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Administrative data

Description of key information

Under the conditions of an OECD 423 compliant GLP study the oral LD50 was found to be larger than 2000 mg/kg in rats.

Under the conditions of an OECD 402 compliant GLP study the dermal LD50 was found to be greater than 2000 mg/kg bw in male and female rats.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
December 17, 2001
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
The test item was homogeneous by visual inspection.
Storage conditions: Room temperature
Batch No.: VFH-2016-08
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Sulzfeld, Germany
- Age at study initiation: approx. 10 weeks
- Weight at study initiation: mean weight test group 1: 189.3g ; mean weight test group 2: 173.3g, mean weight test group 3: 194.7g
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: single housing in Makrolon cages, type III
- Diet: VRF1(P); SDS Special Diets Services, Altrip, Germany
- Water: tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C
- Humidity (%): 30 – 70%
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: structure activity considerations
Doses:
300 and 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: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Additionally, at day of death in animals that died or were sacrificed moribund starting with study day 1. Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. A check for any dead or moribund animals was made at least once each workday.
- Necropsy of survivors performed: yes
Statistics:
Calculations were performed using Microsoft Excel 2003 and checked with a calculator.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality occurred.
Mortality:
none
Clinical signs:
Clinical signs in the first 2000 mg/kg test group revealed in all animals an impaired general state and piloerection from hour 2 until hour 3 or 4 after administration, while dyspnea was noted at hour 2 and persisted in two animals until hour 3.
In the second 2000 mg/kg test group all animals showed impaired general state and piloerection from hour 2 until hour 4 or study day 1 after administration, while dyspnea and cowering position were noticed at hour 2 and persisted in two animals until hour 4 or hour 5.
In all animals of the single 300 mg/kg test group impaired general state and piloerection were observed from hour 0 until hour 4, study day 1 or day 3 after administration, respectively. Dyspnea was noted in all animals from hour 0 or 1 until hour 2 or 3, while a cowering position was seen in these animals from hour1 until hour 2 or 3 after administration.
Body weight:
The body weights of the animals increased within the normal range throughout the study period with 3 exceptions. The body weight of one animal of the second 2000 mg/kg bw. test group and two animals of the single 300 mg/kg bw. test group increased normally during the first observation week but stagnated during the second week. This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth.
Gross pathology:
There were no adverse findings observed at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
LD50 (oral, rat) > 2000 mg/kg bw.
Executive summary:

In an acute oral toxicity study performed according to the Acute Toxic Class Method, doses of 2000 and 300 mg/kg bw of the test item (undiluted or preparations in corn oil Ph. Eur.) were administered by gavage to three test groups of three fasted Wistar rats each (300 mg/kg bw in 3 females and 2000 mg/kg bw in 6 females). The following test substance-related clinical observations were recorded, clinical signs occurred within the first 3 days after administration:

2000 mg/kg (first test group):

No mortality occurred

Impaired general state in all animals

Piloerection in all animals

Dyspnea in all animals

2000 mg/kg (second test group):

No mortality occurred

Impaired general state in all animals

Piloerection in all animals

Dyspnea in all animals

Cowering position in all animals

300 mg/kg (single test group):

No mortality occurred

Impaired general state in all animals

Piloerection in all animals

Dyspnea in all animals

Cowering position in all animals

The body weights of the animals increased within the normal range throughout the study period with 3 exceptions. The body weight of one animal of the second 2000 mg/kg bw. test group and two animals of the single 300 mg/kg bw. test group increased normally during the first observation week but stagnated during the second week. This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth. There were no macroscopic pathological findings in any animal sacrificed at the end of observation period (9 females). The LD50 was greater than 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw

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:
12 - 28.03.2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
February 24, 1987
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: ZH 472 redestilliert.
- Expiration date of the lot/batch: January 2020.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature. The stability of the test item under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
- Homogeneity: The test item was homogeneous by visual inspection.
- Analysis of the test item preparation: Because the test item was used undiluted, no samples were taken.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none. The test item was applied undiluted.

FORM AS APPLIED IN THE TEST (if different from that of starting material) : undiluted.
Species:
rat
Strain:
Wistar
Remarks:
Rat / Wistar / Crl:WI (Han) SPF
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany.
- Females (if applicable) nulliparous and non-pregnant: yes.
- Age at study initiation: Young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks).
- Weight at study initiation: Animals of comparable weight (± 20% of the mean weight).
- Housing: Single housing in Makrolon cage, type III. Bedding: H 15005-29; Ssniff, Spezialdiäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany). Cage enrichment. The animals were housed in fully air-conditioned rooms.
- Diet: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany, ad libitum.
- Water: Tap water ad libitum.
- Acclimation period: Acclimatization period of at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet.
- Identification: Individual identification by cage cards and tail marking.
- Reasons for selection of the test species: Rats were selected since this rodent species is recommended in the respective test guidelines. Wistar rats were selected since there is extensive experience available in the laboratory with this strain of rats.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3°C.
- Humidity (%): 30 – 70% relative humidity.
- Air changes (per hr): Approx. 10.
- Photoperiod (hrs dark / hrs light): 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.).

IN-LIFE DATES: From: 13.03.2018 To: 27.03.2018.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: About 40 cm² covering dorsal and dorsolateral parts of the trunk.
- % coverage: 10% of the body surface.
- Type of wrap if used: semi-occlusive dressing.The test item was covered with an air-permeable dressing (4 layers of absorbent gauze (Ph. Eur. supplied by Lohmann GmbH & Co., KG) and stretch bandage (Fixomull® Stretch (adhesive fleece) supplied by Beiersdorf AG).
- Clipping of the fur: About 24 hours before application.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, rinsing of the application site with warm water.
- Time after start of exposure: 24 hours.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 1.97 mL/kg bw.
- Concentration (if solution): undiluted.

Duration of exposure:
24 hours
Doses:
2000 mg/kg bw / undiluted.
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days.
- Body weight determination: Individual body weights shortly before application (day 0), weekly thereafter and on the last day of observation.
- Clinical observations: Clinical signs for each animal were recorded several times on the day of application and at least once during each workday thereafter.
- Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter and on the last day of observation.
- Mortality: A check for any dead or moribund animals was made at least once each workday, these records are archived by Bioassay.
- Pathology: Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations.

Assessment of skin reactions:
- The evaluation of skin reactions was performed according to Draize, J. H. "Dermal toxicity." Appraisal of the safety of chemicals in foods, drugs and cosmetics (1959): 46-59. Appraisal of the safety of chemicals in foods, drugs and cosmetics. The association of food and drug officials of the United States Austin, Texas:
- Erythema and eschar formation Grading: 0 - No erythema, 1 - Very slight erythema (barely perceptible), 2 - Well- defined erythema, 3 - Moderate to severe erythema, 4 - Severe erythema (beet redness) to eschar formation preventing grading of erythema.
- Edema formation Grading: 0 - No edema, 1 - Very slight edema (barely perceptible), 2 -Slight edema (edges of area well- defined by definite raising), 3 - Moderate edema (raised approx. 1 mm), 4- Severe edema (raised more than 1 mm and extending beyond area of exposure).
- Descriptions of any dermal findings not covered by this scale were recorded.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality occured
Mortality:
No mortality occurred.
Clinical signs:
No systemic clinical signs were observed during clinical examination.
Body weight:
All animals gained weight in a normal range throughout the study period.
Gross pathology:
No macroscopic pathologic abnormalities were noted in any animal examined at the end of the study (5 males and 5 females).
Other findings:
No local effects were observed.
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of this study the median lethal dose (LD50) of the test item after dermal application was found to be greater than 2000 mg/kg bw in male and female rats.
Executive summary:

In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the undiluted test item to the clipped application site (dorsal and dorso-lateral parts of the trunk and covered by semi-occlusive dressing during the 24-hour exposure period. The application area comprised at least 10% of the total body surface). The animals were observed for 14 days.

  • No mortality occurred.
  • Neither signs of systemic toxicity nor local skin effects were observed in the animals.
  • All animals gained weight in a normal range throughout the study period.
  • No macroscopic pathologic abnormalities were noted in any animal examined at the end of the study (5 males and 5 females).

Accordingly, the acute dermal median lethal dose (LD50) was determined to be LD50, dermal, rat > 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw

Additional information

Acute oral toxicity:

In an acute oral toxicity study performed according to the Acute Toxic Class Method (OECD TG 423) and GLP, doses of 2000 and 300 mg/kg bw of the test item (undiluted or preparations in corn oil Ph. Eur.) were administered by gavage to three test groups of three fasted Wistar rats each (300 mg/kg bw in 3 females and 2000 mg/kg bw in 6 females) [BASF, 2018]. The following test substance-related clinical observations were recorded, clinical signs occurred within the first 3 days after administration:

  • 2000 mg/kg (first test group): No mortality occurred, Impaired general state in all animals, Piloerection in all animals, Dyspnea in all animals.
  • 2000 mg/kg (second test group): No mortality occurred, Impaired general state in all animals, Piloerection in all animals, Dyspnea in all animals, Cowering position in all animals.
  • 300 mg/kg (single test group): No mortality occurred, Impaired general state in all animals, Piloerection in all animals, Dyspnea in all animals, Cowering position in all animals.

The body weights of the animals increased within the normal range throughout the study period with 3 exceptions. The body weight of one animal of the second 2000 mg/kg bw test group and two animals of the single 300 mg/kg bw test group increased normally during the first observation week but stagnated during the second week. This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth. There were no macroscopic pathological findings in any animal sacrificed at the end of observation period (9 females).

The LD50(oral,rat) was larger than 2000 mg/kg bw.

Acute dermal toxicity:

In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw of the undiluted test item to the clipped application site (dorsal and dorso-lateral parts of the trunk and covered by semi-occlusive dressing during the 24-hour exposure period [BASF, 2018]. The application area comprised at least 10% of the total body surface). The animals were observed for 14 days.

  • No mortality occurred.
  • Neither signs of systemic toxicity nor local skin effects were observed in the animals.
  • All animals gained weight in a normal range throughout the study period.
  • No macroscopic pathologic abnormalities were noted in any animal examined at the end of the study (5 males and 5 females).

Accordingly, the acute dermal median lethal dose (LD50) was determined to be LD50, dermal, rat > 2000 mg/kg bw.

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 1272/2008. As a result, the substance is not considered to be classified for acute oral or dermal toxicity under Regulation (EC) No. 1272/2008, as amended for the thirteenth time in Regulation (EC) No. 2018/1480.