N,N,N-trimethylanilinium chloride

Endpoint summary

Description of key information

Skin Irritation:

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal.

Hence, it was concluded that the test chemical was Non-Irritating to the skin of female Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

Eye Irritation:

No significant irritation was noted but one of 3 animals died. Hence,the test chemical was considered to be non irritating to the conjunctival sac of rabbits.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Justification for type of information:

data is from experimental reports

Qualifier:

according to guideline

Guideline:

other: OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

To determine the dermal reaction profile of the test chemical in Sprague Dawley rats

GLP compliance:

yes

Specific details on test material used for the study:

- Name of test material (as cited in study report): N,N,N-trimethylanilinium chloride
- Molecular formula : C9H14N.Cl
- Molecular weight : 171.67 g/mol
- Smiles notation: [Cl-].C[N+](C)(C)c1ccccc1
- InChl : InChI=1/C9H14N.ClH/c1-10(2,3)9-7-5-4-6-8-9;/h4-8H,1-3H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
- Batch Number : G27331701
- Laboratory Test Item Code : TAS/122/058
- Manufacturing Date : January; 2017
- Expiry Date : December; 2021
- Consistency : Solid, crystalline
- Activity (Clinical Indication) : Industrial chemical
- Safety Precautions : Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).
- Storage Condition : Ambient Temperature
- Preparation of Test Item : The test item was used as supplied by the Sponsor. Test item was grounded to fine powder prior to application. The particulates were moistened with distilled water before application

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

- Source: National Institute of Biosciences, Pune.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult female rats aged between 8 – 10 weeks old were used.
- Weight at study initiation: weight range of approximately 228.0 to 247.7 grams at initiation of dosing.
Female
Mean : 235.60 g (= 100 %)
Minimum : 228.0 g (- 3.23 %)
Maximum : 247.7 g (+ 5.14 %)
Total No. of animals : 5
- Identification: Each rat was individually identified by the cage number.
- Fasting period before study: No data available
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

Environmental Conditions:
- Temperature (°C): 19.2 to 21.8 degree centigrade
- Humidity (%): 56.0% to 59.1%.
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

not specified

Amount / concentration applied:

Dose range finding study: 200 mg/kg , 1000 mg/kg, 2000 mg/kg
Main study: 2000 mg/kg

Duration of treatment / exposure:

24 hours

Observation period:

14 days

Number of animals:

5 female

Details on study design:

TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.
OBSERVATION TIME POINTS
(indicate if minutes, hours or days) : Dermal reaction was observed daily for study period of 14 days.
SCORING SYSTEM: Draize Method

Irritation parameter:

erythema score

Basis:

mean

Time point:

14 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

14 d

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Evaluation of Dermal Reaction
Sex : Female
Dose Range Finding Study:
Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Main Study:
Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Other effects:

Clinical Signs of Toxicity and Mortality

Sex : Female
Dose Range Finding Study:
Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. The animal survived through the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. The animal survived through the study period of 14 days.
Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. The animal survived through the study period of 14 days.
Main Study:
Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. All animals survived through the study period of 14 days.
Body Weight
Sex : Female
Dose Range Finding Study:
Group I (200 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.74% and 12.62% respectively.
Group I (1000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.82% and 9.68% respectively.
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.09% and 10.75% respectively.
Main Study:
Group II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.53% and 9.31% respectively.
Gross Pathological Findings
Gross pathological examination did not reveal any abnormalities in animals from 200 mg/kg, 1000 mg/kg and 2000 mg/kg dose groups from dose range finding study and main study sacrificed terminally

Interpretation of results:

other: not irritating

Conclusions:

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal.
Hence, it was concluded that the test chemical was Non-Irritating to the skin of female Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

Executive summary:

A study was designed and conducted to determine the dermal reaction profile of the test chemical in Sprague Dawley rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures. 5 female young adult, non-pregnant rats were used for the study. The animals were kept in their cages for at least 5 days prior to administration for acclimatization to the laboratory condition and after acclimatization period, animals were randomly selected.

Approximately 24 hours before application, the hair of each rat was closely clipped from the trunk (dorsal surface and sides from scapular to pelvic area) with an electric clipper, so as to expose at least 10% of the body surface area. The test item was applied directly onto the exposed skin of the animal, taking care to spread the test item evenly over the entire area of approximately 10% of the total body surface area or as much of the area as can reasonably be covered. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item. Following 24 hours of exposure, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.

A single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered at the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals. Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal.

Hence, it was concluded that the test chemical was Non-Irritating to the skin of female Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

data is from authoritative databases

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

Eye irritation test of chemical Trimethylanilinium chloride was conducted in rabbits

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material (as cited in study report): N,N,N-trimethylanilinium chloride
- Molecular formula: C9H14N.Cl
- Molecular weight: 171.67 g/mol
- Smiles notation: [Cl-].C[N+](C)(C)c1ccccc1
- InChl : InChI=1/C9H14N.ClH/c1-10(2,3)9-7-5-4-6-8-9;/h4-8H,1-3H3;1H/q+1;/p-1
- Substance type: Organic
- Physical state: Solid

Species:

rabbit

Strain:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

30 mg powdered

Duration of treatment / exposure:

No data available

Observation period (in vivo):

No data available

Number of animals or in vitro replicates:

3 animals

Details on study design:

Area of exposure: conjunctival sac

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: no data available

Reversibility:

not specified

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

No significant irritation was seen in a test on rabbits eyes in which 30 mg of powdered chemical was instilled into the conjunctival sac but one of 3 animals died.

Interpretation of results:

other: not irritating

Conclusions:

No significant irritation was noted but one of 3 animals died. Hence,the test chemical was considered to be non irritating to the conjunctival sac of rabbits.

Executive summary:

Eye irritation test was performed on rabbits to evaluate the irritation potential of the test chemical The test chemical was instilled into the conjunctival sac of rabbits. 30 mg of powdered test chemical was installed into the conjunctival sac of 3 rabbits (observation period, duration of exposure not mentioned).

No significant irritation was noted but one of 3 animals died. Hence,the test chemical was considered to be non irritating to the conjunctival sac of rabbits.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation:

Various studies have been summarized to determine the level of dermal irritation caused by the test chemical in living organisms. The results include experimental in vitro and in vivo data for the test chemical.

A study was designed and conducted to determine the dermal reaction profile of the test chemical in Sprague Dawley rats. The study was performed as per OECD Guidelines 402 and complying to the GLP procedures. 5 female young adult, non-pregnant rats were used for the study. The animals were kept in their cages for at least 5 days prior to administration for acclimatization to the laboratory condition and after acclimatization period, animals were randomly selected.

Approximately 24 hours before application, the hair of each rat was closely clipped from the trunk (dorsal surface and sides from scapular to pelvic area) with an electric clipper, so as to expose at least 10% of the body surface area. The test item was applied directly onto the exposed skin of the animal, taking care to spread the test item evenly over the entire area of approximately 10% of the total body surface area or as much of the area as can reasonably be covered. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item. Following 24 hours of exposure, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.

A single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered at the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals. Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

The overall irritation score of the substance was determined to be 0 and no erythema and edema (skin irritation) were found at the end of 14 days observation period after patch removal.

Hence, it was concluded that the test chemical was Non-Irritating to the skin of female Sprague Dawley rats under the experimental conditions tested and classified as “Category- Not Classified” as per CLP Classification.

This is supported by the in vitro study performed according to the OECD 439 test guideline to determine the irritation potential of the test chemical. The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article by determining the viability of the tissues following exposure to the test article via MTT. Tissues were exposed to the test article and controls for ~one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay. 

The MTT data show the assay quality controls were met and passed the acceptance of criteria. The Mean % tissue viability compared to negative control (n=3) of the test substance was determined to be 64.7%.

Hence, under the current experimental test conditions it was concluded that test substance was considered to be not irritating to human skin and can thus be classified as “Not Classified'' as per CLP Regulation.

The in vivo and in vitro data are in mutual agreement with each other indicating a strong possibility of the test chemical being not irritating to skin. Hence, the test chemical can be considered to be not irritating to skin. Comparing the above annotations with the criteria of the CLP regulation the test chemical can be classified under the category “Not Classified”.

Eye Irritation:

Various studies have been summarized to determine the extent of ocular damage caused by the test chemical in living organisms. The results include experimental in vitro and in vivo data for the test chemical.

 

An eye irritation test was performed on rabbits to evaluate the irritation potential of the test chemical. The test chemical was instilled into the conjunctival sac of rabbits. 30 mg of powdered test chemical was installed into the conjunctival sac of 3 rabbits (observation period, duration of exposure not mentioned).

No significant irritation was noted but one of 3 animals died. Hence, the test chemical was considered to be non irritating to the conjunctival sac of rabbits.

An in vitro skin irritation study was performed according to the OECD 492 test guideline to assess the ocular irritation potential of the test chemical. The MatTek EpiOcular™ model was used to assess the potential ocular irritation of the test articles by determining the viability of the tissues following exposure to the test article via MTT. "Tissues were exposed to solid test articles and control for approx.6 hours, followed by a 25 minute post-soak and approximately 18 hours recovery after the post-soak. The viability of each tissue was determined by MTT assay.

The MTT data show the assay quality controls were met, passing the acceptance criteria.

The mean % tissue viability of test substance was determined to be 3.4%. Hence, under the experimental test conditions it was concluded that test substance was considered to be irritating to the human eyes and can thus be classified as ‘’Irritating to eyes in Category 2” as per CLP Regulation

These results are supported by the study performed on rabbits to assess the irritation potential of the test chemical. 6 young adult male and female New Zealand White rabbits were used for the study. Volumes of 0.01, 0.03, and 0.10 ml of 0.1% test chemical were instilled into one eye of the rabbits and the untreated eye served as control. Eyes were examined and scored according to the scale by Draize. No signs of irritation observed. Hence, the test chemical can be considered to be not irritating to eyes.

The above results are supported by the study performed according to OECD 405 Guidelines to evaluate the irritation potential of the test chemical.

Rabbits were exposed to undiluted test chemical and effects were observed (duration, dose not specified).

No irritating effects were observed when rabbits were exposed to undiluted test chemical.

Hence, the test chemical can be considered as not irritating to eyes.

Even though the results from the in vitro studies suggest that the test chemical can cause irritation to eyes, but the results from in vivo studies suggest otherwise. Hence, the test chemical can be considered to be not irritating to eyes. Comparing the above annotations with the criteria of the CLP regulation the test chemical can be classified under the category “Not Classified”.

Justification for classification or non-classification

Available studies for the test chemical indicate a possibility that the test chemical can be not irritating to eyes and skin.

Hence, the test chemical can be classified under the category "Not Classified" as per CLP regulation.