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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:
2017
Report Date:
2017

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 purpose of this study was to assess the Toxicological profile of test item to a single administration via oral route to Sprague Dawley rats. This study was designed to determine the acute toxicity at fixed dose levels by oral route of the test item.
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:
- Name of test material (IUPAC name): 1-Phenyl-3-pyrazolidone
- Common name: Phenidone
- Molecular formula: C9H10N2O
- Molecular weight: 162.191 g/mol
- Smiles notation: c1ccc(cc1)N2CCC(=O)N2
- InChl: 1S/C9H10N2O/c12-9-6-7-11(10-9)8-4-2-1-3-5-8/h1-5H,6-7H2,(H,10,12)
- Substance type: Organic

SOURCE OF TEST MATERIAL
- Test Item: 1-phenyl-3-pyrazolidone (CAS No. 92-43-3)
- Source of test material: Sustainability Support Services (Europe) AB, Sweden
- Batch No. of test material: P-17126
- Manufacturing Date: 28/4/2017
- Expiration date of the lot/batch: 28/10/2018
- Consistency: Solid, crystalline powder

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item and prepared formulation(s) were stored at ambient temperature.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Test item was suspended in distilled water. The formulation was prepared fresh on the day of dosing. The test item was administered in the
dose volume of 10 ml/kg body weight

OTHER SPECIFICS:
Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Female rats of the age of approximately 8 to 12 weeks old were used at the commencement of its dosing and its weight were within ± 20% of the mean weight of any animal used for dosing. - Weight at study initiation: Body weights at the start : Body weight range was 195.9 to 202.2 grams.
Female Mean: 198.78 g (= 100 %); Minimum : 195.9 g (- 1.45 %); Maximum : 202.2 g (+ 1.72 %)
- Identification: Each female rat was individually identified by the picric acid marking.
- Fasting period before study: Approximately 16 hours or more.
- Housing: The rats were housed in polycarbonate cages.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.1 to 22.1 degree centigrade.
- Humidity (%): 56.6% to 60.1%.
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 04-07-2017 to 19-09-2017

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Distilled water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 300 mg/kg, 300 mg/kg and 2000 mg/kg

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.
Doses:
Dose Group I : 300 mg/kg
Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg
No. of animals per sex per dose:
Three females were used at each step.
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality and morbidity, until sacrifice.
Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time.
The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.

Body weights: Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.

Gross Pathology: Necropsy was performed on all animals, found dead and sacrificed at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).

Histopathology: Gross pathological examination revealed unabsorbed residual test item in distended stomach in found dead animals from 2000 mg/kg dose group. No gross pathological changes attributable to the treatment were observed in stomach and hence no histopathology was considered.
Statistics:
not specified

Results and discussion

Preliminary study:
not specified
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Group I Step I : Animals treated at the dose level of 300 mg/kg body weight: All animals survived through the study period of 14 days.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight: One animal died at 6 hours after the dosing.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight: Three animals died at 1 hour after the dosing.
Clinical signs:
Group I Step I : Animals treated at the dose level of 300 mg/kg body weight resulted in distension, piloerection, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 2 hours after the dosing. All animals were free of signs of toxicity from day 4 to day 5 after the dosing.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and loss of righting reflex with onset at 30 minutes to 4 hours after the dosing. All surviving animals were free of signs oftoxicity from day 4 to day 5 after the dosing.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight resulted in reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing.
Body weight:
Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.31% and 12.09% respectively.
Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.79% and 12.10% respectively.
Group II Step I (2000 mg/kg) - All animals died at 1 hour, hence, body weight gain could not be calculated.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in found dead animal and animals sacrificed terminally from 300 mg/kg dose group.
Gross pathological examination revealed stomach distended with unabsorbed residual test item in found dead animals from 2000 mg/kg dose group.
Other findings:
not specified

Any other information on results incl. tables

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

Test System : Sprague Dawley Rat

Sex : Female

Group I :   

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

300

Distension

2

2

3

2 hrs. - day 1

2 hrs. - 6 hrs.

0/3

Piloerection

3

1

2,3

2 hrs. - 4 hrs.

2 hrs.

Reduced locomotor activity 

3

1

2

3

30 min. - 6 hrs.

1 hr. - day 3

30 min. - day 1

Ataxic gait

3

1

2,3

30 min. - day 3

30 min. - day 4

Tremors

3

1

2,3

2 hrs.

1 hr. - 2 hrs.

 

Group I : 

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

II

300

Distension

3

4,6

5

1 hr. - 4 hrs.

1 hr. - 6 hrs.

1/3

Piloerection

2

5

6

4 hrs. - day 2

4 hrs. - 6 hrs.

Reduced locomotor activity 

3

4

5

6

30 min. - 4 hrs.

30 min. - 6 hrs.

30 min. - day 3

Ataxic gait

3

4

5

6

30 min. - 4 hrs.

30 min. - day 3

30 min. - day 4

Tremors

3

4,6

5

1 hr. - 4 hrs.

1 hr. - day 2

Loss of righting reflex

1

4

1 hr. - 4 hrs.

 

Group II :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

2000

Reduced locomotor activity 

3

7,8,9

10 min. - 30 min.

 

3/3

Ataxic gait

3

7,8,9

5 min. - 30 min.

 

Tremors

3

7,9

8

10 min. - 30 min.

30 min.

Convulsion

3

7,8,9

30 min.

 

 

Table No.II

Mean Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex : Female

Group I :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

300

Mean

198.23

206.77

4.31

222.20

7.46

12.09

± SD

1.61

1.29

0.69

2.82

0.69

1.11

 

 

Group I :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

II

300

Mean

198.57

206.55

4.79

220.95

6.97

12.10

± SD

2.81

4.03

1.14

3.75

0.27

0.94

 

 

Group II :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

2000

Mean

199.53

-

-

-

-

-

± SD

2.52

-

-

-

-

-

 

 

Table No.III

Summary of Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex : Female

 

Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

300

1, 2, 3

TS

No abnormality detected

 

 Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

300

4

FD

No abnormality detected

5, 6

TS

 

Group II : 

Step No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

7, 8, 9

FD

Stomach : Distended with unabsorbed residual test item.

 FD = Found dead

TS = Terminal Sacrifice

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Under the condition of the study, the acute oral LD50 value of the given test chemical was considered in between 300-2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category 4 (300 – ≤ 2000)” criteria of CLP.
Executive summary:

The reported study was designed and conducted to determine the acute oral toxicity profile of the given test chemical as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) in Sprague Dawley rats.

Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 2 hours after the dosing.

As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and loss of righting reflex with onset at 30 minutes to 4 hours after the dosing. One animal died at 6 hours after the dosing.

One mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing. Three animals died at 1 hour after the dosing.

All surviving animals from 300 mg/kg dose group exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in found dead animal and animals sacrificed terminally from 300 mg/kg dose group. Gross pathological examination revealed stomach distended with unabsorbed residual test item in found dead animals from 2000 mg/kg dose group.

Under the condition of the study, the acute oral LD50 value of the given test chemical was considered in between 300-2000 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category 4 (300 – ≤ 2000)” criteria of CLP.