C16-18-(even numbered)-alkyl fatty acid, compound with C16-18-(even numbered)-alkylamine

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

There is no study avalaible for this substance. A read-across with tallow alkylamines (CAS No. 61790 -33 -8) is applied through an OECD TG 421 in which no reproductive toxicity effect is observed.

Link to relevant study records

Reference

Endpoint:

one-generation reproductive toxicity

Remarks:

based on generations indicated in Effect levels (migrated information)

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Guideline study according to GLP but the reliability has been amended in accordance with 'practical guide 6: How to report read-across and categories' which states that the maximum reliability for a read-across study is 2. The study is considered to be adequate and reliable for the purpose of registration under REACH (Regulation (EC) No. 1907/2006).

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Italia
- Age at study initiation: (P) 8 wks
- Weight at study initiation: (P) Males: 225 - 250 g; Females: 200 - 225 g
- Fasting period before study: no
- Housing: macrolon cages
- Use of restrainers for preventing ingestion (if dermal): no
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2 °C
- Humidity (%): 55 +/- 5 %
- Air changes (per hr): 15 -20
- Photoperiod (hrs dark / hrs light): 12 hours periodically

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 16 h, 7 evenings/week for max. 14 evenings
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
- After successful mating each pregnant female was caged: individually

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

Males: 14 days prior to mating, during mating; 28 days maximum
Females: 14 days prior to mating, during mating, during pregnancy, 3 days post-partum (lactation)
Females, positive smear, no offspring: 14 days prior to mating, during mating, 25 days post-mating
Females, negative smear: 14 days prior to mating, during mating, 25 days post-mating

Remarks:

Doses / Concentrations:
mg/kg body weight per day
Basis:
nominal conc.

Remarks:

Doses / Concentrations:
12.5 mg/kg body weight per day
Basis:
nominal conc.

Remarks:

Doses / Concentrations:
50 mg/kg body weight per day
Basis:
nominal conc.

Remarks:

Doses / Concentrations:
150 mg/kg body weight per day
Basis:

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: based on dose-range-finder in rats
- Rationale for animal assignment (if not random): random
- Other:

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

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

OTHER:

Sperm parameters (parental animals):

Sections of testes were stained with PAS-hematoxylin to allow spermatogenesis to be classified into 14 stages, each stage based primariliy upon the changes of the acrosome and head morphology of the younger generation of spermatids.

Litter observations:

number and sex of pups
stillbirths
live births
postnatal mortality
presence of gross anomalies
weight gain (day 0, day 4)
gross examination of dead pups for external and internal abnormalities

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals on the day after the end of treatment
- Maternal animals: All surviving animals on day 4 of lactation with their pubs

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

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for microscopic examination and weighed, respectively:
- ovaries
- uteri (including horns, cervix, and vagina)
- testes (with special emphasis on stages of spermatogenesis and histopathology of interstitial cell structure)
- epididymides
- accessory sex organs

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring were sacrificed on day 4 of lactation and necropsied

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

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

salivation, hunched posture at mid- and high dose animals, no changes at low-dose animals

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

decrease in mid- and high dose animals, no effect in low-dose animals

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

decrease in mid- and high dose animals, no effect in low-dose animals

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

effects observed, treatment-related

Description (incidence and severity):

Test substance intake: lower mean food consumption in mid- and high-dose animals, no effect in low-dose animals

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

decreased mating and fertility indices in high-dose animals

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
- 150 mg/kg: salivation, hunched posture, soft stools, piloerection; 6/10 male and 5/10 female animals died
- 50 mg/kg: salivation; 1/10 male and 1/10 female animals died
- 12.5 mg/kg: no changes observed
BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
- 150 mg/kg: decreased body weight gain (males and females)
- 50 mg/kg: decreased body weight gain during pre-mating period
- 12.5 mg/kg: slightly decreased body weight gain (males and females)
TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)
- lower food intake in mid and high dose groups (males and females), slightly lower food intake in low dose group during lactation period in females
REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
- mating and fertility indices, mean pre-coital time and parturition were unaffected up to 50 mg/kg/day
ORGAN WEIGHTS (PARENTAL ANIMALS)
- lower absolute weight of epididymides and higher value of testis weight relative to body weight at 150 mg/kg/day
- no differences in ovary weights among the experimental groups
GROSS PATHOLOGY (PARENTAL ANIMALS)
- no pathological changes
HISTOPATHOLOGY (PARENTAL ANIMALS)
- histology of testes, epididymides and ovaries of high dose group animals did not show any compound-related changes
- no changes in testicular staging performed in the PAS-hematoxylin stained sections of control and high dose groups
OTHER FINDINGS (PARENTAL ANIMALS)

Dose descriptor:

NOEL

Effect level:

12.5 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: overall effects

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

no effects up to 50 mg/kg body weight

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

no litters present at highest dose

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

slightly lower pub weight at 50 mg/kg body weight

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

VIABILITY (OFFSPRING)
- no litters present at 150 mg/kg/day
- no effects on postnatal survival up to day 4 of lactation and no effects on sex ration at 50 mg/kg/day

CLINICAL SIGNS (OFFSPRING)
- no litters present at 150 mg/kg/day
- no significant effects up to 50 mg/kg/day

BODY WEIGHT (OFFSPRING)
- no litters present at 150 mg/kg/day
- slightly lower pup body weight seen in 50 mg/kg/day group
- no effects seen at 12.5 mg/kg/day

GROSS PATHOLOGY (OFFSPRING)
- no abnormalities observed in any pub either at birth or at autopsy on day 4 of lactation neither of the 50 or of the 12.5 mg/kg/day group

Dose descriptor:

NOEL

Generation:

F1

Effect level:

12.5 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: overall effects

Reproductive effects observed:

not specified

Conclusions:

Based on findings of general toxicity (death, clinical signs, reduced body weight gain) at daily dosages of 50 mg/kg bw/d a NOEL/systemic toxicity of 12.5 mg/kg bw/d can be derived from the study. Based on the findings of a lower fertility index and a lower conception rate at daily dosages of 50 mg/kg bw/d a NOEL/fertility of 12.5 mg/kg bw/d can be derived from the results of this screening study.

Based on the results of this study, 12.5 mg/kg/day is the NOEL for maternal toxicity and the NOEL for the offspring.

Executive summary:

Groups of 10 rats (Crl:CD (SD) BR) per sex were treated with dosages of 0, 12.5, 50, and 150 mg/kg bw/day of Genamin TA 100 by gavage (administration volume 10 ml/kg bw/day) using sesame oil as a vehicle. Males were treated daily from 14 days prior to mating until the end of the mating period for a maximum of 28 days. Females were treated daily for 14 days before start of the mating period, throughout the same, during pregnancy and until day 3 of lactation. The animals were mated one male with one female.

At daily doses of 150 mg/kg bw 6/10 males and 5/10 females died between day 9 and day 25 of treatment (during the premating and mating period). At daily doses of 50 mg/kg bw 1/10 males and 1/10 females died on day 13 respectively on day 24 of treatment. No animals died at 12.5 mg/kg bw/day, and in the control group 1 female died by accident.

Clinical observations at 150 mg/kg bw/day revealed salivation after treatment, hunched posture and in some cases soft stools and piloerection. The only clinical sign present at 50 mg/kg bw/day was salivation. No changes were seen at 12.5 mg/kg bw/day.

Body weight loss of about 22 g at the 150 mg/kg bw dose group and statistically significant (p > 0.01) lower body weight gain at the 50 mg/kg bw dose group during the premating period together with a lower mean food consumption was observed in the males. Also in the females body weight loss of about 17 g at the 150 mg/kg bw dose group and statistically significant lower body weight gain at the 50 mg/kg bw dose group together with a lower mean food consumption during the premating period was observed.

At sacrifice of the parental animals organ weights of ovaries, testes and epididymides were determined from all experimental groups. Histopathology was carried out on testes, epididymides and ovaries of the controls and of the animals of the 150 mg/kg bw dose group. A statistically significant(p> 0.01) lower absolute and relative weight of epididymides and a statistically significant(p> 0.01) higher value of relative testis weight was observed at 150 mg/kg bw/day. No significant differences were noted in ovary weights among the various experimental groups. Histopathology of testes and epididymides of high dose group animals did not show any compound-related changes. In particular, no changes were seen in the testicular staging performed in the PAS-hematoxylin stained sections of final sacrificed animals of control and of the high dose groups. In the ovaries, moderate increased frequency of atrophic corpora lutea was seen in the animals of the high dose group which died or were sacrificed, compared to the controls, this finding was considered a secondary effect to the decrease in body weight growth induced by treatment in this group.

At the dosages of 150 mg/kg bw/day only 3 females out of 7 mated females had positive vaginal smears and of these only one was pregnant, but with only implantation sites and no live pups. At this dosage level also the mean pre-coital interval was longer (13.4 days) than that of the control group (2.3 days). With regard to the mating index in the control, the 12.5 and the 50 mg/kg bw dose level groups 9/9 (100%), 9/10 (90%) and 9/9 (100%) mated females were sperm positive. With regard to the fertility index 9/9 (100%), 8/10 (80%) and 7/9 (78%) of the mated females became pregnant in the control, the 12.5 and the 50 mg/kg bw dose level groups. Mean pre-coital time and parturition were unaffected in the low and mid dose treated groups.

With regard to the conception rate in the control, the 12.5 and the 50 mg/kg bw dose level groups 9/9 (100%), 8/9 (89%) and 7/9 (78%) of the sperm positive females became pregnant and delivered live pups. Numbers of corpora lutea had not been determined during this study. Thus, any pre-implantation loss was not evaluated. Staining for the presence of implantation sites was only performed with the uteri of apparently non-pregnant females. The mean number of visible implantation sites per dam in the control, the 12.5 and the 50 mg/kg bw dose level groups were 17.6, 13.9 and 15.0. There were no stillborns or litters with only implantations in the control, low, and mid dose groups. The mean number of total pups born per litter was 16.9, 12.3 and 14.3 in the control, the 12.5 and the 50 mg/kg bw/day dose groups. Thus, the mean litter index for post-implantation loss was calculated to 3.4, 9.9 and 4.5% in the control, the low and the mid dose groups.

No effects were noted on the pup sex ratio. No abnormalities were observed in any pup either at birth or at autopsy on day 4 of lactation neither in the 12.5 nor in the 50 mg/kg bw/day treated groups. Lower mean values of live born per litter and of pups per litter alive at day 4 were found for both the low and the intermediate dose groups in comparison to those of the control group, however, the decreases were not dose dependent and the differences were statistically significant for the 12.5 mg/kg bw dose group only. Thus, these findings were considered unlikely to be related to the compound. A slightly lower pup body weight was observed in the 50 mg/kg bw/day treated group in comparison with the control group at birth and at day 4 of lactation but not for the pups of the 12.5 mg/kg bw/day treated group. Based on general toxicity findings a NOEL(systemic toxicity) of 12.5 mg/kg/day can be derived from stis study. Additionally, based on findings of a lower fertility index and a slightly lower conception rate at the mid dose level a NOEL(fertility) of 12 .5 mg/kg body weight per day is deducible.Further, during an oral 28-day study (OECD TG 407) with the same test substance Genamin TA 100 in rats also reproductive organ weights and reproductive organ histopathology of testes and epididymides, as well as organ weights of ovaries and uteri had been investigated. No substance-related findings had been observed during this study on these organs at oral doses up to and including 150 mg/kg bw/day.

Effect on fertility: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

25.6 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The study is GLP compliant and the read-across is valid.

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 registered substance is a salt composed of primary hydrogenated alkylamine and N-hydrogenated tallow fatty acid. As the primary alkyl amine part is much more toxic than the tallow fatty acid part, it is expected that the toxicological behaviour of the substance will not be far different from that of the primary alkylamine compounds or at least not more severe. This has been demonstrated by results obtained with the registered substance in a short-term repeated toxicity study (e.g. 14 days). Thus, assessment of the registered substance can be integrated in the current approach of grouping of primary alkylamines substances. Regarding the reproductive toxicity with the registered substance on fertility, respectively on reproductive function, no experimental studies are available. However, data relevant for hazard assessment with regard to this endpoint, are available from an OECD TG 421 study as well as from repeated oral dose toxicity studies with histopathological investigation of the reproductive organs for related primary alkylamines, which are considered appropriate for extrapolation. In a study according to OECD TG 421 with tallow alkylamines (CAS No. 61790 -33 -8), findings of general toxicity in close accordance to the results from the 28 -day repeated dose toxicity study with the same compound were obtained at identical dose levels. Since severe general toxicity due to the strong irritating / corrosive properties appears to be the predominating substance-related toxic effect, and since no histological compound-related changes were observed on testes, epididymides and ovaries, neither in the OCED 421 nor in the OECD 407 study, even at the highest dose of 150 mg/kg body weight per day tested, a specific substance-related effect on reproduction performance is not deducible. Signs of general toxicity without indications on the reproductive behaviour were also noted at the intermediate dose-level of 50 mg/kg body weight per day. No toxic effects were observed at the low-dose treatment with 12.5 mg/kg body weight per day which therefore can be considered to be a NOAEL (fertility). This conclusion is in line with the evaluation coming from the existing EU risk assessment on primary alkylamines. Additionally, according to the EU risk assessment further testing of primary alkylamines with regard to the endpoint fertility was not considered necessary.

As the registered substance is a salt of primary hydrogenated alkylamine and N-hydrogenated tallow fatty acid with a molar ratio of 1:1, the NOAEL value should be converted as follows: Mw tallow alkylamine = 267, MW salt = 547.5 ; 12.5 x 547.5/267 = 25.6 mg/kg bw/day

Short description of key information:
Reproduction / Developmental Toxicity Screening Test in rats by oral route (OECD 421, GLP) based on read-across from tallow alkyl amine (values converted on mw basis from tallow alkyl amine to the registered salt ): NOAEL parental and F1 = 25.6 mg/kg bw/day.

Justification for selection of Effect on fertility via oral route:
Only available study.

Justification for selection of Effect on fertility via inhalation route:
The registered substance is a solid with no expected inhalable particles and a vapour pressure around 0.0079 Pa at 20°C (value based on read-across from oleylamine). 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, considering the apparently low systemic toxicity of the primary alkylamines, no need for further inhalative testing was seen.

Justification for selection of Effect on fertility via dermal route:
The registered substance as a solid and large salt is not expected to pass the skin. Therefore, the dermal route is not an adapted route of testing.

Effects on developmental toxicity

Description of key information

Developmental Toxicity studies on rabbits  by oral route (OECD 414, GLP) based on read-across from tallow alkyl amine (values converted on mw basis from tallow alkyl amine to the registered salt ): NOAEL maternal toxicity = 6.15 mg/kg bw/day, NOAEL developmental toxicity > 61.5 mg/kg bw/day. OECD 414 with  tallow alkyl amine in rat is also available with higher NOAELs.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

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

Adequacy of study:

key study

Study period:

1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Test procedure according to national guidelines and standards (TSCA Guidelines) including GLP but the reliability has been amended in accordance with 'practical guide 6: How to report read-across and categories' which states that the maximum reliability for a read-across study is 2. The study is considered to be adequate and reliable for the purpose of registration under REACH (Regulation (EC) No. 1907/2006).

Qualifier:

according to guideline

Guideline:

other: EPA regulations, TSCA (40 CFR Part 798.4700, September 1985, and revised edition May 1987)

GLP compliance:

yes

Remarks:

Springborn Laboratories, Inc., Mammalian Toxicology Division, 553 North Broadway, Spencerville, Ohio 45887, USA

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Portage, Michigan
- Age at study initiation: 14 weeks
- Weight at study initiation: 229-316 g
- Housing: Animals were housed individually during acclimatization and while on study, except during cohabitation, in suspended stainless steel wire mesh cages.
- Diet (ad libitum): Purina Certified Rodent Meal #5002
- Water (ad libitum): deionized tap water
- Acclimation period: 19 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): ~22
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12 / 12


IN-LIFE DATES: From: 1989-07-25 To: 1989-08-18

Route of administration:

oral: gavage

Vehicle:

other: Mazola® corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Dose solutions were prepared daily. Appropriate amounts of the test article for each dose group were weighed into volumetric flasks. Mazola® corn oil was added in sufficient quantity to achieve the final concentrations. The flasks were inverted several times to ensure adequate mixture. Dosing solutions were stored under a nitrogen blanket at room temperature.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

On the first day of dosing and near the end of the dosing period, a subsample from each dosing solution, including the control, was taken and analyzed for verification of the test article concentration.

Details on mating procedure:

Female rats determined to be suitable test subjects were cohabitated with proven resident Sprague Dawley Cr1:COBS®CD®BR®VAF/PLUS® male rats. The male rats were of suitable health and were nine to eleven months old. Evidence of mating was determined by the presence of a copulatory plug in the vagina of a sperm positive vaginal smear and was considered day 0 of gestation.

Duration of treatment / exposure:

from gestation day 6 through 15

Frequency of treatment:

single daily doses

Duration of test:

10 days

Remarks:

Doses / Concentrations:
10, 40, 80 mg/kg
Basis:
nominal conc.

No. of animals per sex per dose:

28

Control animals:

yes, concurrent vehicle

Details on study design:

Range-finding study: 50, 100, 150, 250 mg/kg/day
Mortality occurred in the 100, 150 and 250 mg/kg/day groups. Outward clinical signs of toxicity and body weight losses or reduced weight gain occurred at the 50, 100, 150 and 250 mg/kg/day levels. A dose level of 100 mg/kg/day was considered to be excessive for a high dose level of the definitive teratology study due to the induced mortality. Conversely, 50 mg/kg/day did not produce sufficient maternal toxicity to be considered suitable as a high dose level. Thus, 80 mg/kg/day was selected in anticipation of producing sufficient maternal toxicity. Graduated doses of 40 and 10 mg/kg/day were selected as the mid and low dose levels, respectively. A dose level of 40 mg/kg/day was expected to induce minimal maternal toxicity while the 10 mg/kg/day dose level was selected to determine a no effect level for maternal and developmental toxicity.

Maternal examinations:

Animals were examined daily. Clinical signs of toxicity including physical or behavioral abnormalities were recorded. Mortality checks were performedtwice daily (morning and afternoon). During the dosing period, animals were observed for toxic effects between on-half hour and two hours postdosing.

Body weights were recorded on GD 0, 6, 9, 12, 16 and 20. BWC was recorded for GD 0-6. 6-9, 9-12, 12-16, 16-20, 6-16 and 0-20. Net maternal body weight gain (adjusted for gravid uterine weight) was also reported.

Food consumption was measured on GD 0, 6, 9, 12, 16, 20. Food consumption was calculated as g/kg/day and g/animal/day for the following gestation intervals: 0-6, 6-9, 9-12, 12-16, 16-20, 6-16, 0-20.

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:

Fetal morphological examination: external (each fetus), visceral (one-half of the fetuses from each litter), skeletal (one-half of the fetuses from each litter) abnormalities

Statistics:

Analyses were performed by a digital VAX 11/730 computer. Two-tailed tests were utilized unless otherwise indicated for a minimum significance level of 5% comparing the control group to each treatment group. Group comparisons were performed using Dunnett´s test. Count data were analyzed using Chi-square Test for fetal sex ratios, Mann-Whitney U-Test for resorptions, and Fisher´s Exact Test for the number of fetal variations and malformations.

Historical control data:

Cesarean section data, fetal malformation data, fetal variation data were provided.

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
Clinical signs of toxicity were observed at the 40 and 80 mg/kg/day levels. Salient clinical signs included salivation. Dose dependent body weight losses or reduced weight gain, along with a corresponding reduction in food consumption occurred.

Dose descriptor:

NOAEL

Effect level:

10 mg/kg bw/day (nominal)

Basis for effect level:

other: maternal toxicity

Dose descriptor:

NOAEL

Effect level:

> 80 mg/kg bw/day (nominal)

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No treatment-related changes were apparent at any level tested concerning necropsy observations, cesarean section data or fetal morphological examinations.

Dose descriptor:

NOAEL

Effect level:

> 80 mg/kg bw/day (nominal)

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Gross necropsy evaluation of the gastrointestinal tracts from females sacrificed on gestation day 15 revealed no irritative effects.

The oral administration of 40 and 80 mg/kg/day of Oleylamine to pregnant CD rats, induced dose dependent maternal toxicity exhibited by adverse clinical signs, body weight losses and reduced food consumption. A dosage level of 10 mg/kg/day was determined to be a no effect level for maternal toxicity. Oleylamine was neither developmentally toxic nor teratogenic at dosage levels of 10, 40 and 80 mg/kg/day.

Conclusions:

Oral administration of 40 and 80 mg/kg body weight per day of Oleylamine to pregnant CD rats induced dose dependent maternal toxicity exhibited by adverse clinical signs, body weight loss and reduced food consumption. A dosage level of 10 mg/kg body weight per day was determined to be a no effect level for maternal toxicity. Oleylamine was neither developmental toxic nor teratogenic at dosage levels of 10, 40 or 80 mg/kg body weight per day.

Executive summary:

For octadecenylamine ("Oleylamine") a guideline conform teratology study in Sprague Dawley rats has been performed. Prior to initiation of the main study, a range-finding study had been conducted. During the range finding-study, treatment-related deaths had occurred in the 100, 150 and 250 mg/kg bw/day groups. Outward clinical signs of toxicity and body weight loss or reduced weight gain occurred at the 50, 100, 150 and 250 mg/kg bw/day dose levels. A dose level of 100 mg/kg bw/day was considered to be excessive for a high dose level of the main study due to the induced mortality. Conversely, 50 mg/kg bw/day did not produce sufficient maternal toxicity to be considered suitable as a high dose level. Thus, 80 mg/kg bw/day was selected for the main study in anticipation of producing sufficient maternal toxicity.

In the main study groups of 28 pregnant females were treated orally (gavage) with dosages of10, 40 or 80 mg/kg bw/day or with the vehicle (Mazola corn oil) during gestation days 6 to 15. During the study animals were examined daily. Any clinical signs of toxicity including physical or behavioural abnormalities were recorded. Individual body weights and food consumption were recorded on gestation days 0, 6, 9, 12, 16, and 20. Two animals in each group were selected to be sacrificed and necropsied after treatment on gestation day 15 to determine the appearance and severity of gastrointestinal tract irritation. On gestation day 20 caesarean section was performed on all surviving animals. The numbers of viable fetuses, early and late resorptions as well as the number of corpora lutea were recorded. Fetuses were examined for external, visceral and skeletal abnormalities.

All animals survived to scheduled sacrifice. Outward clinical signs of toxicity were observed at the 40 and 80 mg/kg bw/d dose levels. The observations most likely indicated a generalised irritative effect of the test substance as characterised by rales, salivation, unkempt appearance and changes in the amount, colour and consistency of the feces. However, no other signs of treatment-related gastrointestinal irritation or other internal changes were observed at the gestation day 15 and 20 necropsies. More pronounced signs of toxicity were apparent only in the 80 mg/kg bw/d dose group and included emaciation, rough coat and dark red material around the eyes, nose and/or mouth. Similar clinical signs were infrequently noted during the post-dose observations. Dose-dependent body weight loss (during gestation days 6-9) or reduced weight gain (during gestation days 12-16), along with a corresponding reduction in food consumption occurred during the treatment period in the 40 and 80 mg/kg bw/d dose groups. Net body weight gain (adjusted for gravid uterine weight) was also lower at these levels. Following cessation of treatment (days 16-20), increase in weight gain and food consumption were noted at both dose levels. No such effects were observed in the dose group treated with 10 mg/kg bw/d. Caesarean section data obtained from the treated groups did not reveal any meaningful differences (concerning number of corpora lutea, implantation sites, viable fetuses, fetal sex and fetal weight) when compared with the controls. Fetal evaluations of type and frequency of malformations and variations did not reveal any indications for a treatment related teratogenic effect. In summary, oral administration of octadecenylamine to pregnant rats produced dose-dependent maternal toxicity in the 40 and 80 mg/kg bw/d dose groups. No indications of an embryotoxic, fetotoxic or teratogenic effect was observed at any tested level. A NOAEL/maternal toxicity of 10 mg/kg bw/d and a NOAEL/developmental toxicity of>80 mg/kg bw/d can be derived from the study.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

61.5 mg/kg bw/day

Quality of whole database:

Two studies performed according to GLP and exhibited same results.

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

The registered substance is a salt composed of primary hydrogenated alkylamine and N-hydrogenated tallow fatty acid. As the primary alkyl amine part is much more toxic than the tallow fatty acid part, it is expectedthat the toxicological behaviour of the substance will not be far different from that of the primary alkylamine compounds or at least not more severe. This has been demonstrated by results obtained with the registered substance in a short-term repeated toxicity study (e.g. 14 days). Thus, assessment of the registered substance can be integrated in thecurrent approach of grouping of primary alkylamines substances. Experimental studies with the registered substance with respect to the endpoint developmental toxicity are not available. However, data from guideline according prenatal developmental toxicity testing in two species (rats, rabbits) is available for the closely related primary alkylamine (Z)-octacec-9 -enylamine (CAS No. 112 -90 -3). From the rat study a NOAEL (maternal toxicity) of 10 mg/kg body weight per day and a NOAEL (developmental toxicity) of greater 80 mg/kg body weight was derived. In addition, a guideline conform teratology study in rabbits with (Z)-octadec-9 -enylamine revealed general findings of irritation in the gastrointestinal tract and dose-dependent body weight loss or reduced weight gain during the treatment in the 30 mg/kg dose group. Caesarean section data did not reveal any significant differences in reproductive parameters and no indications of an embryotoxic, fetotoxic or teratogenic effect at any tested dose-level. A NOAEL (developmental toxicity) of greater 30 mg/kg body weight per day was retained.

When converted to the corresponding values for the registered salt, the levels become:

- NOAEL maternal toxicity = 6.15 mg/kg bw/day 

- NOAEL embryo-fetal toxicity = 61.5 mg/kg bw/day

 (mw tallow alkyl amine = 267, mw registered substance = 547.5; Correction factor: 547.5/267 = 2.05)

Justification for selection of Effect on developmental toxicity: via oral route:
GLP study with the lowest NOAELs

Justification for selection of Effect on developmental toxicity: via inhalation route:
The registered substance is a solid with no expected inhalable particles and a vapour pressure around 0.0079 Pa at 20°C (value based on read-across from oleylamine). 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, considering the apparently low systemic toxicity of the primary alkylamines, no need for further inhalative testing was seen.

Justification for selection of Effect on developmental toxicity: via dermal route:
The registered substance as a solid and large salt is not expected to pass the skin. Therefore, the dermal route is not an adapted route of testing.

Justification for classification or non-classification

Reproductive toxicity studies on the registered substance are not available. However, data relevant for hazard assessment with respect to the endpoints fertility and developmental toxicity are available for the closely related primary alkylamines (Z)-octadec-9 -enylamine (CAS No. 112 -90 -3) and tallow alkylamines (CAS No. 61790 -33 -8). In line with the existing EU risk assessment on primary alkylamines it is considered appropriate to extrapolate from these data and therefore read-across is considered adequate for hazard assessment. Also in line with the existing EU risk assessment, additional testing is not regarded to be necessary. Thus, according to the results from the available studies classification there is no need to classify the substance with regard to reproductive toxicity.