2-amino-4,6-dinitrophenol

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Data source

Reference

Reference Type:

secondary source

Title:

OPINION ON Picramic acid and sodium picramate COLIPA n° B28

Author:

Scientific Committee on Consumer Safety

Year:

2012

Bibliographic source:

Scientific Committee on Consumer Safety, (SCCS) 18 September 2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Sodium picramate

IUPAC Name:

Sodium picramate

Constituent 2

Reference substance name:

Sodium 2-amino-4,6-dinitrophenoxide

EC Number:

212-603-8

EC Name:

Sodium 2-amino-4,6-dinitrophenoxide

Cas Number:

831-52-7

IUPAC Name:

sodium 2-amino-4,6-dinitrophenolate

Details on test material:

- Name of test material (as cited in study report): Sodium picramate
- Molecular formula (if other than submission substance): C6H4N3NaO5
- Molecular weight (if other than submission substance): 221.10 g/mol
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations): 62.4%

Test animals

Species:

rat

Strain:

other: HanBrl:WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

No data available

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Bi-distilled water

Details on oral exposure:

VEHICLE
- Justification for use and choice of vehicle (if other than water): Water was used.
- Concentration in vehicle: 0, 20, 100 and 250 mg/kg bw/day
- Amount of vehicle (if gavage): 10 ml/kg bw
- Lot/batch no. (if required):
- Purity:

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

14 day

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 20, 100 and 250 mg/kg bw/day
Basis:
no data

No. of animals per sex per dose:

Total no of animals-40
0 mg/kg bw/day -5 male and 5 female.
20 mg/kg bw/day 5 male and 5 female.
100 mg/kg bw/day 5 male and 5 female.
250 mg/kg bw/day 5 male and 5 female.

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data available.

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

FOOD EFFICIENCY: No data available.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water studies): No data available.

OPHTHALMOSCOPIC EXAMINATION: No data available.

HAEMATOLOGY: No data available.

CLINICAL CHEMISTRY: No data available.

URINALYSIS: No data available.

NEUROBEHAVIOURAL EXAMINATION: No data available.

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes,
Histological examinations were performed on organs and tissues from all control and high dose animals, and all gross lesions from all animals were recorded.

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Deaths were observed at the high dose(250 mg/kg bw/day) -two males: Days 5 and 7; three females: Days 1, 3 and 7

Mortality:

mortality observed, treatment-related

Description (incidence):

Deaths were observed at the high dose(250 mg/kg bw/day) -two males: Days 5 and 7; three females: Days 1, 3 and 7

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

The mean absolute body weights were decreased in males treated at the dose of 250 mg/kg bw/day. In females of this dose 250 mg/kg bw/day showed, a non -statistically significant decrease was observed.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

The mean absolute food intake was slightly decreased in males and females treated at the dose of 250 mg/kg bw/day and the mean relative food intake was also decreased in males and females treated at this dose when compared to control rats

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

The macroscopic lesions observed and possibly related to treatment consisted of enlarged spleen observed in 3 male rats and 1 female rat of the 250 mg/kg bw group. They could be correlated to the increase of the weight of the spleen.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Changes in colon as foci, nodules or thickened organ were observed in three male rats and one female rats treated at the dose of 250 mg/kg bw/day and in one female rat treated at the dose of 100 mg/kg bw/day.

Histopathological findings: neoplastic:

not specified

Details on results:

Clinical signs:
In rats treated at the dose of 250 mg/kg bw/day several clinical signs were observed: during the first treatment week, slightly ruffled fur was observed in one female on treatment days 4 and 5 and in an other one on treatment day 3; slight emaciation was observed in four females and two males in the first treatment week and moderate emaciation in a further male on treatment day 6. Decreased spontaneous activity was observed in one female on treatment day 4 and 5 and in one male on treatment day 6. Slightly brown urine was seen in two females and three males on the last day of treatment.

Body weight and weight gain:
The mean body weight gain was decreased in males and females from the high dose group (250 mg/kg bw/day).

Organ weights:
Increased dose-related mean spleen weights and spleen to body ratios were observed in males and females rats treated at 250 mg/kg bw/day. The mean liver to body weight ratio was increased in males treated at the dose of 100 mg/kg bw/day or 250 mg/kg bw/day and in females at the dose of 250 mg/kg bw/day and 100 mg/kg bw/day but at this dose the increase was not statistically significant. An increase of the mean brain to body weight ratio was also observed in male rats at the dose of 250 mg/kg bw/day.

Gross pathology:
Reduced size of testes, epididymis, prostate and seminal vesicles was seen in three males treated at the dose of 250 mg/kg bw/day.

Histopathology:
Thickened cecum was observed in three male rats and foci were seen on the caecum of two males and two females treated at the dose of 250 mg/kg bw/day.

Other:
Other signs observed were discoloration of lung, lung not collapsed and thickened thymus, but they were not considered to be related to the treatment with sodium picramate.

Effect levels

Dose descriptor:

NOAEL

Effect level:

20 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No significant effect was observed in clinical change, mortality, body weight, Food intake, organ weight, grosspathology and histopathology.

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

NOAEL was found to be 20 mg/kg/day for Sodium picramate in Wistar rats by oral (gavage) administration in a 14 day study.

Executive summary:

In a repeated dose toxicity study the effect of Sodium picramate was observed in male and femaleWistarrats for 14 days. The male and female Wistarrats were exposed to Sodium picramate on a daily basis at dose concentration of0, 20, 100 and 250 mg/kg bw/day.Deaths were observed at the high dose [two males: Days 5 and 7; three females: Days 1, 3and 7].Control animals, animals treated at the dose of 20 mg/kg bw/day or 100 mg/kg bw/day showed no treatment related clinical effects.

In rats treated at the dose of 250 mg/kg bw/day several clinical signs were observed:during the first treatment week, slightly ruffled fur was observed in one female on treatment days 4 and 5 and in another one on treatment day 3; slight emaciation was observed in four females and two males in the first treatment week and moderate emaciation in a further male on treatment day 6. Decreased spontaneous activity was observed in one female on treatment day 4 and 5 and in one male on treatment day 6. Slightly brown urine was seen in two females and three males on the last day of treatment. The mean absolute food intake was slightly decreased in males and females treated at the dose of 250 mg/kg bw/day and the mean relative food intake was also decreased in males and females treated at this dose when compared to control rats. The mean absolute body weights were decreased in males treated at the dose of 250 mg/kg bw/day when compared to the control group. In females of this dose group a not statistically significant decrease was observed. The mean body weight gain was decreased in males and females from the high dose group.Increased dose-related mean spleen weights and spleen to body ratios were observed in males and females rats treated at 250 mg/kg bw/day. The mean liver to body weight ratio was increased in males treated at the dose of 100 mg/kg bw/day or 250 mg/kg bw/day andin females at the dose of 250 mg/kg bw/day and 100 mg/kg bw/day but at this dose the increase was not statistically significant. An increase of the mean brain to body weight ratio was also observed in male rats at the dose of 250 mg/kg bw/day. The macroscopic lesions observed and possibly related to treatment consisted of enlarged spleen observed in 3 male rats and 1 female rat of the 250 mg/kg bw group. They could be correlated to the increase of the weight of the spleen and were considered related to the treatment. Reduced size of testes, epididymes, prostate and seminal vesicles was seen in three males treated at the dose of 250 mg/kg bw/day. Changes in colon as foci, nodules or thickened organ were observed in three male rats and one female rats treated at the dose of 250 mg/kg bw/day and in one female rat treated at the dose of 100 mg/kg bw/day. Thickened caecum was observed in three male rats and foci were seen on the caecum of two males and two females treated at the dose of 250 mg/kg bw/day. Other signs observed were discoloration of lung, lung not collapsed and thickened thymus, but they were not considered to be related to the treatment with sodium picramate. Therefore NOAEL was considered to be 20 mg/kg/day for Sodium picramate inWistarrats by oral (gavage) for 14 days.