C16-18-(even numbered, C18-unsaturated)-alkylamines acetates

Endpoint summary

Description of key information

Only limited data are available for the primary alkylamine C16-18-(even numbered)-alkylamines with regard to repeated dose toxicity. However, 14-day extended dose-range-finder screening studies, have illustrated clear consistencies in the toxicity profile within alkylamine acetates on one side and their corresponding alkyl amines on the other side. Therefore the 28-day oral toxicity test with C16-18-(even numbered, saturated and unsaturated)- alkylamines can be used for hazard and risk characterization purposes based on read-across principles. Groups of five male and female rats received the test item by oral gavage at dose levels of 0, 3.25, 12.5 or 50 mg/kg body weight per day for a period of 28 days. At a dose of 50 mg/kg body weight per day clinical signs such as gait abnormalities, reduction in body weight gain and clinical pathology findings indicating mild toxic effects on the liver and kidneys were found. Effects observed at the mid dose level (12.5 mg/kg) were slight reduction in growth. At the low dose group of 3.25 mg/kg body weight per day no effects were observed. Hence, the NOAEL of this study was placed at 3.25 mg/kg body weight per day. This value is considered to be conservative but valid also for C16-18-(even numbered)-alkylamine acetates and will be used for all relevant exposures by route-to-route extrapolation for the registration substance.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study according to GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

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: Harlan Winkelmann GmbH, Borchen, Germany
- Age at study initiation: approx. 6 weeks
- Fasting period before study: no
- Housing: transparent macrolon cages (type IV)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 50 +/- 20 %
- Air changes (per hr): 12 - 15
- Photoperiod (hrs dark / hrs light): 12 hours periodically

Route of administration:

oral: gavage

Vehicle:

other: sesame oil

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

28 applications in 29 days

Frequency of treatment:

daily

Remarks:

Doses / Concentrations:
0 mg/kg body weight per day (control)
Basis:
actual ingested

Remarks:

Doses / Concentrations:
3.25 mg/kg body weight per day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
12.5 mg/kg body weight per day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
50 mg/kg body weight per day
Basis:
actual ingested

No. of animals per sex per dose:

5 male and 5 female

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: preliminary dose-range-finding study
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: recovery group
- Post-exposure recovery period in satellite groups: 14 days
- Section schedule rationale (if not random): random

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily for mortality, once daily for clinical signs

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once daily

BODY WEIGHT: Yes
- Time schedule for examinations: before treatment, then twice weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption was determined continously two times per week

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at termination and after recovery period
- Anaesthetic used for blood collection: Yes (ketamin-hydrochloride + xylazine)
- Animals fasted: No
- How many animals: all

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after blood sampling for haematological testing, animals were killed and exsanguinated
- Animals fasted: No
- How many animals: all

URINALYSIS: Yes
- Time schedule for collection of urine:urine collection overnight from day 25 to day 26
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes: food and water was withdrawn during this period

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: once before first treatment, then once a week
- Dose groups that were examined: all
- Examinations: at all examinations: changes in appearance, occurence of secretions/excretions, autonomic activity (lacrimation, salivation, nasal discharge, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture, response to handling, presence of clonic/tonic movements, tremor, other abnormal motor movements (excessive grooming, repetitive circling, other stereotypes), bizarre behaviour, defecation, urination
- Battery of functions tested: at termination of the study:sensory activity, grip strengt, motor activity

Sacrifice and pathology:

GROSS PATHOLOGY: Yes: macroscopic examinations: skin, orifices, eyes, teeth, oral mucosa, internal organs
HISTOPATHOLOGY: Yes (macroscopic and microscopic examinations): adrenals, bone marrow (sternum), brain with medulla oblongata, epididymides, heart, small intestine (jejunum), large intestine (colon) kidneys, liver, lungs, lymph nodes (mandibular, iliac), nerve (sciatic), ovaries with oviducts, prostate, seminal vesicle, spinal cord (cervical) spleen, stomach, testes, thymus, thyroid gland with parathyroids, trachea with larynx, urinary bladder, uterus, nasal cavities

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

stilted gait, no unscheduled deaths

Mortality:

mortality observed, treatment-related

Description (incidence):

stilted gait, no unscheduled deaths

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

decreased for high-dose males and females, and mid-dose males

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

increased neutrophils in high-dose animals

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

slightly increased ASAT and ALAT acitivity in high-dose males

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
- no unscheduled deaths throughout the study
- 50 mg/kg/day: motility impairment from 2nd/3rd week in 2 males, 1 female (main group) and in 4 females (with recovery) in the recovery group;
respiratory sounds (1 female, day 13)
- 12.5 mg/kg/day: no changes
- 3.25 mg/kg/day: no changes


BODY WEIGHT AND WEIGHT GAIN
- 50 mg/kg/day: approx. 10% decrease in body weight in males from day 11 onwards for about; the effect showed a tendency of recovery. Mean body weight was decreased in females from day 22 onwards, with clear subsequent recovery. Mean overall body weight gain over the study period was moderately affected.
- 12.5 mg/kg/day: Slight decrease (<5%) of mean body weight and mean overall body weight gain in males from day 22 onwards; the effects were considered not to be toxicologically relevant, since they were marginal and not recorded in females.
- 3.25 mg/kg/day: no significant changes


FOOD CONSUMPTION AND COMPOUND INTAKE
remained unaffected in all groups


HAEMATOLOGY
- 50 mg/kg/day: males: slightly increased haematocrit, slightly decreased reticulocyte counts, subsequent recovery; females: similar tendency without statistical significance. Slightly increased white blood cell counts with shift towards neutrophils in males and females (not stat. significant but
outside range of historical controls). All findings being reversible.
- 12.5 mg/kg/day: no significant changes
- 3.25 mg/kg/day: no significant changes


CLINICAL CHEMISTRY
50 mg/kg/day: increased total bilirubin in males and females, slightly increased urea nitrogen in females and ASAT and ALAT activity in males.
12.5 mg/kg/day: increased total bilirubin and slightly increased urea nitrogen in females.
3.25 mg/kg/day: increased total bilirubin in females

URINALYSIS
remained unaffected in all groups

NEUROBEHAVIOUR
- Open field observations, assessment of sensory function, and forelimb and hindlimb grip strength not influenced in all groups.
- Negative trend in number of movements in males, but not significant and not present in females. Effect within the range of historical (inhouse)
controls, and thus, this was not considered to be related to treatment.

ORGAN WEIGHTS
- 50 mg/kg/day: Statistically significantly decrease of heart and brain weights in females and tendencially in males. Increase in relative adrenal weight (main group) and relative spleen weight (recovery group) in males. Since the effects were related to decreased terminal body weight and no histologically correlate was found, they are regarded to be no organ specific effects.
- 12.5 mg/kg/day: no changes
- 3.25 mg/kg/day: no changes

GROSS PATHOLOGY
No gross pathology findings attributable to administration of test compound in all dose groups.

HISTOPATHOLOGY: NON-NEOPLASTIC
No histopathological findings which could be related to the administration of the test compound in all test groups. Sporadically observed changes occurred at an incidence and severity historically seen for rats of this age and strain.

HISTOPATHOLOGY: NEOPLASTIC
No neoplastic chages observed in any dose group.

Dose descriptor:

NOAEL

Effect level:

3.25 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effetcs

Critical effects observed:

not specified

Conclusions:

Based on the test results the study director concluded that repeated administration of genamin OL 100 D at the high dose level of 50 mg/kg body weight per day induced general clinical signs, impairment of body weight gain and mild clinical pathology findings without histopathology correlates. It is concluded that the NOAEL from this valid oral 28-day study is 3.25 mg/kg body weight per day.

Executive summary:

Groups of five male and female SD-rats received octadecenylamine (Genamin OL 100 D, vehicle sesame oil) by oral gavage at dose levels of 0,3.25,12.5 or 50 mg/kg bw/day for a period of 28 days. On day 29 animals were necropsied. In the control and high dose groups, additional five male and females were examined and necropsied after a recovery period of 14 days. The study design and examinations conducted were in full accordance to the EU method B.7 (OECD 407) for subacute oral toxicity (including neurobehavioral observation and functional observation battery testing).

In a preliminary study on dose-range finding three males and females received the test substance at doses of 25, 100 and 400 mg/kg bw/d over a period of 14 days and surviving animals were necropsied on day 15. After administration of 400 mg/kg bw/d one male and one female died at days 4 and 7, respectively. The other animals of this dose group were killed for animal welfare reasons. The animals of the 100 mg/kg bw/d group showed clinical signs of impaired motility and respiration. The male animals were clearly more sensitive than the female animals. After administration of 25 mg/kg bw/d no symptoms were observed except of one female rat, which showed uncoordinated gait at study day 2. The body weight gains of animals exposed to doses of 25 and 100 mg/kd bw/d were impaired. Necropsy of the descendent and prior killed animals showed changes in the stomach and intestinal mucosa. The animals of 100 mg/kg bw/d showed reddening of the stomach mucosa. No macroscopically visible changes were observed in the 25 mg/kg dose groups.

In the main study, treatment resulted in no unscheduled deaths throughout the study. Behaviour and state of health remained unaffected by the administration of the test compound in the low and mid dose groups. Clinical findings in the high dose group (2 males and 5 females out of 10) from the 2nd or 3rd week onwards comprised impairments of motility (stilted and/or uncoordinated gait) and lasted until the end of treatment, with subsequent recovery. In addition, respiratory sounds were noted in one high dose female only on study day 13. No opacity of the refracting media of the eyes, changes of the oral mucosa, or impairment of dental growth was observed. No abnormal neurobehavior was observed in any group.

Mean body weight was significantly lower for high dose males from study day 11 and for high dose females from day 22 until the end of treatment, and remained to be different as compared to the controlat the end of recovery period. Also mean body weights for mid dose males were significantly lower from day 22 onwards to the end of treatment.

Compared to control animals, mean body weight gain was significantly lower at the end of treatment for high dose males (-19%), high dose females (-20%) and for mid dose males (-15%). A dose-dependent, small (-9%, statistically non-significant) reduction of body weight gain was also reported for the low dose males. The weight gain was normalised at the end of the recovery for high dose males or even higher in the high dose female group indicating a tendency to recovery (no recovery conducted for the mid dose group). Food consumption remained unaffected throughout the study in all dose groups.

Hematology findings in high dose groups included significantly increased hematocrit and decreased reticulocyte counts (males only) and slightly increased white blood cell counts with a shift towards increased neutrophils (both genders), all findings being reversible. Clinical chemistry changes comprised significantly increased total bilirubin for the high dose group, slightly increased urea nitrogen for mid and high dose females and very slightly increased ASAT and ALAT activity in the liver of high dose males. Urinalysis remained unaffected in all dose groups. Likewise the urine sediment was unobtrusive for control and high dose group animals.

No anatomic pathology correlates (organ weights, macroscopy, microscopy) of toxicological significance were detected.

In conclusion, repeated administration of Genamin OL 100 D (octadecenylamine) at a dose of 50 mg/kg bw/day induced clinical signs as gait abnormalities, reduction in body weight gain and clinical pathology findings indicating mild toxic effects on the liver and kidneys. The only treatment-related effects observed at the mid-dose level were reduction in growth and increased urinary concentration of urea nitrogen. By standard requirements on screening for neurotoxicology given for this type of study, stilted gait or uncoordinated gait was not associated with any other symptom of altered neurobehavior or neurotoxicity and may be discussed as being of unspecific nature. In the preliminary study, macroscopic findings in the gastrointestinal mucosa were observed in early deaths and in animals receiving 100 mg/kg bw/d, but were absent in animals at a dosage of 25 mg/kg bw/d from the 14-day study and in dose groups receiving 50 mg/kg bw/d.

Except for growth reduction, full recovery of findings was seen at the end of recovery period.

Based on the significantly reduced body growth at 12.5 mg/kg bw/d observed in the present 28-day study, the NOAEL of 3.25mg/kg bw/d was derived.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

3.25 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

For the alkylamines acetates C16-18-(even numbered, C18-unsaturated)-alkylamines acetates and C16-18-(even numbered)-alkylamines acetates 14 day extended dose-range-finder studies are available which exhibited very comparable toxicity profiles which, moreover, are also very similar to respective data from the analogous primary alkylamines. Based hereupon, read-across from analogue primary alkylamines is scientifically valid and robust based not only on structural, functional, metabolic considerations, but also based on close toxicological similarities. The chosen guideline conform 28-day repeated oral dose toxicity study is according to GLP and provides sufficient and valid information for hazard identification and classification also for the alkylamines acetates. No derivations and/or confounders have been identified. Klimisch rating provides reliability score of 1 meaning reliability without restrictions. The data base is considered to be sufficient to conclude on the labelling and classification of the registration substance.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

The available data allows`for route-to-route extrapolation.

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

The available data allows`for route-to-route extrapolation.

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

The available data allows for route-to-route extrapolation and/or expert judgement.

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

The available data allows for expert judgement concerning this endpoint.

Additional information

There are no human data available on repeated dose toxicity of the test item. Likewise no data from repeated dose studies are available for the inhalative or dermal route of exposure.

For the oral route of exposure, information from 14 -day dose-range-finder studies in rats on the registration substance C16 -18 -(even numbered, C18 -unsaturated)-alkylamine acetates as well as on the analogous unsaturated C16 -18 -(even numbered)-alyklamine acetates are available, which demonstrates good comparability of the results within saturated and unsaturated primary alkylamine acetates. A comparison with the toxicity profile obtained for the analoguous primary alkylamine C16 -18 -(even numbered, C18 -unsaturated)-alkylamines, also reveals a good correlation with regard to the general toxicity and effect profile, target organs and toxicokinetic behaviour.Therefore a 28-day oral toxicity test with C16-18-(even numbered, saturated and unsaturated)- alkylamines can be used for hazard and risk characterization purposes using read-across principles. This approach is scientifically justified because of close structural, functional, metabolic and demonstrated toxicological similarities between source and target molecules.

Groups of five male and female rats received the test item by oral gavage at dose levels of 0, 3.25, 12.5 or 50 mg/kg body weight per day for a period of 28 days. At a dose of 50 mg/kg body weight per day clinical signs such as gait abnormalities, reduction in body weight gain and clinical pathology findings indicating mild toxic effects on the liver and kidneys were found. Effects observed at the mid-dose level (12.5 mg/kg) were slight reduction in growth. At the low dose group of 3.25 mg/kg body weight per day no effects were observed. Hence, the NOAEL of this study was placed at 3.25 mg/kg body weight per day. This value is considered to be conservative but valid also for C16-18-(even numbered, C18 -unsaturated)-alkylamine acetates and will be used for all relevant inhalation and or dermal exposures by route-to-route extrapolation for the registration substance although the following limited data with dermal application exists for primary alkylamines .

Octadecenylamine (oleylamine) was dermally applied using occlusive conditions on two 5-days periods with an intermediate 2-day non dosing-period to groups of 4 male and 4 female Sprague-Dawley rats at concentrations of 0, 0.3, 1.5 and 3.0% (v/v) in mineral oil (corresponding to doses of 0, 12.5, 62.5 and 125 mg/kg bw/d) in order to more closely reproduce conditions of human exposure to the test substance. Due to excessive tissue destruction indicated by sloughing, scores of moderate to severe erythema, scabbing, hardening of the skin, and sensitivity to touch, the dosing at the intermediate and high dose levels (1.5 and 3.0%) was discontinued on Day 9. These animals were subsequently sacrificed on Day 10. All remaining rats survived until scheduled sacrifice. Concentrations of 1.5 and 3.0% produced moderate to severe irritation (erythema scores 2-4), which in some instances progressed to hardening and sloughing of the skin. In the 0.3 % group, erythema scores of 1 to 2 were observed, indicating mild to moderate irritation, and flaking of the outer layers of the epidermis was observed. An increased sensitivity in females to the irritant effects of the test substance as compared to males was observed. No other treatment-related irritant effects or clinical signs were observed. Individual group comparisons revealed that body weights in both the 1.5 and 3% groups were significantly lower than controls. Females in the 3.0% group showed a mean weight loss during the first week of the study, although this finding was not significant. Food consumption during the first week of study was also reduced significantly in the 1.5% group males when expressed as total food consumed. No significant difference was noted when expressed on a per weight basis. The study provides additional data on the toxicity of repeated dermal dosing, including severe irritation, of this test substance at concentrations of 0.3, 1.5 and 3.0%. A NOAEL for local dermal effects (irritation) could not be derived and a LOAEL of 0.3% (v/v) was established. No NOAEL for systemic toxicity was derivable because of lack of histomorphology data from other organs and tissues. However, no macroscopic indication of systemic toxicity was observed.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
A repeated dose toxicity study from oleyl alkylamine [C16-18-(even numberd, saturated, unsaturated)-alkyl amines] is used based on read-across considerations. This is justified based on structural, functional, metabolic and toxicological similarities as demonstrated in the read-across rational provided (see also chapter 13). The chosen study is a guideline conform test according to GLP with a Klimisch rating of 1. Read-across from this study allows proper endpoint related classification of the registration substance.

Repeated dose toxicity: via oral route - systemic effects (target organ) digestive: stomach; digestive: other

Justification for classification or non-classification

The results from the available GLP conform screening studies with repeated exposure of the registration substance revealed comparable toxicity profiles for primary alkylamine acetates and the corresponding primary alkylamines. Based hereupon, data from a guideline conform repeated dose toxicity study according to OECD TG 407 is used for labelling and classification purposes of C16 -18 -(even numbered, C18 -unsaturated)-alkylamine acetates. This study revealed clinical signs such as gait abnormalities, reduction in body weight gain and clinical pathology findings indicating mild toxic effects on the liver and kidneys at a dose of 50 mg/kg body weight. Effects observed at the mid dose level (12.5 mg/kg) were slight reduction in growth. At the low dose group of 3.25 mg/kg body weight per day no effects were observed. Hence, the NOAEL of this study was placed at 3.25 mg/kg body weight per day.

Based on ECHA guidance on the application of the CLP criteria (version 3.0), substances are classified as specific target organ toxicants following repeated exposure by the use of expert judgment on the basis of the weight of all evidence available, including the use of recommended guidance values. In this respect, the evidence from studies with repeated dose toxicity studies indicate that the submission substance has the potential to be harmful to human health following repeated exposure. In this respect, identical classification proposals from the Committee for Risk Assessment (RAC) based on Regulation (EC) No 1272/2008 (CLP) exist for primary alkylamines. Taking into account the respective RAC opinion for C16 -18 -(even numbered, saturated and unsaturated)-alkylamines as well as for C16 -18 -(even numbered, C18 -unsaturated)-alkylamines which are used as source molecules for read-across, the registration substance C16 -18 -(even numbered, C18 -unsaturated)-alkylamine acetates should be classified as

Xn; R48/22 Harmful: Danger of serious damage to health by prolonged exposure if swallowed (according to DSD)

Warning; STOT-RE Category 2 - H373: May cause damage to organs (gastro-intestinal tract, liver, immune system) through prolonged or repeated exposure via the oral route (according to CLP)