Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 
LD50 (male and female rat): 1824 mg/kg bw (Val 2, non-GLP, comparable to OECD 401, BASF XXII/42, 1972).
Acute dermal toxicity:
LD50 (male and female rat): > 5000 mg/kg bw; (Val 1, GLP, OECD 402, Bioassay, 11-BF-DT112, 2012).

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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to guideline study with acceptable restrictions (no data on test substance purity, reduced observation period, limited documentation).
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
(no data on test substance purity, reduced observation period, limited documentation)
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Gassner, Germany
- Mean body weight at study initiation: males 267 g, females 187 g

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 16% (G/V)

MAXIMUM DOSE VOLUME APPLIED: 15.6 mL/kg bw

Doses:
640, 800, 1000, 1250, 1600, 2000 and 2500 µL/kg bw (corresponding to approx. 730, 912, 1136, 1423, 1824, 2280, 2859 mg/kg bw; calculation based on density of 1.14 g/cm³)
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 7 days
- Frequency of observations and weighing: Body weight was determined before the beginning of the study for dose calculation.
Observation of clinical signs was several times on the day of administration and once daily afterwards with the exception of weekends and holidays.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs
Sex:
male/female
Dose descriptor:
LD50
Effect level:
ca. 1 824 mg/kg bw
Based on:
test mat.
Mortality:
730 and 912 mg/kg bw: no deaths occured;
1136 mg/kg bw: 1/10 animals (males: 1/5 animals died 7 days after treatment; females: 0/5 animals died).
1423 mg/kg bw: 1/10 animals (males: 1/5 animals died within 48 hours after treatment; females: 0/5 animals died).
1824 mg/kg bw: 5/10 animals (males: 1/5 animals died within 48 hours after treatment, 3/5 animals were dead at the end of the observation period of 7 days; females: 1/5 animals died within 48 hours after treatment, 2/5 animals were dead at the end of the observation period of 7 days).
2280 mg/kg bw: 5/10 animals (males: 3/5 animals died within 48 hours after treatment, 4/5 animals were dead at the end of the observation period of 7 days; females: 1/5 animals died within 48 hours after treatment, 1/5 animals were dead at the end of the observation period of 7 days).
2859 mg/kg bw: 10/10 animals (males: 2/5 animals died within 24 hours after treatment, 5/5 animals died within 48 hours after treatment; females: 2/5 animals died within 24 hours after treatment, 5/5 animals died within 48 hours after treatment).

Clinical signs:
On the first study day accelerated respiration, crouched position, reddened, partly closed eyes. On the following observation days crouched position, partly abdominal position, intermittent breathing and reddened eyes. After the 5th day of observation animals were without findings.
Gross pathology:
Perished animals: 1136, 1423, 1824, 2280 mg/kg bw: kidney and liver: acute congestive hyperemia; dilatation of the heart; thymic petechiae; slight lung edema; liver: loam-yellow-grey with peripheral lobular pattern (peripheral fatty degeneration, necroses).
Sacrificed animals: no abnormalities were noted at necropsy of animals sacrificed at the end of the study.

Executive summary:

The non-GLP acute oral toxicity study was performed according to a standardized BASF test protocol. In principle this test protocol is similar to the the OECD Guideline 401: Wistar rats (5/sex/dose) were administered a single oral dose (gavage) of the test item (analytical purity: unknown) at dose levels of 640, 800, 1000, 1250, 1600, 2000 and 2500 mg/kg body weight (bw). The test item was applied as aquaeous solution at a concentration of 16% w/v and at a maximum dose volume of 15.6 mL/kg bw.

The animals were observed for a post-dosing period of 7 days. Subsequently, all surviving animals were killed and like animals that died during the study subjected to gross pathology.

Mortality occurred in the high dose group within 24 hours after application of the test item and symptoms reported were described as accelerated respiration, crouched position, reddened, partly closed eyes. On following observation days crouched position, partly abdominal position, intermittent breathing and reddened eyes were observed. From the 6th day of observation animals were without findings.

No data were available on body weight gain. Gross necropsy findings in rats that died included acute congestive hyperemia, dilatation of the heart, thymic petechiae, slight lung edema, loam-yellow-grey liver with peripheral lobular pattern. No abnormalities were noted at necropsy of animals sacrificed at the end of the study.

The acute oral toxicity study is acceptable (reliability 2), though not fully compliant with the actual guideline requirements for an oral toxicity test (OECD 401) in rats (incomplete characterisation of the test substance - purity not given in the study report, observation period shortened - 7 instead of 14 days).

Under the conditions of the study the LD50 of the test item after oral application was determind to be 1824 mg/kg bw.

 

 

Classification of the test item for acute oral toxicity as Xn, R22 according EU Directive 67/548/EC as well as Cat 4, H302, Harmful if swallowed according to the CLP Regulation 1272/2008 is warranted.  

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 824 mg/kg bw

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:
From October 18, 2011 to November 15, 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
adopted February 24, 1987
Deviations:
no
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008 of 30 May 2008, PartB: Methods for the determination of toxicity and other health effects: Acute Toxicity (Dermal); Official Journal of the European Union, No. L 142
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
adopted August 1998
Deviations:
no
Qualifier:
according to
Guideline:
other: Japan MAFF Testing Guideline of 12 Nosan No. 8147
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
30.03.2011
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Test species / strain / quality: Rat / Wistar / Crl:WI (Han) SPF
- Source: Charles River Wiga GmbH, Sulzfeld, Germany
- Age at study initiation: Young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks); female animals were nulliparous and non-pregnant
- Mean body weight (± SD) at study initiation: Males: 236.4 ± 8.85; Females : 202.8 ± 3.83
- Housing: single, Makrolon cage, 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 ± 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Clipping of the fur: about 24 hours before administration
- Area of exposure: dorsal and dorsolateral parts of the trunk; about 40 cm²
- % coverage: at least 10% of the body surface
- Type of wrap if used: 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).

REMOVAL OF TEST SUBSTANCE
- Washing: 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): 4.33 mL/kg bw
- Concentration: undiluted
- Constant volume or concentration used: yes
Duration of exposure:
24 hours
Doses:
5000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
• Body weight determination: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.
• Clinical observations: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals.
• Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), several times until the last day of observation.
• Mortality: A check for any dead or moribund animals was made at least once each workday.
- Necropsy of survivors performed: yes
•Assessment of skin reactions
The evaluation of skin reactions was performed according to Draize, J.H. (1959): Appraisal of the safety of chemicals in foods, drugs and cosmetics. The association of food and drug officials of the United States Austin, Texas
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 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:
Males: The mean body weight increased within the normal range throughout the study period.
Females: The mean body weight did not adequately increase during the first post-exposure observation week, probably due to the bandage procedure, but increased during the second week within the normal range.
Gross pathology:
No macroscopic pathologic abnormalities were noted in the animals (5 males and 5 females) examined on the last day of observation.
Other findings:
Local effects:
Males: In four male animals well-defined erythema (grade 2) was noted on study day 1 and 2, decreased to very slight erythema (grade 1) on study day 5 and persisted in these animals up to study day 9, 12 or 14.
In the fifth animal only very slight erythema (grade 1) was noted from study day 1 until study day 12.
Additionally, incrustations were observed in two animals from study day 8 until study day 14.
Orange discoloration of the fur was observed in two other males from study day 5 until study day 14.
Females: In three female animals well-defined erythema (grade 2) was observed on study day 1 and persisted in two of these animals until study day 9 or 13. Well-defined erythema decreased in these two animals to very slight (grade 1) and was seen until study day 14. In the third animal well-defined erythema increased to moderate erythema (grade 3) on study day 2, persisted until study day 9 and decreased to well-defined erythema again from day 12 to day 14.
The two other animals showed very slight erythema on study day 1 and 2, which increased to well-defined erythema from study day 5 until day 9. In two females very slight edema (grade 1) was noted on study day 1 or 2 only. In a further animal slight edema (grade 2) was noted on study day 2, decreased so very
slight (grade 1) on study day 5 and 6 but increased to slight again on study day 7 up to study day 9. On study day 12 very slight edema was noted again in this animal. Furthermore incrustations and scaling were noted in the three females which showed edema from study day 5 until study day 14. In two of these animals orange discoloration of the fur was noted from study day 5 until study day 14 also.

Table 1: Mean body weight

Mean

SD

Day 0

 

Males

236.4

8.85

Females

202.8

3.83

Day 7

 

Males

262.6

13.16

Females

206.0

3.32

Day 14

 

Males

291.8

17.50

Females

215.8

9.28

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
Value:
5 000 mg/kg bw

Additional information

There are valid in vivo data available for the assessment of the acute oral and dermal toxicity of the test item.

Oral

In a non-GLP acute oral toxicity study which was performed comparable to OECD 401, Wistar rats (5/sex/dose) were administered a single oral dose (gavage) of the test item at dose levels of 640, 800, 1000, 1250, 1600, 2000 and 2500 mg/kg body weight (BASF SE, 1972). The study is acceptable for the assessment of acute oral toxicity (reliability 2), though not fully conforming to current requirements, e.g. no GLP, incomplete characterisation of the test substance, observation period shortened (7 instead of 14 days).

The test item was applied as aqueous solution at a concentration of 16% w/v and at a maximum dose volume of 15.6 mL/kg body weight.

The animals were observed for a post-dosing period of 7 days. Subsequently, all surviving animals were sacrificed and subjected to gross pathology, as were any animals that died. Based on the observed mortality a LD50 of approximately 1824 mg/kg body weight was derived. Gross necropsy findings included macroscopical liver changes.

A supporting acute oral toxicity screening assay with rats was performed according to OECD test guideline 423 (BASF SE, 2001). With the limit dose of 2000mg/kg bw no mortality was observed in 3 male and 3 female rats.

Dermal

In an acute dermal toxicity study (Limit test, GLP) conducted according to OECD Guideline 402, a single dermal dose of 5000 mg/kg body weight of the undiluted test item was applied for 24 h to the clipped skin (dorsal and dorso-lateral parts of the trunk) (Bioassay, 2012). The application area comprised at least 10% of the total body surface area. At the end of the exposure period the application site was rinsed with water and the animals were observed for 14 days.

No mortality and no signs of systemic toxicity occurred. Very slight to moderate erythema (grade 1 to 3), very slight to slight edema (grade 1 to 2), incrustations, scaling and orange discoloured fur were observed. No macroscopic pathologic abnormalities were noted in the animals examined at the end of the study.

The acute dermal toxicity study is valid to assess acute dermal toxicity (reliability 1), and does fully satisfy the guideline requirements for a dermal toxicity test (OECD 402) in rats.

Accordingly, the acute dermal median lethal dose (LD50) in rats was determined to be > 5000 mg/kg body weight.

Inhalation

In an acute Inhalation Hazard Test 3 rats per sex were exposed to saturated vapours of the test substance at 20 and 50°C (BASF SE, 1972). No analytical determination of the test atmosphere was performed. This assay is indicating, that saturated vapours of the test substance do not result in mortality or severe signs of toxicity, even at elevated temperatures.

According to column 2 of REACH Annex VIII and with respect to the acute toxicity, at least one further route of exposure (dermal, inhalation) must be provided in addition to the oral route. The further route has to be selected according to the possible route of human exposure during manufacture, handling and use of the test article.

For this substance the dermal route of exposure was identified as the most relevant for human worker exposure during manufacture, handling and use. Due to the very low vapour pressure of the test article (3.7E-03 Pa at 25°C) a significant human exposure to vapour can not be anticipated at room temperature.

Summarized an acute inhalation toxicity study does not need to be performed due to (1) the physicochemical characteristic mentioned above (low vapor pressure), (2) the moderate or low general toxic potential observed on the oral (LD50 acute oral 1824 mg/kg bw) and dermal route of exposure (LD50 acute dermal > 5000 mg/kg bw), and (3) no signs of toxicity observed in a preliminary inhalation risk assay with saturated vapors of the test substance.

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available experimental test data for oral and dermal toxicity are reliable and suitable for the purpose of classification under Directive 67/548/EEC. As a result the substance is considered to be classified for acute oral toxicity (Xn, R22) and not warranted to be classified for dermal toxicity under Directive 67/548/EEC.

 

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data for oral and dermal toxicity are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. As a result the substance is considered to be classified for acute oral toxicity (Cat 4, H302, Harmful if swallowed) and not warranted to be classified for dermal toxicity under Regulation (EC) No.1272/2008.