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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:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental 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 (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Tetrabutylammonium hydrogen sulphate
- Molecular formula :C16H36N.HO4S
- Molecular weight : 339.53g/mol
- Smiles notation: C([N+](CCCC)(CCCC)CCCC)CCC.S([O-])(=O)(=O)O
- InChl : 1S/C16H36N.H2O4S/c1-5-9-13-17(14-10-6-2,15-11-7-3)16-12-8-4;1-5(2,3)4/h5-16H2,1-4H3;(H2,1,2,3,4)/q+1;/p-1
- Substance type: Organic
- Physical state: Solid
- Purity as per Certificate of Analysis: 100.03%
- Lot No. : L199101607
- Manufactured date : JUL-2016
- Expiry Date : JUN-2021
- pH : 1.73 at 26°C
- Density : 0.448 g/cm3 at 25.9°C
- Storage conditions : Ambient (+15 to +25°C)

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals 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:165.34 g to 210.05 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): 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 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 (%): 66 to 68%
- Air changes (per hr): air conditioned with adequate fresh air supply (between 13.1 and13.2 air changes/hour)
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

IN-LIFE DATES: From: 24 July 2018 To: 11 September 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.
- Amount of vehicle (if gavage):dose volume was 10 mL/kg body weight
Doses:
G1 (FTS) - 300 mg/kg
G1 (STS) - 300 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
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
Sex:
female
Dose descriptor:
LD50
Effect level:
1 000 mg/kg bw
Based on:
test mat.
Mortality:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: There were no mortality/morbidity in FTS and STS till the termination.
G2 - [2000 mg/kg body weight - Treatment (FTS)]: There were two mortalities observed at 0.5 hours post-dose observation.
Clinical signs:
G1 - [300 mg/kg body weight - Treatment (FTS and STS)]: There were no clinical signs in FTS and STS till the termination.
G2 - [2000 mg/kg body weight - Treatment (FTS)]: 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)]: The body weights of a survived rat 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

G2

(FTS)

2000

 

Rw313

F

177.04

185.29

8.25

191.87

14.83

NA

NA

 

2/3

 

66.67

Rw314

F

193.57

-

-

-

-

193.29

1 (12.24 PM)

Rw315

F

165.34

-

-

-

-

165.22

1 (12.24 PM)

F: Female        FTS: First 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

11:51 AM

to

11:54 AM

Rw307

F

190.06

1.90

N

N

N

N

N

Rw308

F

194.65

1.95

N

N

N

N

N

Rw309

F

174.59

1.75

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

 

Rw307

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw308

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw309

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       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

12:10 PM

to

12:12 PM

Rw310

F

210.05

2.10

N

N

N

N

N

Rw311

F

198.68

1.99

N

N

N

N

N

Rw312

F

196.13

1.96

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

 

Rw310

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw311

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw312

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     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

11:52 AM

to

11:54 AM

Rw313

F

177.04

1.77

N

N

N

N

N

Rw314

F

193.57

1.94

016

-

-

-

-

Rw315

F

165.34

1.65

016

-

-

-

-


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

 

Rw313

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rw314

F

-

-

-

-

-

-

-

-

-

-

-

-

-

-

NAD

Rw315

F

-

-

-

-

-

-

-

-

-

-

-

-

-

-

NAD

F: Female           FTS: First treatment step        N: Normal   016: Dead       min: minutes   kg: kilograms        mL: millilitre NAD: No Abnormality Detected      

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Based on the results of the present study, the LD50 value of the test chemical was considered to be 1000 mg/kg body weight. Thus, the test chemical was classified in “Category 4 (300 – ≤ 2000)” criteria of CLP.
Executive summary:

The acute oral toxicity study was conducted to assess the toxicological profile of the test chemical as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) in Wistar rats. 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 continued at the dose of 2000 mg/kg body weight (G2-FTS). The two rats (Rw314 and Rw315) were found dead at 0.5 hours post dose observations, hence as per the scheme - Annex 2c of the guideline OECD 423, 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 survived rats was unaffected by the test chemical. There were no gross pathological changes at necropsy, hence histopathology was not performed. Based on the results of the present study, the LD50 value of the test chemical was considered to be 1000 mg/kg body weight. Thus, the test chemical was classified in “Category 4 (300 – ≤ 2000)” criteria of CLP.