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Acute Toxicity: oral

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Administrative data

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.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
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:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
12H-phthaloperin-12-one
EC Number:
230-049-5
EC Name:
12H-phthaloperin-12-one
Cas Number:
6925-69-5
Molecular formula:
C18H10N2O
IUPAC Name:
12H-isoindolo[2,1-a]perimidin-12-one
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material : 12H-phthaloperin-12-one
- Common name : C.I. Solvent Orange 60
- Molecular formula : C18H10N2O
- Molecular weight : 270.29 g/mol
- Smiles notation : O=C1N2c3c4c(cccc4N=C2c2ccccc12)ccc3
- InChl : 1S/C18H10N2O/c21-18-13-8-2-1-7-12(13)17-19-14-9-3-5-11-6-4-10-15(16(11)14)20(17)18/h1-10H
- Substance type : Organic
- Physical state : Solid
Specific details on test material used for the study:
- Purity as per Certificate of Analysis:97%
- Lot No.:20120720
- Manufactured date:DEC-2013
- Retest date:20 June 2027
- pH:4.69 at 26.8°C
- Density:1.176 g/cm3 at 27°C
- Storage conditions:Ambient (+15 to +25°C)
- SAFETY PRECAUTIONS: Gloves, cap and face mask were used in addition to protective body garments and shoes, to ensure adequate personal health and safety and to avoid inhalation and skin contact with the test item.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Geniron Biolabs Pvt. Ltd. Bengaluru
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10 to 12 Weeks
- Weight at study initiation: 177.51 g to 202.61 g
- Identification:By rat accession number. Identification of individual rats was by cage card and turmeric colour body marking. The rat accession number was 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:Three rats/cage were housed (except during exposure period and up to observation period of 48 hours wherein housed individually) in the standard polysulfone cages (Size: 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 an enrichment object and were changed along with the cage at least once a week.
- Diet (e.g. ad libitum):Hypro Rat & Mice pellet feed
- 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 six days for G1-FTS, eight days for G1-STS and twelve days for G2-FTS before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 24°C
- Humidity (%): 65 to 67%
- Air changes (per hr): air conditioned with adequate fresh air supply (12.4 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

IN-LIFE DATES: From: 03 July 2018 To: 09 July 2018

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 300 & 2000 mg/kg body weight.
- Amount of vehicle (if gavage):dose volume was 20 mL/kg
Doses:
G1 (FTS) - 300 mg/kg
G1 (STS) - 300 mg/kg
G2 (FTS) - 2000 mg/kg
G2 (STS) - 2000 mg/kg
G2 (FTS) - 2000 mg/kg
No. of animals per sex per dose:
G1 (FTS) - 300 mg/kg - 3
G1 (STS) - 300 mg/kg - 3
G2 (FTS) - 2000 mg/kg - 3
G2 (STS) - 2000 mg/kg - 3
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 - At each step, the animals were observed five times on test day 1 (day of administration) i.e. at 30 minutes and four times at hourly intervals and once daily during days 2 to 15 post administration. Observations 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. Attention was directed to the observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma and all observed clinical signs were recorded. Body weights - The body weights were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration).
- Necropsy of survivors performed: yes, the rats surviving to the end of the observation period were euthanised by using isoflurane anaesthesia and subjected to detailed necropsy.
- Other examinations performed: Gross pathological findings were recorded and reported. Microscopic examination was not carried out as no gross pathological changes were observed.
Statistics:
not specified

Results and discussion

Preliminary study:
not specified
Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no other details available
Mortality:
G1 (FTS) - 300 mg/kg - There were no pre-terminal deaths observed.
G1 (STS) - 300 mg/kg - There were no pre-terminal deaths observed.
G2 (FTS) - 2000 mg/kg - No deaths
G2 (STS) - 2000 mg/kg - No deaths
Clinical signs:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: There were no clinical sings and pre-terminal deaths.
G2 - [2000 mg/kg body weight - Treatment (FTS and STS)]: There were no clinical signs observed in any of the rats.
Body weight:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: The body weights of all the rats increased throughout the observation period.
G2 - [2000 mg/kg body weight - Treatment (FTS and STS)]: The body weights of all the rats increased throughout the observation period.
Gross pathology:
There were no gross pathological changes at necropsy.
Other findings:
not specified

Any other information on results incl. tables

TABLE 1.      Body weight, body weight change and pre-terminal deaths

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Body weight (g)

Day of Death

 (Time of Death)

No. dead/

 No. tested

Pre-terminal deaths

(%)

Initial

(Day 1)

8thday

Weight change

(day 8 – Initial)

15thday

Weight change          

(day 15 – Initial)

At Death

G1

(FTS)

300

 

Rw271

F

183.00

189.24

6.24

196.41

13.41

NA

NA

 

0/3

 

0

Rw272

F

191.08

198.16

7.08

203.67

12.59

NA

NA

Rw273

F

192.26

201.49

9.23

208.12

15.86

NA

NA

G1

(STS)

300

 

Rw274

F

194.47

201.67

7.2

208.38

13.91

NA

NA

 

0/3

 

0

Rw275

F

202.61

209.28

6.67

215.83

13.22

NA

NA

Rw276

F

191.21

198.96

7.75

204.71

13.50

NA

NA

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Body weight (g)

Day of Death

 (Time of Death)

No. dead/

 No. tested

Pre-terminal deaths

(%)

Initial

(Day 1)

8thday

Weight change

(day 8 – Initial)

15thday

Weight change          

(day 15 – Initial)

At Death

G2

(FTS)

2000

 

Rw277

F

190.07

198.12

8.05

204.96

14.89

NA

NA

 

0/3

 

0

Rw278

F

189.94

199.61

9.67

207.21

17.27

NA

NA

Rw279

F

177.51

186.32

8.81

194.68

17.17

NA

NA

G2

(STS)

2000

 

Rw280

F

187.43

195.82

8.39

203.78

16.35

NA

NA

 

0/3

 

0

Rw281

F

186.19

197.13

10.94

206.18

19.99

NA

NA

Rw282

F

180.49

189.78

9.29

198.19

17.70

NA

NA

F: Female        FTS: First Treatment Step            STS: Second Treatment Step             NA: Not Applicable           

 

APPENDIX 1.      Individual clinical signs, dose administration and necropsy findings

Group and

Dose

(mg/kg

body weight)

Date and

time of administration

Rat

No.

Sex

Body

weight

(Day 1)

(g)

Total volume administered

(mL)

Day of observation

Day 1

30 min

1hour

2 hours

3 hours

4 hours

G1

(FTS)

300

 

 

03 July 2018 and

11:30 AM

to

11:33 AM

Rw271

F

183.00

3.66

N

N

N

N

N

Rw272

F

191.08

3.82

N

N

N

N

N

Rw273

F

192.26

3.85

N

N

N

N

N

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Day of observation

Necropsy

Findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G1

(FTS)

300

 

Rw271

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw272

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw273

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female            FTS: First treatment step              N: Normal            min: minutes         mg: milligrams            kg: kilograms       

mL: millilitre         055 (b): Recumbent;            043 (1): Ataxia – slight;              NAD: No Abnormality Detected 

 

APPENDIX 2 contd. Individual clinical signs, dose administration and necropsy findings               

Group and

Dose

(mg/kg

body weight)

Date and

time of administration

Rat

No.

Sex

Body

weight

(Day 1)

(g)

Total volume administered

(mL)

Day of observation

Day 1

30 min

1hour

2 hours

3 hours

4 hours

G1

(STS)

300

 

 

05 July 2018 and

11:48 AM

to

11:50 AM

Rw274

F

194.47

3.89

N

N

N

N

N

Rw275

F

202.61

4.05

N

N

N

N

N

Rw276

F

191.21

3.82

N

N

N

N

N

  

 

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Day of observation

Necropsy

Findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G1

(STS)

300

 

Rw274

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw275

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw276

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female        STS: Second treatment step               N: Normal              min: minutes       mg: milligrams                      kg: kilograms             

mL: millilitre         043 (1): Ataxia – slight;             NAD: No Abnormality Detected           

 

APPENDIX 2 contd. Individual clinical signs, dose administration and necropsy findings         

Group and

Dose

(mg/kg

body weight)

Date and

time of administration

Rat

No.

Sex

Body

weight

(Day 1)

(g)

Total volume administered

(mL)

Day of observation

Day 1

30 min

1hour

2 hours

3 hours

4 hours

G2

(FTS)

2000

 

 

10 July 2018 and

11:33 AM

to

11:35 AM

Rw277

F

190.07

3.80

N

N

N

N

N

Rw278

F

189.94

3.80

N

N

N

N

N

Rw279

F

177.51

3.55

N

N

N

N

N

        

 

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Day of observation

Necropsy

Findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G2

(FTS)

2000

 

Rw277

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw278

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw279

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female         FTS: First treatment step                 min: minutes   mg: milligrams                       kg: kilograms     mL: millilitre      

055 (b): Recumbent;    016: Dead;              NAD: No Abnormality Detected

 


Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
Acute oral toxicity study was conducted in order assess the toxicological profile of the test item on wistar rats. Based on the results, The LD50 of test chemical was determined to be 5000 mg/kg ,Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.
Executive summary:

The acute oral toxicity study with test chemical in Wistar rats was conducted to assess the toxicological profile of the test item. The dose formulation was prepared by using Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) three female rats (G1-FTS) at the dose of 300 mg/kg body weight. There were no clinical signs of toxicity and pre-terminal deaths observed. As all the rats survived at this step, the test was confirmed with three additional female animals with the same dose of 300 mg/kg body weight (G1-STS). There were no clinical signs of toxicity and pre-terminal deaths observed at this step.Based on the scheme - Annex 2c of the guideline OECD 423, the test was repeated at the dose of 2000 mg/kg body weight (G2-FTS). There were no clinical signs of toxicity observed. Hence, the test was confirmed with three additional female rats at the same dose of 2000 mg/kg body weight (G2-STS) as per the scheme - Annex 2c of the guideline OECD 423. There were no clinical signs of toxicity and pre-terminal deaths observed at this confirmatory test. Hence, the further dosing was stopped.The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Gross necropsy was performed for all the rats at termination. The body weight growth of all rats was unaffected by the test item. There were no gross pathological changes at necropsy, hence histopathology was not performed.Based on the results of the present study, the LD50of test item is 5000 mg/kg body weight or infinity as per LD50cut-off value,Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.