Registration Dossier

Diss Factsheets

Administrative data

Description of key information

 Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on the study conducted on rats for the given test chemical. The study concluded that the LD50 value is 1049 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:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to low vapour pressure of the test chemical. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on the study conducted on rats for the given test chemical. The study 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
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Principles of method if other than guideline:
The objective of this acute oral toxicity study was to assess the toxicological profile of the test item when administered to rats by a single oral gavage.
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
no
Specific details on test material used for the study:
- Name of test material: (2-methoxy ethyl) benzene
- Molecular formula: C9H12O
- Molecular weight: 136.193 g/mole
- Smiles : COCCc1ccccc1
- Inchl: InChI=1/C9H12O/c1-10-8-7-9-5-3-2-4-6-9/h2-6H,7-8H2,1H3
- Substance type: Organic
- Physical state: Clear colourless, mobile liquid, almost insoluble in water
- Purity as per Certificate of Analysis : 99.7302%
- Lot No. : PE0201/18
- Manufactured date : Jan 2018
- Expiry Date : Dec 2022
- pH : 3.98 at 26.8°C
- Density : 0.952g/cm3
- Storage conditions : Ambient (+18 to +36ºC)
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-vivo Biosciences,Karnataka,India
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10 to 12 weeks
- Weight at study initiation: 207.5 to 226.5 g
- Identification:By rat accession number, cage card and turmeric colour body marking. The rat accession numbers were allotted during the course of the study. The temporary body marking during acclimatization period was done with crystal violet.
- Fasting period before study: rats were fasted for approximately 16 to 18 hours
- Housing:Rats were housed individually in standard polysulfone cages (Size: L 425 x B 266 x H 185 mm), with stainless steel top grill
- Diet (e.g. ad libitum): Hypro Rat & Mice pellet feed, ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier, Mumbai, ad libitum
- Acclimation period: the animals were acclimatized for six to twenty two days (Steps 1 to 7) under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 25°C
- Humidity (%):64 to 68%
- Air changes (per hr): air conditioned with adequate fresh air supply (between 13.13 and 14.1 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

IN-LIFE DATES: From: 05 December 2018 To: 05 January 2019
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2000 and 550 mg/kg bw.
- Amount of vehicle (if gavage):2.1 mL/kg body weight to attain the dose of 2000 mg/kg body weight (Step 1, 3, 5 and 7) and 0.58 mL/kg body weight to attain the dose of 550 mg/kg body weight (Step 2, 4, 6) respectively.

MAXIMUM DOSE VOLUME APPLIED: 2.1 mL/kg body weight
Doses:
Step 1 :2000 mg/kg body weight
Step 2 :550 mg/kg body weight
Step 3 :2000 mg/kg body weight
Step 4 :550 mg/kg body weight
Step 5: 2000 mg/kg body weight
Step 6 :550 mg/kg body weight
Step 7 :2000 mg/kg body weight
No. of animals per sex per dose:
TOTAL: 7 animals
Step 1 :2000 mg/kg body weight :1
Step 2 :550 mg/kg body weight:1
Step 3 :2000 mg/kg body weight:1
Step 4 :550 mg/kg body weight:1
Step 5: 2000 mg/kg body weight:1
Step 6 :550 mg/kg body weight:1
Step 7 :2000 mg/kg body weight:1
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Clinical signs and pre-terminal deaths:The animals were observed five times on test day 1 (day of administration) i.e., at 30 minutes and 1, 2, 3 and 4 hours after dose administration and once daily during days 2 to 15. Additional observation was done for Rw2351, Rw2353 and Rw2357.Observation was included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. All observed clinical signs were recorded.
- Body weights:Individual body weights of animals were recorded on test day 1 (Pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration) and at death.
- Necropsy of survivors performed: yes, At the end of the observation period all the surviving rats were euthanized by using deep isoflurane anaesthesia and the pre-terminally dead animals were also subjected to detailed necropsy by an experienced prosector. Microscopic examination was not carried out as no gross pathological changes were observed.
Statistics:
not specified
Preliminary study:
As per the Material Safety Data Sheet provided by the Sponsor, the acute oral LD50 of test chemical in rats is 2292.314 mg/kg body weight. Hence the study was initiated with the starting dose of 2000 mg/kg body weight (Step 1). The rat died and as per the AOT425 Statistical program, the six rats (Step 2 to Step 7) were administered with dose of 550, 2000, 550, 2000, 550 and 2000 mg/kg sequentially. The stopping criteria was met with 5 reversals occurred in 6 consecutive animals tested (as per AOT statistical program).Dose progression and stopping criteria was calculated using a dedicated software program (Acute Oral Toxicity (Guideline 425) Statistical Program) provided by the EPA.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
1 049 mg/kg bw
Based on:
test mat.
95% CL:
>= 550 - <= 2 000
Remarks on result:
other: 50% mortality was observed
Mortality:
2000 mg/kg body weight – Step 1: animal found dead on during day 3 observation.
2000 mg/kg body weight – Step 3: animal found dead on during end of treatment day 1 observation.
2000 mg/kg body weight – Step 5: animal found dead on 4th hour post dose observation.
2000 mg/kg body weight – Step 7: animal found dead on day 2.
550 mg/kg body weight – Step 2, 4 and 6: no pre-terminal deaths were observed.
Clinical signs:
2000 mg/kg body weight – Step 1: Slight ataxia was observed during 4th hour post dose to end of the treatment day1 and recumbent, slight lacrimation, dyspnoea on day 2 observation.
2000 mg/kg body weight – Step 3: Slight ataxia was observed 3rd hour post dose. Recumbent and dyspnoea 4th hour post dose.
2000 mg/kg body weight – Step 5: Moderate tremors and gasping were observed during 3rd hour post dose.
2000 mg/kg body weight – Step 7: Hypoactivity observed 3rd hour to 4th hour post dose, Recumbent was observed on end of the treatment day 1.
550 mg/kg body weight – Step 2, 4 and 6: There were no clinical signs of toxicity.
Body weight:
2000 mg/kg body weight – Step 1, 3, 5 and 7: The body weight of all the dead rats decreased at death.
550 mg/kg body weight – Step 2, 4 and 6: All rats gained body weight throughout the observation period when compared to their initial weight.
Gross pathology:
There were no abnormalities detected at necropsy.
Other findings:
not specified
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Based on the results, the acute oral LD50 value for the given test chemical is considered to be 1049 mg/kg body weight (approximate 95% confidence interval is 550 to 2000) in female wistar rats.The test item is classified as “Category 4 (300-2000 mg/kg bw)” as per the criteria of CLP.
Executive summary:

The acute oral toxicity study was conducted to assess the toxicological profile of the given test item (2-methoxy ethyl) benzene (CAS no.: 3558 -60 -9, E.C. no.: 222 -619 -7) as per OECD Guideline 425 ( Acute Oral Toxicity -Up-and-Down Procedure, adopted- 3rd october 2008), in Wistar rats to produce toxicity when a single dose was administered via the oral route.The undiluted test item was administered as a single oral gavage to rats fasted for 16 to 18 hours.In Preliminary test,the acute oral LD50 of test chemical in rats was considered to be 2292.314 mg/kg body weight. Hence the study was initiated with the starting dose of 2000 mg/kg body weight (Step 1). The rat died and as per the AOT425 Statistical program, the six rats (Step 2 to Step 7) were administered with dose of 550, 2000, 550, 2000, 550 and 2000 mg/kg sequentially. The stopping criteria was met with 5 reversals occurred in 6 consecutive animals tested.Dose progression and stopping criteria was calculated using a dedicated software program (Acute Oral Toxicity (Guideline 425) Statistical Program) provided by the EPA.Clinical signs and pre-terminal deaths, Body weights and Necropsy were observed in treated animals.At dose 2000 mg/kg body weight (Step 1) Slight ataxia was observed during 4th hour post dose to end of the treatment day1 and recumbent, slight lacrimation, dyspnoea on day 2 were observed and found dead on during day 3 observation.At dose 2000 mg/kg body weight (Step 3) Slight ataxia was observed 3rd hour post dose with recumbent and dyspnoea 4th hour post dose and found dead on during end of treatment day 1 observation.At dose 2000 mg/kg body weight (Step 5) moderate tremors and gasping were observed during 3rd hour post dose.At dose 2000 mg/kg body weight (Step 7) hypoactivity observed 3rd hour to 4th hour post dose with respect to recumbent which was observed on end of the treatment day 1 and found dead on day 2.At dose 550 mg/kg body weight (Step 2, 4 and 6) there were no clinical signs of toxicity and no pre-terminal deaths were observed.Changes in body weight were seen at dose 2000 mg/kg body weight (Step 1, 3, 5 and 7) and 550 mg/kg body weight (Step 2, 4 and 6). There were no abnormalities detected at necropsy.Based on the results of the present study, the LD50 value for the given test chemical is considered to be 1049 mg/kg (approximate 95% confidence interval is 550 to 2000).Therefore, The test item is classified as “Category 4 (300-2000 mg/kg bw)” as per the criteria of CLP. This test was performed according to GLP.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 049 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from experimental study report.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
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 study report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
The objective of this acute dermal toxicity study was to assess the toxicological profile of the test item on application as a single semi-occlusive dermal application to rats.
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
- Name of test material: Benzene(-2-methoxyethyl)
- Molecular formula: C9H12O
- Molecular weight: 136.193 g/mole
- Smiles : COCCc1ccccc1
- Inchl: InChI=1/C9H12O/c1-10-8-7-9-5-3-2-4-6-9/h2-6H,7-8H2,1H3
- Substance type: Organic
- Physical state: Clear colourless, mobile liquid, almost insoluble in water
- Purity as per Certificate of Analysis: 99.7302%
- Lot No. : PE0201/18
- Manufactured date : Jan 2018
- Expiry Date : Dec 2022
- pH :6.81
- Density :0.952g/cm3
- Storage conditions : Ambient (+18 to +36ºC)
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-vivo Biosciences,Bengaluru,Karnataka, India
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 weeks
- Weight at study initiation: Females:226.2 to 233.5 g
- Identification:By rat accession number. Identification of individual rats is by cage card and crystal violet and turmeric body markings. The temporary body marking during acclimatization period was done with crystal violet. The rat accession numbers were allotted during the course of the study and was included in raw data and reported.
- Housing: Animals were housed individually in standard polysulfone cages (Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top grill having facilities for pelleted food and drinking water in polycarbonate bottle.Additionally, polycarbonate rat huts were placed inside the cage as enrichment objects and were changed along with the cage once a week. Bedding: Steam sterilized corn cob was used and changed once a week along with the cage.
- Diet (e.g. ad libitum):Rat & Mice pellet feed, ad libitum
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier, ad libitum
- Acclimation period: the rats were acclimatized for six to fourteen days before treatment for dose range finding and main study respectively under standard laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):20 to 25°C
- Humidity (%): 64 to 68%
- Air changes (per hr): Air conditioned with adequate fresh air supply (13.3 to 14.1 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle

IN-LIFE DATES: From:05 December 2018 To: 02 January 2019
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: clipped skin of dorsolateral thoracic surface of the skin was clipped (approximately 10 x 8 cm) with an electric clipper (Aesculap - Germany).
- % coverage: 10% of the body surface
- Type of wrap if used: The applied area was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The dressing was removed and the applied area was washed with deionized water and wiped dry using clean towel.
- Time after start of exposure:24 hours
Duration of exposure:
24 hours
Doses:
DRF G1 - 200 mg/kg
DRF G2 - 1000 mg/kg
DRF G3 - 2000 mg/kg
Main G3 - 2000 mg/kg
No. of animals per sex per dose:
DRF G1 - 200 mg/kg - 1
DRF G2 - 1000 mg/kg - 1
DRF G3 - 2000 mg/kg - 1
Main G3 - 2000 mg/kg - 2
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
- Clinical examination and pre-terminal deaths:The animals were observed for clinical signs and pre-terminal deaths (mortality) once during first 30 minutes after application, and at hourly
intervals for 6 hours after application on the day of treatment (day 1) and once daily during Days 2 to 15. In addition, the site of applied area was observed for skin reactions at 24, 48
and 72 hours after removal of test chemical using the Draize criteria.All rats were observed for changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Body weights -Individual body weights of animals were recorded on test days 1 (Pre-application), 8 (7 days post application), and 15 (14 days post application).
- Necropsy of survivors performed: yes,at the end of the observation period, all rats were euthanised and exsanguinated under isoflurane anesthesia and subjected to detailed necropsy by an experienced prosector and the findings were recorded.
- Other examinations performed: Microscopic examination was not carried out as no gross pathological changes were observed.
Statistics:
not specified
Preliminary study:
There was no information available on the toxicity of the test item. Hence, a starting dose of 200 mg/kg body weight was selected and tested in 1 female rat (dose range finding study). As there was no mortality at this dose range finding study as per Annexure 1 of the guideline the dose range finding study was continued with 1 female rat (dose range finding study) at the next higher dose of 1000 mg/kg body weight. There was no mortality, the dose range finding study was continued with 1 female rat (dose range finding study) at the next higher dose of 2000 mg/kg body weight. There was no mortality; the test was continued with the main study with 2 animals at the dose of 2000 mg/kg body weight to confirm the classification. There was no test item-related mortality.The subsequent dosing was done 2 to 3 days after the previous dosing.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
There were no pre-terminal deaths (mortality) observed during the study.
Clinical signs:
There were no clinical signs observed during the study. There were no skin reactions at the site of application at 24,48 and 72 hours after test patch removal (as per draize method).
Body weight:
All rats gained body weight throughout the observation period.
Gross pathology:
No abnormality was detected at necropsy.
Other findings:
not specified

Table 1. Individual body weight, body weight changes and pre-terminal deaths

Group and

Dose

(mg/kg body

weight)

Rat No.

Sex

Body weight (g)

Pre-terminal

Initial deaths

Initial (Day 1 - at

treatment)

8thday

Weight change

(day 8 – Initial)

15thday

Weight change

(day 15 – Initial)

G1 and

200

DRF

Rw2363

F

229.0

236.3

7.3

248.6

19.6

0

G2 and

1000

DRF

Rw2364

F

233.5

239.8

6.3

248.9

15.4

0

G3 and

2000

DRF

Rw2365

F

226.2

238.6

12.4

247.6

21.4

0

G3 and

2000

Main study

Rw2366

F

226.2

239.5

13.3

250.7

24.5

0

Rw2367

F

232.1

244.4

12.3

252.3

20.5

0

 

DRF: Dose Range Finding F: Female

 

Table 2: Individual test item application, clinical signs, skin reaction and necropsy findings 

Dose range finding study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Volume

(mL)

applied

Observations and skin reaction

Days

1

2

3

4

5

30 min

1 h

2 h

3 h

4 h

5 h

6 h

*

Er @

Ed @

*

Er @

Ed @

*

Er @

Ed @

G1 and

200

DRF

11 December

2018

and

10:41 AM

Rw2363

F

229.0

0.05

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

 

Group &

Dose

(mg/kg

body weight)

Rat

Number

Sex

Observation

Necropsy

findings

Days

6

7

8

9

10

11

12

13

14

15

G1 and

200

DRF

Rw2363

F

N

N

N

N

N

N

N

N

N

N

NAD

F: Female  N: Normal  h: hour   min: minutes   NAD: No abnormality detected   Er: Erythema   Ed: Edema

Score 0: No Erythema / Edema

 

Table 2 contd.: Individual test item application, clinical signs, skin reaction and necropsy findings

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Volume

(mL)

applied

Observations and skin reaction

Days

1

2

3

4

5

30 min

1 h

2 h

3 h

4 h

5 h

6 h

*

Er @

Ed @

*

Er @

Ed @

*

Er @

Ed @

G2 and

1000

DRF

14 December

2018

and

10:40 AM

Rw2364

F

233.5

0.25

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

Group &

Dose

(mg/kg

body weight)

Rat

Number

Sex

Observation

Necropsy

findings

Days

6

7

8

9

10

11

12

13

14

15

G2 and

1000

DRF

Rw2364

F

N

N

N

N

N

N

N

N

N

N

NAD

F: Female  N: Normal  h: hour   min: minutes   NAD: No abnormality detected   Er: Erythema   Ed: Edema

Score 0: No Erythema / Edema

 

Table 2 contd.: Individual test item application, clinical signs, skin reaction and necropsy findings

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Volume

(mL)

applied

Observations and skin reaction

Days

1

2

3

4

5

30 min

1 h

2 h

3 h

4 h

5 h

6 h

*

Er @

Ed @

*

Er @

Ed @

*

Er @

Ed @

G3 and

2000

DRF

17 December

2018

and

11:10 AM

Rw2365

F

226.2

0.48

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

 

Group &

Dose

(mg/kg

body weight)

Rat

Number

Sex

Observation

Necropsy

findings

Days

6

7

8

9

10

11

12

13

14

15

G3 and

2000

DRF

Rw2365

F

N

N

N

N

N

N

N

N

N

N

NAD

 

F: Female  N: Normal  h: hour   min: minutes   NAD: No abnormality detected   Er: Erythema   Ed: Edema

Score 0: No Erythema / Edema

 

Table 2 contd. Individual test item application, clinical signs, skin reaction and necropsy findings

 

Main study

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Volume

(mL)

applied

Observations and skin reaction

Days

1

2

3

4

5

30 min

1 h

2 h

3 h

4 h

5 h

6 h

*

Er @

Ed @

*

Er @

Ed @

*

Er @

Ed @

G3 and

2000

(Main)

19 December

2018

and

11:13 to 11:

Rw2366

F

226.2

0.48

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Rw2367

F

232.1

0.49

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

 

Group &

Dose

(mg/kg

body weight)

Rat

Number

Sex

Observation

Necropsy

findings

Days

6

7

8

9

10

11

12

13

14

15

G3 and

2000

(Main)

Rw2366

F

N

N

N

N

N

N

N

N

N

N

NAD

Rw2367

F

N

N

N

N

N

N

N

N

N

N

NAD

F: Female N: Normal h: hour min: minutes NAD: No abnormality detected Er: Erythema Ed: Edema

Score 0: No Erythema / Edema

*: Clinical signs; @: Skin scoring as per Draize method (approximately 24, 48 and 72 hours) after test patch removal

Interpretation of results:
other: Not classified
Conclusions:
Based on the present study results, the acute dermal LD50 value of the given test chemical is >2000 mg/kg body weight in female Wistar rats.
Executive summary:

The acute dermal toxicity study was conducted by using the given test chemical (2 -methoxy ethyl) benzene (CAS no.: 3558 -60 -9, E.C. no.: 222 -619 -7) as per OECD Guideline 402 (Acute Dermal Toxicity) in Wistar rats was tested with 200 (G1-DRF), 1000 (G2-DRF) and 2000 (G3-DRF) mg/kg with 1 female each for the dose range finding study and 2 female for main study (G3). Approximately 24 hours before treatment, the hair on the dorsolateral thoracic surface of the skin was clipped (approximately 10 x 8 cm) with an electric clipper (Aesculap - Germany). Based on the individual body weight, the test item at the doses of 200, 1000 and 2000 mg/kg body weight, the undiluted test item was applied directly to the clipped skin of the animal to cover about 10% of the body surface of the animal (semi-occlusive). The area of application was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was for 24 hours. After the 24 hours contact period, the dressing was removed, and the applied area was washed with deionized water and wiped dry using clean towels.All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the treatment site was observed at 24, 48 and 72 hours after removal of test item using the Draize criteria. There were no clinical signs and pre-terminal deaths (mortality) observed during the study. There were no skin reactions at the site of application at 24, 48 and 72 hours after test patch removal (as per draize method). All rats gained body weight throughout the observation period.No abnormality was detected at necropsy.Based on the present study results, the acute dermal LD50 value of the given test chemical is >2000 mg/kg body weight in female Wistar rats. 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”.

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

Additional information

Acute oral toxicity: 

In different studies, test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often the studies are based on in vivo experimental data in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

The acute oral toxicity study was conducted to assess the toxicological profile of the given test item as per OECD Guideline 425 ( Acute Oral Toxicity -Up-and-Down Procedure) in Wistar rats to produce toxicity when a single dose was administered via the oral route.The undiluted test item was administered as a single oral gavage to rats fasted for 16 to 18 hours.In Preliminary test,the acute oral LD50 of test chemical in rats was considered to be 2292.314 mg/kg body weight. Hence the study was initiated with the starting dose of 2000 mg/kg body weight (Step 1). The rat died and as per the AOT425 Statistical program, the six rats (Step 2 to Step 7) were administered with dose of 550, 2000, 550, 2000, 550 and 2000 mg/kg sequentially. The stopping criteria was met with 5 reversals occurred in 6 consecutive animals tested.Dose progression and stopping criteria was calculated using a dedicated software program (Acute Oral Toxicity (Guideline 425) Statistical Program) provided by the EPA.Clinical signs and pre-terminal deaths, Body weights and Necropsy were observed in treated animals.At dose 2000 mg/kg body weight (Step 1) Slight ataxia was observed during 4th hour post dose to end of the treatment day1 and recumbent, slight lacrimation, dyspnoea on day 2 were observed and found dead on during day 3 observation.At dose 2000 mg/kg body weight (Step 3) Slight ataxia was observed 3rd hour post dose with recumbent and dyspnoea 4th hour post dose and found dead on during end of treatment day 1 observation.At dose 2000 mg/kg body weight (Step 5) moderate tremors and gasping were observed during 3rd hour post dose.At dose 2000 mg/kg body weight (Step 7) hypoactivity observed 3rd hour to 4th hour post dose with respect to recumbent which was observed on end of the treatment day 1 and found dead on day 2.At dose 550 mg/kg body weight (Step 2, 4 and 6) there were no clinical signs of toxicity and no pre-terminal deaths were observed.Changes in body weight were seen at dose 2000 mg/kg body weight (Step 1, 3, 5 and 7) and 550 mg/kg body weight (Step 2, 4 and 6). There were no abnormalities detected at necropsy.Based on the results of the present study, the LD50 value for the given test chemical is considered to be 1049 mg/kg (approximate 95% confidence interval is 550 to 2000).Therefore, The test item is classified as “Category 4 (300-2000 mg/kg bw)” as per the criteria of CLP.

In another study,acute oral toxicity study was conducted by using the given test chemical in male and female rats at the dose concentration range of 3100-5500 mg/kg bw.Animals were observed for mortality. 50% mortality was observed at 4100 mg/kg bw in treated animals.Hence, the LD50 value was considered to be 4100 mg/kg bw, with 95% confidence limits of 3100-5500 mg/kg bw, when male and female rats was treated with the given test chemical via oral route.

Thus, based on the above experimeantal Guideline study of test chemical, it can be concluded that LD50 value is between 300-2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation,test chemical can be classified as “Category IV” for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to low vapour pressure of the test chemical. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

Acute Dermal toxicity:

In different experimental studiestest chemicalhave been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats and rabbits fortest chemical. The studies are summarized as below -

 

The acute dermal toxicity study was conducted by using the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Wistar rats was tested with 200 (G1-DRF), 1000 (G2-DRF) and 2000 (G3-DRF) mg/kg with 1 female each for the dose range finding study and 2 female for main study (G3). Approximately 24 hours before treatment, the hair on the dorsolateral thoracic surface of the skin was clipped (approximately 10 x 8 cm) with an electric clipper (Aesculap - Germany). Based on the individual body weight, the test item at the doses of 200, 1000 and 2000 mg/kg body weight, the undiluted test item was applied directly to the clipped skin of the animal to cover about 10% of the body surface of the animal (semi-occlusive). The area of application was covered with cotton gauze (size: Females: 8 x 5 cm of 6 ply) and it was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was for 24 hours. After the 24 hours contact period, the dressing was removed, and the applied area was washed with deionized water and wiped dry using clean towels.All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the treatment site was observed at 24, 48 and 72 hours after removal of test item using the Draize criteria. There were no clinical signs and pre-terminal deaths (mortality) observed during the study. There were no skin reactions at the site of application at 24, 48 and 72 hours after test patch removal (as per draize method). All rats gained body weight throughout the observation period.No abnormality was detected at necropsy.Based on the present study results, the acute dermal LD50 value of the given test chemical is >2000 mg/kg body weight in female Wistar rats. 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”.

In another study, acute dermal toxicity study was conducted by using the given test chemical in rabbits at the dose concentration range of 2440-6460 mg/kg bw. Animals were observed for mortality. 50% mortality was observed at 3970 mg/kg bw in treated animals. Hence, the LD50 value was considered to be 3970 mg/kg bw, with 95% confidence limits of 2440-6460 mg/kg bw, when rabbits were treated with the given test chemical by dermal application.

Thus, based on the above summarised studies ontest chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation,test chemicalcannot be classified for acute dermal toxicity.

Justification for classification or non-classification

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