Registration Dossier

Administrative data

Description of key information

ORAL

Acute oral LD50 was determined to be LD50 3330 mg/kg (male) and 3031 mg/kg (female) in the rat; study conducted in accordance with OECD 401; Potokar (1984a)

DERMAL

Acute dermal LD50 was determined to be >2000 mg/kg bw (male/female); rabbit; study conducted in accordance with EU Method B3; Kästner (1984)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17 April 1984 to 27 August 1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, equivalent to a valid guidelines and the study was conducted under GLP conditions. The study was performed with test material being used to support the substance on the basis of read-across.
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, WIGA GmbH, Sulzfeld, Germany
- Fasting period before study: animals were fasted for 16 hours before dose administration and up to 3 hours following dose administration
- Housing: 5 animals per cage, by dose group
- Housing: Makrolon 3 cages
- Diet: Altromin standard diet 1324, Altromin GmbH, 4937 Lage, Germany ad libitum
- Water: tap water ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: approximately 21 °C
- Humidity: approximately 51 %
- Photoperiod: 12 hours dark / 12 hours light

Route of administration:
oral: gavage
Vehicle:
other: Arachidic oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20 % (2000 mg/kg), 25.1 % (2510 mg/kg), 31.6 % (3160 mg/kg), 39.8 % (3980 mg/kg)
- Amount of vehicle (if gavage): 10 mL/kg
Doses:
2000, 2510, 3160 and 3980 mg/kg
No. of animals per sex per dose:
10 males and 10 females per dose group
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations for symptoms and mortality were performed several times on the day of dose administration, then twice daily thereafter. Body weights were recorded on the day before dosing, on the day of dosing and then on days 2, 7 and 14
- Necropsy of survivors performed: yes; an assessment of the internal organs was performed.
Statistics:
According to the Behrens - Reed- Muench method, Drug and Chemical Toxicology (1981) 4: 297-305
Sex:
male
Dose descriptor:
LD50
Effect level:
3 339 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 912 - <= 3 829
Sex:
female
Dose descriptor:
LD50
Effect level:
3 031 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 499 - <= 3 677
Mortality:
Seven 3980 mg/kg males died on day 1, a further male died on day 2. Two days after dose administration four 3160 mg/kg males and one 2510 mg/kg male died. All of the male animals dosed at 2000 mg/kg survived until the end of the study. Two days following dose administration nine 3980 mg/kg females, five 3160 mg/kg females and two 2510 mg/kg females died. One 2000 mg/kg female died seven days following dose administration. All remaining animals survived until the end of the study (see table 1 for further information).
Clinical signs:
Clinical signs that were recorded during the study included unkempt coat, reduced activity, atony, emaciation, prone position and increased respiratory frequency (see Table 3 for further information).
Body weight:
All animals gained weight during the study. There was a slight depression of body weight following dose administration (see table 2 for further information).
Gross pathology:
Pathological findings included gastroesophageal erosions, pinky red spleen and hyperaemia at the knee joints in the 3980 mg/kg dose group; pinky red spleen, hyperemia at the knee joints in the 3160 mg/kg dose group and diarrhoea pulmonary oedema and hyperemia at the knee joints in the 2510 mg/kg dose group.

Table 1: Mortality

Dose group (mg/kg)

Males

Females

3980

3160

2510

2000

3980

3160

2510

2000

Dead animals after                      1 h

0

0

0

0

0

0

0

0

1 d

7

0

0

0

0

0

0

0

2 d

8

4

1

0

9

5

2

0

7 d

8

4

1

0

9

5

2

1

14 d

8

4

1

0

9

5

2

1

Table 2: Body Weights

Dose group (mg/kg)

Males

Females

3980

3160

2510

2000

3980

3160

2510

2000

1 d

212

212

217

209

177

186

181

180

start of testing

198

197

202

196

164

168

167

168

2 d

168

195

201

212

156

162

163

172

7 d

206

254

248

253

188

205

199

197

14 d

274

312

300

302

217

230

236

216

Table 3: Clinical Signs

Males

Dose group (mg/kg)

3980

3160

2510

2000

Unkempt coat

30 min - 3 h (10)

30 min - 3 h (10)

30 min (1)

Reduced activity

1 - 3 h (10)

1 d (1)

In prone position

1 d (1)

Increased respiratory frequency

1 d (1)

Found dead

1 d (7)

2 d (4)

2 d (1)

2 d (1)

Females

Dose group (mg/kg)

3980

3160

2510

2000

Unkempt coat

30 min - 3 h (10)

30 min - 3 h (10)

Reduced activity

1-3 h (1)

3.5-6.5 h (1)

1-3 h (10)

1 d (2)

Atony, Emaciation

1 d (1)

In prone position

1 d (1)

1-3 h (1)

1 d (2)

Increased respiratory frequency

1 d (1)

1 d (1)

Found dead

1 d (1)

2 d (5)

2 d (2)

2 d (1)

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the test, the acute oral LD50 was determined to be 3339 mg/kg in males rats and 3031 mg/kg in female rats.
Executive summary:

The acute oral toxicity of the test material was determined under GLP conditions to the standardised guideline OECD 401. During the study 10 male and 10 female rats were dosed orally, by gavage, with a single administration of test material in arachidic oil. Animals were dosed at 2000, 2510, 3160 and 3980 mg/kg and were observed for a period of 14 days post administration for mortality and clinical signs; body weights were also recorded. Fourteen days following test material administration animals were sacrificed and an assessment of the internal organs was performed. Under the conditions of the study seven 3980 mg/kg males died one day following dose administration, a further male died on day 2. Two days following dose administration four 3160 mg/kg males and one 2510 mg/kg male died. All of the male animals dosed at 2000 mg/kg survived until the end of the study. Two days following dose administration nine 3980 mg/kg females, five 3160 mg/kg females and two 2510 females mg/kg died. One 2000 mg/kg female died seven days after dose administration. All remaining animals survived until the end of the study. The LD50 of the test material was subsequently determined to be 3339 mg/kg in males and 3031 mg/kg in females.

The test material is considered sufficiently similar to (z)-3-hexenyl salicylate on the basis of read-across such that the results from this study can be used to address the acute oral toxicity of (z)-3-hexenyl salicylate.

Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached (section 13) for full details.

It is proposed to read-across to another salicylate substance in order to fulfil data requirements.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Please see ''Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.

Source substance: cyclohexyl salicylate (EC 400-410-3, CAS 25485 88-5).
Target substance: (z)-3-hexenyl salicylate (EC 265-745-8, CAS 65405-77-8)

3. ANALOGUE APPROACH JUSTIFICATION
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.

(z)-3-hexenyl salicylate (the target substance) and the read-across substance cyclohexyl salicylate (source substance) have been characterised using the categories and databases present in the OECD QSAR Toolbox. From the profiling, it can be seen that the two substances share structural similarities and also ‘mechanistic action’ similarities which are both general and endpoint specific. The main difference observed is structural. The target substance ((z)-3-hexenyl salicylate) contains an acyclic hexenyl chain and the source substance (cyclohexyl salicylate) contains a cyclic hexyl group.
The output from the OECD QSAR Toolbox shows that the profiles of (z)-3-hexenyl salicylate and cyclohexyl salicylate are sufficiently similar such that any available data from the source substance can be used to address the following endpoints in the REACH registration dossier for (z)-3-hexenyl salicylate:

4. DATA MATRIX
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.
Reason / purpose:
read-across source
Sex:
male
Dose descriptor:
LD50
Effect level:
3 339 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 912 - <= 3 829
Sex:
female
Dose descriptor:
LD50
Effect level:
3 031 mg/kg bw
Based on:
test mat.
95% CL:
>= 2 499 - <= 3 677
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the test, the acute oral LD50 was determined to be 3339 mg/kg in males rats and 3031 mg/kg in female rats.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
3 031 mg/kg bw
Quality of whole database:
The key study was performed under GLP conditions and was performed in line with a standardised guideline. The study was assigned a reliability score of 2 in accordance with the criteria for assessing data quality as defined in Klimisch (1997) on the basis that the test material is a read-across substance.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Study initiated 27 November 1984 (application of test material)
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, equivalent to a valid guidelines and the study was conducted under GLP conditions. The study was performed with test material being used to support the substance on the basis of read-across.
Qualifier:
equivalent or similar to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rabbit
Strain:
other: Small Russian Chbb: HM
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Karl Thomae GmbH Biberach
- Weight at study initiation: 2286 g (males); 2156 g (females)
- Diet: Altromin standard diet 2023 ad libitum
- Water: tap water ad libitum
- Acclimation period: 4 days for females, males had been held in the laboratory for two months prior to test initiation (20.09.1984).

ENVIRONMENTAL CONDITIONS
- Temperature: 20-21 °C:
- Humidity: 45-50 %:
- Photoperiod: 12 hrs dark / 12 hrs light
Type of coverage:
occlusive
Vehicle:
other: aqueous suspension of 2 % carboxymethyl cellulose and 0.5 % cremophor
Details on dermal exposure:
TEST SITE
- Area of exposure: 150-170 cm²
- % coverage: approximately 10 %
- Type of wrap if used: double-layer gauze bandage covered with polyethylene film and then an elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the bandages were removed and resides of the applied preparation washed off
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4g/kg

VEHICLE
- Concentration (if solution): The test material was prepared as a 50 % preparation.
- Justification for choice of vehicle: 50 % preparation of the test material was prepared with an aqueous suspension of 2 % carboxymethyl cellulose and 0.5 % cremophor with equal parts of the diluting agent and test material mixed together. The test material was dosed in a vehicle as neat applications of the test material demonstrated poor adherence to the skin and restlessness in the test animals in preliminary tests.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Five
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 1, 2, 3, 4 and 6 hours after application and then twice daily (intoxicating effects); end of application and 2, 3, 6, 10 and 14 days after application (local reactions assessed according to the Draize scale); 1 hour before application and 24 hours, 7 and 14 days after application (body weights)
- Necropsy of survivors performed: yes and internal organs assessed.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
None.
Clinical signs:
In three females and one male, a slight reduction in body temperature was recorded during the initial six hours after application. Within the six hours after application, all animals exhibited slightly reduced activity. This was not observed after the removal of the bandage. Immediately after application, all animals exhibited slightly increased respiratory frequency. This was resolved by one hour after application. Twenty-four hours after the removal of the bandage, one male and four females, and after a further 24 hours, three males and all females, exhibited scratches and bite marks on the area of treated skin which they were considered to have inflicted on themselves. No general symptoms of intoxication were observed, 24, 48 and 72 hours, 6, 8, 9, 10, 13 and 14 days after application.
Body weight:
Slight weight loss occurred during application and no significant weight gain was considered to have occurred (see table 2). The weight loss was attributed to the limited mobility caused by the bandage.
Gross pathology:
The application area was variable in hairiness, but featured no special characteristics; this was likewise the case for the hypoderm and corresponding lymph nodes. The G.I. tract was on the whole well-filled, the respiratory tract and the parenchyma organs did not feature any special characteristics. The same was true for the reproductive organs. The pathological/anatomical findings for all test animals 14 days after treatment did not yield any peculiarities.
Other findings:
LOCAL REACTIONS
While the test material was being spread onto the skin, a slight erythema occurred across the entire area of treated skin, which could be assessed according to the Draize scale as being classification 1 for erythema. Oedema of the skin was not determined. Immediately after the bandage was removed and the residues of the applied preparation were washed off, a distinct erythema was determined for all animals, however no swellings of the skin were observed. Twenty-four hours after the bandage was removed, an intensification of the erythema was observed for one female. For one male, a reduction in the reactions was determined. However, for the remaining animals, the reactions remained constant in their intensity. Five days after application, the acute symptoms of inflammation had completed subsided for all of the animals, however severe desquamation of the treated skin was apparent, in particular in the back area. This desquamation only healed towards the end of testing (from the 12th test day).

Table 2: Bodyweights

     Body weight (g)   
 Animal no.  Sex  before application  24 h after application  7 d after application  14 d after application
 298  F  2200  2170  2240  2200
 300 F 2130   2080  2150  2100
 303  F  2100  2110  2150  2200
 305  F  2170  2160  2190  2120
 306  F  2180  2160  2200  2200
 197  M  2250  2180  2220  2160
 198  M  2380  2300  2300  2260
 199  M  2250  2230  2280  2160
 200  M  2250  2270  2260  2160
 202  M  2300  2330  2290  2190

Table 2: Skin reactions (assessed according to the Draize scale)

Animal no.

Sex

Immediately

Hours after application

Days after application

1

24

48

120

6 to 9

12

13

Reaction

E

O

E

O

E

O

E

O

E

O

E

O

E

O

E

O

298

F

2

0

2

0

2

0

2

0

0*

0

0*

0

0*

0

0*

0

300

F

2

0

2

0

2

0

2

0

0*

0

0*

0

0*

0

0*

0

303

F

2

0

2

0

3

0

2

0

0*

0

0*

0

0*

0

0*

0

305

F

2

0

2

0

2

0

1

0

0*

0

0*

0

0*

0

0*

0

306

F

2

0

2

0

2

0

2

0

0*

0

0*

0

0*

0

0*

0

197

M

2

0

1

0

1

0

1

0

0*

0

0*

0

0*

0

0*

0

198

M

2

0

2

0

2

0

1

0

0*

0

0*

0

0*

0

0*

0

199

M

2

0

2

0

2

0

1

0

0*

0

0*

0

0*

0

0*

0

200

M

2

0

2

0

2

0

1

0

0*

0

0*

0

0*

0

0*

0

202

M

2

0

2

1

2

0

2

0

0*

0

0*

0

0*

0

0*

0

* Distinct desquamation

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the test, the acute dermal LD50 of the test material was determined to be >2000 mg/kg bw in the rabbit.
Executive summary:

The acute dermal toxicity of the test material was determined under GLP conditions to the standardised guideline EU Method B3. During the study 5 male and 5 female rats were dosed dermally with a single administration of a 50% preparation of the test material. Animals were dosed at 2000 mg/kg and were observed for a period of 14 days post administration for mortality, body weights and clinical signs. Fourteen days following test material administration animals were sacrificed and an assessment of the internal organs was performed. Under the conditions of the study no mortality was observed. Limited clinical signs were seen and local reactions included erythema classified as 1 on the Draize scale up to five days after application however severe desquamation of the treated skin was apparent, in particular in the back area which only healed towards the end of testing. The LD50 of the test material was determined to be >2000 mg/kg bw.

The test material is considered sufficiently similar to (z)-3-hexenyl salicylate on the basis of read-across such that the results from this study can be used to address the acute dermal toxicity of (z)-3-hexenyl salicylate.

Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached (section 13) for full details.

It is proposed to read-across to another salicylate substance in order to fulfil data requirements.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Please see ''Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.

Source substance: cyclohexyl salicylate (EC 400-410-3, CAS 25485 88-5).
Target substance: (z)-3-hexenyl salicylate (EC 265-745-8, CAS 65405-77-8)

3. ANALOGUE APPROACH JUSTIFICATION
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.

(z)-3-hexenyl salicylate (the target substance) and the read-across substance cyclohexyl salicylate (source substance) have been characterised using the categories and databases present in the OECD QSAR Toolbox. From the profiling, it can be seen that the two substances share structural similarities and also ‘mechanistic action’ similarities which are both general and endpoint specific. The main difference observed is structural. The target substance ((z)-3-hexenyl salicylate) contains an acyclic hexenyl chain and the source substance (cyclohexyl salicylate) contains a cyclic hexyl group.
The output from the OECD QSAR Toolbox shows that the profiles of (z)-3-hexenyl salicylate and cyclohexyl salicylate are sufficiently similar such that any available data from the source substance can be used to address the following endpoints in the REACH registration dossier for (z)-3-hexenyl salicylate:

4. DATA MATRIX
Please see 'Justification for read-across to support the REACH registration of (z)-3-hexenyl salicylate' document attached for full details.
Reason / purpose:
read-across source
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the test, the acute dermal LD50 of the test material was determined to be >2000 mg/kg bw in the rabbit.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
The key study was performed under GLP conditions and was performed in line with a standardised guideline. The study was assigned a reliability score of 2 in accordance with the criteria for assessing data quality as defined in Klimisch (1997) on the basis that the test material is a read-across substance.

Additional information

Acute oral toxicity

The acute oral toxicity of the test material was determined under GLP conditions to the standardised guideline OECD 401. During the study 10 male and 10 female rats were dosed orally, by gavage, with a single administration of test material in arachidic oil. Animals were dosed at 2000, 2510, 3160 and 3980 mg/kg and were observed for a period of 14 days post administration for mortality and clinical signs; body weights were also recorded. Fourteen days following test material administration animals were sacrificed and an assessment of the internal organs was performed. Under the conditions of the study seven 3980 mg/kg males died one day following dose administration, a further male died on day 2. Two days following dose administration four 3160 mg/kg males and one 2510 mg/kg male died. All of the male animals dosed at 2000 mg/kg survived until the end of the study. Two days following dose administration nine 3980 mg/kg females, five 3160 mg/kg females and two 2510 females mg/kg died. One 2000 mg/kg female died seven days after dose administration. All remaining animals survived until the end of the study. The LD50 of the test material was subsequently determined to be 3339 mg/kg in males and 3031 mg/kg in females. The test material is considered sufficiently similar to (z)-3-hexenyl salicylate on the basis of read-across and so the results from this study can be used to address the acute oral toxicity of (z)-3-hexenyl salicylate.

Acute inhalation toxicity

In line with Column 2, point 8.5.2, Annex VIII of Regulation 1907/2006, an acute inhalation study does not need to be performed as the substance has a low vapour pressure and the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be unlikely to occur. The acute toxicity endpoint has been addressed by assessing the toxicity via the oral and dermal routes, which is more appropriate when considering the properties of this substance.

Acute dermal toxicity

The acute dermal toxicity of the test material was determined under GLP conditions to the standardised guideline EU Method B3. During the study 5 male and 5 female rats were dosed dermally with a single administration of a 50 % preparation of the test material. Animals were dosed at 2000 mg/kg and were observed for a period of 14 days post administration for mortality, body weights and clinical signs. Fourteen days following test material administration animals were sacrificed and an assessment of the internal organs was performed. Under the conditions of the study no mortality was observed. Limited clinical signs were seen and local reactions included erythema classified as 1 on the Draize scale up to five days after application however severe desquamation of the treated skin was apparent, in particular in the back area which only healed towards the end of testing. The LD50 of the test material was determined to be >2000 mg/kg bw. The test material is considered sufficiently similar to (z)-3-hexenyl salicylate on the basis of read-across such that the results from this study can be used to address the acute dermal toxicity of (z)-3-hexenyl salicylate.

Justification for selection of acute toxicity – oral endpoint

The key study was selected on the basis that it was performed in line with a standardised guideline and under GLP conditions. The study was performed on a suitable analogue which was deemed sufficiently similar to the registered substance and the results read-across to fulfil the data requirement. The supporting study was in agreement with the results of the key study, and thus confirms that the key study is considered to be suitable for classification and risk assessment purposes.

Justification for selection of acute toxicity – dermal endpoint

The key study was selected on the basis that it was performed in line with a standardised guideline and under GLP conditions. The study was performed on a suitable analogue which was deemed sufficiently similar to the registered substance and the results read-across to fulfil the data requirement. The supporting study was in agreement with the results of the key study, and thus confirms that the key study is considered to be suitable for classification and risk assessment purposes.

Justification for classification or non-classification

According to Regulation EC 1272/2008 (CLP), the substance does not require classification for acute toxicity;

Acute oral toxicity: The reported LD50 results are above the cut-off criteria for CLP classification (>2000 mg/kg bw)

Acute dermal toxicity: The reported LD50 results are above the cut-off criteria for CLP classificaiton (>2000 mg/kg bw)

STOT-SE: In acute oral and dermal studies any clinical effects observed were considered not to support classification for STOT-SE. Minor effects noted in the key acute oral study were mainly noted at dose levels above the CLP guidance value range for STOT-SE Cat 2 classification.

Aspiration toxicity: Classification not required as the substance is not a hydrocarbon and