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Description of key information

Based on the data available on acute oral toxicity in rats, according to OECD 423, the LD50 of the test substance was determined as higher than 2000mg/kg bw and thus the test item is considered to be not classified.

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:
2016-02-29 to 2016-05-02
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Young adult rats, 9 weeks old in group 1 and group 2
- Weight at study initiation: 227-233 g
- Fasting period before study: The day before treatment the animals were fasted. The food but not water was withheld overnight. Animals were weighed before the application and the food was given back 3 hours after the treatment.
- Housing: 3 animals/sex/cage; cage type: Type II polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets.
- Diet: The animals received ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum. The food is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.
- Water: Animals received tap water from watering bottles ad libitum. The drinking water is periodically analysed and is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.
- Acclimation period: 13 days in first step and 14 days in second step.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 - 70 %
- Air changes: above 10 air exchanges/hour by central air-condition system.
- Photoperiod: Artificial light, from 6 am. to 6 pm.
Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Lot/batch no.: 12J110516

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: The starting dose was selected on the basis of the available information about the test item.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed individually after dosing once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h, after the treatment and once per day for 14 days thereafter. The body weight were recorded on day 0 (shortly before the treatment), on day 7 and on day 15 on all animals with a precision of 1 g, respectively.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The test item did not induce mortality following a single oral administration to female rats at a dose of 2000 mg/kg bw. All female rats survived the performed treatment until the end of the 14-day observation period.
Clinical signs:
In group 1 and 2 treated with 2000 mg/kg bw no systemic toxic symptoms were observed throughout the 14-day post-treatment period.
Body weight:
In group 1 and 2 (2000 mg/kg bw) the mean body weight and body weight gain of the animals corresponded to their species and age throughout the study.
Gross pathology:
All animals treated with 2000 mg/kg bw dose of test item survived until the scheduled necropsy on Day 15. Internal necropsy finding as pale kidneys was observed in animal No.: 1169 of group 2. This alteration could not be related to the test item toxic effect, but was regarded an individual variation. Most likely the observation is a congenital anomaly. Severe hydrometra was found in two females (No.: 1169, 1172) of the group 2. Hydrometra is physiological finding and connected to the cycle of the animals. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals.
Interpretation of results:
GHS criteria not met
Conclusions:
The method used is not intended to allow for the calculation of a precise LD50 value. However for this acute oral toxicity study with the test item in rats the determined LD50 is above 2000 mg/kg bw. Thus, the test item is ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as not classified.
Executive summary:

An acute oral toxicity study was carried out using the class method according to OECD guideline 423. The starting dose was selected on the basis of the available information about the test item. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level. Therefore, treatment with 2000 mg/kg bw was repeated on further three female rats. Again, no animal died in the second step, thus, no further testing was required. The stopping criteria of Annex 2d of OECD Guideline No. 423 (presented in Appendix 7) were met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on 15th day after the treatment. No lethality was noted following oral administration of a single dose of 2000 mg/kg bw. In the first and second step at a dose level of 2000 mg/kg bw no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal. The body weight development was normal in all animals. All organs of the animals treated with 2000 mg/kg bw dose proved to be free of treatment related gross pathological changes.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
2 000 mg/kg bw
Quality of whole database:
GLP and guideline study

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016-03-09 to 2016-05-11
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Version / remarks:
2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
1998
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Toxi-Coop Zrt. 1103 Budapest, Cserkesz u. 90
- Females nulliparous and non-pregnant: yes
- Age at study initiation: young adult rats
- Weight at study initiation: preliminary study: 219-241 g; main study males: 257-286 g; females: 248-285 g
- Housing: During acclimatisation: 3 animals/sex/cage During the study: animals were housed individually. Type II polypropylene/polycarbonate; rat type cages with a solid floor, stainless steel wire covers and self-feeding baskets.
- Diet: ssniff® SM R/M-Z+H complete diet produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany, ad libitum
- Water: tap water from watering bottles ad libitum
- Acclimation period: preliminary study: 13 days; main study males: 6 days; females 27 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 30 - 70
- Air changes (per hr): above 10 air exchanges/hour by central air-condition system.
- Photoperiod (hrs dark / hrs light): Artificial light, from 6 am. to 6 pm.
Type of coverage:
semiocclusive
Vehicle:
polyethylene glycol
Details on dermal exposure:
TEST SITE
- Area of exposure: back
- % coverage: approx. 10 %
- Type of wrap if used: plastic wrap

REMOVAL OF TEST SUBSTANCE
- Washing: with body temperature water
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount applied: 2000, 300, 50, 5 mg/kg bw
- Concentration: 400, 60, 10, 1 mg/mL
- Constant volume or concentration used: yes

VEHICLE
- Lot/batch no.: 12J110516
Duration of exposure:
24 h
Doses:
Preliminary study: 5, 50, 300, 2000 mg/kg bw
Main study: 2000 mg/kg bw
No. of animals per sex per dose:
Preliminary study: 2 females
Main study: 5 males/females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days in main study
- Frequency of observations and weighing: inspection for signs of morbidity and mortality were made twice daily; weighing on day 0, 5 and 15; animals were observed individually 1 h and 5 h after dosing, and once each day for 14 days thereafter.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ appearance
Preliminary study:
No mortality observed up to high dose. The data of main study were evaluated only.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred after the 24-hour dermal exposure to test item in Crl(WI)Br male and female rats during the study.
Clinical signs:
General symptoms: No behavioural changes or systemic toxic signs were noted during the study.

Dermal irritation symptom as erythema and other signs as dry skin, desquamation and wounds were observed on the treatment site. Well defined erythema (score 2) appeared in all males and females. It was detectable between Day 1 and Day 3 in males and between Day 1 and Day 6 in females. A slight redness (score +1) was observed between Day 2 and Day 5 in males and between Day 5 and Day 14 in females. Dry skin and desquamation were found in all animals. Dry skin occurred between Day 1 and Day 7 in males and between Day 1 and Day 8 in females. Desquamation was detected between Day 1 and Day 2 in males and between Day 1 and Day 8 in females. Wounds were recorded in all females between Day 6 and Day 14. All males were free of symptoms between Day 8 and Day 14 and three females (No.: 1194, 1192, 1193) became free of symptoms between Day 9 and Day 14.
Body weight:
Mean body weight development was within the normal range for male animals of this strain and age. A body weight loss was observed in female No.: 1195 treated with 2000 mg/kg bw test item between Day 0 and Day 7. The body weight loss was very slight (approx. 1.1 %).
Gross pathology:
All animals survived until the scheduled necropsy on Day 15.
Internal macroscopic changes were observed. A pale liver was detected in male animal No.: 1219, and pale kidneys were found in two male animals (No.: 1219, 1221). These alterations could not be related to the test item toxic effect, but were regarded an individual variation. Most likely the observations are a congenital anomaly.
Severe hydrometra was observed in two females (No.: 1195, 1196). Hydrometra is a physiological finding and connected to the cycle of the animal.
No macroscopic alterations due to the systemic toxic effects of the test item were found.
Interpretation of results:
GHS criteria not met
Conclusions:
In this acute dermal toxicity study with the test item the obtained acute dermal LD50 value was greater than 2000 mg/kg bw in Crl(WI)Br male and female rats. According to Regulation (EC) No 1272/2008 and UN GHS, the test item has not been classified into any category. On the other hand, it is to be noted that the test item caused dermal irritation response on the site of administration.
Executive summary:

An acute dermal toxicity study was performed with test item in Crl(WI)Br rats, in compliance with OECD Guideline No. 402 and OPPTS 870.1200. A limit test was carried out. A single group of male and female animals (n=5 animals/sex) was exposed to test item at 2000 mg/kg bw by dermal route. The test item was applied in undiluted form and left in contact with the skin for 24 hours, followed by a 14-day observation period.

The results of the study were summarised as follows:

No mortality occurred after the 24-hour dermal exposure to the test item in male and female rats during the study.

Neither male nor female animals treated with 2000 mg/kg bw of the test item showed behavioural changes and no systemic toxic signs were noted during the study. The test item caused dermal irritation symptoms as well defined and slight erythema in both sexes, between Day 1 and Day 5 in males and between Day 1 and Day 14 in females. Other dermal irritation symptom as dry skin, desquamation were recorded between Day 1 and Day 7 in males and between Day 1 and Day 8 in females and wounds were found in females between Day 6 and Day 14, as well. Mean body weight development was within the normal range for male animals of this strain and age. Slight body weight loss was observed in one female on first week. It was not evaluated as a toxic effect of the test item. No macroscopic alterations of organs and tissues referred to the systemic toxic effect of the test item were seen during the necropsy.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
GLP and guideline study

Additional information

Acute toxicity oral:

An acute oral toxicity study was carried out using the class method according to OECD guideline 423. The starting dose was selected on the basis of the available information about the test item. The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level. Therefore, treatment with 2000 mg/kg bw was repeated on further three female rats. Again, no animal died in the second step, thus, no further testing was required. The stopping criteria of Annex 2d of OECD Guideline No. 423 (presented in Appendix 7) were met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out on 15th day after the treatment. No lethality was noted following oral administration of a single dose of 2000 mg/kg bw. In the first and second step at a dose level of 2000 mg/kg bw no clinical symptoms were observed on the day of the treatment and during the 14-day observation period, the general state and behaviour of experimental animals were normal. The body weight development was normal in all animals. All organs of the animals treated with 2000 mg/kg bw dose proved to be free of treatment related gross pathological changes. The method used is not intended to allow for the calculation of a precise LD50 value. However for this acute oral toxicity study with the test item in rats the determined LD50 is above 2000 mg/kg bw. Thus, the test item is ranked into classes of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423 as not classified.

Acute toxicity dermal:

An acute dermal toxicity study was performed with test item in Crl(WI)Br rats, in compliance with OECD Guideline No. 402 and OPPTS 870.1200. A limit test was carried out. A single group of male and female animals (n=5 animals/sex) was exposed to test item at 2000 mg/kg bw by dermal route. The test item was applied in undiluted form and left in contact with the skin for 24 hours, followed by a 14-day observation period.

The results of the study were summarised as follows:

No mortality occurred after the 24-hour dermal exposure to the test item in male and female rats during the study.

Neither male nor female animals treated with 2000 mg/kg bw of the test item showed behavioural changes and no systemic toxic signs were noted during the study. The test item caused dermal irritation symptoms as well defined and slight erythema in both sexes, between Day 1 and Day 5 in males and between Day 1 and Day 14 in females. Other dermal irritation symptom as dry skin, desquamation were recorded between Day 1 and Day 7 in males and between Day 1 and Day 8 in females and wounds were found in females between Day 6 and Day 14, as well. Mean body weight development was within the normal range for male animals of this strain and age. Slight body weight loss was observed in one female on first week. It was not evaluated as a toxic effect of the test item. No macroscopic alterations of organs and tissues referred to the systemic toxic effect of the test item were seen during the necropsy. In this acute dermal toxicity study with the test item the obtained acute dermal LD50 value was greater than 2000 mg/kg bw in Crl(WI)Br male and female rats. According to Regulation (EC) No 1272/2008 and UN GHS, the test item has not been classified into any category. On the other hand, it is to be noted that the test item caused dermal irritation response on the site of administration.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on acute oral and dermal toxicity, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighth time in Regulation (EU) No 2016/918.