Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016-11-03 - 2017-01-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Version / remarks:
2001
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: The test item should be stored at room temperature (20 ± 5ºC). It should be protected from humidity and water.

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Cmdb: WI; outbred
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: the husbandry of laboratory animals of the Experimental Medicine Centre at the Medical University in Białystok, kept behind the breeding barrier (number in the register of units entitled to the husbandry of laboratory animals: 0043). The Health Certificate issued by the Experimental Medicine Centre at the Medical University in Białystok confirmed good health of the animals.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation / Weight at study initiation: One 9-week-old animal weighing 173.0 g (dose of 300 mg/kg b.w.), and one 11-week-old animal weighing 185.0 g (dose of 2000 mg/kg b.w.) was used in the preliminary experiment. Four 10-week-old animals whose average body weight was 193.5 g (dose of 2000 mg/kg b.w.) were used in the main experiment.
- Fasting period before study: yes, about 19 hours before the administration of the test item
- Housing: The animals were kept in plastic cages covered with wire bar lids. The dimensions of the cages were 58 x 37 x 21 cm (length x width x height). In the preliminary experiment, the animals were caged individually. In the main experiment, there were four animals in one cage. Autoclaved, and additionally UV-irradiated wood shavings were used as bedding. The environment of the animals was enriched by placing wooden blocks and nesting materials for laboratory animals in the cages.
- Diet (e.g. ad libitum): The animals were given ad libitum access to "Labofeed H Standard" standard laboratory fodder (lot numbers: 2/16) produced by Zofia Połczyńska Wytwórnia Pasz “Morawski”, Kcynia.
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 – 23 °C
- Humidity (%): 26 – 65%
- Air changes (per hr): about 13 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark (artificial fluorescent lighting)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 60 resp 400 mg/l
- Amount of vehicle (if gavage): 0.5 ml/100g bw

MAXIMUM DOSE VOLUME APPLIED: 0.5 ml/100g bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Since no data were available, the preliminary experiment started with the administration of the test item at a dose of 300 mg/kg b.w. to one animal.
Doses:
300 or 2000 mg/kg bw
No. of animals per sex per dose:
1 (300 mg/kg), 5 (2000 mg/kg)
Control animals:
no
Remarks:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The evaluation of general condition of the animals, i.e. the observation of all animals for morbidity and mortality was conducted twice a day or once a day (on days off) during the 14-day experiment.
The detailed clinical observations were performed 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.
Body weights of the animals were determined on days 0 (directly before the administration of the test item), 7, and 14 (before euthanasia).
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.

Results and discussion

Preliminary study:
Clinical signs
Following single administration of the test item at a dose of 300 mg/kg b.w. to one animal used in the preliminary experiment, no signs of toxicity were noticed. The animal survived the experiment.
Following single administration of the test item at a dose of 2000 mg/kg b.w. to the next animal used in the preliminary experiment, the following changes were observed: rounded back on the administration day, and from the 1st to the 2nd day, slight decrease of locomotor activity on the administration day, and from the 1st to the 2nd day, animal could be very easy to catch on the administration day, and dejection on the administration day. The animal survived the experiment.

Body weights of the animals
During the 14-day experiment, the body weight gain was observed in both animals.

Gross examinations
The both animals did not exhibit any pathological changes.
Effect levels
Key result
Sex:
female
Dose descriptor:
discriminating dose
Effect level:
2 000 mg/kg bw
Based on:
test mat.
Mortality:
Following single administration of the test item at a dose of 2000 mg/kg b.w. to four animals used in the main experiment, the animals survived the experiment.
Clinical signs:
Following single administration of the test item at a dose of 2000 mg/kg b.w. to four animals used in the main experiment, no signs of toxicity were noticed in two animals (no. 2, and no. 4). Rounded back, slight decrease of locomotor activity, and bristled coat was observed in two animals (no. 3, and no. 5) on the administration day. The animals survived the experiment.
Body weight:
During the 14-day experiment, the body weight gain was observed in the animals.
Gross pathology:
The animals did not exhibit any pathological changes.
Other findings:
none stated

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The study was conducted under GLP according to OECD guideline 420 on the registered substance itself. The method is to be considered scientifically reasonable with no deficiencies in documentation or any deviations. Hence, the results can be considered as reliable to assess the acute toxicity of the test substance in rats.
On the grounds of the obtained results, the test item, i.e. 3-(2-Benzothiazolylthio)-propanesulfonic acid, sodium salt (ZPS) can be classified to the following categories:
- category 5 – according to the Globally Harmonized System (GHS),
- is beyond categorization – according to the Commission Regulation (EU) No. 286/2011 of March 10, 2011 amending, for the purposes of its adaptation to technical and scientific progress, Regulation (EC) No. 1272/2008 of the European Parliament and of the Council on classification, labelling, and packaging of substances and mixtures.
Executive summary:

The acute oral toxicity study based on the fixed dose method was performed according to OECD 420 (GLP) in order to obtain information on health hazards likely to arise from a single oral exposure to the test item, i.e. 3-(2-Benzothiazolylthio)-propanesulfonic acid, sodium salt (ZPS).

Since no data were available, the preliminary experiment started with the administration of the test item at a dose of 300 mg/kg b.w. to one animal, Wistar female rats (Cmdb: WI; outbred). Because the animal survived after the administration of the test item at a dose of 300 mg/kg b.w., the second animal was treated with the test item at a dose of 2000 mg/kg b.w.

On the grounds of the preliminary experiment results, four animals used in the main experiment were given the test item at a dose of 2000 mg/kg b.w. (the animal from the preliminary experiment which had been given the dose of 2000 mg/kg b.w. was included in the main experiment).

Before the administration, the animals had been fasted for about 19 hours. The test item in the form of an aqueous solution in a volume of 0.5 mL/100 g b.w. was administered with a metal stomach tube.

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.

Following single administration of the test item at a dose of 300 mg/kg b.w. to one animal used in the preliminary experiment, no signs of toxicity were noticed. The animal survived the experiment.

Following single administration of the test item at a dose of 2000 mg/kg b.w. to the next animal used in the preliminary experiment, changes in the body posture, locomotor activity, reaction of animal for catching, and reaction to sound stimuli were observed. The animal survived the experiment.

Following single administration of the test item at a dose of 2000 mg/kg b.w. to four animals used in the main experiment, no signs of toxicity were noticed in two animals (no. 2, and no. 4). Changes in body posture, locomotor activity, and bristled coat were observed in two animals (no. 3, and no. 5). The animals survived the experiment.

During the 14-day experiment, the body weight of all the animals increased.

Gross examinations did not reveal any pathological changes in the examined animals.

 

Dose of the test item (mg/kg bw)

300

2000

Preliminary experiment

Preliminary experiment

Main experiment

Mortality

0/1

0/1

0/4

Clinical signs

no changes

rounded back, slight decrease of locomotor activity, animal could be very easy to catch, dejection

in two animals (no. 2, 4): no changes; in two animals (no. 3, 5): rounded back, slight decrease of locomotor activity, bristled coat

 

On the grounds of the obtained results, the test item, i.e. 3-(2-Benzothiazolylthio)-propanesulfonic acid, sodium salt (ZPS) can be classified to the following categories:

- category 5 – according to the Globally Harmonized System (GHS),

- is beyond categorization – according to the Commission Regulation (EU) No. 286/2011 of March 10, 2011 amending, for the purposes of its adaptation to technical and scientific progress, Regulation (EC) No. 1272/2008 of the European Parliament and of the Council on classification, labelling, and packaging of substances and mixtures.