N-[3-(dimethylamino)propyl]stearamide

Endpoint summary

Description of key information

- Subacute (14 day dose-range finding study) repeated dose toxicity study oral (gavage), rat Crl:WI(Han)) m/f (similar to OECD TG 407, GLP), dose levels: 0, 50, 200, 500 mg/kg bw/d; no NOAEL derived from this dose range finding study

- Subchronic (90 day) repeated dose toxicity study, dermal, rabbit (New Zealand White) m/f (similar to OECD TG 411 GLP), dose levels: 5, 200 mg/kg bw/d: NOAEL = 200 mg/kg bw/d

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

Remarks:

other: 14 d dose range finding study

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2012-05-09 to 2012-05-30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Version / remarks:

30 May 2008

Deviations:

no

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

03 October 2008

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Crl:WI(Han)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: app. 6 weeks
- Housing: group housing of 3 animals per sex in Macrolon cages with sterilised sawdust as bedding material and paper as cage-enrichment
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24°C
- Humidity (%): 40-70%
- Air changes (per hr): 15/h
- Photoperiod (hrs dark / hrs light):12/12

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
formulations were placed on a magnetic stirrer during dosing;
Formulations (w/w) were prepared daily within 6 hours prior to dosing and were homogenised to a visually acceptable level. Adjustment was made for specific gravity of the vehicle.

Dose volume: 5 mL/kg bw, actual dose volumes were calculated weekly according to the latest body weight

VEHICLE
- Justification for use and choice of vehicle (if other than water): Based on trial formulations performed at testing laboratory and on information from the sponsor.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

No chemical analysis of formulations were performed in the 14-day range finding study. Instead, analysis on formulations will be performed as part of the subsequent repeated dose toxicity study.

Duration of treatment / exposure:

14 d

Frequency of treatment:

daily

Remarks:

Doses / Concentrations:
0, 50, 200, 500 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

3

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Initially, one dose group at 500 mg/kg bw/d was initiated (next to a concurrent control group). Based on the results obtained at 500 mg/kg bw/d, two additional doses at 50 and 200 mg/kg bw/d were added one week later.
- Rationale for animal assignment (if not random): animals were allocated at random, with all animals within ± 20% of the sex mean
- Rationale for selecting satellite groups: no satellite groups were used in the dose range finding study

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least twice daily; animals showing pain, distress or discomfort, which was considered not transient in nature or was likely to become more severe, were sacrificed for humane reasons
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least once daily from start of treatment onwards, detailed clinical observations were made in all animals at 0-15 minutes, 1 hour (±15 minutes) and 3 hours (± 30 minutes) after dosing.
BODY WEIGHT: Yes
- Time schedule for examinations: Days 1, 4, 7, 10 and 14.

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- over days 1-4, 4-7, 7-10 and 10-14

WATER CONSUMPTION: Yes
- Time schedule for examinations: Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked in table [No.1] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: immediately prior to scheduled post mortem examination
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked in table [No.1] were examined.

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table 2)
HISTOPATHOLOGY: Yes (see table 2)

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not examined

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

not specified

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
All animals at 500 mg/kg bw/d were sacrificed for humane reasons between Days 6 and 8.
No mortality occurred at 50 and 200 mg/kg bw/d.

500 mg/kg bw/d:
- lethargy, hunched posture, labored respiration, abdominal swelling, piloerection, chromodacryorrhoea, a lean appearance and/or ptosis from day 4 of treatment onwards

200 mg/kg bw/d:
- all animals showed piloerection on two days of week 2 only

50 mg/kg bw/d:
- no clinical signs were noted

Salivation seen after dosing among all animals at 200 and 500 mg/kg bw/d on a few days of treatment was considered to be a physiological response rather than a sign of systemic toxicity considering the nature and minor severity of the effect and its time of occurrence (i.e. after dosing).

BODY WEIGHT AND WEIGHT GAIN
500 mg/kg bw/d
- two females showed weight loss between days 1 and 4 (2 or 5% compared to day 1), followed by a slight weight gain between days 4 and 7. One female and all males at 500 mg/kg bw/d showed a reduced weight gain throughout the treatment period.

At 50 and 200 mg/kg, body weights and body weight gain remained in the same range as controls over the study period (the animals used for dosing at 50 and 200 mg/kg bw/d were from a different batch of delivery, and hence the starting body weight on day 1 was different to that for control animals and animals at 500 mg/kg bw/d).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
500 mg/kg bw/d:
- food consumption before or after correction for body weight was reduced in both genders over days 1-4 and 4-7

200 mg/kg bw/d:
- food consumption before or after correction for body weight was similar to controls over the study period

50 mg/kg bw/d:
- food consumption before or after correction for body weight was similar to controls over the study period

HAEMATOLOGY
The following changes in haematology parameters distinguished treated from control animals and from normal range levels encountered for rats of this age and strain:
- Slightly lower red blood cell counts in males at 50 and 200 mg/kg bw/d (no clear dose related trend).
- Higher reticulocyte counts in males at 50 and 200 mg/kg bw/d (no clear dose related trend).

CLINICAL CHEMISTRY
The following changes in clinical biochemistry parameters distinguished treated animals from control animals and from normal range levels encountered for rats of this age and strain:
-higher alanine aminotransferase activity (ALAT) in two males at 50 mg/kg bw/d, and two males and one female at 200 mg/kg bw/d
-Higher alkaline phosphatase activity (ALP) in one female at 200 mg/kg bw/d
-Higher potassium level in males at 50 and 200 mg/kg bw/d


ORGAN WEIGHTS
Spleen and thymus weights of females at 200 mg/kg bw/d appeared slightly increased compared to the control group.

The apparent lower liver, spleen and heart weight of males at 200 mg/kg bw/d was ascribed to a slightly lower terminal body weight as the ratio to body weight for these organs was similar to control levels.

GROSS PATHOLOGY
500 mg/kg bw/d:
- reduced size of seminal vesicles, prostate, epididymides, spleen and/or thymus, gelatinous contents in the gastro-intestinal tract, small intestines and/or caecum, gastro-intestinal tract distended with gas, and emaciated appearance

200 mg/kg bw/d:
- no abnormalities

50 mg/kg bw/d:
- no abnormalities

HISTOPATHOLOGY: NON-NEOPLASTIC
500 mg/kg bw/d:
-Thymus: Lymphoid atrophy in 2/2 females (moderate).
-Stomach: Hyperplasia of the forestomach in 3/3 males and 3/3 females at slight or moderate degree, inflammation of the forestomach in 3/3 males and 3/3 females up to moderate degree, ulceration of the forestomach in 1/3 males and 1/3 females at slight degree.
-Duodenum: Hyperplasia of the villi in 2/3 males and 2/3 females up to slight degree.
-Jejunum: Hyperplasia of the villi in 1/3 females at minimal degree.
-Mesenterial lymph node: Foamy macrophages and sinusoidal dilation in 3/3 males and 3/3 females at slight or moderate degree, and congestion/erythrophagocytosis in 1/3 males and 2/3 females at minimal degree.
-Testes: Absence of spermiation (massive degree) and degeneration of spermatids in 3/3 males up to slight degree.
-Epididymides: Oligospermia in 2/2 males at massive degree and seminiferous cell debris in 2/2 males at slight degree.
-Prostate: Reduced contents in 2/2 males at slight degree.
-Seminal vesicles: Reduced contents in 2/2 males at moderate degree.

No treatment-related microscopic abnormalities were noted at 50 and 200 mg/kg bw/d.

Dose descriptor:

other: based on the results, doses were selected for OECD TG 421 study

Basis for effect level:

other: see 'Remark'

Remarks on result:

not measured/tested

Remarks:

Effect level not specified (migrated information)

Critical effects observed:

not specified

All animals at 500 mg/kg bw/d were sacrificed for humane reasons between Days 6 and 8, and showed lethargy, hunched posture, laboured respiration, abdominal swelling, piloerection, chromodacryorrhoea, a lean appearance and/or ptosis from Day 4 of treatment onwards. All animals showed weight loss or reduced body weight gain and reduced food consumption during the treatment period. Necropsy findings at 500 mg/kg bw/d primarily consisted of gelatinous contents in the gastro-intestinal tract or parts thereof, and emaciation. The main cause for moribundity at this dose level was forestomach ulceration and/or hyperplasia of the squamous epithelium of the forestomach. Other histopathological changes noted at this dose level included:

-     Lymphoid atrophy of the thymus in 2/2 females examined, correlating to a reduced size of the thymus at necropsy.

-     Hyperplasia and inflammation (3/3 males and 3/3 females) and ulceration (1/3 males and 1/3 females) of the forestomach.

-     Hyperplasia of the villi in the duodenum (2/3 males and 2/3 females) and jejunum (1/3 females).

-     Foamy macrophages and sinusoidal dilation (3/3 males and 3/3 females) and congestion/ erythrophagocytosis (1/3 males and 2/3 females) in the mesenterial lymph node.

-     Absence of spermiation and degeneration of spermatids in the testes in 3/3 males, oligospermia and seminiferous cell debris in the epididymides, and reduced contents in the prostate and seminal vesicles in 2/2 males examined, which corresponded to a reduced size of seminal vesicles, prostate and epididymides at necropsy.

 

At 50 and 200 mg/kg bw/d, no mortality occurred. All animals at 200 mg/kg bw/d showed piloerection on two days of Week 2 only, whilst no clinical signs were noted at 50 mg/kgbw/d. Body weights, body weight gain and food intake remained in the same range as controls over the study period at these dose levels.

 

At 50 and 200 mg/kg bw/d, haematological changes consisted of slightly lower red blood cell and higher reticulocyte counts in males. No clear dose related trend was noted for these changes, which were generally slight in nature. Clinical biochemistry changes consisted of higher alanine aminotransferase activity in two males at 50 mg/kg bw/d, and two males and one female at 200 mg/kg bw/d, higher alkaline phosphatase activity in one female at 200 mg/kg bw/d, and higher potassium level in males at 50 and 200 mg/kg bw/d.

 

At 50 and 200 mg/kg bw/d, no abnormalities were noted at necropsy. Spleen and thymus weights of females at 200 mg/kg bw/d appeared slightly increased compared to the control group. No treatment-related histopathological changes were noted at 50 and 200 mg/kg bw/d.

Conclusions:

In a 14 d dose range finding subacute toxicity study, Stearic acid 3-(dimethylaminopropyl)amide was administered to 3 Crl:WI(Han) rats/sex/dose orally via gavage at dose levels of 0, 50, 200 and 500 mg/kg bw/day.
All animals of the highest dose group were sacrificed for humane reasons based on clinical signs. The main cause for moribundity was forestomach ulceration and/or hyperplasia of the squamous epithelium of the forestomach.
In the 50 and 200 mg/kg bw/d dose groups, no mortality occurred, and no or only minor clinical signs were observed. No treatment related effects were observed at necropsy or histopathology.

Executive summary:

In a 14 d dose range finding study according to OECD guideline 407, adopted 03 October 2008, and EU method B.7, May 2008, Stearic acid 3-(dimethylaminopropyl)amide was administered to 3Crl:WI(Han) rats/sex/dose orally via gavage at dose levels of 0, 50, 200 and 500 mg/kg bw/day.

All animals in the 500 mg/kg bw/d dose group were sacrificed for humane reasons between days 6 and 8. They showed lethargy, hunched posture, laboured respiration, abdominal swelling, piloerection, chromodacryorrhoea, a lean appearance and/or ptosis from day 4 of treatment onwards. All animals showed weight loss or reduced body weight gain and reduced food consumption during the treatment period. Necropsy findings at 500 mg/kg bw/d primarily consisted of gelatinous contents in the gastro-intestinal tract or parts thereof, and emaciation. The main cause for moribundity at this dose level was forestomach ulceration and/or hyperplasia of the squamous epithelium of the forestomach. Other histopathological changes noted at this dose level included: lymphoid atrophy of the thymus, correlating to a reduced size of the thymus at necropsy; hyperplasia and inflammation of the forestomach; hyperplasia of the villi in the duodenum and jejunum; foamy macrophages and sinusoidal dilation and congestion/ erythrophagocytosis in the mesenterial lymph node; absence of spermiation and degeneration of spermatids in the testes, oligospermia and seminiferous cell debris in the epididymides, and reduced contents in the prostate and seminal vesicles, which corresponded to a reduced size of seminal vesicles, prostate and epididymides at necropsy.

 

At 50 and 200 mg/kg bw/d, no mortality occurred. All animals at 200 mg/kg bw/d showed piloerection on two days of week 2 only, whilst no clinical signs were noted at 50 mg/kg bw/d. Body weights, body weight gain and food intake remained in the same range as controls over the study period at these dose levels.

 

At 50 and 200 mg/kg bw/d, haematological changes consisted of slightly lower red blood cell and higher reticulocyte counts in males. No clear dose related trend was noted for these changes, which were generally slight in nature. Clinical biochemistry changes consisted of higher alanine aminotransferase activity in two males at 50 mg/kg bw/d, and two males and one female at 200 mg/kg bw/d, higher alkaline phosphatase activity in one female at 200 mg/kg bw/d, and higher potassium level in males at 50 and 200 mg/kg bw/d.

 

At 50 and 200 mg/kg bw/d, no abnormalities were noted at necropsy. Spleen and thymus weights of females at 200 mg/kg bw/d appeared slightly increased compared to the control group. No treatment-related histopathological changes were noted at 50 and 200 mg/kg bw/d.

 

Based on these results, dose leves were selected for the reproduction/developmental toxicity screening test according to OECD guideline.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Study duration:

subacute

Species:

other: rat; animals in the highest dose group hat to be killed due to humane reasons; no NOAEL derived from this dose range finding study

Quality of whole database:

dose range finding study similar to OECD guideline, GLP

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From March 14, 1984 to June 13, 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

Only 2 dose levels tested while guideline recommends at least 3 doses; 5 animals/sex/group against standard 10 animals/sex/group; vehicle control (30/70 EtOH/water) not tested; clinical chemistry not performed. Test material was applied only for 4 hours.

GLP compliance:

yes

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hazleton Research Animal
- Age at study initiation: Not reported
- Weight at study initiation: 1970 – 2593 g
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
No information on environmental conditions is provided by the study report. However, standard operating procedure of the test facility was followed for environmental conditions.

IN-LIFE DATES: From: March 14, 1984 To: June 13, 1984

Type of coverage:

open

Vehicle:

other: 30/70 Ethanol/water

Details on exposure:

TEST SITE
- Area of exposure: Area on the back of each animals, from shoulder to rump, approximately 15 cm wide (intact skin)
- % coverage: Not reported
- Time intervals for shavings or clippings: The animals were clipped as needed while the test is in progress.

METHOD OF APPLICATIONS: Appropriate concentration of test material or control substance was applied evenly over the clipped area using a syringe.

REMOVAL OF TEST SUBSTANCE
- Washing: The treated skin was washed with tepid water and gently blotted dry with disposable paper towels or equivalent.
- Time after start of exposure: 4 hours

TEST MATERIAL:
- Amount(s) applied: 2.0 mL/kg/day (each animal received a constant dosage volume based upon its most recent bodyweight).
- Concentration: 0 (distilled water), 0.25 and 10% w/v of the test substance in 30%/70% ethanol/water.
- Constant concentration used: Yes
- Rate of preparation of dosing solution: Dosing solutions were prepared fresh weekly.
- Storage temperature of dosing solution: Stock solutions were stored at ambient temperature and humidity upon receipt and during the entire length of the study. Test solutions after preparation were stored in refrigerator. Each day’s dosing solution was equilibrated to room temperature prior to dosing.

VEHICLE: 30%/70% ethanol/water

USE OF RESTRAINERS FOR PREVENTING INGESTION: Yes, collars were used to restrain animals from oral ingestion.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

100 mL aliquot of each week’s dosing solution was sampled on the day of preparation and sent back to sponsor for analysis. A 100 mL aliquot of the distilled water control was sampled and sent for analysis on the first and last sampling. All samples were shipped under ambient conditions in glass containers.
The formulations were shown to be accurately prepared for this study. Details on analytical results are provided in the study report.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Once daily (5 days/week)

Remarks:

Doses / Concentrations:
0, 0.25 and 10 % (w/v) (equivalent to 0, 5 and 200 mg/kg bw/day) of the test substance
Basis:
nominal per unit area

No. of animals per sex per dose:

5 animals/sex/dose group

Control animals:

other: yes (distilled water)

Details on study design:

- Dose selection rationale: Not reported
- Selection of animals for experiment: Based on acclimation body weight, general health and pre-dose haematological and biochemical parameters, rabbits were selected for allocation to 3 groups. Prior to final assignment to the study, a veterinary examination was conducted to ensure that the selected rabbits were healthy.
- Rationale for animal assignment: Animals were randomized into groups by weight and sex with 5 males and 5 females in each group by weight and sex.
- Assignment of animals: The animals were assigned into following experimental groups in the study:
Group 1: Control (Distilled water (2 mL/kg))
Group 2: 0.25% w/v in 30%/70% ethanol/water (equivalent to 5 mg/kg/day)
Group 3: 10% w/v in 30%/70% ethanol/water (equivalent to 200 mg/kg/day)
All animals were dosed at a constant dosage volume of 2 mL/kg/day.

Observations and examinations performed and frequency:

Mortality/Morbidity: Yes
- Time schedule: Once daily

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily.
- Cage side observations included: Signs of ill health and abnormalities.

DETAILED CLINICAL OBSERVATIONS: Not reported

DERMAL IRRITATION: Yes, immediately prior to treatment each day (after Day 1), the skin sites were assessed on a numerical basis according to Draize skin reaction scoring system. The scoring scale for skin reactions is provided in the study report.

BODY WEIGHT: Yes
- Time schedule for examinations: All rabbits were weighed prior to dosing and subsequently at weekly intervals throughout the study.

FOOD CONSUMPTION: No

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes, blood samples were obtained from the central ear artery and were collected into tubes containing appropriate amounts of anticoagulant (EDTA).
- Time schedule for collection of blood: 7 days prior to the start of the study and at termination.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All animals
- Parameters examined: Complete blood counts including a differential white blood cell count.

CLINICAL CHEMISTRY: No

Sacrifice and pathology:

SACRIFICE: Yes, all the surviving animals were sacrificed with an IV overdose of sodium pentobarbital.

GROSS PATHOLOGY: Yes, complete necropsy was performed on all animals and macroscopic appearance of the tissues was recorded.

HISTOPATHOLOGY: Yes, tissues collected at necropsy were processed for microscopic evaluations.
-Samples of the following tissues from all rabbits collected for histological examination: Lung, heart, aorta, tongue, esophagus, thyroid (parathyroid), submandibular lymph node, ileocecocolic lymph node, anterior mesenteric lymph node, stomach, liver, gallbladder, duodenum, jejunum, ileum, cecum, colon, rectum, urinary bladder, kidneys, reproductive tracts, adrenal glands, thymus, spleen, pancreas, bone marrow, skin, brain, spinal cord, sciatic nerve, submandibular salivary gland, pituitary, eye. For paired tissues (except kidney and lungs), the left side was taken for processing and the right side was placed into a save jar for possible later histology.
All tissues were fixed in 10% neutral buffered formalin with the exception of the following tissues which were fixed in a glutaraldehyde/formalin fixative:
Anterior mesenteric lymph nodes, ileocecocolic lymph node, submandibular lymph node, spleen, adrenals, duodenum, jejunum, ileum, cecum, colon, rectum.

Other examinations:

ORGAN WEIGHT: The following organs from each sacrificed animal, were dissected free of fat and weighed: Adrenals, kidneys, liver and gross observations.

Statistics:

Initial body weights, body weight changes, hematology, and organ/body weight ratios were statistically analyzed by R D Bruce and B Stinson of Statistical resources using Procter & Gamble’s computer program B8944.

Clinical signs:

no effects observed

Dermal irritation:

effects observed, treatment-related

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY: No mortality or test related toxicity was observed throughout the study excepting slight conjunctivitis in one animal of control and two animals of Group 2 (5 mg/kg/day) animals. This was attributed to the continuous restraint which prevented the animals from grooming themselves.
The collars of one control animal, one Group 2 (5 mg/kg/day) animal and one Group 3 (200 mg/kg/day) animal were removed temporarily because of sores on the necks. Some animals removed their collars overnight and they were collared again the next day.

BODY WEIGHT AND WEIGHT GAIN: No treatment related changes in body weight and body weight gain were observed during the study.

HAEMATOLOGY: Statistically significant increases in the WBC values were noted in the Group 3 (200 mg/kg/day) animals. In addition, there was an increase in platelet values from baseline to necropsy of the Group 2 (5 mg/kg/day) animals. The changes in WBC of the Group 3 animals were attributed to the chronic stress of collaring and not considered to be test substance related. The significant increase in platelet values of Group 2 animals were a result of low baseline values.

ORGAN WEIGHTS: No test related biologically significant changes were noted in the absolute and relative liver, kidney and adrenal weight determinations.

GROSS & HISTO PATHOLOGY (NON-NEOPLASTIC):
Treated skin on the back of rabbits in both the treated groups had a dry hair coat with an accumulation of test material on the surface at necropsy. Except the treatment site skin, no test related gross effects were observed. Histopathological examinations revealed minimal acanthosis and hyperkeratosis in the treatment sites skin of all treated groups. The incidence and severity were similar in both groups. Incidental non treatment related histopathological changes were noted in several other tissues such as brain, liver, kidney, prostate and pancreas. However, these were considered to be incidental, and not treatment related.

OTHER FINDINGS: Local Dermal Reactions:
Treatment of animals with 0.25% test material (5 mg/kg/day) in 30/70% ethanol/water produced moderate or slight erythema, slight edema, slight desquamation and slight fissuring. At 10% (200 mg/kg/day), moderate erythema, slight edema, slight desquamation and slight fissuring were produced.

Dose descriptor:

NOAEL

Remarks:

(systemic)

Effect level:

>= 200 mg/kg bw/day

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Overall effects

Dose descriptor:

LOAEL

Remarks:

(local effects)

Effect level:

5 mg/kg bw/day

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: dermal irritation

Critical effects observed:

not specified

Conclusions:

Dermal application of Stearamidopropyldimethyl amine (SAPDMA) to hair clipped skin of New Zealand White rabbits at concentrations of 0 (distilled water), 0.25 and 10% w/v of the test substance in 30%/70% ethanol/water (equivalent to 0, 5 and 200 mg/kg bw/day) for 13 weeks revealed a systemic NOAEL of ≥10% w/v (equivalent to 200 mg/kg bw/day).

Executive summary:

The subchronic toxicity study of Stearamidopropyldimethyl amine (SAPDMA) was performed following methods comparable to the OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study).

Male and female New Zealand White rabbits, obtained from Hazleton Research Animals were used in the study. The body weight range of animals at study initiation was 1970 – 2593 g. Animals were acclimated to laboratory conditions for a period of 7 days prior to test initiation.

Test solutions were prepared fresh weekly in distilled, 30%/70% ethanol/water for each group. Post acclimation, animals were randomized into treatment/control groups (each consisting of 5 animals/sex).

The test solution was applied to intact skin of rabbits once daily, 5 days/week for thirteen consecutive weeks at the following dose levels:

Group 1(Control): Distilled water

Group 2: 0.25% w/v in 30%/70% ethanol/water (equivalent to 5 mg/kg/day)

Group 3: 10% w/v in 30%/70% ethanol/water (equivalent to 200 mg/kg/day) 

All animals (test and control) were dosed at a constant dosage volume of 2 mL/kg/day

Appropriate concentration of test material was applied evenly on the back of each animal, from shoulder to rump (approximately 15 cm wide) using a syringe.

Animals were observed for gross toxicity daily. The skin at the treatment sites was graded prior to each day’s treatment. Body weights were taken prior to initiation and then weekly until termination. Blood was taken prior to initiation and at termination for hematological examinations. After treatment, animals were sacrificed by an IV overdose of sodium pentobarbital. Adrenals, kidneys and liver were dissected free of fat and weighed. Tissue samples (including treated skin samples) were collected for histological examinations.

No mortality or test related gross toxicity was observed throughout the study except slight conjunctivitis in one control animal and two Group 2 (5 mg/kg/day) animals. This was attributed to the continuous restraint which prevented the animals from grooming themselves. The collars of one control animal, one Group 2 (5 mg/kg/day) animal and one Group 3 (200 mg/kg/day) animal were removed temporarily because of sores on the necks. Some animals removed their collars overnight and they were collared again the next day.

Treatment of animals with 0.25% test material (5 mg/kg/day) in 30/70% ethanol/water produced moderate or slight erythema, slight edema, slight desquamation and slight fissuring. At 10% (200 mg/kg/day), moderate erythema, slight edema, slight desquamation and slight fissuring were produced. No treatment related changes in body weight and body weight gain were observed during the study. No test related biologically significant changes were noted in the absolute and relative liver, kidney and adrenal weight determinations.

Statistically significant increases in the WBC values were noted in the Group 3 (200 mg/kg/day) animals. In addition, there was an increase in platelet values from baseline to necropsy of the Group 2 (5 mg/kg/day) animals. The changes in WBC of the Group 3 animals was attributed to the chronic stress of collaring and not considered to be test substance related. The significant increase in platelet values of Group 2 animals were a result of low baseline values.

Treated skin on the backs of rabbits in both treated groups had a dry hair coat with an accumulation of test material on the surface at necropsy. Histopathological examinations revealed minimal acanthosis and hyperkeratosis at the treatment sites of all treated groups. The incidence and severity were similar in both groups. Incidental non treatment related histopathological changes were noted in several other tissues such as brain, liver, kidney, prostate and pancreas. However, these were considered to be incidental, and not treatment related.

Based on above it can be concluded that, no systemic toxicity associated with the sub chronic percutaneous exposure to test material was observed at any dose levels.

Dermal application of Stearamidopropyldimethyl amine (SAPDMA) to hair clipped skin of New Zealand White rabbits at concentrations of 0 (distilled water), 0.25 and 10% w/v of the test substance in 30%/70% ethanol/water (equivalent to 0, 5 and 200 mg/kg bw/day)for 13 weeks revealed a systemic NOAEL of ≥10% w/v (equivalent to 200 mg/kg bw/day).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

200 mg/kg bw/day

Study duration:

subchronic

Species:

other: rabbit; highest dose tested

Quality of whole database:

similar to OECD guideline study, GLP

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From March 14, 1984 to June 13, 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

Only 2 dose levels tested while guideline recommends at least 3 doses; 5 animals/sex/group against standard 10 animals/sex/group; vehicle control (30/70 EtOH/water) not tested; clinical chemistry not performed. Test material was applied only for 4 hours.

GLP compliance:

yes

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Hazleton Research Animal
- Age at study initiation: Not reported
- Weight at study initiation: 1970 – 2593 g
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
No information on environmental conditions is provided by the study report. However, standard operating procedure of the test facility was followed for environmental conditions.

IN-LIFE DATES: From: March 14, 1984 To: June 13, 1984

Type of coverage:

open

Vehicle:

other: 30/70 Ethanol/water

Details on exposure:

TEST SITE
- Area of exposure: Area on the back of each animals, from shoulder to rump, approximately 15 cm wide (intact skin)
- % coverage: Not reported
- Time intervals for shavings or clippings: The animals were clipped as needed while the test is in progress.

METHOD OF APPLICATIONS: Appropriate concentration of test material or control substance was applied evenly over the clipped area using a syringe.

REMOVAL OF TEST SUBSTANCE
- Washing: The treated skin was washed with tepid water and gently blotted dry with disposable paper towels or equivalent.
- Time after start of exposure: 4 hours

TEST MATERIAL:
- Amount(s) applied: 2.0 mL/kg/day (each animal received a constant dosage volume based upon its most recent bodyweight).
- Concentration: 0 (distilled water), 0.25 and 10% w/v of the test substance in 30%/70% ethanol/water.
- Constant concentration used: Yes
- Rate of preparation of dosing solution: Dosing solutions were prepared fresh weekly.
- Storage temperature of dosing solution: Stock solutions were stored at ambient temperature and humidity upon receipt and during the entire length of the study. Test solutions after preparation were stored in refrigerator. Each day’s dosing solution was equilibrated to room temperature prior to dosing.

VEHICLE: 30%/70% ethanol/water

USE OF RESTRAINERS FOR PREVENTING INGESTION: Yes, collars were used to restrain animals from oral ingestion.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

100 mL aliquot of each week’s dosing solution was sampled on the day of preparation and sent back to sponsor for analysis. A 100 mL aliquot of the distilled water control was sampled and sent for analysis on the first and last sampling. All samples were shipped under ambient conditions in glass containers.
The formulations were shown to be accurately prepared for this study. Details on analytical results are provided in the study report.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Once daily (5 days/week)

Remarks:

Doses / Concentrations:
0, 0.25 and 10 % (w/v) (equivalent to 0, 5 and 200 mg/kg bw/day) of the test substance
Basis:
nominal per unit area

No. of animals per sex per dose:

5 animals/sex/dose group

Control animals:

other: yes (distilled water)

Details on study design:

- Dose selection rationale: Not reported
- Selection of animals for experiment: Based on acclimation body weight, general health and pre-dose haematological and biochemical parameters, rabbits were selected for allocation to 3 groups. Prior to final assignment to the study, a veterinary examination was conducted to ensure that the selected rabbits were healthy.
- Rationale for animal assignment: Animals were randomized into groups by weight and sex with 5 males and 5 females in each group by weight and sex.
- Assignment of animals: The animals were assigned into following experimental groups in the study:
Group 1: Control (Distilled water (2 mL/kg))
Group 2: 0.25% w/v in 30%/70% ethanol/water (equivalent to 5 mg/kg/day)
Group 3: 10% w/v in 30%/70% ethanol/water (equivalent to 200 mg/kg/day)
All animals were dosed at a constant dosage volume of 2 mL/kg/day.

Observations and examinations performed and frequency:

Mortality/Morbidity: Yes
- Time schedule: Once daily

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily.
- Cage side observations included: Signs of ill health and abnormalities.

DETAILED CLINICAL OBSERVATIONS: Not reported

DERMAL IRRITATION: Yes, immediately prior to treatment each day (after Day 1), the skin sites were assessed on a numerical basis according to Draize skin reaction scoring system. The scoring scale for skin reactions is provided in the study report.

BODY WEIGHT: Yes
- Time schedule for examinations: All rabbits were weighed prior to dosing and subsequently at weekly intervals throughout the study.

FOOD CONSUMPTION: No

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes, blood samples were obtained from the central ear artery and were collected into tubes containing appropriate amounts of anticoagulant (EDTA).
- Time schedule for collection of blood: 7 days prior to the start of the study and at termination.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All animals
- Parameters examined: Complete blood counts including a differential white blood cell count.

CLINICAL CHEMISTRY: No

Sacrifice and pathology:

SACRIFICE: Yes, all the surviving animals were sacrificed with an IV overdose of sodium pentobarbital.

GROSS PATHOLOGY: Yes, complete necropsy was performed on all animals and macroscopic appearance of the tissues was recorded.

HISTOPATHOLOGY: Yes, tissues collected at necropsy were processed for microscopic evaluations.
-Samples of the following tissues from all rabbits collected for histological examination: Lung, heart, aorta, tongue, esophagus, thyroid (parathyroid), submandibular lymph node, ileocecocolic lymph node, anterior mesenteric lymph node, stomach, liver, gallbladder, duodenum, jejunum, ileum, cecum, colon, rectum, urinary bladder, kidneys, reproductive tracts, adrenal glands, thymus, spleen, pancreas, bone marrow, skin, brain, spinal cord, sciatic nerve, submandibular salivary gland, pituitary, eye. For paired tissues (except kidney and lungs), the left side was taken for processing and the right side was placed into a save jar for possible later histology.
All tissues were fixed in 10% neutral buffered formalin with the exception of the following tissues which were fixed in a glutaraldehyde/formalin fixative:
Anterior mesenteric lymph nodes, ileocecocolic lymph node, submandibular lymph node, spleen, adrenals, duodenum, jejunum, ileum, cecum, colon, rectum.

Other examinations:

ORGAN WEIGHT: The following organs from each sacrificed animal, were dissected free of fat and weighed: Adrenals, kidneys, liver and gross observations.

Statistics:

Initial body weights, body weight changes, hematology, and organ/body weight ratios were statistically analyzed by R D Bruce and B Stinson of Statistical resources using Procter & Gamble’s computer program B8944.

Clinical signs:

no effects observed

Dermal irritation:

effects observed, treatment-related

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY: No mortality or test related toxicity was observed throughout the study excepting slight conjunctivitis in one animal of control and two animals of Group 2 (5 mg/kg/day) animals. This was attributed to the continuous restraint which prevented the animals from grooming themselves.
The collars of one control animal, one Group 2 (5 mg/kg/day) animal and one Group 3 (200 mg/kg/day) animal were removed temporarily because of sores on the necks. Some animals removed their collars overnight and they were collared again the next day.

BODY WEIGHT AND WEIGHT GAIN: No treatment related changes in body weight and body weight gain were observed during the study.

HAEMATOLOGY: Statistically significant increases in the WBC values were noted in the Group 3 (200 mg/kg/day) animals. In addition, there was an increase in platelet values from baseline to necropsy of the Group 2 (5 mg/kg/day) animals. The changes in WBC of the Group 3 animals were attributed to the chronic stress of collaring and not considered to be test substance related. The significant increase in platelet values of Group 2 animals were a result of low baseline values.

ORGAN WEIGHTS: No test related biologically significant changes were noted in the absolute and relative liver, kidney and adrenal weight determinations.

GROSS & HISTO PATHOLOGY (NON-NEOPLASTIC):
Treated skin on the back of rabbits in both the treated groups had a dry hair coat with an accumulation of test material on the surface at necropsy. Except the treatment site skin, no test related gross effects were observed. Histopathological examinations revealed minimal acanthosis and hyperkeratosis in the treatment sites skin of all treated groups. The incidence and severity were similar in both groups. Incidental non treatment related histopathological changes were noted in several other tissues such as brain, liver, kidney, prostate and pancreas. However, these were considered to be incidental, and not treatment related.

OTHER FINDINGS: Local Dermal Reactions:
Treatment of animals with 0.25% test material (5 mg/kg/day) in 30/70% ethanol/water produced moderate or slight erythema, slight edema, slight desquamation and slight fissuring. At 10% (200 mg/kg/day), moderate erythema, slight edema, slight desquamation and slight fissuring were produced.

Dose descriptor:

NOAEL

Remarks:

(systemic)

Effect level:

>= 200 mg/kg bw/day

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Overall effects

Dose descriptor:

LOAEL

Remarks:

(local effects)

Effect level:

5 mg/kg bw/day

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: dermal irritation

Critical effects observed:

not specified

Conclusions:

Dermal application of Stearamidopropyldimethyl amine (SAPDMA) to hair clipped skin of New Zealand White rabbits at concentrations of 0 (distilled water), 0.25 and 10% w/v of the test substance in 30%/70% ethanol/water (equivalent to 0, 5 and 200 mg/kg bw/day) for 13 weeks revealed a systemic NOAEL of ≥10% w/v (equivalent to 200 mg/kg bw/day).

Executive summary:

The subchronic toxicity study of Stearamidopropyldimethyl amine (SAPDMA) was performed following methods comparable to the OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study).

Male and female New Zealand White rabbits, obtained from Hazleton Research Animals were used in the study. The body weight range of animals at study initiation was 1970 – 2593 g. Animals were acclimated to laboratory conditions for a period of 7 days prior to test initiation.

Test solutions were prepared fresh weekly in distilled, 30%/70% ethanol/water for each group. Post acclimation, animals were randomized into treatment/control groups (each consisting of 5 animals/sex).

The test solution was applied to intact skin of rabbits once daily, 5 days/week for thirteen consecutive weeks at the following dose levels:

Group 1(Control): Distilled water

Group 2: 0.25% w/v in 30%/70% ethanol/water (equivalent to 5 mg/kg/day)

Group 3: 10% w/v in 30%/70% ethanol/water (equivalent to 200 mg/kg/day) 

All animals (test and control) were dosed at a constant dosage volume of 2 mL/kg/day

Appropriate concentration of test material was applied evenly on the back of each animal, from shoulder to rump (approximately 15 cm wide) using a syringe.

Animals were observed for gross toxicity daily. The skin at the treatment sites was graded prior to each day’s treatment. Body weights were taken prior to initiation and then weekly until termination. Blood was taken prior to initiation and at termination for hematological examinations. After treatment, animals were sacrificed by an IV overdose of sodium pentobarbital. Adrenals, kidneys and liver were dissected free of fat and weighed. Tissue samples (including treated skin samples) were collected for histological examinations.

No mortality or test related gross toxicity was observed throughout the study except slight conjunctivitis in one control animal and two Group 2 (5 mg/kg/day) animals. This was attributed to the continuous restraint which prevented the animals from grooming themselves. The collars of one control animal, one Group 2 (5 mg/kg/day) animal and one Group 3 (200 mg/kg/day) animal were removed temporarily because of sores on the necks. Some animals removed their collars overnight and they were collared again the next day.

Treatment of animals with 0.25% test material (5 mg/kg/day) in 30/70% ethanol/water produced moderate or slight erythema, slight edema, slight desquamation and slight fissuring. At 10% (200 mg/kg/day), moderate erythema, slight edema, slight desquamation and slight fissuring were produced. No treatment related changes in body weight and body weight gain were observed during the study. No test related biologically significant changes were noted in the absolute and relative liver, kidney and adrenal weight determinations.

Statistically significant increases in the WBC values were noted in the Group 3 (200 mg/kg/day) animals. In addition, there was an increase in platelet values from baseline to necropsy of the Group 2 (5 mg/kg/day) animals. The changes in WBC of the Group 3 animals was attributed to the chronic stress of collaring and not considered to be test substance related. The significant increase in platelet values of Group 2 animals were a result of low baseline values.

Treated skin on the backs of rabbits in both treated groups had a dry hair coat with an accumulation of test material on the surface at necropsy. Histopathological examinations revealed minimal acanthosis and hyperkeratosis at the treatment sites of all treated groups. The incidence and severity were similar in both groups. Incidental non treatment related histopathological changes were noted in several other tissues such as brain, liver, kidney, prostate and pancreas. However, these were considered to be incidental, and not treatment related.

Based on above it can be concluded that, no systemic toxicity associated with the sub chronic percutaneous exposure to test material was observed at any dose levels.

Dermal application of Stearamidopropyldimethyl amine (SAPDMA) to hair clipped skin of New Zealand White rabbits at concentrations of 0 (distilled water), 0.25 and 10% w/v of the test substance in 30%/70% ethanol/water (equivalent to 0, 5 and 200 mg/kg bw/day)for 13 weeks revealed a systemic NOAEL of ≥10% w/v (equivalent to 200 mg/kg bw/day).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEL

5

Study duration:

subchronic

Species:

rabbit

Quality of whole database:

similar to OECD guideline study, GLP

Additional information

In a 14 d dose range finding study according to OECD guideline 407, adopted 03 October 2008, and EU method B.7, May 2008, Stearic acid 3-(dimethylaminopropyl)amide was administered to 3 Crl:WI(Han) rats/sex/dose orally via gavage at dose levels of 0, 50, 200 and 500 mg/kg bw/day.

All animals in the 500 mg/kg bw/d dose group were sacrificed for humane reasons between days 6 and 8. They showed lethargy, hunched posture, laboured respiration, abdominal swelling, piloerection, chromodacryorrhoea, a lean appearance and/or ptosis from day 4 of treatment onwards. All animals showed weight loss or reduced body weight gain and reduced food consumption during the treatment period. Necropsy findings at 500 mg/kg bw/d primarily consisted of gelatinous contents in the gastro-intestinal tract or parts thereof, and emaciation. The main cause for moribundity at this dose level was forestomach ulceration and/or hyperplasia of the squamous epithelium of the forestomach. Other histopathological changes noted at this dose level included: lymphoid atrophy of the thymus, correlating to a reduced size of the thymus at necropsy; hyperplasia and inflammation of the forestomach; hyperplasia of the villi in the duodenum and jejunum; foamy macrophages and sinusoidal dilation and congestion/ erythrophagocytosis in the mesenterial lymph node; absence of spermiation and degeneration of spermatids in the testes, oligospermia and seminiferous cell debris in the epididymides, and reduced contents in the prostate and seminal vesicles, which corresponded to a reduced size of seminal vesicles, prostate and epididymides at necropsy.

At 50 and 200 mg/kg bw/d, no mortality occurred. All animals at 200 mg/kg bw/d showed piloerection on two days of week 2 only, whilst no clinical signs were noted at 50 mg/kg bw/d. Body weights, body weight gain and food intake remained in the same range as controls over the study period at these dose levels.

At 50 and 200 mg/kg bw/d, haematological changes consisted of slightly lower red blood cell and higher reticulocyte counts in males. No clear dose related trend was noted for these changes, which were generally slight in nature. Clinical biochemistry changes consisted of higher alanine aminotransferase activity in two males at 50 mg/kg bw/d, and two males and one female at 200 mg/kg bw/d, higher alkaline phosphatase activity in one female at 200 mg/kg bw/d, and higher potassium level in males at 50 and 200 mg/kg bw/d.

At 50 and 200 mg/kg bw/d, no abnormalities were noted at necropsy. Spleen and thymus weights of females at 200 mg/kg bw/d appeared slightly increased compared to the control group. No treatment-related histopathological changes were noted at 50 and 200 mg/kg bw/d. The results from this study were used to select the doses for the reproduction/developmental screening study. No reliable NOAEL could be derived from this dose-range-finding study. Due to the small number of animals, the relevance of the clinical biochemistry changes could not be fully justified.

 

A repeated dose 91-day dermal toxicity study comparable to OECD guideline 411 (reliable with restrictions), is available for Stearic acid 3-(dimethylaminopropyl)amide. The test substance was dermally administered to 5 New Zealand White rabbits/sex/group at dose levels of 0, 5 and 200 mg/kg bw/d for 4 h daily followed by rinsing, 5 times weekly. The animals were collared throughout the study to prevent oral ingestion of the test substance.

No test item related gross toxicity was observed throughout the study except slight conjunctivitis in 2/10 animals in the 5 mg/kg bw/d dose group. Animals of the 5 mg/kg bw/d dose group showed moderate or slight erythema, slight edema, slight desquamation and slight fissuring.

At 200 mg/kg bw/d moderate erythema, slight edema, slight desquamation and slight fissuring were noted. Body weight was not influenced by the treatment.

Statistically significant increases in white blood cell counts were noted in animals treated with 200 mg/kg bw/d. However, this was attributed to the chronic stress of collaring and not considered to be test item related.

An increase in platelet values at necropsy compared to baseline values was observed in animals treated with 5 mg/kg bw/d, which was considered to be the result of low baseline values.

No test item related changes in organ weights were observed. Histopathologic examination showed no test item related findings except from acanthosis and hyperkeratosis of the skin at the treatment sites.

In conclusion, Stearic acid 3-(dimethylaminopropyl)amide did not show systemic toxicity up to and including 200 mg/kg bw/d (highest dose administered). The dermal 90 d NOAEL in rabbit was 200 mg/kg bw/d in this study.

This dermal toxicity study in the rabbit is acceptable and is considered to be equivalent to the corresponding OECD guideline although minor deviations are reported: Only 2 dose levels were tested while the guideline recommends at least 3 doses; only 5 animals/sex/dose were used instead of 10 as recommended in the later implemented guideline; no vehicle control (30/70 EtOH/water) was tested, and no clinical chemistry was performed. These deviations are reflected by setting the reliability to 2.

This study used the appropriate route (dermal) and duration (subchronic) compared to the expected most relevant route of exposure in humans (dermal). Thus, it is considered to be relevant for hazard assessment.

There are no data gaps for the endpoint repeated dose toxicity. No human data are available. However, there is no reason to believe that these results from rat and rabbits would not be applicable to humans.

Justification for classification or non-classification

Based on the available data - an oral 14 day dose range finding study as well as a dermal 90 day repeated dose toxicity study, Stearic acid 3-(dimethylaminopropyl)amide does not need to be classified and labelled according to the CLP Regulation (EC) No 1272/2008 and Directive 67/548/EEC with respect to repeated dose toxicity.