N-[3-(dimethylamino)propyl]stearamide monoacetate

Endpoint summary

Description of key information

Three studies are available for this endpoint. The test item NDPSa (H-31339, Cas n.:13282-70-7).

Acute Tox. Oral:

K1, Oral, OECD TG 425, 14-days up-and-down procedure on Rats; LD50= 1403 mg/kg, M. N. Fallers, B.A. (2015)

Acute Tox. Inhalation:

K1, OECD TG 403, Rat 4-hr inhalation aerosol, LC50 > 4 mg/L (maximum practically-attainable atmospheric concentration), LC50 > 0.66 mg/L (solids aerosol concentration), Sheung P.Ng. (2015)

Acute Tox. Dermal:

K1, OECD B.3, Rat 24 hr, LD50 >5000 mg/kg, M.N. Fallers, B.A. (2015)

 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2014-10-17 to 2015-02-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)

Deviations:

no

GLP compliance:

yes

Test type:

up-and-down procedure

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Remarks:

CRL

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable): based on consistently acceptable health status and on extensive experience with the strain at DuPont Haskell.
- Source: Charles River Laboratories International, Inc., Raleigh, North Carolina, U.S.A.
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Rationale for use of males (if applicable):n/a
- Age at study initiation: 8-11 weeks old on the day of dosing.
- Weight at study initiation: n/a
- Fasting period before study: 16-18 hours prior to dosing.
- Housing:individually in solid-bottom caging with bedding and appropriate species- specific enrichment.
- Historical data:n/a
- Diet (e.g. ad libitum): PMI® Nutrition International, LLC Certified Rodent LabDiet® 5002, ad libitum.
- Water (e.g. ad libitum):ad libitum except as noted in section E. Dosing.
- Acclimation period:6-day quarantine period.
- Microbiological status when known: Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead, and other contaminants. Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers.
- Method of randomisation in assigning animals to test and control groups: A software package (AOT425StatPgm)a was used to determine the dose progression.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26°C
- Humidity (%): 30-70%
- Air changes (per hr): n/a
- Photoperiod (hrs dark / hrs light): Animal rooms were artificially illuminated (fluorescent light) on an approximate 12-hour light/dark cycle.

IN-LIFE DATES: From:21 October 2014 To: 3 December 2014

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

0.1% Tween 80 (V/V) in deionized water

Details on oral exposure:

VEHICLE
- Concentration in vehicle:(1:1)
- Amount of vehicle (if gavage):0.1%
- Justification for choice of vehicle:n/a
- Lot/batch no. (if required): n/a
- Purity: n/a

Doses:

175, 550, 1750, 5000, and 2000 mg/kg.
The rats were dosed at a volume of 10, 17.5, 20, or 40 mL per kg of body weight.

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:daily observation and weighted on days -1,1,8 and 15 or on the day of the sacrifice.
- Necropsy of survivors performed: yes
- Clinical signs including body weight: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:n/a

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

1 403 mg/kg bw

Based on:

test mat.

Mortality:

All animals dosed at 5000 or 2000 mg/kg died within a day after dosing

Clinical signs:

diarrhoea

Body weight:

other body weight observations

Gross pathology:

Gross findings were present in 4 female rats; 2 found dead and 2 sacrificed prior to scheduled sacrifice. The 2 found dead rats were administered 2000 mg/kg and had brown/green staining of the skin of the perineum. Of the 2 rats sacrificed prior to scheduled sacrifice, one was administered 2000 mg/kg and had brown staining about the perineum, yellow/clear fluid in the small intestine and cecum as well as whole body thinness (presumed dehydration). The other rat was administered 5000 mg/kg and had brown staining of the skin of the perineum. These observations are non-specific and not indicative of target organ toxicity. No other gross findings were observed.

Other findings:

- 5000 or 2000 mg/kg groups, preceding death.:
cold to touch, dehydration, eyelid ptosis, hunched posture, hypoactive, moribund status, decreased muscle tone, pale, high posture, prostrate, soiled skin or fur, and wet fur.
- At 1750 mg/kg:
diarrhea, soiled skin or fur, and wet fur between test days 2 and 3.
- At 550 mg/kg:
diarrhea and soiled skin or fur in 2/3 animals between test days 1 and 3.

Test Sequence	Group ID	Dose (mg/kg)	Short-term Result	Long-term Result
1	1	175	O	O
2	2	550	O	O
3	3	1750	O	O
4	4	5000	X	X
5	5	2000	X	X
6	6	550	O	O
7	7	2000	X	X
8	8	550	O	O
9	9	2000	X	X

X Died

O Survived

 

Summary of Long-Term Results

 

Dose (mg/kg)	O	X	Total
175	1	0	1
550	3	0	3
1750	1	0	1
2000	0	3	3
5000	0	1	1
All Doses	5	4	9

 Statistical Estimate based on long term outcomes: the LD50 is 1403 mg/kg.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Under the conditions of this study, the oral LD50 for H-31339 was 1403 mg/kg for female rats.
According to the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS), Fifth revised edition 2013, H-31339 is classified in Category 4.

Executive summary:

H-31139 was administered by oral gavage to fasted female rats at a dose of 175, 550, 1750, 5000, or 2000 mg/kg. The rats were dosed one at a time at approximately 48-hour intervals. All rats were observed for mortality, body weight effects, and clinical signs for 14 days after dosing. The rats were necropsied to detect grossly observable evidence of organ or tissue damage.

All animals dosed at 5000 or 2000 mg/kg died within a day after dosing; clinical abnormalities preceding death included abnormal gait, cold to touch, dehydration, diarrhea, eyelid ptosis, hunched posture, hypoactive, moribund status, decreased muscle tone, pallor, high posture, prostrate, salivation, soiled skin or fur, and wet fur. At 1750 mg/kg, there were no incidents of mortality or overall body weight losses; clinical abnormalities included diarrhea, soiled skin or fur, and wet fur between test days 2 and 3. At 550 mg/kg, there were no incidents of mortality or overall body weight losses; clinical abnormalities included diarrhea and soiled skin or fur in 2/3 animals between test days 1 and 3. At 175 mg/kg there were no incidents of mortality, overall body weight losses, or clinical abnormalities.

Gross findings were present in 4 female rats; 2 found dead and 2 sacrificed prior to scheduled sacrifice. The 2 found dead rats were administered 2000 mg/kg and had brown/green staining of the skin of the perineum. Of the 2 rats sacrificed prior to scheduled sacrifice, one was administered 2000 mg/kg and had brown staining about the perineum, yellow/clear fluid in the small intestine and cecum as well as whole body thinness (presumed dehydration). The other rat was administered 5000 mg/kg and had brown staining of the skin of the perineum. These observations are non-specific and not indicative of target organ toxicity. No other gross findings were observed.

Under the conditions of this study, the oral LD50 for H-31339 was 1403 mg/kg for female rats.

In accordance with the provisions of Directive 67/548/EEC amended by COMMISSION DIRECTIVE 2001/59/EC of 6 August 2001, Annex VI, H-31339 is assigned the symbol “Xn” and the risk phrase “Harmful if swallowed.”

According to the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS), Fifth revised edition 2013, H-31339 is classified in Category 4.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 403 mg/kg bw

Quality of whole database:

The materials/methods and results are described in detail and are sufficient for evaluation.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2015-03-09 To 2015-07-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Remarks:

CRL

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable): n/a
- Source: Charles River Laboratories International, Inc., Raleigh, North Carolina, U.S.A.
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Rationale for use of males (if applicable):n/a
- Age at study initiation: 7 weeks old.
- Weight at study initiation: Male between 371 and 395 grams; Females 231 and 268 grams at the time of exposure.
- Fasting period before study:n/a
- Housing:housed individually in solid bottom caging with bedding and enrichment.
- Historical data:n/a
- Diet (e.g. ad libitum): PMI® Nutrition International, LLC Certified Rodent LabDiet® 5002, ad libitum.
- Water (e.g. ad libitum):tap water were available ad libitum.
- Acclimation period:6 days prior testing.
- Microbiological status when known: Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead, and other contaminants. Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers.
- Method of randomisation in assigning animals to test and control groups: n/a

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26°C
- Humidity (%): 30-70%
- Air changes (per hr): 28 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle.


IN-LIFE DATES: From: 9 March 2015 To: 23 March 2015

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

water

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:Spraying Systems Company® nebulizer
- Exposure chamber volume: 34 L.
- Method of holding animals in test chamber:restrained in perforated stainless steel cylinders with conical nose pieces. The restrainers were inserted into a polymethylmethacrylate faceplate attached to the exposure chamber so that the nose of each animal extended into the exposure chamber.
- Source and rate of air (airflow):
- Method of conditioning air:
- System of generating particulates/aerosols:
- Method of particle size determination:
- Treatment of exhaust air:
- Temperature, humidity, pressure in air chamber:

TEST ATMOSPHERE
- Brief description of analytical method and equipment used: The test substance was metered into the nebulizer with a Cole-Palmer Masterflex® console drive pump. High-pressure air, metered into the nebulizer by a Brooks model 5850E mass flow controller, carried the resulting atmosphere into the exposure chamber. Chamber concentrations of test substance were controlled by varying the test substance feed rate to the nebulizer.
The test atmosphere was exhausted through a high-capacity particle filter cartridge prior to discharge into the fume hood.
- Samples taken from breathing zone: yes
- Time needed for equilibrium of exposure concentration before animal exposure:

VEHICLE
- Composition of vehicle (if applicable): test material diluted 10 times by volume within distilled water.
- Concentration of test material in vehicle (if applicable): 5 mg/L
- Justification of choice of vehicle: n/a
- Lot/batch no. (if required): n/a
- Purity: n/a

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution:Samples to determine aerosol size distribution (mass median aerodynamic diameter, geometric standard deviation, and percent percent aerosols less than 1, 3, and 10 μm diameter.
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): The MMAD (GSD) were 4.1 μm (2.5), 3.0 μm (2.5), and 3.9 μm (2.4) which are regarded as respirable.

Duration of exposure:

4 h

Concentrations:

maximum practically-attainable atmospheric concentration of 4.0 ± 0.30 mg/L (diluted)

No. of animals per sex per dose:

one group of 5 male and 5 female (nulliparous and non-pregnant)

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing:
- Necropsy of survivors performed: yes/no
- Clinical signs including body weight : yes.
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Statistics:

Upon completion of the exposures, CITADS sample results were transferred to the Camile Inhalation Reporting and Analysis System (CIRAS), which collated sample calculations.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4 mg/L air

Based on:

test mat. (dissolved fraction)

Remarks:

diluted

Exp. duration:

4 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

All animals survived the 4-hour exposure to 4.0 mg/L (diluted) H-31339 but 2 (of 5) male rats and 1 female rat died during the 14-day recovery period. The fractional mortalities were 2/5 for male rats and 1/5 for female rats.

Clinical signs:

bodyweight loss

Body weight:

- greater than 10% body weight loss: n/a
- lower than 10% body weight loss:maximum lost 13% .
- other body weight observations:
Five (of 5) male rats lost 6.6-13% of their body weights on test day 2. One male was later found dead on the same day and another male was found dead on day 3. One of the 3 surviving males lost 7.7 and 1.7% of his body weight on test day 3 and 4, respectively. Three (of 5) female rats lost 12-13 % of their body weights on test day 2 and one of them was later found dead on the same day. Two of the 4 surviving females lost <7% of their body weights on test day 3 and only 1 female lost 4.1% on test day 4. The weight variation in animals used on an exposure did not exceed ± 20% of the mean weight of each sex.
All other animals gain weights and no abnormalities were detected during the rest of the 14-day recovery period.

Gross pathology:

Gross findings were present in 3 male and 4 female rats administered 4.0 mg/L of diluted test substance: 2 found dead males, 1 found dead female and the remainder at scheduled sacrifice. Five of these animals (2 males and 3 females) had multifocal to diffuse dark discoloration of the lung, often concentrating in the cranial portion of the lung. One found dead male additionally had expanded lungs and cloudy discoloration of the left eye. The two found dead males had red staining on their faces, and the found dead female had thick, tan discoloration in the trachea. The observations are presumed related to test substance exposure. No other gross findings were observed.

Mortality

 

Sex	Mean Atmospheric Concentration (mg/L)	Mortality (deaths/exposed)
Male	4.0	2/5
Female	4.0	1/5

 

Summary of Clinical Observations and Mortality in Male Rats

 

Found dead
Number of Observations	2
Number of Animals	2
Days from – to	2  3
Scheduled sacrifice
Number of Observations	3
Number of Animals	3
Days from – to	15  15
Breathing - laboured
Number of Observations	8
Number of Animals	3
Days from – to	1  4
Breathing – lung noise
Number of Observations	3
Number of Animals	1
Days from – to	2  4
Breathing - slow
Number of Observations	1
Number of Animals	1
Days from – to	2  2
Discharge - red
Number of Observations	10
Number of Animals	5
Days from – to	1  4
Gasping
Number of Observations	6
Number of Animals	3
Days from – to	1  4
Discoloured fur - yellow
Number of Observations	1
Number of Animals	1
Days from – to	2  2

 

Summary of Clinical Observations and Mortality in Female Rats

 

 

Found dead
Number of Observations	1
Number of Animals	1
Days from – to	2  2
Scheduled sacrifice
Number of Observations	4
Number of Animals	4
Days from – to	15  15
Breathing – laboured
Number of Observations	7
Number of Animals	3
Days from – to	1  4
Breathing – lung noise
Number of Observations	1
Number of Animals	1
Days from – to	4  4
Discharge - red
Number of Observations	9
Number of Animals	5
Days from – to	1  4
Gasping
Number of Observations	7
Number of Animals	3
Days from – to	1  4
Discoloured fur – yellow
Number of Observations	5
Number of Animals	2
Days from – to	2  4

Interpretation of results:

GHS criteria not met

Conclusions:

While an exposure atmosphere of ≥ 5 mg/L could not be obtained due to the waxy property of the test substance, 4.0 mg/L H-31339 was the highest feasible airborne concentration that could be achieved and required a 10X dilution of the test substance with distilled water.
Under the conditions of this study, the 4-hour inhalation median lethal concentration (LC50) for (diluted) H-31339 in male and female rats was > 4.0 mg/L, the maximum practically-attainable atmopheric concentration, and > 0.66 mg/L solids aerosol concentration.

Executive summary:

The objective of this study was to evaluate the acute inhalation toxicity of H-31339 when administered as an aerosol for a single, 4-hour, nose-only exposure to one group of 5 male and
5 female (nulliparous and non-pregnant) Crl:CD(SD) rats. H-31339 used for this study was a white solid with a purity of 96% by analysis. Due to the waxy property, the test substance was diluted 10 times (by volume) with distilled water to facilitate the generation of an aerosol exposure atmosphere. The test atmosphere was generated by aerosolization of H-31339 in air using a nebulizer and the airborne concentration of H-31339 was determined by gravimetric analysis. During a 14-day recovery period, all animals were weighed and observed for mortality and clinical signs of toxicity. A gross pathological examination was performed on all animals at the scheduled necropsy or shortly after they were found dead.

During the exposure, rats were exposed to a maximum practically-attainable atmospheric concentration of 4.0 ± 0.30 mg/L (diluted) H-31339 (mean ± standard deviation) as determined by gravimetric analysis of wet filters. The solids aerosol concentration of the same atmosphere, determined by desiccated filter weights, was 0.66 ± 0.030 mg/L. The mass median aerodynamic diameter and geometric standard deviation, MMAD (GSD), was determined 3 times for the atmosphere and measured 4.1 μm (2.5), 3.0 μm (2.5), and 3.9 μm (2.4).

All animals survived the 4-hour exposure to 4.0 mg/L (diluted) H-31339 but 2 (of 5) male rats and 1 female rat died during the 14-day recovery period. The fractional mortalities were 2/5 for male rats and 1/5 for female rats.

All animals displayed normal startle response during the exposure (test day 1), and no abnormalities were observed. Clinical signs of toxicities including labored breathing, slow breathing, lung noise, gasping, and/or discolored fur were observed in 3 male and 4 female rats during test days 1-4. Five (of 5) male rats lost 6.6-13% of their body weights on test day 2. One male was later found dead on the same day and another male was found dead on day 3. One of the 3 surviving males lost 7.7 and 1.7% of his body weight on test day 3 and 4, respectively. Three (of 5) female rats lost 12-13 % of their body weights on test day 2 and one of them was later found dead on the same day. Two of the 4 surviving females lost <7% of their body weights on test day 3 and only 1 female lost 4.1% on test day 4. All other animals gain weights and no abnormalities were detected during the rest of the 14-day recovery period.

Gross findings were present in 3 male and 4 female rats administered 4.0 mg/L of diluted test substance: 2 found dead males, 1 found dead female and the remainder at scheduled sacrifice. Five of these animals (2 males and 3 females) had multifocal to diffuse dark discoloration of the lung, often concentrating in the cranial portion of the lung. One found dead male additionally had expanded lungs and cloudy discoloration of the left eye. The two found dead males had red staining on their faces, and the found dead female had thick, tan discoloration in the trachea. The observations are presumed related to test substance exposure. No other gross findings were observed.

While an exposure atmosphere of ≥ 5 mg/L could not be obtained due to the waxy property of the test substance, 4.0 mg/L H-31339 was the highest feasible airborne concentration that could be achieved and required a 10X dilution of the test substance with distilled water.

Under the conditions of this study, the 4-hour inhalation median lethal concentration (LC50) for (diluted) H-31339 in male and female rats was > 4.0 mg/L, the maximum practically-attainable atmopheric concentration, and > 0.66 mg/L solids aerosol concentration.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

> 0.66 mg/L air

Physical form:

other: solid

Quality of whole database:

The materials/methods and results are described in detail and are sufficient for evaluation.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2014-10-16 To 2015-01-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

Test type:

fixed dose procedure

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Remarks:

CRL The Crl:CD(SD) rat was selected based on consistently acceptable health status and on extensive experience with the strain at DuPont Haskell.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable)
- Source: Charles River Laboratories International, Inc., Raleigh, North Carolina, U.S.A.
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Rationale for use of males (if applicable) n/a
- Age at study initiation: 9 weeks old.
- Weight at study initiation: n/a
- Fasting period before study: n/a
- Housing:housed individually in solid-bottom caging with bedding and appropriate species- specific enrichment.
- Historical data:n/a
- Diet (e.g. ad libitum): PMI® Nutrition International, LLC Certified Rodent LabDiet® 5002, ad libitum,
- Water (e.g. ad libitum):tap water, ad libitum.
- Acclimation period:6-day quarantine period.
- Microbiological status when known:Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead, and other contaminants. Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers.
- Method of randomisation in assigning animals to test and control groups:n/a

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26 °C
- Humidity (%): 30-70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): artificially illuminated (fluorescent light) on an approximate 12-hour light/dark cycle.

IN-LIFE DATES: From: 2014-10-23 To: 2014-11-06

Type of coverage:

semiocclusive

Vehicle:

water

Remarks:

deionized water.

Details on dermal exposure:

TEST SITE
- Area of exposure: scapular to the lumbar region.37 square centimeters.
- % coverage: 5 cm x 7.4 cm.
- Type of wrap if used: 2-ply gauze patch, then wrapped with stretch gauze bandage and self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, with paper towels soaked in soap and warm water, and the skin was dried.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5000 mg/Kg bw.
- For solids, paste formed: yes

VEHICLE
The aliquot of test substance designated for an animal was moistened with approximately 4 mL of deionized water for male rats and 3 mL of deionized water for female rats to form a thick paste.

Duration of exposure:

24 hours

Doses:

- 5000 mg/kg bw; single dose.

No. of animals per sex per dose:

5 males and 5 females.

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing:daily for clinical signs and toxicity, days 1, 8, and 15 for body weight.
- Necropsy of survivors performed: no
- Clinical signs including body weight: yes.
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:examined to detect grossly observable evidence of organ or tissue damage.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

There were no instances of mortality at 5000 mg/kg.

Gross pathology:

No gross lesions were present in the rats at necropsy.

Other findings:

On test days 2 and 3, one female exhibited ulceration on the treated area of skin, which was resolved by test day 4. Clinical signs of hyperkeratosis, desquamation, or epidermal scaling were observed in all animals between test days 3 and 8. The clinical observation of scabbing on the back of one female was present from test day 12-14. At final euthanasia, no clinical abnormalities were present.
At the beginning of the observation period (24 hours after exposure), on test day 2, edema was observed on 5/5 males and 4/5 females, with scores of 1 or 2. All edema was resolved by test day 3. Erythema was observed in all animals between test days 2 and 3, with scores of 1, 2, or 3. From tests 4-9, 1/5 females exhibited erythema scores of 1 or 2.
There were no overall (test day 1-15) body weight losses among any animals.

Individual Erythema Scores Males. Dose: 5000 mg/kg

 

Days relative to start date	2	3	4	5	6	7	8	9	12	13	14	15
N. animal
4975	3	1	0	0	0	0	0	0	0	0	0	0
4976	3	2	0	0	0	0	0	0	0	0	0	0
4977	3	1	0	0	0	0	0	0	0	0	0	0
4978	3	1	0	0	0	0	0	0	0	0	0	0
4979	3	2	0	0	0	0	0	0	0	0	0	0

 

 

 

Individual Erythema Scores Females. Dose: 5000 mg/kg

 

Days relative to start date	2	3	4	5	6	7	8	9	12	13	14	15
N. animal
4980	3	3	2	2	1	1	1	1	0	0	0	0
4981	1	1	0	0	0	0	0	0	0	0	0	0
4982	2	2	0	0	0	0	0	0	0	0	0	0
4983	2	0	0	0	0	0	0	0	0	0	0	0
4984	3	2	0	0	0	0	0	0	0	0	0	0

 

 

 

Individual Edema Scores Males. Dose: 5000 mg/kg

 

Days relative to start date	2	3	4	5	6	7	8	9	12	13	14	15
N. animal
4975	2	0	0	0	0	0	0	0	0	0	0	0
4976	2	0	0	0	0	0	0	0	0	0	0	0
4977	2	0	0	0	0	0	0	0	0	0	0	0
4978	2	0	0	0	0	0	0	0	0	0	0	0
4979	2	0	0	0	0	0	0	0	0	0	0	0

 

 

 

 

Individual Edema Scores Females. Dose: 5000 mg/kg

 

Days relative to start date	2	3	4	5	6	7	8	9	12	13	14	15
N. animal
4980	1	0	0	0	0	0	0	0	0	0	0	0
4981	0	0	0	0	0	0	0	0	0	0	0	0
4982	1	0	0	0	0	0	0	0	0	0	0	0
4983	1	0	0	0	0	0	0	0	0	0	0	0
4984	2	0	0	0	0	0	0	0	0	0	0	0

 

 

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the dermal LD50 for H-31339 was greater than 5000 mg/kg of body weight when applied to the skin of male and female rats for 24 hours.
In accordance with the provisions of Directive 67/548/EEC amended by COMMISSION DIRECTIVE 2001/59/EC of 6 August 2001, Annex VI, classification is not required.
According to the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS), Fifth revised edition 2013, classification is not required.

Executive summary:

A single dose of H-31339 was applied to the shaved, intact skin of 5 male and 5 female rats at a dose level of 5000 mg/kg of body weight. The application site was covered with a semi-occlusive dressing for 24 hours, after which the test substance was removed. The rats were observed for 14 days following application. The rats were necropsied to detect grossly observable evidence of organ or tissue damage at the end of the 15-day test period.

There were no instances of mortality.

On test days 2 and 3, one female exhibited ulceration on the treated area of skin, which was resolved by test day 4. Clinical signs of hyperkeratosis, desquamation, or epidermal scaling were observed in all animals between test days 3 and 8. The clinical observation of scabbing on the back of one female was present from test day 12-14. At final euthanasia, no clinical abnormalities were present.

At the beginning of the observation period (24 hours after exposure), on test day 2, edema was observed on 5/5 males and 4/5 females, with scores of 1 or 2. All edema was resolved by test day 3. Erythema was observed in all animals between test days 2 and 3, with scores of 1, 2, or 3. From tests 4-9, 1/5 females exhibited erythema scores of 1 or 2.

There were no overall (test day 1-15) body weight losses among any animals. No gross lesions were present in the rats at necropsy.

Under the conditions of this study, the dermal LD50 for H-31339 was greater than 5000 mg/kg of body weight when applied to the skin of male and female rats for 24 hours.

In accordance with the provisions of Directive 67/548/EEC amended by COMMISSION DIRECTIVE 2001/59/EC of 6 August 2001, Annex VI, classification is not required.

According to the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS), Fifth revised edition 2013, classification is not required.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

> 5 000 mg/kg bw

Quality of whole database:

The materials/methods and results are described in detail and are sufficient for evaluation.

Additional information

Oral route: 

H-31139 was administered by oral gavage to fasted female rats at a dose of 175, 550, 1750, 5000, or 2000 mg/kg. The rats were dosed one at a time at approximately 48-hour intervals. All rats were observed for mortality, body weight effects, and clinical signs for 14 days after dosing. The rats were necropsied to detect grossly observable evidence of organ or tissue damage.

All animals dosed at 5000 or 2000 mg/kg died within a day after dosing; clinical abnormalities preceding death included abnormal gait, cold to touch, dehydration, diarrhea, eyelid ptosis, hunched posture, hypoactive, moribund status, decreased muscle tone, pallor, high posture, prostrate, salivation, soiled skin or fur, and wet fur. At 1750 mg/kg, there were no incidents of mortality or overall body weight losses; clinical abnormalities included diarrhea, soiled skin or fur, and wet fur between test days 2 and 3. At 550 mg/kg, there were no incidents of mortality or overall body weight losses; clinical abnormalities included diarrhea and soiled skin or fur in 2/3 animals between test days 1 and 3. At 175 mg/kg there were no incidents of mortality, overall body weight losses, or clinical abnormalities.

Gross findings were present in 4 female rats; 2 found dead and 2 sacrificed prior to scheduled sacrifice. The 2 found dead rats were administered 2000 mg/kg and had brown/green staining of the skin of the perineum. Of the 2 rats sacrificed prior to scheduled sacrifice, one was administered 2000 mg/kg and had brown staining about the perineum, yellow/clear fluid in the small intestine and cecum as well as whole body thinness (presumed dehydration). The other rat was administered 5000 mg/kg and had brown staining of the skin of the perineum. These observations are non-specific and not indicative of target organ toxicity. No other gross findings were observed.

Under the conditions of this study, the oral LD50 for H-31339 was 1403 mg/kg for female rats.

 

Justification for selection of Acute toxicity – oral route - endpoint.

Key 1 study.

 

Inhalation route: 

The objective of this study was to evaluate the acute inhalation toxicity of H-31339 when administered as an aerosol for a single, 4-hour, nose-only exposure to one group of 5 male and
5 female (nulliparous and non-pregnant) Crl:CD(SD) rats. H-31339 used for this study was a white solid with a purity of 96% by analysis. Due to the waxy property, the test substance was diluted 10 times (by volume) with distilled water to facilitate the generation of an aerosol exposure atmosphere. The test atmosphere was generated by aerosolization of H-31339 in air using a nebulizer and the airborne concentration of H-31339 was determined by gravimetric analysis. During a 14-day recovery period, all animals were weighed and observed for mortality and clinical signs of toxicity. A gross pathological examination was performed on all animals at the scheduled necropsy or shortly after they were found dead.

During the exposure, rats were exposed to a maximum practically-attainable atmospheric concentration of 4.0 ± 0.30 mg/L (diluted) H-31339 (mean ± standard deviation) as determined by gravimetric analysis of wet filters. The solids aerosol concentration of the same atmosphere, determined by desiccated filter weights, was 0.66 ± 0.030 mg/L. The mass median aerodynamic diameter and geometric standard deviation, MMAD (GSD), was determined 3 times for the atmosphere and measured 4.1 μm (2.5), 3.0 μm (2.5), and 3.9 μm (2.4).

All animals survived the 4-hour exposure to 4.0 mg/L (diluted) H-31339 but 2 (of 5) male rats and 1 female rat died during the 14-day recovery period. The fractional mortalities were 2/5 for male rats and 1/5 for female rats.

All animals displayed normal startle response during the exposure (test day 1), and no abnormalities were observed. Clinical signs of toxicities including labored breathing, slow breathing, lung noise, gasping, and/or discolored fur were observed in 3 male and 4 female rats during test days 1-4. Five (of 5) male rats lost 6.6-13% of their body weights on test day 2. One male was later found dead on the same day and another male was found dead on day 3. One of the 3 surviving males lost 7.7 and 1.7% of his body weight on test day 3 and 4, respectively. Three (of 5) female rats lost 12-13 % of their body weights on test day 2 and one of them was later found dead on the same day. Two of the 4 surviving females lost <7% of their body weights on test day 3 and only 1 female lost 4.1% on test day 4. All other animals gain weights and no abnormalities were detected during the rest of the 14-day recovery period.

Gross findings were present in 3 male and 4 female rats administered 4.0 mg/L of diluted test substance: 2 found dead males, 1 found dead female and the remainder at scheduled sacrifice. Five of these animals (2 males and 3 females) had multifocal to diffuse dark discoloration of the lung, often concentrating in the cranial portion of the lung. One found dead male additionally had expanded lungs and cloudy discoloration of the left eye. The two found dead males had red staining on their faces, and the found dead female had thick, tan discoloration in the trachea. The observations are presumed related to test substance exposure. No other gross findings were observed.

While an exposure atmosphere of ≥ 5 mg/L could not be obtained due to the waxy property of the test substance, 4.0 mg/L H-31339 was the highest feasible airborne concentration that could be achieved and required a 10X dilution of the test substance with distilled water.

Under the conditions of this study, the 4-hour inhalation median lethal concentration (LC50) for (diluted) H-31339 in male and female rats was > 4.0 mg/L, the maximum practically-attainable atmopheric concentration, and > 0.66 mg/L solids aerosol concentration.

 

Justification for selection of Acute toxicity – inhalation route - endpoint.

Key 1 study.

 

Dermal route:

A single dose of H-31339 was applied to the shaved, intact skin of 5 male and 5 female rats at a dose level of 5000 mg/kg of body weight. The application site was covered with a semi-occlusive dressing for 24 hours, after which the test substance was removed. The rats were observed for 14 days following application. The rats were necropsied to detect grossly observable evidence of organ or tissue damage at the end of the 15-day test period.

There were no instances of mortality.

On test days 2 and 3, one female exhibited ulceration on the treated area of skin, which was resolved by test day 4. Clinical signs of hyperkeratosis, desquamation, or epidermal scaling were observed in all animals between test days 3 and 8. The clinical observation of scabbing on the back of one female was present from test day 12-14. At final euthanasia, no clinical abnormalities were present.

At the beginning of the observation period (24 hours after exposure), on test day 2, edema was observed on 5/5 males and 4/5 females, with scores of 1 or 2. All edema was resolved by test day 3. Erythema was observed in all animals between test days 2 and 3, with scores of 1, 2, or 3. From tests 4-9, 1/5 females exhibited erythema scores of 1 or 2.

There were no overall (test day 1-15) body weight losses among any animals. No gross lesions were present in the rats at necropsy.

Under the conditions of this study, the dermal LD50 for H-31339 was greater than 5000 mg/kg of body weight when applied to the skin of male and female rats for 24 hours.

In accordance with the provisions of Directive 67/548/EEC amended by COMMISSION DIRECTIVE 2001/59/EC of 6 August 2001, Annex VI, classification is not required.

According to the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS), Fifth revised edition 2013, classification is not required.

 

Justification for selection of Acute toxicity – dermal route - endpoint.

Key 1 Study

Justification for classification or non-classification

Harmonised classification:

The substance does not have a harmonised classification according to the Regulation (EC) No. 1272/2008 (CLP). 

 

Self-classification:

Based on the available information on the registered substance, self-classification is proposed according to GHS and CLP:

GHS Category 4 for the Oral route.

GHS not classified for the Inhalation Route.

GHS not classified for Dermal Route.