4-mesyl-2-nitrotoluene

Endpoint summary

Description of key information

Oral: Male LD50=1426 (971-2479) mg/kg, Female LD50=933 (692-1259) mg/kg, rat, OECD 401, Johnson 1999
Dermal: LD50>2000 mg/kg, rat, OECD 402, Lees 1999
Inhalation: LC50>5400 mg/m3, rat, OECD 403, Rattray 2005

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:

29 April - 21 July 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, no restrictions, fully adequate for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: Alpk:APfSD

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: approximately 8-12 weeks
- Weight at study initiation: 266-404 g (males), 193-263 g (females)
- Fasting period before study: overnight prior to dosing
- Housing: maximum of 5 per cage, sexes separately in cages suitable for rats of this strain and weight range expected during the course of the study
- Diet: RMI ad libitum (except during overnight fast)
- Water: mains water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 30-70%
- Air changes: minimum of 15 per hr
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 2 June 1999 To: 21 July 1999

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

1%

Details on oral exposure:

Dose levels prepared by adjusting concentration of the dosing preparations. The volume of the dose was calculated for each animal according to its weight at the time of dosing. A standard volume of 10 mL/kg bodyweight of the dosing preparation was dosed.

Doses:

200, 500, 1000 or 2000 mg/kg bodyweight

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observed twice on day 1 and once per day thereafter. Weighed prior to fasting (day -1), immediately before dosing (day 1) and on days 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: none

Statistics:

The acute oral median lethal dose was estimated from mortality data by logistic regression using nominal dose values. Confidence limits calculated using a likelihood ratio interval (Williams DA, 1986. Interval Estimation of the Median Lethal Dose. Biometrics 42, 641-645).

Sex:

male

Dose descriptor:

LD50

Effect level:

1 426 mg/kg bw

Based on:

test mat.

Remarks:

2479

95% CL:

971

Sex:

female

Dose descriptor:

LD50

Effect level:

933 mg/kg bw

Based on:

test mat.

Remarks:

1259

95% CL:

692

Mortality:

200 or 500 mg/kg - no deaths; 1000 mg/kg - 1 male and 3 females killed in extremis on day 2; 2000 mg/kg - 4 males and all 5 females found dead or killed in extremis between days 1 and 3.

Clinical signs:

other: 200 mg/kg - signs of slight systemic toxicity (including tip-toe gait, lachrymation, salivation, subdued behaviour, upward curvature of spine and breathing irregular) with complete recovery by day 2. 500 mg/kg - signs of slight or moderate toxicity (inclu

Gross pathology:

No treatment-related findings at 500 mg/kg. In the other groups there was a reduction in testes and epididymes of some males, changes to the stomach/abnormal stomach contents and pallor of liver and kidneys, speckling of the thymus and non-specific staining around the nose and mouth.

Table 1: Mortality data

Dose Level	Day Number	Number of Deaths
(mg/kg)		Male	Female
200	Total at day 15	0/5	0/5
500	Total at day 15	0/5	0/5
1000	2	1	3
	Total at day 15	1/5	3/5
2000	1	0	1
	2	3	4
	3	1	0
	Total at day 15	4/5	5/5

 

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute oral LD50 of the test substance was estimated to be 1426 mg/kg (95% confidence limits 971-2479) to male rats and 933 mg/kg (95% confidence limits 692-1259) to female rats.

Executive summary:

Groups of 5 male and 5 female Alpk:APfSD rats received a single oral dose of 200, 500, 1000 or 2000 mg/kg of the test substance. The animals were assessed daily for the following 14 days for any signs of systemic toxicity and their bodyweights recorded at intervals throughout the study. Animals in extremis and those surviving to the end of the study were killed and, together with those found dead, subjected to a macroscopic examination post mortem. Surviving animals showed signs of slight to moderate toxicity which were resolving towards the end of the experimental period. Findings at post mortem were restricted to males dosed with 1000 mg/kg and above and included reduced testicular and epididymal weights. The acute oral LD₅₀of the test substance was estimated to be 1426 mg/kg (95% confidence limits 971-2479) to male rats and 933 mg/kg (95% confidence limits 692-1259) to female rats.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

933 mg/kg bw

Quality of whole database:

The available data are considered to be relevant, reliable and adequate for risk assessment, classification and labelling of the substance.

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:

17 February - 28 October 2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, no restrictions, fully adequate for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Alpk:APfSD (Wistar-derived)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: approximately 8 weeks
- Weight at study initiation: 345±16.6 g (males), 240.8±12.3 g (females)
- Fasting period before study: none
- Housing: 5 per cage, sexes separately in cages suitable for rats of this strain and weight range expected during the course of the study
- Diet: RMI ad libitum (except during exposure)
- Water: mains water ad libitum (except during exposure)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3°C
- Humidity: 30-70%
- Air changes: minimum of 15 per hr
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 14 October 2005 To: 28 October 2005

Route of administration:

inhalation

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: PERSPEX exposure chamber covered with an aluminium cone and standing on an aluminium base
- Exposure chamber volume: approximately 9.2 L (in this study 2 chambers were connected giving a total chamber volume of approximately 27.6 L)
- Method of holding animals in test chamber: polycarbonate restraining tubes
- Source and rate of air: clean, dried and filtered air, at least 10 air-changes per hour. Flow rates 20 L/min.
- System of generating particulates/aerosols: Rotating Brush Generator (RBG)
- Method of particle size determination: Marple Cascade Impactor
- Temperature, humidity, pressure in air chamber: 21.2-21.3°C; 5-7%, slightly higher than ambient

TEST ATMOSPHERE
- Brief description of analytical method used: HPLC
- Samples taken from breathing zone: yes

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

5 mg/L

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observed prior to study start, frequently during exposure, at end of exposure period and once per day thereafter. Weighed prior to study start (day -1), days 1, 8 and 15
- Necropsy of survivors performed: yes
- Other examinations performed: none

Statistics:

None (limit test, no mortalities)

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.41 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: limit test, no mortalities

Mortality:

none

Clinical signs:

other: During exposure, abnormalities generally associated with restraint (wet fur and chromodacryorrhea) were seen and all animals had substance around the snout. Immediately after exposure wet fur, salivation, stains around the nose and chromodacryorrhea were

Body weight:

All animals gained weight by day 8 and continued to gain weight until the end of the study.

Gross pathology:

No abnormalities were seen.

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Nose-only exposure for 4 hours to a particulate concentration of 5.41 mg/L of the test substance resulted in no deaths and no adverse effects. The acute inhalation LC50 is in excess of 5.41 mg/L.

Executive summary:

A group of 5 male and 5 female Alpk:APfSD (Wistar-derived) rats were exposed nose-only for a single 4-hour period to the test substance at a target particulate concentration of 5 mg/L. Test atmospheres were analysed for particulate concentration and the test substance. Following exposure, the animals were retained for 14 days without treatment. Clinical observations and bodyweights were recorded throughout the study and at the end of the scheduled period the animals were killed and subjected to a gross examination post mortem. The achieved test atmosphere had the following characteristics: achieved particulate concentration = 5.41 mg/L; MMAD 7.02 and 6.06 µm; GSD 1.77 and 1.74. Nose-only exposure for 4-hours to a particulate concentration of 5.41 mg/L resulted in no deaths and no adverse effects. The acute inhalation LC₅₀is in excess of 5.41 mg/L.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available data are considered to be relevant, reliable and adequate for risk assessment, classification and labelling of the substance.

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:

11 September - 10 October 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, no restrictions, fully adequate for assessment

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

Qualifier:

according to guideline

Guideline:

EPA OPP 81-2 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: Alpk:APfSD

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: young adults
- Weight at study initiation: 237-270 g (males), 191-218 g (females)
- Fasting period before study: none
- Housing: individually in cages suitable for rats of this strain and weight range expected during the course of the study
- Diet: PCD ad libitum
- Water: mains water ad libitum
- Acclimation period: at least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21±2°C
- Humidity: 40-70%
- Air changes: approximately 25-30 per hr
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 25 September 1995 To: 10 October 1995

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorso-lumbar region (approximately 10 cm x 5 cm shaved the day prior to application) and test substance applied to approximately 1/3 shaved area)
- Type of wrap if used: 4-ply gauze (approximately 7 x 7 cm), covered by plastic film (approximately 7 x 7 cm), held in position using adhesive bandage (approximately 25 x 7.5 cm), secured with two pieces of PVC tape (each approximately 2.5 x 20 cm)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed using clean swabs of cotton wool soaked in clean warm water and dried with clean tissue paper
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 28.4-32.3 mg/cm2 for males and 22.9-26.1 mg/cm2 for females
- For solids, paste formed: yes (with 0.5 mL deionised water)

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observed prior to study start, once between 1-4 hours after application and once daily thereafter. Bodyweights recorded immediately before dosing (day 1) and on days 3, 4, 8 and 15
- Necropsy of survivors performed: yes

Statistics:

none (limit test, no mortalities)

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: limit test, no mortalities

Mortality:

none

Clinical signs:

other: none

Gross pathology:

no treatment-related findings

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute dermal LD50 of the test substance is in excess of 2000 mg/kg to male and female rats.

Executive summary:

A group of five male and five female Alpk:APfSD (Wistar-derived) rats received a single dermal application of 2000 mg/kg of the test substance. The rats were assessed daily for the following 14 days for any signs of systemic toxicity and their bodyweights were recorded at intervals throughout the study. At the end of the study all the rats were killed and given a macroscopic examination post mortem. None of the animals died and there were no signs of systemic toxicity or skin irritation. All animals showed an overall weight gain during the study. There were no treatment related abnormalities at post mortem. The acute dermal LD₅₀was in excess of 2000 mg/kg to male and female rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available data are considered to be relevant, reliable and adequate for risk assessment, classification and labelling of the substance.

Additional information

Oral toxicity

Groups of 5 male and 5 female Alpk:APfSD rats received a single oral dose of 200, 500, 1000 or 2000 mg/kg of the test substance (Johnson 1999). The animals were assessed daily for the following 14 days for any signs of systemic toxicity and their bodyweights recorded at intervals throughout the study. Animals in extremis and those surviving to the end of the study were killed and, together with those found dead, subjected to a macroscopic examination post mortem. Surviving animals showed signs of slight to moderate toxicity which were resolving towards the end of the experimental period. Findings at post mortem were restricted to males dosed with 1000 mg/kg and above and included reduced testicular and epididymal weights. The acute oral LD50 of the test substance was estimated to be 1426 mg/kg (95% confidence limits 971-2479 mg/kg) to male rats and 933 mg/kg (95% confidence limits 692-1259 mg/kg) to female rats.

Dermal toxicity 

A group of five male and five female Alpk:APfSD (Wistar-derived) rats received a single dermal application of 2000 mg/kg of the test substance (Lees 1999). The rats were assessed daily for the following 14 days for any signs of systemic toxicity and their bodyweights were recorded at intervals throughout the study. At the end of the study all the rats were killed and given a macroscopic examination post mortem. None of the animals died and there were no signs of systemic toxicity or skin irritation. All animals showed an overall weight gain during the study. There were no treatment related abnormalities at post mortem. The acute dermal LD50 was in excess of 2000 mg/kg to male and female rats.

Inhalation toxicity 

A group of 5 male and 5 female Alpk:APfSD (Wistar-derived) rats were exposed nose-only for a single 4-hour period to the test substance at a target particulate concentration of 5 mg/L (Rattray 2005). Test atmospheres were analysed for particulate concentration and the test substance. Following exposure, the animals were retained for 14 days without treatment. Clinical observations and bodyweights were recorded throughout the study and at the end of the scheduled period the animals were killed and subjected to a gross examination post mortem. The achieved test atmosphere had the following characteristics: achieved particulate concentration = 5.41 mg/L; MMAD 7.02 and 6.06 µm; GSD 1.77 and 1.74. Nose-only exposure for 4 hours to a particulate concentration of 5.41 mg/L resulted in no deaths and no adverse effects. The 4-hour acute inhalation LC50 was in excess of 5.41 mg/L.

Conclusion 

The above OECD guideline studies were conducted under GLP for acute oral, dermal and inhalation endpoints. The results are considered to be relevant, reliable and adequate for risk assessment, classification and labelling of the substance. The available data are considered to be conclusive.

Justification for selection of acute toxicity – oral endpoint
Only one study was available for acute oral toxicity.

Justification for selection of acute toxicity – inhalation endpoint
Only one study was available for acute inhalation toxicity. The LC50 value was greater than 5400 mg/m3, which was the highest dose in the test.

Justification for selection of acute toxicity – dermal endpoint
Only one study was available for acute dermal toxicity. The LD50 value was greater than 2000 mg/kg, which was the highest dose in the test.

Justification for classification or non-classification

Acute oral toxicity

Acute oral toxicity, LD50 = 933 mg/kg bw, leads to the classification of the substance as Category 4, H302, Harmful if swallowed, according to the criteria in Regulation (EC) No. 1272/2008, Annex I, Part 3, 3.1.2.

Acute inhalation toxicity

The acute inhalation LC50 of the substance is >5410 mg/m3. As a result the substance does not meet the criteria for classification according to Regulation (EC) No. 1272/2008, Annex I, Part 3, 3.1.2.

Acute dermal toxicity

The acute dermal toxicity of the substance is >2000 mg/kg. As a result the substance does not meet the criteria for classification according to Regulation (EC) No. 1272/2008, Annex I, Part 3, 3.1.2.