N-(2-(4-amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate

Endpoint summary

Description of key information

Acute oral toxicity (equivalent to OECD 401), rat: 75 mg/kg bw > LD50 < 300 mg/kg bw

Acute dermal toxicity (according to OECD 402, GLP compliant), rat: > 2000 mg/kg bw

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:

28 Jan-11 Feb 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

1981

Deviations:

no

Principles of method if other than guideline:

- Principle of test: Test doses were determined based on an initial "viability test" conducted at 5000 mg/kg bw. Doses administered via gavage were 1200, 300, and 75 mg/kg bw, based on initial fasted body weights to groups of four young male rats per dose level, which were then observed for mortality and clinical signs of toxicity daily for up to 7 days following administration.
- Short description of test conditions: Standard acute oral toxicity testing under controlled laboratory conditions, at three dose levels, with necropsy 7 days after administration. Dead animals were necropsied as soon as possible to try to ascertain the cause of death.
- Parameters analysed / observed: Body weights, food consumption, water consumption, clinical observations of toxicity and mortality.

GLP compliance:

no

Remarks:

pre-GLP

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

other: CRJ-F344

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Labouratories Japan, Inc.
- Age at study initiation: 6-wk
- Weight at study initiation: 105.5 ± 5.17 g
- Fasting period before study: from 16 h before dosing to 6 h after administration
- Housing: individually in five-modular stainless steel cages with barrier system
- Diet: solid feed NMF, Oriental Yeast Co., Ltd., autoclave sterilized, ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1
- Humidity (%): 55 ± 5
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 28 Jan 1982 To: 11 Feb 1982

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 7.5, 30 and 120 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw

Dose range-finding study:
- Rationale for the selection of the doses: A "viability test" was conducted at 5000 mg/kg bw

Doses:

75, 300 and 1200 mg/kg bw

No. of animals per sex per dose:

4 males/dose

Control animals:

other: one control group was tested for all 9 test compounds

Details on study design:

- Duration of observation period following administration: 7 days after dosing
- Frequency of observations and weighing: Observations for clinical signs of toxicity and mortality were conducted on the day of dosing immediately after dosing, 1, 2, 4, 6 and 8 h post-dose, and at the same time each morning on days 1 to 7 after dosing. Food consumption and water consumption were measured on Days 1, 2, 3, 5 and 7 after dosing. Body weights were measured on the day of dosing (fasted, prior to administration), and 1, 2, 3, 5 and 7 days after dosing.
- Necropsy of survivors performed: yes

Statistics:

Means with SD were calculated for body weights, food consumption and water consumption.

Preliminary study:

A viability test was conducted on 9 test articles via oral administration of 5000 mg/kg bw using rats, prior to conduct of this LD50 study.

Key result

Dose descriptor:

LD50

Effect level:

> 75 - < 300 mg/kg bw

Mortality:

75 mg/kg bw: 0/4 males died
300 mg/kg be: 3/4 males died
1200 mg/kg bw: 3/4 males died

Clinical signs:

other: 75 mg/kg bw: piloerection in 4/4 males (2-6 h post-dose) 300 mg/kg bw: piloerection and lacrimation in 4/4 males (2 h post-dose); convulsion, cyanosis, respiratory depression, and death in 2/4 males (day of dose); cyanosis, piloerection and lacrimation in

Gross pathology:

At the necropsies of the animals found dead during the study period, atrophy of the stomach and exfoliation of gastric mucosa were noted. During the necropsies conducted at termination of the study, no abnormalities were found.

Other findings:

Food consumption
75 mg/kg bw: similar trend with control group
300 and 1200 mg/kg bw: decreased food consumption throughout the study compared to control group

Water consumption
75 mg/kg bw: similar trend with control group
300 and 1200 mg/kg bw: decreased water consumption through Day 2 post-dose, then recovering to the control group level throughout the remainder of the study

 Mortality and Clinical Signs for CD-3 Acute Oral Toxicity Study in Rats

Dose [mg/kg bw]	Mortality	Clinical Signs
	N^a	N^a
Males
0	0/4	0/4
75	0/4	4/4
300	3/4	4/4
1200	3/4	4/4

^aNumber of animals dosed

Interpretation of results:

other: CLP/EU GHS Category 3 (H301) according to Regulation (EC) No 1272/2008

Conclusions:

The test substance CD-3 when administered at a dose level of 75 mg/kg bw produced no deaths in 4 dosed animals, however at a dose level of 300 mg/kg bw, 3 of 4 dosed animals died.
Therefore, in this study an LD50 value greater than 75 mg/kg bw but smaller than 300 mg/kg bw was found for CD-3, giving a Acute Tox. Hazard Category 3 classification.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

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:

7 Oct - 12 Nov 1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

1987

Deviations:

yes

Remarks:

occlusive dressing instead of semi-occlusive dressing

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Sprague-Dawley® rats [SAS:V AF®(SD)] obtained from SASCO, Inc., Stone Ridge (Kingston), NY
- Age at study initiation: male rats were 8 weeks of age and female rats were 10 weeks of age
- Weight at study initiation: male rats weighed 206 to 220 grams and female rats weighed 210 to 214 grams
- Housing: Animals were housed in an American Association for Accreditation of Laboratory Animal Care-accredited vivarium·in accordance with the Guide for the Care and Use of Laboratory Animals, singly housed in suspended, stainless-steel, wire mesh cages. Cages and racks were washed once a week. Absorbent paper, used to collect excreta, was changed at least three times a week.
- Diet: Certified Rodent Diet (Purina Rodent Chow #5002, pellets) was available ad libitum.
- Water: ad libitum through an automatic watering system. The source of the water was the local public water system. There have been no contaminants identified in periodic water analyses that would be expected to interfere with the conduct of the study. Semi-annual analyses of water are maintained on file within the testing laboratory.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23
- Humidity (%): 48-62
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: The hair was removed from an area of the dorsal skin with an electric clipper. A single
dose of the test substance was placed in contact with the skin using a fiber pad.
- Type of wrap if used: occlusive wrap

REMOVAL OF TEST SUBSTANCE
- Washing: At the end of the exposure period, any residual test material was removed with running water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw, The test substance was administered as a solid moistened thoroughly with water.

VEHICLE
- Amount(s) applied: The test substance was administered as a solid moistened thoroughly with water.

Duration of exposure:

24 hours
At the end of the exposure period, any residual test material was removed with running water.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 male and 5 female rats per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Body weights were measured on Days O (prior to treatment), 7, and 14.
- Necropsy of survivors performed: yes, All animals were euthanatized and necropsied at the completion of the 14-day observation period.
- Other examinations performed: Animals were observed at least once during the exposure period, and once each day thereafter for the duration of the experiment. Observations included, but were not limited to, examination of the hair, skin, eyes, mucous membranes, motor activity, feces, urine, respiratory system, circulatory system, autonomic nervous system, central nervous system, and behavior patterns.

Statistics:

No statistical procedures were required during the study.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

In the dermal toxicity study, no mortality was noted.

Clinical signs:

other: Discoloration (Black) of the hair was observed immediately adjacent to the application site in all rats (male and female).

Gross pathology:

Treatment-related changes seen at necropsy were limited to discoloration (black) of the hair adjacent to the application site of all animals. Hydrometra was an incidental finding noted in one female rat. No tissue was collected for microscopic examination.
No signs of organ toxicity were observed. All rats survived until scheduled sacrifice at the end of the observation period.

Table 1: Overview Results

Dose	Number of rats (male/female)	Number of Deaths after 14 days (male/female)
2000 mg/kg bw	5/5	0/0

Interpretation of results:

other: no classification required according to Regulation (EC) 1272/2008.

Conclusions:

The acute dermal median lethal dose (LD50) of the test substance in male and female Sprague-Dawley strain rats was > 2000 mg/kg bw.
The available data on acute dermal toxicity of the test substance do not indicate requirement of classification according to Regulation (EC) 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

Additional information

Acute toxicity: oral

An acute oral toxicity study is available for the substance, N-(2-(4-Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate, which was conducted similar to OECD TG 401. In this non-GLP study groups of four male rats were administered the test material at doses of 75, 300, or 1200 mg/kg bw via gavage (NBR, 1982). The animals were observed for mortality, clinical signs of toxicity and body weight change after administration. No mortality occurred in animals dosed with 75 mg/kg bw; however, in both the 300 and 1200 mg/kg bw dose groups, 3 out of 4 animals died. Piloerection was observed in all treated animals. Animals dosed with 300 or 1200 mg/kg bw also exhibited the following clinical signs of toxicity: convulsion, cyanosis, respiratory depression, lacrimation, salvation, hunched posture, hypothermia and respiratory paralysis. Decreased body weight, as well as decreased food and water consumption, also were observed at 300 and 1200 mg/kg bw. In those animals that were found dead, atrophy of the stomach and exfoliation of gastric mucosa were noted. In summary, the LD50 was established at 75 mg/kg bw > LD50 < 300 mg/kg bw.

Acute toxicity: dermal

A GLP compliant acute dermal toxicity study is available for the substance, N-(2 -(4 -Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate, which was conducted according to OECD TG 402 (Shepard, 1997). In this study groups of 5 male and 5 female Sprague-Dawley strain rats were administered the test substance at a dose of 2000 mg/kg bw dermally (solid, moistened with water). A single dose of the test substance was placed in contact with the skin using a fiber pad and an occlusive wrap to hold the test material in place for 24 hours. At the end of the exposure period, any residual test material was removed with running water. The animals were observed for 14 days for mortality, clinical signs of toxicity and body weight change after administration. No mortality occurred at a dose of 2000 mg/kg bw. The only treatment-related changes seen at necropsy were limited to discoloration (black) of the hair adjacent to the application site of all animals. Hydrometra was an incidental finding noted in one female rat. The LD50 for males and females was established at > 2000 mg/kg bw.

Justification for classification or non-classification

The harmonized classification for N-(2 -(4 -Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate is acute toxicity category 4 with the hazard statement H302, according to Annex VI of (EC) No 1272/2008 (CLP Regulation, Index number 612-134-00-2).

Based on the summarized data, N-(2 -(4 -Amino-N-ethyl-m-toluidino)ethyl)methanesulphonamide sesquisulphate is classified as category 3 with the hazard statement H301 ‘Toxic if swallowed’ according to Regulation (EC) 1272/2008.

The available data on acute dermal toxicity of the test substance do not indicate requirement of classification according to Regulation (EC) 1272/2008 and are therefore conclusive but not sufficient for classification.