.48

2

1.49

9*

6.34

2

2.2

Summary of Fetal Skeletal Examination

Observations

Group

Control

Low Dose

Mid Dose

High Dose

Dose

0 mg/kg bw

1.25 mg/kg bw

5 mg/kg bw

20 mg/kg bw

No. Of litters evaluated

23

24

23

16

No. Of pups evaluated

147

147

152

99

 

A

B

A

B

A

B

A

B

Supernumerary rib-14th T (B)

6

4.08

10

6.8

24*

15.79

4

4.04

Supernumerary rib-14th T (R)

2

1.36

8

5.44

8

5.26

6*

6.06

Large-NasoFrontal Suture

0

0.0

1

0.68

5*

3.29

0

0.0

IO-4th Sternerbum

0

0.0

5*

3.4

1

0.66

0

0.0

Split-Interparietal

15

10.2

13

8.84

18

11.84

19*

19.19

Small-Hyoid

0

0.0

2

1.36

3

1.97

3*

3.03

Conclusions:
Based on the findings, the NOAEL (No observed adverse effect level) for maternal toxicity is believed to be 1.25 mg/ kg body weight based on clinical observations seen in some animals at 5 mg/kg, whereas the NOAEL for embryo-fetal toxicity is believed to be 20 mg/ kg body weight.
Executive summary:

This Prenatal developmental toxicity study of N-Oleyl-1,3-diaminopropane was conducted in pregnant female Wistar rats to detect the possible adverse effect on pregnant females and embryofetal development when administered by oral gavage from respective gestation day 0 to 19.

Nulliparous and non pregnant females were mated with males (2:1 ratio) and divided into four groups based on their body weights on day of positive vaginal smears (GD 0). Four groups of presumed pregnant females were dosed daily by oral gavage with 1.25, 5 and 20 mg/kg body weight per day of N-Oleyl-1,3-diaminopropane at dose volume of 4 mL/kg body weight. Control animals were handled identically as treated groups and received vehicle in similar volume as treated groups. The test item formulation was prepared freshly and dose volumes were adjusted based on the most recent body weight measurement. Animals were examined daily for the clinical signs and mortality. Body weight and food consumption was measured on various gestation days. The treated and control females were sacrificed on respective gestation day 20.

Followed by the gross necropsy evaluation of the females, the uteri and ovaries were removed, weighed and examined for number of implantations, resorptions (early and late) live and dead fetuses. Fetuses were identified by colour strings, sexed and weighed. All fetuses were observed for the external abnormalities, half of the fetuses for the visceral abnormalities, craniofacial examination and remain half of the litter for skeletal abnormalities. Uteri of the non pregnant females were processed with 0.5 % ammonium sulphide solution and checked for the early embryonic deaths if any.

 

Results:

Test item related clinical signs were observed in high dose females during the entire treatment period. Also effects on clinical observations were observed in animals of the MD group.

However, there were four decedents in this study [1 in MD group (Animal 62) and 3 in HD group (Animals 80, 82 and 98)]. Animal no. 82 was euthanised for humane reason. The death of two animals (Animals 62 and 98) were considered due to gavaging error and other two (Animals 80 and 82) might be considered due to toxicity. 

Statistically significant decrease was observed for body weight and body weight change throughout the gestation period in HD group.  Statistically significant decrease in overall food consumption was observed in HD group compared to corresponding control. Statistical analysis of prenatal data revealed significance in prenatal parameters like gravid uterus weight and adjusted maternal weight in HD group compared to corresponding controls. No other prenatal parameters like No. of corpora lutea, implantations percent preimplantation loss, group mean number of live fetuses, early resorptions, late resorptions, total resorptions, group mean number of female fetuses, sex ratio (M/F) and percent post implantation loss showed statistical deviation compared to corresponding controls.

 

Statistically significant difference was observed for group male litter weight (LD group), sex ratio (LD and MD groups) and total number of male fetuses (LD group) compared with controls. These findings were not attributed to toxicity as no dose related pattern was observed. Decrease in pregnancy rate was observed in HD group (69.56%) as compared to LD (96%), MD (95.8%), and control (95.8%).

Few gross external abnormalities were seen in fetuses among the control and treatment groups. Typical external findings noted were protruding tongue, malrotated limbs, micrognathia, edematous neck, small neck and hematoma (localised).But no statistical deviation was observed for these above findings except for hematoma which were localised and did not show dose related pattern.

Internal observation of the viscera by free hand micro discussion technique revealed range of visceral abnormalities in all groups including control. However, the statistical analysis revealed differences for findings viz., hemorrhagic kidney-bilateral (HD group), convoluted ureter-bilateral (HD group), dilated renal pelvis-left side (MD group), split thymus (MD), small spleen (MD group). Most of the above findings (except for hemorrhagic kidney and convoluted ureter in HD group) were not attributed to toxicity due to lack of dose dependent effect.

Craniofacial examination by razor blade serial sectioning technique revealed no statistical significant difference for any of the findings observed in treatment and control groups. 

Skeletal examination of the Alizarin red stained fetuses revealed a range of abnormalities which were of a type or which occurred at an incidence in both treatment and control groups. The statistical difference observed for supernumerary 14th rib-right side-bilaterally (MD group) and right (HD group), large naso-frontal suture (MD group), incomplete ossification of 4th sternebrum (LD group), split interparietal (HD group) and small hyoid (HD group) were attributed to toxicity, but the statistical difference observed for other anomalies were not considered of toxicological relevance due to lack of dose related pattern.

The terminally sacrificed animals belonging to the HD group revealed incidences of few lesions at necropsy, which were as gas filled stomach and intestine, whitish spots on adrenals, discoloured liver, small spleen and thymus, enlarged adrenals, bloody lung, discoloured heart, bloody lung. The finding like dark coloured food rest in caecum observed in most of the animals of control and treatment groups cannot be considered as toxicity related and in most of the animals this finding observed was not reported.

 

Based on the findings, the NOAEL (No observed adverse effect level) for maternal toxicity is believed to be 1.25 mg/ kg body weight based on clinical observations seen in some animals at 5 mg/kg, whereas the NOAEL for embryo-fetal toxicity is believed to be 20 mg/ kg body weight.

Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From 07 July 2004 to 02 February 2005
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Cross-reading to data available on other diamines and polyamines is acceptable on the basis of identical alkyl-diamine structure, resulting to the same functional groups with similar properties leading to common biological activity, and common metabolic degradation. Further information on the applicability of the read-across from various diamines to Oleyl-diamine can be obtained from the document "Category polyamines - 20170518.pdf" added to IUCLID Ch. 13.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rabbit
Strain:
Himalayan
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: LPT Laboratory of Pharmacology and Toxicology KG, Hamburg, Germany; branch facility: LPT Löhndorf
- Age at study initiation: 4-5 months
- Weight at study initiation: 2.37-3.26 kg
- Housing: Individually in breeding cages with wire floor (surface of ca. 0.2 m2)
- Diet: Ssniff K-Z V2323, ad libitum
- Water: Ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12/ 12
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The test substance preparations were prepared freshly every day. The test substance was suspended in the vehicle to appropriate dose levels.

VEHICLE
- Amount of vehicle (if gavage): 2 mL/kg bw
- Concentration in vehicle: 3, 4.5 and 10 mg/mL
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analytical determinations of the test substance in aqueous solution and in test substance-carrier were performed.
Details on mating procedure:
- Impregnation procedure: Cohoused
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation: Until copulation was ascertained by observation
- Verification of same strain and source of both sexes: Yes
- Proof of pregnancy: Signs of copulation referred to as Day 0 of pregnancy
Duration of treatment / exposure:
Days 6-28 of gestation
Frequency of treatment:
Daily
Duration of test:
Until Day 29 of gestation
Remarks:
Doses / Concentrations:
0, 6, 9 and 20 mg/kg bw/day (pure substance)
Basis:
actual ingested
No. of animals per sex per dose:
20 animals per group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on the results of a dose-range finding study (NOEL of 7.5 mg/kg bw/day, based on reduced food intake and local intolerance reactions).
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once daily
- Cage side observations included: Behavioral changes, reaction to treatment or illness. In addition animals were observed for viability early in the morning and in the afternoon.

BODY WEIGHT: Yes
- Time schedule for examinations: Daily, starting at Day 0 of gestation

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation Day # 29
- Organs examined: Internal organs
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: Placenta weight was determined.
Fetal examinations:
- External examinations: Yes, all per litter
- Soft tissue examinations: Yes, all per litter
- Skeletal examinations: Yes, all per litter
- Head examinations: Yes, all per litter
Statistics:
The following statistical analysis was used:
For numeric values: Bartlett chi-square test, Dunnett test (for homogeneous variances), Student's t-test (for heterogeneous variances).
For the comparison of classification measurements the Fisher's exact test was employed.
Analytical determinations of Lonzabac in aqueous solution and in test item-carrier were performed.
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
Maternal toxicity was indicated at 20 mg/kg/day by the death of two dams, by marginally reduced body weight and food intake during the last days of pregnancy. At 20 mg/kg/day, the body weight was marginally reduced during the last gestion days. There was a reduction in food consumption at 20 mg/kg/day from gestation Day 16 onwards, which was up to 36% below the control value (p = 0.05).
Necropsy revealed an irritation of the gastrointestinal tract in 5 dams and an increased incidence of resorptions.
Dose descriptor:
NOAEL
Effect level:
9 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
- At 20 mg/kg bw/day there was a marginal increased incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity.
- No test substance-related influence was detected on the prenatal foetal development. The incidences of foetal malformations, variations and retardations were similar in all groups. Under the conditions of the study no teratogenic effects (as indicated by the incidences of foetal malformations, variations and retardations) were found up to 20 mg/kg bw/day of the test substance.
Dose descriptor:
NOAEL
Effect level:
9 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: embryotoxicity
Dose descriptor:
NOAEL
Effect level:
> 20 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
Under the conditions of the study, the test substance was not considered to be a developmental toxicant. The NOAELs for maternal effects and embryotoxicity was set at 9 mg a.i./kg bw/day and the NOAEL for teratogenicity was set at 20 mg a.i./kg bw/day respectively.
Executive summary:

A pre-natal developmental toxicity study was conducted to assess the developmental or teratogenic potential of N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine in Himalayan rabbits according to the OECD Guideline 414 and in compliance with GLP.

 

Groups of 20 pregnant females received the substance by gavage at dose levels of 0, 6, 9 and 20 mg a.i./kg bw/day during Days 6 -28 of gestation. Animals were sacrificed on gestation Day 29.

Maternal toxicity was indicated at 20 mg a.i./kg bw/day by the death of two dams and by marginally reduced body weight and food intake during the last days of pregnancy. Necropsy revealed an irritation of the gastrointestinal tract in 5 dams and an increased incidence of resorptions. Marginal increased incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity was observed at 20 mg a.i./kg bw/day. No substance-related influence was detected on the pre-natal foetal development. The incidences of foetal malformations, variations and retardations were similar in all groups. As a result, the NOAELs for both maternal and embryotoxicity were set at 9 mg a.i./kg bw/day and the NOAEL for teratogenicity was set at 20 mg a.i./kg bw/day.

Under the conditions of the study, the test substance was not considered to be a developmental toxicant.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
20 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The study is GLP compliant and has Klimisch score 1.
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

A study according to the OECD 414 guideline and under GLP conditions is available with (Z)-N-9-octadecenyl-1,3-diaminopropane (Oleyl-diamine).

This study was performed by dosing pregnant rats from day 0-19 of gestation with either 0, 1.25, 5 or 20 mg/kg bw of (Z)-N-9-octadecenyl-1,3-diaminopropane. The animals were then sacrificed and the fetuses subject to external, visceral and skeletal examination. There was a slight increase in visceral and skeletal variations. No malformations were observed. Neither of the variations are considered to be of toxicological relevance as there were no dose-response pattern. Maternal toxicity including deaths in the high dose group, reduced bodyweight gain and reduced pregnancy rate were observed. The NOAEL (No observed adverse effect level) for maternal toxicity is believed to be 1.25 mg/ kg body weight based on clinical observations seen in some animals at 5 mg/kg, whereas the NOAEL for embryo-fetal toxicity is believed to be 20 mg/ kg body weight.

 

Cross-reading to data available on other diamines and polyamines is acceptable on the basis of identical alkyl-diamine structure, resulting to the same functional groups with similar properties leading to common biological activity, and common metabolic degradation. Further information on the applicability of the read-across from various diamines to Oleyl-diamine can be obtained from the document "Category polyamines - 20170518.pdf" added to IUCLID Ch. 13.

 

On overall, there are various reproduction screening studies, a full 2-generation study and various developmental toxicity studies available, covering the range of Polyamines, which show a clear lack for concern for developmental toxicity. This includes a developmental toxicity study in rabbits on N-(3-aminopropyl)-N-

dodecylpropane-1,3-diamine, resulting to a NOAEL maternal of 9 mg/kg bw, a NOAEL teratology > 20 mg/kg bw, and a NOAEL development also of 9 mg, based on incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity seen at 20 mg.

 

The table below further presents available data from reproduction toxicity studies in rat on various polyamines:

Sub-group

Alkyl chain

Study

Results NOAEL (in mg/kg bw/d)

Diamines

Oleyl

Development rat (OECD 414)

No effects on pre/post implantation rate, late/early resorptions, corpora lutea or number of live fetuses were seen in this study.

There was a slight increase in visceral and skeletal variations. No malformations were observed.

NOAEL development: 20 mg

Triamines

Coco

OECD 422

NOAEL reproduction and development: 30 mg (highest dose since all animals were sacrificed at 100 mg/kg/day, and hence no potential effect on reproductive/developmental parameters could be determined at this dose level.)

 

Tallow

Development rat (OECD 414)

NOAEL maternal = 30 mg/kg based on signs of toxicity and lower BW seen from 60 mg/kg.

NOAEL development: 30 mg/kg, based on the observation of pale adrenals in 7 foetuses, and signs of retarded skeletal ossification seen at 60 and 120 mg/kg.

Y-triamines

C12

Development rat (OECD 414)

NOAEL maternal = 7.5 mg

NOAEL development = 22.5 mg, based on secondary effects due to maternal toxicity observed in the foetuses (increase in incidence of early embryonic deaths and slightly decreased mean foetal weight seen at 60 mg).

 

 

Development rabbit (OECD 414)

NOAEL maternal = 9 mg

NOAEL teratology > 20 mg

NOAEL development = 9 mg, based on incidence of resorptions suggesting early embryonic deaths secondary to maternal toxicity seen at 20 mg.

 

 

2-Gen reproduction (OECD 416)

NOAEL maternal, F1 and F2: 9 mg. Based on reduced body weight gain, clinical signs of reaction to treatment at higher levels. There were no obvious adverse effects on mating and littering performance at any of the levels tested.

Tetramines

Tallow

OECD 422

NOAEL reproduction and development: 100 mg (highest dose since all animals were sacrificed at 300 mg/kg/day, and hence no potential effect on reproductive/developmental parameters could be determined at this dose level.)

 

Further studies for investigation of developmental toxicity are not indicated on the basis of the following argumentation:

- No indications for specific concerns for developmental toxicity:

The 28-day study with N-Oleyl-1,3-diaminopropane (Oleyl diamine) resulted to significant toxicity (mortality) at the highest dose of 20 mg/kg bw/day with effects on weight of body and organs without histological changes. This is below 30 mg/kg bw and therefore also results to classification STOT-RE Cat.1. No adverse effects on reproductive organs were identified. A developmental toxicity study was performed by dosing pregnant rats from day 0-19 of gestation with either 0, 1.25, 5 or 20 mg/kg bw of Oleyl diamine, resulting to maternal toxicity including deaths in the high dose group, reduced bodyweight gain and reduced pregnancy rate. No effects on pre/post implantation rate, late/early resorptions, corpora lutea or number of live fetuses were seen in this study. At fetal evaluations no malformations were observed. A slight increase in visceral and skeletal variations was seen. As these showed no dose-response pattern this was not considered to be of toxicological relevance. The NOAEL (No observed adverse effect level) for maternal toxicity was 1.25 mg/ kg bw based on clinical observations seen in some animals at 5 mg/kg, whereas the NOAEL for embryo-fetal toxicity is believed to be 20 mg/ kg body weight. Available data indicate a relative low NOAEL for systemic toxicity, but even at levels causing maternal mortality, no significant developmental toxicity was observed in rats.

 

- Toxicological mode of actions:

Physicochemical properties indicate that the substance is a cationic surfactant structure which leads to high adsorptive properties to negatively charged surfaces as cellular membranes. The apolar tails easily dissolve in the membranes, whereas the polar head causes disruption and leakage of the membranes leading to cell damage or lysis of the cell content. As a consequence, the whole molecule will not easily pass membrane structures. Cytotoxicity at the local site of contact through disruption of cell membrane is considered the most prominent mechanism of action for toxic effects. With such mechanism of action the substance is not expected to easily pass membranes and placenta. The repeated dose studies results to effects on the small intestine and mesenteric lymph nodes as most critical effect. Taking into consideration the relatively strong corrosive effects of this substance and the route of administration, this probably reflects a point-of-first-contact effect.

 

With these considerations on the non-specific toxic mode of action consisting of local cytotoxicity at point-of-first-contact, species differences are not very likely. Further, as no reproductive effects are observed at the highest dose levels in the rats leading even to mortality, shows a low concern for specific developmental toxicity. This lack of developmental toxicity in rats, in combination with the non-specific mode of action of local cytotoxicity at the point of contact, an additional developmental study in an second species would not be scientifically justified.

 

- Limited exposures:

Oleyl diamine is a liquid/paste with a vapour pressure less than 0.0015 Pa at 20°C (value is an overestimation as it is based on read-across from shorter chain C12-14-diamine). Also the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalation route will be unlikely to occur. 

 

Furthermore, use and handling of the substance is only in industrial and professional setting, and the classification of the substance as severely corrosive requires risk management methods which eliminate the risk potential exposures via the skin.

Mode of Action Analysis / Human Relevance Framework

Based on structure and mechanism of cytotoxicity, reproduction toxicity is not expected. The observed effects are local, reflecting a point-of-first-contact effect.

In physiological circumstances, the diamines have a cationic surfactant structure (nitrogens are fully protonated)which leads to high adsorptive properties to negatively charged surfaces as cellular membranes. The apolar hydrophobic tails are pushed out of solution and easily dissolve in the lipid bilayer, whereas the polar head causes disruption and leakage of the membranes leading to cell damage or lysis of the cell content. As a consequence, the whole molecule will not easily pass membrane structures. Cytotoxicity through disruption of cell membrane at exposure site will occur rather than absorption over the cell membrane. Consequently, significant uptake followed by placental transfer is not expected to occur.

Justification for classification or non-classification

Adequate study data is only available for developmental toxicity from a valid OECD 414 guideline compliant study, showing no specific concerns for developmental toxicity. Further information from repeated dose studies up to 90-days OECD 408 performed on C12/14-diamine showed no indication of concern with respect to reproductive organs evaluated, and also the developmental toxicity study did not show effects on reproductive parameters.

In addition, there is no consumer exposure to (Z)-N-9-octadecenyl-1,3-diaminopropane and manufacture and industrial/professional use are highly controlled and occupational measurements of exposure to workers are available limiting the possibility of exposures.

 

However, as a firm conclusion from a study with this compound is lacking, no definite conclusion might be drawn for classification purposes.

But based on limited exposures by dermal route (substance is severely irritating/ corrosive) or by inhalation (very low vapour pressure), as well as lack of indication for concerns regarding reproductive toxicity from repeated dose studies and a developmental toxicity study there are no concerns and further testing is not indicated.