[No public or meaningful name is available]

Endpoint summary

Description of key information

ORAL
LD50 > 2000 mg/kg bw, female rat, OECD 420, EU Method B.1 bis, Bradshaw (2009)
DERMAL
LD50 > 2000 mg/kg bw, male/female rat, OECD 402, Zelenák (2013)

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:

12 February 2009 to 3 March 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of Signature: 15/10/2007; Date of Inspection: 21/08/2007

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Sex: Female
- Strain: Wistar (HsdRccHan: WIST)
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 166-176 g
- Fasting period before study: overnight
- Housing: suspended solid-floor polypropylene cages furnished with wood flakes.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 °C.
- Humidity (%): 30-70%
- Air changes (per hr): at least 15 per hour.
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES: From: Day 0 To: Day 14

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: Arachis Oil BP was used because the test material did not dissolve/suspend in distilled water.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

CLASS METHOD
- Rationale for the selection of the starting dose: Using available information on the toxicity of the test material, 2000 mg/kg was chosen as the starting dose.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

5 females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: Clinical observations were made ½, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: Yes
- Other examinations performed: Morbidity & mortality checks were made twice daily.

Statistics:

The test material will be classified according to Annex 3 of the OECD Guidelines for Testing of Chemicals No. 420 "Acute Oral Toxicity - Fixed Dose Method" (adopted 17 December 2001) as shown in the attached flow charts.
Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of the toxic effects. If possible, the signs of evident toxicity were described. Evident toxicity refers to the toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probable mortality. Effects on bodyweights and abnormalities noted at necropsy were also identified.
Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test material was made.

Preliminary study:

There was no mortality or signs of systemic toxicity observed in the preliminary study (2000 mg/kg bw). In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of animals was treated at the same dose level (2000 mg/kg bw).

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Confidence limits not stated.

Mortality:

There were no deaths.
See Table 1.

Clinical signs:

other: Signs of systemic toxicity noted during the day of dosing in one animal were hunched posture, lethargy and decreased respiratory rate. There were no other signs of systemic toxicity noted. See Table 1.

Gross pathology:

Individual necropsy findings are given in Table 3.
No abnormalities were noted at necropsy.

Table 1: Individual Clinical Observations and Mortality Data

Dose Level mg/kg	Animal Number and Sex	Effects Noted After Dosing (Hours)				Effects Noted During Period After Dosing (Days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	1-0 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-0 Female	HLRd	HLRd	HLRd	HRd	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-1 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-2 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	2-3 Female	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

0 = No signs of systemic toxicity

H = Hunched posture

L = Lethargy

Rd = decreased respiratory rate

Table 2: Individual Bodyweights and Bodyweight Changes

Dose Level mg/kg	Animal Number and Sex	Bodyweight (g) at Day			Bodyweight Gain (g) During Week
		0	7	14	1	2
2000	1-0 Female	167	180	193	13	13
	2-0 Female	166	167	181	1	14
	2-1 Female	168	186	197	18	11
	2-2 Female	176	201	217	25	16
	2-3 Female	169	185	196	16	11

 

Table 3: Individual Necropsy Findings

Dose Level mg/kg	Animal Number and Sex	Time of Death	Macroscopic Observations
2000	1-0 Female	Killed Day 14	No abnormalities detected
	2-0 Female	Killed Day 14	No abnormalities detected
	2-1 Female	Killed Day 14	No abnormalities detected
	2-2 Female	Killed Day 14	No abnormalities detected
	2-3 Female	Killed Day 14	No abnormalities detected

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute oral median lethal dose (LD50) of the test material in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

Executive summary:

The study was performed to assess the acute oral toxicity of the test material in the Wistar strain rat. The method was designed to meet the requirements of the following: OECD 420 and EU Test Method B1bis.

Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a suspension in arachis oil BP, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

During the 14 day observation period there were no recorded deaths. Signs of systemic toxicity noted during the day of dosing in one animal were hunched posture, lethargy and decreased respiratory rate. There were no other signs of systemic toxicity noted. All animals showed expected gains in bodyweight. No abnormalities were noted at necropsy.

The acute oral median lethal dose (LD₅₀) of the test material in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The study was performed in accordance with standardised guidelines and under GLP conditions and was assigned a reliability score of 1 in accordance with the criteria for assessing data quality as described in Klimisch et al. (1997). The quality of the database is therefore considered to be high.

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:

30 January 2013 to 13 February 2013

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted on structural analogue in compliance with GLP and agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of relevant results.

Justification for type of information:

See Read-Across Justification in Section 13.

Reason / purpose for cross-reference:

other: Target record

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: CRL:(WI) (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Young adults
- Weight at study initiation: 219 to 248 g
- Housing: Individually in type II polypropylene/polycarbonate cages
- Diet: ad libitum
- Water: municipal tap water ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Humidity: 30 to 70 % (relative)
- Air changes: 15 to 20 air changes per hour
- Photoperiod: 12 hour light/dark cycle (light from 06:00 to 18:00)

IN-LIFE DATES:
From: 24 January 2013
To: 13 February 2013

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Back. Animals were shaved 24 hours prior to exposure
- % coverage: Approximately 10 %
- Type of wrap if used: Sterile gauze pads held in place by a patch with an adhesive hypoallergenic plaster and the entire trunk of the animal wrapped in a semi-occlusive plastic wrap.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Water (at body temperature)
- Time after start of exposure: 24 hours

TEST MATERIAL
- For solids, paste formed: Yes; the test material was moistened with water to ensure good contact with the skin

Duration of exposure:

24 hours

Doses:

2000 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: Clinical observations were performed on the day of treatment at 1 and 5 hours after application and once each day thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. Body weights were recorded on Day 0 (prior to test material administration), and on days 7 and 14.
- Necropsy of survivors performed: Yes. All animals were anaesthetised with RELEASE 300 mg/mL inj. A.U.V and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

other: No clinical signs, including local dermal signs, were observed after treatment with the test material or during the 14-day observation period.

Gross pathology:

There was no evidence of any macroscopic changes at necropsy.

Table 1: Bodyweight and Bodyweight Gain for Males

Animal No.	Bodyweight (g)			Bodyweight Gain (g)
	Day 0	Day 7	Day 14	Days 0 to 7	Days 7 to 14	Days 0 to 14
9419	237	292	333	55	41	96
9420	248	292	334	44	42	86
9421	245	287	313	42	26	68
9422	234	281	329	47	48	95
9423	234	284	334	50	50	100
Mean	239.6	287.2	328.6	47.6	41.4	89.0
Standard Deviation	6.5	4.9	9.0	5.1	9.4	12.8

 

Table 2: Bodyweight and Bodyweight Gain for Females

Animal No.	Bodyweight (g)			Bodyweight Gain (g)
	Day 0	Day 7	Day 14	Days 0 to 7	Days 7 to 14	Days 0 to 14
9424	230	252	262	22	10	32
9425	227	256	272	29	16	45
9426	243	260	273	17	13	30
9427	220	240	275	20	35	55
9428	219	232	251	13	19	32
Mean	227.8	248.0	266.6	20.2	18.6	38.8
Standard Deviation	9.7	11.7	10.1	6.0	9.8	10.8

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of this study, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw in male and female rats.

Executive summary:

The acute dermal toxicity of the test material was determined in male and female CRL:(WI) rats in accordance with the standardised guideline OECD 402 under GLP conditions.

The study was performed as a limit test. 2000 mg/kg bw was applied to the shaved back of 5 animals of each sex (equating to around 10 % of the surface area of the skin) and covered with a semi-occlusive dressing. After 24 hours, the test site was washed with water and the animals were observed for any reactions over a fourteen day observation period. Bodyweights were recorded at days 0, 7 and 14. At the end of the study, all surviving animals were submitted for necropsy.

There was no mortality throughout the study and none of the animals demonstrated any clinical signs of toxicity. Bodyweights and bodyweight gains were not affected by test material administration. All animals were found to be normal at necropsy.

Under the conditions of this study, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw in male and female rats.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

The study was performed in accordance with standardised guidelines and under GLP conditions and was assigned a reliability score of 2 in accordance with the criteria for assessing data quality as described in Klimisch et al. (1997). The quality of the database is therefore considered to be high.

Additional information

ORAL

In the key study (Bradshaw 2009), the actue oral toxicity of the test material was determined in accordance with the standardised guidelines OECD 420 and EU Test Method B1 bis, using the fixed dose method.

Following a sighting test at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a suspension in arachis oil BP, at a dose level of 2000 mg/kg bw. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy. A total of 5 female Wistar rats were exposed to the test material during this study.

During the 14 day observation period no deaths were noted. Signs of systemic toxicity noted during the day of dosing in one animal were hunched posture, lethargy and decreased respiratory rate. There were no other signs of systemic toxicity noted. All animals showed expected gains in bodyweight. No abnormalities were noted at necropsy.

Under the conditions of the test the LD50 was determined to be > 2000 mg/kg in female rats.

INHALATION

In accordance with column 2, point 8.5.2 of Annex VIII of the Regulation (EC) No. 1907/2006 (REACH), testing via the inhalatory route is appropriate if it is a likely route of exposure to humans. As the acute toxicity of the test material has been addressed by the two most likely routes of exposure (oral and dermal), testing for this endpoint is therefore omitted.

DERMAL

In the key study (Zelenák 2013) the acute dermal toxicity of a structural analogue of the registered material was determined in male and female CRL:(WI) rats in accordance with the standardised guideline OECD 402 under GLP conditions. The study was therefore assigned a reliability score of 2 in accordance with the principles for assessing data quality as defined in Klimisch et al. (1997).

The study was performed as a limit test. 2000 mg/kg bw was applied to the shaved back of 5 animals of each sex (equating to around 10 % of the surface area of the skin) and covered with a semi-occlusive dressing. After 24 hours, the test site was washed with water and the animals were observed for any reactions over a fourteen day observation period. Bodyweights were recorded at days 0, 7 and 14. At the end of the study, all surviving animals were submitted for necropsy.

There was no mortality throughout the study and none of the animals demonstrated any clinical signs of toxicity. Bodyweights and bodyweight gains were not affected by test material administration. All animals were found to be normal at necropsy.

Under the conditions of this study, the acute dermal median lethal dose (LD50) was determined to be > 2000 mg/kg bw in male and female rats.

Justification for selection of acute toxicity – oral endpoint
A single good quality study was available for evaluation.

Justification for selection of acute toxicity – dermal endpoint
A single good quality study conducted on a structural analogue of the registered material was available for evaluation.

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No. 1272/2008, the substance does not require classification with respect to acute toxicity via the oral and dermal route.