Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on experimental studies conducted on rats for the test chemical. The LD50 value is between 300-2000 mg/kg bw for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified as “Category IV” for acute oral toxicity.

Acute Inhalation Toxicity:

In accordance with column 2 of Annex VIII, this end point was considered for waiver since the vapour pressure of 4,4'-bis(dimethylamino)-4''-(methylamino)trityl alcohol is very low (0.00000000052 Pa) and thus there is no possibility of exposure by the inhalation route in this case. Also, considering that the particle size of this chemical ranges between 53 to 250 micrometer in size; there is no possibility of inhalable dust particles (size usually in nano meters) being generated during the use.

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on experimental study report conducted on rats for the test chemical. The studies concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental study report
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
The aim of this study was to assess the toxicity potential of test chemical after single oral administration in rats.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Bharat Serum and Vaccines Limited, Mumbai, India.
- Age at study initiation:9 - 11 weeks at the time of dosing.
- Health Status :Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
- Weight at study initiation:Minimum: 178 g and Maximum: 211 g (Individual body weights were within ± 6% prior to treatment after overnight fasting)-.Fasting period:Rats were fasted for 16-18 hrs
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding:All cages were provided with corn cobs.
- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle:All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum):All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum.
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period:Animal nos. 1-3 were acclimatized for six days, 4-6 for nine days and 7-9 for seven days prior to administration of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 20.00 °C and Maximum: 23.60 °C
- Humidity (%):Minimum: 37.40% and Maximum: 61.80%
- Air changes (per hr):More than 12 changes per hour.
- Photoperiod (hrs dark / hrs light):12:12

Route of administration:
oral: unspecified
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Amount of vehicle (if gavage):10 ml
- Justification for choice of vehicle:Corn oil was selected as a vehicle because test item was not soluble in distilled water.
- Lot/batch no. (if required):MKBD4650
- Purity:N/A

MAXIMUM DOSE VOLUME APPLIED:10 ml/kg body weight.

Doses:
G1 = 2000 mg/kg bw
G2 = 300 mg/kg bw
No. of animals per sex per dose:
9 female rats
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Daily
- Necropsy of survivors performed: yes
At the end of 14 day observation period, all the survived rats were euthanised by overdose of CO2 for external and internal observations.
- Other examinations performed:
Clinical Observation
After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all the surviving animals were observed once a day during the 14 day observation period.

Body weight
All surviving rats were weighed on days 0 (prior to dosing), 7 and 14. Animals were weighed immediately after found dead.

other:
Mortality
All the surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.
Statistics:
not specified
Preliminary study:
not specified
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality was observed
Mortality:
At 2000 mg/kg, all the three animals (animal nos. 1-3) were found dead on day 1 post dosing, whereas no mortality was observed in the animals treated with 300 mg/kg dose throught out the 14 days observation period.
Clinical signs:
At 2000 mg/kg, animal nos. 1 and 3 were observed normal at 30 minutes, 1, 2, 3 and 4 hours post dosing, severe abdominal breathing and sternal recumbency on day 1 followed by found dead. Animal no. 2 observed normal at 30 minutes, 1, 2, 3 and 4 hours post dosing, severe lethargy, moderate abdominal breathing and lateral recumbency on day 1 followed by found dead. At 300 mg/kg, all the six animals (animal nos. 4-9) were observed normal throughout the experimental period.
Body weight:
Mean body weight of the animals treated with 300 mg/kg was observed with gain on day 7 and 14, as compared to day 0. At 2000 mg/kg, all the found dead animals were observed with decreased body weight as compared to day 0.
Gross pathology:
At 2000 mg/kg, all three animals (animal nos. 1-3) were observed with blue colour around perineal area, whreas no external gross pathological changes were observed in the animals treated with 300 mg/kg dose.
At 2000 mg/kg, animal nos. 1 and 3 were observed with moderate red discoloration of all the lobes of lungs, severe dark colour observed in all the lobes of liver, moderate dark colour observed in the kidney, blue tinched urine observed in the urinary bladder and blue discoloration of test item observed in the stomach and intestine. Animal no. 2 was observed with severe red discoloration of all the lobes of lungs, severe dark colour observed in all the lobes of liver, moderate dark colour observed in the kidney, blue tinched urine observed in the urinary bladder and blue discoloration of test item observed in the stomach and intestine, whreas no internal gross pathological changes were observed in the animals treated with 300 mg/kg dose.
Other findings:
not specified

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

 

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Found Dead

Day

0-7

Day

0-14

1

G1/ 2000

194

 -

 -

173

 -

 -

2

190

 -

 -

169

 -

 -

3

190

 -

 -

166

 -

 -

4

G2/ 300

211

240

250

 -

13.74

18.48

5

200

224

231

 -

12.00

15.50

6

207

229

234

 -

10.63

13.04

7

197

216

211

 -

9.64

7.11

8

182

189

202

 -

3.85

10.99

9

178

195

205

 -

9.55

15.17

Key:- = Not applicable

 

  

Table 2: Summary of Animal Body Weight (g) and Body Weight Changes (%)

 

Sex:Female

Group/ Dose (mg/kg)

Rats Body Weight (g)

Body Weight Changes (%)

Day 0

Day 7

Day 14

0-7

0-14

G1/ 2000

Mean

191.33

 -

 -

 -

 -

SD

2.31

 -

 -

 -

 -

n

3

 -

 -

 -

 -

G2/ 300

Mean

195.83

215.50

222.17

9.90

13.38

SD

13.29

19.89

19.07

3.36

3.97

n

6

6

6

6

6

Keys:- = Not applicable, SD = Standard Deviation, n = Number of Animals


Table 3: Individual Animal Clinical Signs and Symptoms

 

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Hours (Day 0)

1/2

1

2

3

4

1

G1/ 2000

1

1

1

1

1

2

1

1

1

1

1

3

1

1

1

1

1

4

G1/ 300

 

1

1

1

1

1

5

1

1

1

1

1

6

1

1

1

1

1

7

1

1

1

1

1

8

1

1

1

1

1

9

1

1

1

1

1

 

Animal No.

Group/ Dose (mg/kg)

Days post dosing

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1

G2/ 2000

4+++

155

2

-

-

-

-

-

-

-

-

-

-

-

-

-

2

99+++4++

98

2

-

-

-

-

-

-

-

-

-

-

-

-

-

3

4+++

155

2

-

-

-

-

-

-

-

-

-

-

-

-

-

4

G2/ 300

1

1

1

1

1

1

1

1

1

1

1

1

1

1

5

1

1

1

1

1

1

1

1

1

1

1

1

1

1

6

1

1

1

1

1

1

1

1

1

1

1

1

1

1

7

1

1

1

1

1

1

1

1

1

1

1

1

1

1

8

1

1

1

1

1

1

1

1

1

1

1

1

1

1

9

1

1

1

1

1

1

1

1

1

1

1

1

1

1

Keys:- = Not applicable, 1 = Normal, 2 = Found dead, 4 = Abdominal breathing, 98 = Lateral recumbency, 99 = Lethargy, 155 = Sternal recumbency,++= Moderate,+++= Severe


Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The lethal concentration LD50 value for acute oral toxicity test was considered to be >300-<2000 mg/kg bw,when nine female wisatr rats were treated with test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).
Executive summary:

The acute oral toxicity profile of test chemical in nine wistar rats at dose concentration of 300 mg/kg and 2000 mg/kg bw.corn oil was used as vehicle.This study was performed as per OECD No. 423. Nine female Wistar rats were selected for acute oral toxicity study. The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing, with food withheld but drinking water providedad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs.Three rats of group G1 were dosed with starting dose of 2000 mg/kg body weight, as the animals were found dead (on day 1 post dosing). Hence three animals of the group G2 were dosed with 300 mg/kg body weight and no mortality was observed, therefore another three animals of same group G2 were dose with 300 mg/kg and no mortality was observed. Hence, further dosing was stopped.Body weights were re­corded on day 0 (prior to dosing) 7 and 14 and of found dead. Mean body weight of the animals treated with 300 mg/kg was observed with gain on day 7 and 14, as compared to day 0.At 2000 mg/kg, all the found dead animals were observed with decreased body weight as compared to day 0. At 2000 mg/kg, animal nos. 1 and 3 were observed normal at 30 minutes, 1, 2, 3 and 4 hours post dosing, severe abdominal breathing and sternal recumbency on day 1 followed by found dead. Animal no. 2 observed normal at 30 minutes, 1, 2, 3 and 4 hours post dosing, severe lethargy, moderate abdominal breathing and lateral recumbency on day 1 followed by found dead. At 300 mg/kg, all the six animals (animal nos. 4-9) were observed normal throughout the experimental period.At 2000 mg/kg, all three animals (animal nos. 1-3) were observed with blue colour around perineal area, whreas no external gross pathological changes were observed in the animals treated with 300 mg/kg dose.At 2000 mg/kg, animal nos. 1 and 3 were observed with moderate red discoloration of all the lobes of lungs, severe dark colour observed in all the lobes of liver, moderate dark colour observed in the kidney, blue tinched urine observed in the urinary bladder and blue discoloration of test item observed in the stomach and intestine. Animal no. 2 was observed with severe red discoloration of all the lobes of lungs, severe dark colour observed in all the lobes of liver, moderate dark colour observed in the kidney, blue tinched urine observed in the urinary bladder and blue discoloration of test item observed in the stomach and intestine, whreas no internal gross pathological changes were observed in the animals treated with 300 mg/kg dose.The acute oral LD50value of test chemical was >300 - ≤2000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the test chemicals, it infers that test chemical exhibit acute oral toxicity in “Category 4” LD50 > 300 to ≤ 2000 mg/kg body weight.

 

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
1 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from study report.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Quality of whole database:
Waiver

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental study report
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
The objective of the study was to assess the dermal toxicity of test chemical after single dose application by dermal route in rats.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:In-House Bred at sa-Ford, Animal Facility.
- Age at study initiation:N/A
- Health Status:Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
- Weight (Prior to Treatment):Male:Minimum: 232 g and Maximum: 265 g , Female:Minimum: 248 g and Maximum: 261 g
- Fasting period before study:N/A
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding : All cages were provided with corn cobs.
- Room Sanitation : The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle : All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum):All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum.
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period:All animals were acclimatized to the test conditions for 6 days prior to administration of the test item.
- Randomization : Animals were selected manually. No computer generated randomization program was used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 20.40 °C and Maximum: 23.10 °C
- Humidity (%):Minimum: 38.40% and Maximum: 58.70%
- Air changes (per hr):More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):12:12

Type of coverage:
semiocclusive
Vehicle:
other: distilled water
Details on dermal exposure:
TEST SITE
- Area of exposure:The test item was applied uniformly over clipped dorsal area of rat skin.
- % coverage:Approximately 10% body surface area of rat.
- Type of wrap if used:The porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done):The residual test item was removed by using distilled water.
- Time after start of exposure:24-hour.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):A limit dose of 2000 mg/ kg body weight of test item was applied.
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit):0.2 ml distilled water
- Concentration (if solution):N/A
- Lot/batch no. (if required):N/A
- Purity:N/A
Duration of exposure:
24 hrs
Doses:
2000 mg/kg body weight.
No. of animals per sex per dose:
10 (Five per sex)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Daily
- Necropsy of survivors performed: yes
At the end of 14 day observation period, all the surviving rats were euthanised by overdose of CO2 and subjected to gross pathology examination, for external and internal observations.
- Other examinations performed:
- Clinical signs : After test item administration, individual animals were frequently observed at 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all animals were observed once a day during the 14 day observation period.

- Body weight: All rats were weighed on days 0 (prior to dosing), 7 and 14.

other:
- Local Signs/Skin Reactions
All animals were observed once daily during days 1-14 (in common with clinical signs).

- Mortality
Animals were observed twice daily for any mortality during the experimental period.
Statistics:
No statistical analysis was performed since the study was terminated with limit test.
Preliminary study:
not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality was observed
Mortality:
No mortality was observed at limit dose of 2000 mg/kg body weight of test item during the 14 day observation period.
Clinical signs:
No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except mild alopecia was observed in female animal no. 9 on day 9 and 10.
Body weight:
The mean body weight of male and female animals was observed with gain compared to day 0 throughout the experiment.
Gross pathology:
The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality
Other findings:
not specified

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

 

Dose:2000 mg/ kg bodyweight                                                                                                         

Animal No.

Sex

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day 0-7

Day 0-14

1

Male

260

263

279

1.15

6.08

2

257

252

263

-1.95

4.37

3

265

260

277

-1.89

6.54

4

260

258

278

-0.77

7.75

5

232

262

260

12.93

-0.76

6

Female

251

260

273

3.59

5.00

7

248

251

259

1.21

3.19

8

256

261

260

1.95

-0.38

9

259

258

265

-0.39

2.71

10

261

260

267

-0.38

2.69

 

 

 

Table 2: Individual Animal Clinical Signs and Symptoms

 

Dose:2000 mg/kg body weight

Animal No.

Sex

Hour(s) - Day 0

Day

1

2

3

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1

Male

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

4

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

5

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

6

Female

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

7

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

8

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

9

1

1

1

1

1

1

1

1

1

1

1

1

12+

12+

1

1

1

1

10

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

Keys: 1 = Normal, 12 = Alopecia, + = Mild


Table 3: Individual Animal Mortality Record

 

Dose:2000 mg/kg body weight

       Animal No.

Sex

Days of Observation (0 to 14)

Morning Observations

Evening Observations

1

Male

No mortality and morbidity

No mortality and morbidity

2

No mortality and morbidity

No mortality and morbidity

3

No mortality and morbidity

No mortality and morbidity

4

No mortality and morbidity

No mortality and morbidity

5

No mortality and morbidity

No mortality and morbidity

6

Female

No mortality and morbidity

No mortality and morbidity

7

No mortality and morbidity

No mortality and morbidity

8

No mortality and morbidity

No mortality and morbidity

9

No mortality and morbidity

No mortality and morbidity

10

No mortality and morbidity

No mortality and morbidity


Interpretation of results:
other: not classified
Conclusions:
The LD50 value was considered to be >2000 mg/kg bw,when groups of 5 male and female wistar rats were semiocclusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).
Executive summary:

The acute dermal toxicity profile of test chemical in groups of 5 male and female wistar rats.This study was performed as per OECD No.402. Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment. On test day 0, anamount oftestitem moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area.This gauze patch was covered with a semi-occlusive dressing.The dressing was wrapped around the abdomen and anchored with non-irritating adhesive tape.After the 24-hour application period, the dressings were removed and theskin was gently wiped with distilled water.The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were re­corded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically.No mortality was observed in any animal till the end of the experimental period. No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except mild alopecia was observed in female animal no. 9 on day 9 and 10.The mean body weight of male and female animals was observed with gain compared to day 0 throughout the experiment.The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Hence,The LD50 value was considered to be >2000 mg/kg bw,when male and female wistar rats were semiocclusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from study report.

Additional information

Acute oral toxicity:

The acute oral toxicity profile of test chemical in nine wistar rats at dose concentration of 300 mg/kg and 2000 mg/kg bw.corn oil was used as vehicle.This study was performed as per OECD No. 423. Nine female Wistar rats were selected for acute oral toxicity study. The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing, with food withheld but drinking water providedad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs.Three rats of group G1 were dosed with starting dose of 2000 mg/kg body weight, as the animals were found dead (on day 1 post dosing). Hence three animals of the group G2 were dosed with 300 mg/kg body weight and no mortality was observed, therefore another three animals of same group G2 were dose with 300 mg/kg and no mortality was observed. Hence, further dosing was stopped.Body weights were re­corded on day 0 (prior to dosing) 7 and 14 and of found dead. Mean body weight of the animals treated with 300 mg/kg was observed with gain on day 7 and 14, as compared to day 0.At 2000 mg/kg, all the found dead animals were observed with decreased body weight as compared to day 0. At 2000 mg/kg, animal nos. 1 and 3 were observed normal at 30 minutes, 1, 2, 3 and 4 hours post dosing, severe abdominal breathing and sternal recumbency on day 1 followed by found dead. Animal no. 2 observed normal at 30 minutes, 1, 2, 3 and 4 hours post dosing, severe lethargy, moderate abdominal breathing and lateral recumbency on day 1 followed by found dead. At 300 mg/kg, all the six animals (animal nos. 4-9) were observed normal throughout the experimental period.At 2000 mg/kg, all three animals (animal nos. 1-3) were observed with blue colour around perineal area, whreas no external gross pathological changes were observed in the animals treated with 300 mg/kg dose.At 2000 mg/kg, animal nos. 1 and 3 were observed with moderate red discoloration of all the lobes of lungs, severe dark colour observed in all the lobes of liver, moderate dark colour observed in the kidney, blue tinched urine observed in the urinary bladder and blue discoloration of test item observed in the stomach and intestine. Animal no. 2 was observed with severe red discoloration of all the lobes of lungs, severe dark colour observed in all the lobes of liver, moderate dark colour observed in the kidney, blue tinched urine observed in the urinary bladder and blue discoloration of test item observed in the stomach and intestine, whreas no internal gross pathological changes were observed in the animals treated with 300 mg/kg dose.The acute oral LD50value of test chemical was >300 - ≤2000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the test chemicals, it infers that test chemical exhibit acute oral toxicity in “Category 4” LD50 > 300 to ≤ 2000 mg/kg body weight.

Thus, based on the above summarised study on test chemical, all rats were died at 2000 mg/kg bw, hence the LD100 was considered to be 2000 mg/kg bw. Considering this value, the LD50 value can be assumed to be 1000 mg/kg bw. Therefore, comparing this value with the criteria of CLP regulation, the given test chemical can be classified in “Category 4 (300 – ≤ 2000)” for acute oral toxicity.

Acute Inhalation Toxicity:

In accordance with column 2 of Annex VIII, this end point was considered for waiver since the vapour pressure of 4,4'-bis(dimethylamino)-4''-(methylamino)trityl alcohol is very low (0.00000000052 Pa) and thus there is no possibility of exposure by the inhalation route in this case. Also, considering that the particle size of this chemical ranges between 53 to 250 micrometer in size; there is no possibility of inhalable dust particles (size usually in nano meters) being generated during the use.

Acute Dermal Toxicity:

The acute dermal toxicity profile of test chemical in groups of 5 male and female wistar rats.This study was performed as per OECD No.402. Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment. On test day 0, anamount oftestitem moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area.This gauze patch was covered with a semi-occlusive dressing.The dressing was wrapped around the abdomen and anchored with non-irritating adhesive tape.After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water.The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on testday 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were re­corded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically.No mortality was observed in any animal till the end of the experimental period. No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except mild alopecia was observed in female animal no. 9 on day 9 and 10.The mean body weight of male and female animals was observed with gain compared to day 0 throughout the experiment.The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Hence,The LD50 value was considered to be >2000 mg/kg bw,when male and female wistar rats were semiocclusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female wistar rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is between 300-2000 mg/kg bw for acute oral and >2000 mg/kg bw for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified as “Category IV” for acute oral toxicity and cannot be classified for acute dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.