Registration Dossier

Administrative data

Description of key information

Key study: Test method OECD 420. GLP study. No signs of toxicity were observed following single administration of the test item at a dose of 2000 mg/kg bw in rats. The discriminating dose was determined to be 2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
9 October 2014 - 4 November 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Name of test material (as cited in study report): 4-amino-5-(ethylsulphonyl)-o-anisic acid
- Physical state: Solid
- Analytical purity: 99.81%
- Lot/batch No.: MP1032.31

Species:
rat
Strain:
Wistar
Remarks:
(Crl: WI(Han); outbred)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 11 week-old
- Weight at study initiation: average body weight:197.8 g
- Fasting period before study: about 19 hours before the administration of the test item, restored 3 hours after the administration
- Housing: plastic cages covered with wire bar lids, 58 x 37 x 21 cm, with UV-sterilized wood shavings as bedding.
- Diet (e.g. ad libitum): ad libitum, "Murigran" standard granulated laboratory food.
- Water (e.g. ad libitum): ad libitum, tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-24 ºC
- Humidity (%): 40-85%
- Air changes (per hr): 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 60 mg/mL (dose of 300 mg/kg bw) or 400 mg/mL (dose of 2000 mg/kg bw)
- Amount of vehicle (if gavage): 1 mL

MAXIMUM DOSE VOLUME APPLIED: 0.5 mL/100 g bw

Rationale for the selection of the starting dose: The test item at a single dose of 300 mg/kg bw was administered to one animal. The starting dose for the sighting study was selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg bw. Since no data was available, the sighting study commenced with the administration of the test item at a dose of 300 mg/kg bw to one female rat. No signs of toxicity were observed and the animal survived the experiment. A dose of 2000 mg/kg bw was administered to a second rat. No signs of toxicity were observed and the animal survived the experiment. On the grounds of the sighting study results, the dose of 2000 mg/kg b.w. was selected to be used in the main study.
Doses:
Sighting study: 300 and 2000 mg/kg bw
Main study: 2000 mg/kg bw
No. of animals per sex per dose:
2 females in the sighting study.
4 females in the main study.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: on days 0 (directly before the administration of the test item), 7, and 14 (before euthanasia).
- Necropsy of survivors performed: yes
- Other examinations performed:
General condition: observation of all animals for morbidity and mortality: twice a day or once a day (on days off) during the 14-day experiment.
Detailed clinical observations: on the day of the test item administration (day 0), i.e. 10, 30, and 60 minutes after the administration and then at hourly intervals up to the 5th hour after the administration. From the 1st to the 14th day of the experiment, the detailed clinical observations were performed once a day.
Gross examinations: After the 14-day observation period, all animals were euthanized by intraperitoneal administration of morbital at a dose of 200 mg/kg bw and subjected to gross examinations. The detailed gross examinations comprised the observation of the external body surface, all natural apertures, and the cranial, thoracic, and abdominal cavities with their contents.
Preliminary study:
No signs of toxicity were observed and the animals survived the experiment.
Key result
Sex:
female
Dose descriptor:
discriminating dose
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Mortality:
All animals survived the experiment.
Clinical signs:
No signs of toxicity were observed.
Body weight:
During the 14-day experiment, body weights of all animals increased.
Gross pathology:
No pathological changes were stated.

Table No.1:

Dose of the test item

(mg/kg b.w.)

300

2000

Sighting study

Sighting study

Main study

Mortality

0/1

0/1

0/4

Clinical signs

not found

not found

not found

Table No.2: Clinical signs – overall list.

 

 

 

                       Animal number

Dose

(mg/kg b.w.)

Day after the

administration

Number

of living

animals

1*

2

3

4

5

 

 

 

 

 

 

300

0

1

NC

-

-

   -

  -

1

1

NC

-

-

   -

  -

2

1

NC

-

-

   -

  -

3

1

NC

-

-

   -

  -

4

1

NC

-

-

   -

  -

5

1

NC

-

-

   -

  -

6

1

NC

-

-

   -

  -

7

1

NC

-

-

   -

  -

8

1

NC

-

-

   -

  -

9

1

NC

-

-

   -

  -

10

1

NC

-

-

   -

  -

11

1

NC

-

-

   -

  -

12

1

NC

-

-

   -

  -

13

1

NC

-

-

   -

  -

14

1

NC

-

-

   -

  -

 

 

 

 

 

 

 

2000

0

5

NC

NC

NC

NC

NC

1

5

NC

NC

NC

NC

NC

2

5

NC

NC

NC

NC

NC

3

5

NC

NC

NC

NC

NC

4

5

NC

NC

NC

NC

NC

5

5

NC

NC

NC

NC

NC

6

5

NC

NC

NC

NC

NC

7

5

NC

NC

NC

NC

NC

8

5

NC

NC

NC

NC

NC

9

5

NC

NC

NC

NC

NC

10

5

NC

NC

NC

NC

NC

11

5

NC

NC

NC

NC

NC

12

5

NC

NC

NC

NC

NC

13

5

NC

NC

NC

NC

NC

14

5

NC

NC

NC

NC

NC

*the female from the sighting study

NC:no changes

Table No.3: Body weights of the animal - overall list

 

 

Day of the experiment / Body weight

 

Dose

(mg/kg b.w.)

Animal

number

0

7

14

Body weight

gain(g)

(014)

300

1*

209

240

236

27

 

 

 

2000

1*

202

234

229

27

2

199

215

227

28

3

199

219

230

31

4

201

213

229

28

5

188

207

213

25

* the female from the sighting study

Table No.4: Detailed clinical signs: Female Number 1 , 3000mg/kg bw

Parameter

                                      Day 0 of observation

10 min

30 min

1h

2h

3h

4h

5h

 

 

 

 

 

 

 

Locomotor

system,

behaviour,

reactions to

stimuli

1. Body posture

0

0

0

0

00

0

0

2. Gait

0

0

0

0

00

0

0

3. Locomotor activity

0

0

0

0

00

0

0

4. Involuntary

movement-clonic

0

0

0

0

00

0

0

5. Involuntary

movement-tonic

 

0

0

0

0

00

0

0

6.Reaction to being

caught

0

0

0

0

00

0

0

7. Vocalization

0

0

0

0

00

0

0

8. Reaction to sound

stimuli

0

0

0

0

00

0

0

9. Other

0

0

0

0

00

0

0

 

 

 

Skin and hair

Skin colour

0

0

0

0

00

0

0

2. Edema

0

0

0

0

00

0

0

3. Epidermis

0

0

0

0

00

0

0

4. Coat

0

0

0

0

00

0

0

5. Skin changes

0

0

0

0

00

0

0

6. Other

0

0

0

0

00

0

0

 

 

 

 

 

Eyes and

eyelids

1. Ocular discharge

0

0

0

0

00

0

0

2. Exophthalmos

0

0

0

0

00

0

0

3. Corneal opacity

0

0

0

0

00

0

0

4. Eyelids

0

0

0

0

00

0

0

5. Other

0

0

0

0

00

0

0

 

 

Respiratory

system

1. Respiration

0

0

0

0

00

0

0

2. Discharge from the

nostrils

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

 

Digestive

system

1. Salivation

0

0

0

0

00

0

0

2. Diarrhea

0

0

0

0

00

0

0

3. Bleeding from the

anus

0

0

0

0

00

0

0

4. Other

0

0

0

0

00

0

0

 

Urinary

system

1. Urination

0

0

0

0

00

0

0

2. Hematuria

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

Reproductive system

1. Pathological

discharge from the

reproductive tract

0

0

0

0

00

0

0

 

2. Other

0

0

0

0

00

0

0

Table No.5:Detailed obseravation, dosage 300mg/kg b.w, female No.1

Date of euthanasia: 23.10.2014

Fasting: 18 hours

Animal age: 10 weeks

Parameter

                                      Day of observation

1

2

3

4

5

6

7

 

 

 

 

 

 

 

Locomotor

system,

behaviour,

reactions to

stimuli

1. Body posture

0

0

0

0

00

0

0

2. Gait

0

0

0

0

00

0

0

3. Locomotor activity

0

0

0

0

00

0

0

4. Involuntary

movement-clonic

0

0

0

0

00

0

0

5. Involuntary

movement-tonic

 

0

0

0

0

00

0

0

6.Reaction to being

caught

0

0

0

0

00

0

0

7. Vocalization

0

0

0

0

00

0

0

8. Reaction to sound

stimuli

0

0

0

0

00

0

0

9. Other

0

0

0

0

00

0

0

 

 

 

Skin and hair

Skin colour

0

0

0

0

00

0

0

2. Edema

0

0

0

0

00

0

0

3. Epidermis

0

0

0

0

00

0

0

4. Coat

0

0

0

0

00

0

0

5. Skin changes

0

0

0

0

00

0

0

6. Other

0

0

0

0

00

0

0

 

 

 

 

 

Eyes and

eyelids

1. Ocular discharge

0

0

0

0

00

0

0

2. Exophthalmos

0

0

0

0

00

0

0

3. Corneal opacity

0

0

0

0

00

0

0

4. Eyelids

0

0

0

0

00

0

0

5. Other

0

0

0

0

00

0

0

 

 

Respiratory

system

1. Respiration

0

0

0

0

00

0

0

2. Discharge from the

nostrils

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

 

Digestive

system

1. Salivation

0

0

0

0

00

0

0

2. Diarrhea

0

0

0

0

00

0

0

3. Bleeding from the

anus

0

0

0

0

00

0

0

4. Other

0

0

0

0

00

0

0

 

Urinary

system

1. Urination

0

0

0

0

00

0

0

2. Hematuria

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

Reproductive system

1. Pathological

discharge from the

reproductive tract

0

0

0

0

00

0

0

 

2. Other

0

0

0

0

00

0

0

Parameter

                                      Day of observation

8

9

10

11

12

13

14

 

 

 

 

 

 

 

Locomotor

system,

behaviour,

reactions to

stimuli

1. Body posture

0

0

0

0

00

0

0

2. Gait

0

0

0

0

00

0

0

3. Locomotor activity

0

0

0

0

00

0

0

4. Involuntary

movement-clonic

0

0

0

0

00

0

0

5. Involuntary

movement-tonic

 

0

0

0

0

00

0

0

6.Reaction to being

caught

0

0

0

0

00

0

0

7. Vocalization

0

0

0

0

00

0

0

8. Reaction to sound

stimuli

0

0

0

0

00

0

0

9. Other

0

0

0

0

00

0

0

 

 

 

Skin and hair

Skin colour

0

0

0

0

00

0

0

2. Edema

0

0

0

0

00

0

0

3. Epidermis

0

0

0

0

00

0

0

4. Coat

0

0

0

0

00

0

0

5. Skin changes

0

0

0

0

00

0

0

6. Other

0

0

0

0

00

0

0

 

 

 

 

 

Eyes and

eyelids

1. Ocular discharge

0

0

0

0

00

0

0

2. Exophthalmos

0

0

0

0

00

0

0

3. Corneal opacity

0

0

0

0

00

0

0

4. Eyelids

0

0

0

0

00

0

0

5. Other

0

0

0

0

00

0

0

 

 

Respiratory

system

1. Respiration

0

0

0

0

00

0

0

2. Discharge from the

nostrils

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

 

Digestive

system

1. Salivation

0

0

0

0

00

0

0

2. Diarrhea

0

0

0

0

00

0

0

3. Bleeding from the

anus

0

0

0

0

00

0

0

4. Other

0

0

0

0

00

0

0

 

Urinary

system

1. Urination

0

0

0

0

00

0

0

2. Hematuria

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

Reproductive system

1. Pathological

discharge from the

reproductive tract

0

0

0

0

00

0

0

 

2. Other

0

0

0

0

00

0

0

Gross lesions: not found

Table NO.6:Detailed opbservations, dose 2000mg/kg b.w, FEMALE NO.1,2,3,4 and 5

Fasting: 19 hours

Animal age: 11 weeks

Parameter

                               Day 0 of observation

10 min

30 min

1h

2h

3h

4h

5h

 

 

 

 

 

 

 

Locomotor

system,

behaviour,

reactions to

stimuli

1. Body posture

0

0

0

0

00

0

0

2. Gait

0

0

0

0

00

0

0

3. Locomotor activity

0

0

0

0

00

0

0

4. Involuntary

movement-clonic

0

0

0

0

00

0

0

5. Involuntary

movement-tonic

 

0

0

0

0

00

0

0

6.Reaction to being

caught

0

0

0

0

00

0

0

7. Vocalization

0

0

0

0

00

0

0

8. Reaction to sound

stimuli

0

0

0

0

00

0

0

9. Other

0

0

0

0

00

0

0

 

 

 

Skin and hair

Skin colour

0

0

0

0

00

0

0

2. Edema

0

0

0

0

00

0

0

3. Epidermis

0

0

0

0

00

0

0

4. Coat

0

0

0

0

00

0

0

5. Skin changes

0

0

0

0

00

0

0

6. Other

0

0

0

0

00

0

0

 

 

 

 

 

Eyes and

eyelids

1. Ocular discharge

0

0

0

0

00

0

0

2. Exophthalmos

0

0

0

0

00

0

0

3. Corneal opacity

0

0

0

0

00

0

0

4. Eyelids

0

0

0

0

00

0

0

5. Other

0

0

0

0

00

0

0

 

 

Respiratory

system

1. Respiration

0

0

0

0

00

0

0

2. Discharge from the

nostrils

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

 

Digestive

system

1. Salivation

0

0

0

0

00

0

0

2. Diarrhea

0

0

0

0

00

0

0

3. Bleeding from the

anus

0

0

0

0

00

0

0

4. Other

0

0

0

0

00

0

0

 

Urinary

system

1. Urination

0

0

0

0

00

0

0

2. Hematuria

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

Reproductive system

1. Pathological

discharge from the

reproductive tract

0

0

0

0

00

0

0

 

2. Other

0

0

0

0

00

0

0

Table No.7:

Detailed observations, dose 2000mg/kg b.w, FEMALE NO.1,2,3,4 and 5

Parameter

                               Day  of observation

1

2

3

4

5

6

7

 

 

 

 

 

 

 

Locomotor

system,

behaviour,

reactions to

stimuli

1. Body posture

0

0

0

0

00

0

0

2. Gait

0

0

0

0

00

0

0

3. Locomotor activity

0

0

0

0

00

0

0

4. Involuntary

movement-clonic

0

0

0

0

00

0

0

5. Involuntary

movement-tonic

 

0

0

0

0

00

0

0

6.Reaction to being

caught

0

0

0

0

00

0

0

7. Vocalization

0

0

0

0

00

0

0

8. Reaction to sound

stimuli

0

0

0

0

00

0

0

9. Other

0

0

0

0

00

0

0

 

 

 

Skin and hair

Skin colour

0

0

0

0

00

0

0

2. Edema

0

0

0

0

00

0

0

3. Epidermis

0

0

0

0

00

0

0

4. Coat

0

0

0

0

00

0

0

5. Skin changes

0

0

0

0

00

0

0

6. Other

0

0

0

0

00

0

0

 

 

 

 

 

Eyes and

eyelids

1. Ocular discharge

0

0

0

0

00

0

0

2. Exophthalmos

0

0

0

0

00

0

0

3. Corneal opacity

0

0

0

0

00

0

0

4. Eyelids

0

0

0

0

00

0

0

5. Other

0

0

0

0

00

0

0

 

 

Respiratory

system

1. Respiration

0

0

0

0

00

0

0

2. Discharge from the

nostrils

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

 

Digestive

system

1. Salivation

0

0

0

0

00

0

0

2. Diarrhea

0

0

0

0

00

0

0

3. Bleeding from the

anus

0

0

0

0

00

0

0

4. Other

0

0

0

0

00

0

0

 

Urinary

system

1. Urination

0

0

0

0

00

0

0

2. Hematuria

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

Reproductive system

1. Pathological

discharge from the

reproductive tract

0

0

0

0

00

0

0

 

2. Other

0

0

0

0

00

0

0

Parameter

                                 Day  of observation

8

9

10

11

12

13

14

 

 

 

 

 

 

 

Locomotor

system,

behaviour,

reactions to

stimuli

1. Body posture

0

0

0

0

00

0

0

2. Gait

0

0

0

0

00

0

0

3. Locomotor activity

0

0

0

0

00

0

0

4. Involuntary

movement-clonic

0

0

0

0

00

0

0

5. Involuntary

movement-tonic

 

0

0

0

0

00

0

0

6.Reaction to being

caught

0

0

0

0

00

0

0

7. Vocalization

0

0

0

0

00

0

0

8. Reaction to sound

stimuli

0

0

0

0

00

0

0

9. Other

0

0

0

0

00

0

0

 

 

 

Skin and hair

Skin colour

0

0

0

0

00

0

0

2. Edema

0

0

0

0

00

0

0

3. Epidermis

0

0

0

0

00

0

0

4. Coat

0

0

0

0

00

0

0

5. Skin changes

0

0

0

0

00

0

0

6. Other

0

0

0

0

00

0

0

 

 

 

 

 

Eyes and

eyelids

1. Ocular discharge

0

0

0

0

00

0

0

2. Exophthalmos

0

0

0

0

00

0

0

3. Corneal opacity

0

0

0

0

00

0

0

4. Eyelids

0

0

0

0

00

0

0

5. Other

0

0

0

0

00

0

0

 

 

Respiratory

system

1. Respiration

0

0

0

0

00

0

0

2. Discharge from the

nostrils

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

 

Digestive

system

1. Salivation

0

0

0

0

00

0

0

2. Diarrhea

0

0

0

0

00

0

0

3. Bleeding from the

anus

0

0

0

0

00

0

0

4. Other

0

0

0

0

00

0

0

 

Urinary

system

1. Urination

0

0

0

0

00

0

0

2. Hematuria

0

0

0

0

00

0

0

3. Other

0

0

0

0

00

0

0

 

Reproductive system

1. Pathological

discharge from the

reproductive tract

0

0

0

0

00

0

0

 

2. Other

0

0

0

0

00

0

0

Table No.8: Body weight and date of euthanasia

Animal

                          Weight

0                                7                            14

Day of euthanasia

1

202

234

229

28.10.2014

2

199

215

227

4.11.2014

3

199

219

230

4.11.2014

4

201

213

229

4.11.2014

 5

188 

 207

213 

 4.11.2014

Interpretation of results:
GHS criteria not met
Remarks:
Not classified
Conclusions:
No signs of toxicity were observed following single administration of the test item at a dose of 2000 mg/kg bw in rats. The discriminating dose was determined to be 2000 mg/kg bw.
Executive summary:

The acute oral toxicity study based on a fixed dose method was performed according to OECD Guideline 420. The sighting study in which the test item at a dose of 300 and 2000 mg/kg bw was administered by gavage to one animal and then to the second animal did not show any signs of toxicity. On the grounds of these results, four animals were given the test item at a dose of 2000 mg/kg bw in the main test. After the administration of the test item, the animals were observed for 14 days. General and detailed clinical observations were conducted daily during the entire experiment. Body weights of the animals were determined on days 0 (directly before the administration of the test item), 7, and 14. After the 14-day observation period, the animals were euthanized and subjected to a detailed gross examination. All animals survived the experiment. After the experiment, body weights of all animals increased. Gross examinations did not reveal any pathological changes.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
Klimisch score = 1. GLP study.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Key study: The acute oral toxicity study based on a fixed dose method was performed according to OECD Guideline 420. On the grounds of the sighting study results, four animals (plus one of the sighting study) were given the test item at a dose of 2000 mg/kg bw in the main test. After the administration of the test item, the animals were observed for 14 days. Then, the animals were euthanized and subjected to a detailed gross examination. All animals survived the experiment. The body weights of all animals increased. Gross examinations did not reveal any pathological changes.

Justification for classification or non-classification

Based on available data, the substance is not classified according to CLP Regulation (EC) no. 1272/2008.