Registration Dossier

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:
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

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- IUPAC Name: 3-nitrobenzoic acid
- InChI: 1S/C7H5NO4/c9-7(10)5-2-1-3-6(4-5)8(11)12/h1-4H,(H,9,10)
- Smiles: c1(cc(ccc1)[N+](=O)[O-])C(=O)O
- Molecular formula :C7H5NO4
- Molecular weight :167.1195 g/mol
- Substance type:Organic
- Physical state:Yellow crystalline powder
- Purity as per Certificate of Analysis:98.0%
- Lot No.:S51551512
- Manufactured date:Dec 2015
- Retest date:Nov 2020
- pH:4.05 at 24 °C
- Density:1.465 g/cm3 @20°C
- Storage conditions:Ambient (+18 to +36 °C )

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Vivo Bio Tech Ltd, Telangana
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 Weeks
- Weight at study initiation: 142.08 g to 167.18 g
- Identification:By rat accession number. Identification of individual rats was by cage card and turmeric colour body markings. 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: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): 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 five days for G1-FTS, ten days for G1-STS, thirteen days for G2-FTS and sixteen days for G2-STS. Animals were observed once daily during acclimatization period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 59 to 67%
- Air changes (per hr): air conditioned with adequate fresh air supply (12.9 to 14.0 air changes/hour).
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

IN-LIFE DATES: From: 30 March 2018 To: 12 July 2018

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Milli-Q water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 300 & 2000 mg/kg body weight (G1 and G2– First and second treatment steps)
- Amount of vehicle (if gavage):10 mL/kg

DOSAGE PREPARATION (if unusual): A required quantity (g) of test item was weighed in mortar and mixed using pestle by adding small volume of Milli-Q water until a uniform suspension was obtained. The mixture was quantitatively transferred to a measuring
cylinder. Further, a small volume of vehicle was added to the mortar and rinsed with the vehicle, all the rinsing was quantitatively transferred into the measuring cylinder. The final volume was made up to the mark with Milli-Q water to get the desired test item concentration (mg/mL).
Doses:
G1 (FTS) - 300 mg/kg
G1 (STS) - 300 mg/kg
G2 (FTS) - 2000 mg/kg
G2 (STS) - 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. 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
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]:There were no pre-terminal deaths.
G2 - [2000 mg/kg body weight - Treatment (FTS and STS)]: There were no pre-terminal deaths.
Clinical signs:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: There were no clinical sings observed in any of the rats.
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

 

Rm8749

F

155.28

202.69

47.41

215.76

60.48

NA

NA

 

0/3

 

0

Rm8750

F

165.21

184.97

19.76

200.32

35.11

NA

NA

Rm8751

F

142.08

166.44

24.36

178.43

36.35

NA

NA

G1

(STS)

300

 

Rm8752

F

162.46

180.12

17.66

172.30

9.84

NA

NA

 

0/3

 

0

Rm8753

F

151.04

177.88

26.84

188.70

37.66

NA

NA

Rm8754

F

143.05

154.52

11.47

172.67

29.62

NA

NA

F: Female        FTS: First Treatment Step            STS: Second Treatment Step                   NA: Not Applicable  0: No deaths 

TABLE 1 contd.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

G2

(FTS)

2000

 

Rm8755

F

150.86

159.21

8.35

178.35

27.49

NA

NA

 

0/3

 

0

Rm8756

F

152.97

163.02

10.05

172.08

19.11

NA

NA

Rm8757

F

151.21

161.78

10.57

176.14

24.93

NA

NA

G2

(STS)

2000

 

Rm8758

F

167.18

178.85

11.67

185.87

18.69

NA

NA

 

0/3

 

0

Rm8759

F

156.58

164.03

7.45

178.69

22.11

NA

NA

Rm8760

F

147.12

159.81

12.69

168.22

21.10

NA

NA

F: Female        FTS: First Treatment Step            STS: Second Treatment Step                   NA: Not Applicable    0: No deaths

Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
Based on the results of the present study, the LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute oral toxicity. CLP criteria "Not Classified".
Executive summary:

The acute oral toxicity study was conducted to assess the toxicological profile of the given test chemical in Wistar rats as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

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. Necropsy was performed for all the rats at termination. All survived rats gained weight during experimental period. There were no gross pathological changes at necropsy.

Based on the results of the present study, the acute oral LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute oral toxicity. CLP criteria "Not Classified".