Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: LD50 >3,000mg/kg in rat for both males and females
Acute inhalation waived
Acute dermal toxicity: LD50>2000mg/kg in rat

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:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP study in accordance with EU Method B.1. To address toxicological endpoints as part of the REACH registration of Benzyl Salicylate (Target Substance) it is proposed to read-across to Cyclohexyl Salicylate (Source Substance). The use of read-across works within the spirit of REACH and the stated aim of the legislation to reduce animal testing where possible. The Target Substance and Source Substance have been characterised using the categories and databases present in the OECD [Q]SAR 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. Therefore read across is justified.
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
- Age at study initiation: not reported
- Weight at study initiation: average body weight of males was 209 to 217 g, average body weight of females was 164 to 168 g
- Fasting period before study: yes, 16 hours prior to and 3 hours after exposure
- Housing: 5 animals per Makrolon 3 cage, softwood bedding
- Diet (e.g. ad libitum): Altromin rodent diet 1324 ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 51
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours darkness
Route of administration:
oral: gavage
Vehicle:
peanut oil
Remarks:
DAB 8
Details on oral exposure:
VEHICLE
- Concentration in vehicle: depending on dose level 20 to 39.8% (g/v)
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: not reported
- Lot/batch no. (if required): 30809, Caesar and Loretz, Hilden

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg
Doses:
2000, 2510, 3160 and 3980 mg/kg
No. of animals per sex per dose:
10 males/10 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observations for clinical signs and mortality several times on day of application, then 2 times per day for observation period; body weight was recorded one day before application, on day of application, and 2, 7 and 14 days after application
- Necropsy of survivors performed: yes
Statistics:
According to Behrens and Reed-Muench (1981), Drug and Chemical Toxicology 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:
see attached table on mortality
Clinical signs:
Scrubby posture, lowered activity, increased breathing frequency, face-down position, atony
Body weight:
No significant findings reported
Gross pathology:
2510 mg/kg dose group: diarrhoe, lung oedema, hyperemia in the area of the knee joints
3160 mg/kg dose group: reddish spleen, hyperemia in the area of the knee joints
3980 mg/kg dose group: suspected erosion of the forestomach, reddish spleen, hyperemia in the area of the knee joints (pineal gland of the tibiae, femora and patella)

Table: Mortality of animals

Cumulative number of dead male animals (n= 10 per dose group)

Time point of observation

3980 mg/kg

3160 mg/kg

2510 mg/kg

2000 mg/kg

1 hour

0

0

0

0

1 day

7

0

0

0

2 days

8

4

1

0

7 days

8

4

1

0

14 days

8

4

1

0

Cumulative number of dead female animals (n= 10 per dose group)

Time point of observation

3980 mg/kg

3160 mg/kg

2510 mg/kg

2000 mg/kg

1 hour

0

0

0

0

1 day

0

0

0

0

2 days

9

5

2

0

7 days

9

5

2

1

14 days

9

5

2

1

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LD50 value of cyclohexyl salicylate was greater than 2000 mg/kg in an acute oral toxicity study with male and female Sprague-Dawley rats.
Executive summary:

The oral acute toxicity of the substance cyclohexyl salicylate to young adult male and female Sprague-Dawley rats was studied under GLP in accordance with EU Method B.1. The substance was dissolved in peanut oil DAB 8 and administered to rats after a fasting period of 16 hours at a volume of 10 mL/kg once by oral gavage at doses of 2000, 2510, 3160 and 3980 mg/kg. Ten male and ten female animals per dose group were then observed for a period of 14 days. The mortality rate was found to be dose-dependent: 8 males and 9 females in the group receiving 3980 mg/kg were found dead on day 2 after dosing, whereas 4 males and 5 females in the group receiving 3160 mg/kg were found dead on day 2 after dosing. One male and two females receiving 2510 mg/kg and no male and one female receiving 2000 mg/kg were found dead during the observation period. Clinical and pathological findings in animals found dead included scrubby posture, lowered activity, increased breathing frequency, face-down position, atony, diarrhoe, lung oedema, hyperemia in the area of the knee joints (pineal gland of the tibiae, femora and patella), suspected erosion of the forestomach, reddish spleen. The calculated LD50 value was 3339 mg/kg in males and 3031 mg/kg in females.

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Limited methodology reported, study pre-dates GLP.
Principles of method if other than guideline:
Test material fed to rats at 3 concentrations and observed for 7 d.
GLP compliance:
no
Remarks:
study predates GLP
Limit test:
yes
Species:
rat
Strain:
not specified
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: No data
- Age at study initiation: No data
- Weight at study initiation: 100 g to 200 g
- Fasting period before study: 18 hrs
- Housing: No data
- Diet (e.g. ad libitum): No data
- Water (e.g. ad libitum): No data
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data
- Humidity (%): No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data
Route of administration:
oral: gavage
Vehicle:
not specified
Doses:
1250 mg/kg bw, 2500 mg/kg bw, 5000 mg/kg bw
No. of animals per sex per dose:
6 males, 0 females, per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 7 days
- Frequency of observations: Daily
- Necropsy of survivors performed: No
- Other examinations performed: Clinical signs
Statistics:
Thompson moving average
Sex:
male
Dose descriptor:
LD50
Effect level:
2 227 mg/kg bw
Based on:
test mat.
95% CL:
> 1 925 - < 2 580
Mortality:
1250 mg/kg bw 0/6; 2500 mg/kg bw 4/6; 5000 mg/kg bw 6/6
Clinical signs:
Depression
Body weight:
No data
Gross pathology:
No data
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
3 000 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

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP study in accordance with EU Method B.3. To address toxicological endpoints as part of the REACH registration of Benzyl Salicylate (Target Substance) it is proposed to read-across to Cyclohexyl Salicylate (Source Substance). The use of read-across works within the spirit of REACH and the stated aim of the legislation to reduce animal testing where possible. The Target Substance and Source Substance have been characterised using the categories and databases present in the OECD [Q]SAR 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. Therefore read across is justified.
Qualifier:
according 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: "Kleinrusse" Chbb: HM
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Karl Thomae GmbH, Biberach
- Age at study initiation: young adult
- Weight at study initiation: 2286 g (males), 2156 g (females)
- Housing: individually in rabbit batteries
- Diet (e.g. ad libitum): Altromin rabbit food 2023 ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: female animals were acclimatised for four days, male animals were maintained at the laboratory for about 2 months

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 21
- Humidity (%): 45 to 50
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light
Type of coverage:
occlusive
Vehicle:
other: aqueous suspension with 2% carboxymethyl cellulose and 0.5% Cremophor
Details on dermal exposure:
TEST SITE
- Area of exposure: back and on the sides
- % coverage: about 10% of skin surface area
- Type of wrap if used: application area was covered by gauze and polyethylene foil that was fixed with Leukosilk strips; the whole area was then wrapped with an elastic bandange (Acrylastik Kompressionsbinde) with acrylate adhesive

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed-off with water
- Time after start of exposure: after removal of cover following 24 hours after application

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): between 8.4 and 9.5 g of the preparation containing the test substance at a level of 50% was applied to the skin of the animals by brushing the preparation onto the skin
VEHICLE
- Amount(s) applied (volume or weight with unit): the substance was applied as an aqueous solution containing 2% carboxymethylcellulose and 0.5% Cremophor
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 females, 5 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: Directly and 1, 2, 3, 4 and 6 hours after application, then twice daily
- Frequency of weighing: animals were weighed one hour before application and 1, 7 and 14 days after application
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: skin reaction
Statistics:
not applicable
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Slightly reduced activity during the first six hours after application; slightly reduced body temperature in three females and one male during first six hours after application
Body weight:
Slight body weight reduction after application
Gross pathology:
No findings were reported.
Other findings:
Skin reactions: Slight reddening of skin after application (erythema score of 1 according to Draize); clear signs of erythema in all animals after removal of coverage; signs of erythema were reversible within 5 days, but scaling was observed until test day 12
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The LD50 value in an acute dermal toxicity study with rabbits was LD50 > 2000 mg/kg bw and the substance is not classified for acute dermal toxicity according to the CLP regulation.
Executive summary:

The acute toxicity of the substance Cyclohexyl salicylate was studied under GLP in accordance with the EU Method B.3. Five female and five male young adult rabbits of the strain "Kleinrusse" were used in the test. Females and males had body weights of 2156 g and 2286 g, respectively. The substance was applied to the shave skin of the back and on the sides (application area about 10% of the total skin surface area) in form of a preparation, which was an aqueous suspension containing 50% of the test substance, 2% carboxymethylcellulose and 0.5% Cremophor. The preparation was brushed onto the skin and animals were then exposed to the material under occlusion for 24 hours. After this exposure period, the coverage was removed and remaining test substance was washed-off with water. None of the animals died after application or during the 14-day observation period. A slight body weight reduction was observed after application. Animals exhibited signs of skin irritation (erythema persisted for up to 5 days, scaling occurred and persisted until test day 12). No clinical signs were observed after patch removal. No pathological findings were made at necropsy following the 14-day observation period. The acute dermal toxicity study resulted in a LD50 value > 2000 mg/kg bw.

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Study is well conducted but in its design not sufficient to cover the endpoint.
Principles of method if other than guideline:
3 groups of 3 rabbits exposed to doses of test material for 24 hrs and observed for 7 d. Clinical chemistry examined in survivors at 5 d.
GLP compliance:
no
Remarks:
Study predates GLP
Test type:
standard acute method
Limit test:
yes
Species:
rabbit
Strain:
other: Albino
Sex:
not specified
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
- Age at study initiation:
- Weight at study initiation: Approximately 1,500 to 2,000 g.
- Fasting period before study:
- Housing:
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period:

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
- Humidity (%):
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):
Type of coverage:
other: Bandages used so either occlusive or semiocclusive.
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure:
- % coverage:
- Type of wrap if used:

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Constant volume or concentration used: yes/no
- For solids, paste formed: yes/no

VEHICLE
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:
Duration of exposure:
24 hrs
Doses:
5,000 mg/kg bw, 10,000 mg/kg bw and 20,000 mg/kg bw.
No. of animals per sex per dose:
3 animals of unspecified sex per dose.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 7 d
- Frequency of observations: Daily
- Necropsy of survivors performed
- Other examinations performed: clinical chemistry of surviving animals examined on day 5.
Statistics:
LD₅₀ calculated using the method of Weil (reference: Biometrics Vol. 8 issue 3, page 249, 1952-09).
Sex:
not specified
Dose descriptor:
LD50
Effect level:
14 150 mg/kg bw
Based on:
test mat.
95% CL:
> 4 560 - < 43 860
Mortality:
5,000 mg/kg bw - 0/3; 10,000 mg/kg bw 1/3; 20,000 mg/kg bw 2/3.
Clinical signs:
10,000 mg/kg bw and 20,000 mg/kg bw were depressed and showed slow respiration on removal of bandages. Among animals that died, intoxication persisted and gradually deepened to severe depression, loss of righting reflex, coma and death. Survivors appeared essentially normal by day 5.

5,000 mg/kg bw appeared normal and alert during the 7 d observation period.
Body weight:
No data
Gross pathology:
No significant gross pathology was noted in any of the animals, whether they died during the study or survived to termination.
Other findings:
HAEMATOLOGY AND CLINICAL CHEMISTRY
- For full results please refer to the attached supporting information.
- Apparent low haemoglobin value was noted for the survivor treated at 20,000 mg/kg bw.
- All other values were within normal limits.
Conclusions:
The LD₅₀ of the test material benzyl salicylate was determined to be 14,150 mg/kg bw.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Acute oral toxicity:

The oral acute toxicity of the Source Substance, cyclohexyl salicylate, to young adult male and female Sprague-Dawley rats was studied under GLP in accordance with EU Method B.1. The LD50 value of cyclohexyl salicylate was greater than 2000 mg/kg in an acute oral toxicity study with male and female Sprague-Dawley rats.

Limited data to support lack of acute toxicity via the oral route is available on Benzyl salicylate (Target Substance) and the available results support the read across proposal that the substance is not classified with regards to acute oral toxicity in accordance with regulation (EC) No 1272/2008.

Acute inhalation toxicity:

Testing by the inhalation route in accordance with column 2 of Annex VIII of 1907/2006/EC, is considered to be inappropriate as exposure of humans via inhalation is unlikely taking into account the vapour pressure of the substance (0.0104 Pa at 25°C) and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size.

Acute dermal toxicity:

The acute toxicity of the Source Substance, Cyclohexyl salicylate was studied under GLP in accordance with the EU Method B.3. Five female and five male young adult rabbits of the strain "Kleinrusse" were used in the test.

The LD50 value in an acute dermal toxicity study with rabbits was LD50 > 2000 mg/kg bw and the substance is not classified for acute dermal toxicity according to the CLP regulation.

Limited data to support lack of acute toxicity via the dermal route is available on Benzyl salicylate (Target substance) and the available results support the read across proposal that the substance is not classified with regards to acute dermal toxicity in accordance with regulation (EC) No 1272/2008.

To address toxicological endpoints as part of the REACH registration of Benzyl Salicylate (Target Substance) it is proposed to read-across to Cyclohexyl Salicylate (Source Substance).

The use of read-across works within the spirit of REACH and the stated aim of the legislation to reduce animal testing where possible.

TheTarget Substance and Source Substance have been characterised using the categories and databases present in the OECD [Q]SAR 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.

For classification and labelling purposes this indicates that the structural analogue is relatively harmless and would not, in itself, be classified in accordance with Regulation (EC) No 1272/2008 (CLP). The registration substance is considered to have similar properties and is therefore not classified in accordance with CLP.


Justification for selection of acute toxicity – oral endpoint
The oral acute toxicity of (the Source Substance), cyclohexyl salicylate, to young adult male and female Sprague-Dawley rats was studied under GLP in accordance with EU Method B.1. The LD50 value of cyclohexyl salicylate was greater than 2000 mg/kg in an acute oral toxicity study with male and female Sprague-Dawley rats.
Klimisch4-rated data to support lack of acute toxicity via the oral route is available on the Target Substance and the available results support the read across proposal that the substance is not classified with regards to acute oral toxicity in accordance with regulation (EC) No 1272/2008.

Justification for selection of acute toxicity – dermal endpoint
The LD50 value of the Source Substance, cyclohexyl salicylate, in an acute dermal toxicity study with rabbits was LD50 > 2000 mg/kg bw and the substance is not classified for acute dermal toxicity according to the CLP regulation.
Limited data to support lack of acute toxicity via the dermal route is available on the Target substance and the available results support the read across proposal that the substance is not classified with regards to acute dermal toxicity in accordance with regulation (EC) No 1272/2008.

Justification for classification or non-classification

No deaths occurred in all animals administered a dose of 2000mg/kg of the structural analogue, Cyclohexyl Salicylate via the oral and dermal route of exposure.

To address toxicological endpoints as part of the REACH registration of Benzyl Salicylate (Target Substance) it is proposed to read-across to Cyclohexyl Salicylate (Source Substance).

The use of read-across works within the spirit of REACH and the stated aim of the legislation to reduce animal testing where possible.

The Target Substance and Source Substance have been characterised using the categories and databases present in the OECD [Q]SAR 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.

For classification and labelling purposes this indicates that the structural analogue is relatively harmless and would not, in itself, be classified in accordance with Regulation (EC) No 1272/2008 (CLP). The registration substance is considered to have similar properties and is therefore not classified in accordance with CLP.