Reaction mass of (3R,3aS,6R,7R,8aS)-3,6,8,8-tetramethyloctahydro-1H-3a,7-methanoazulen-6-yl rel-acetate and rel-(1aR,4aR,8aR)-2,4a,8,8-tetramethyl-1,1a,4,4a,5,6,7,8-octahydrocyclopropa[d]naphthalene and rel-(3R,3aS,7S,8aS)-3,6,8,8-tetramethyl-2,3,4,7,8,8a-hexahydro-1H-3a,7-methanoazulene

Endpoint summary

Description of key information

Acute oral is >5000 mg/kg bw based on read across from Cedryl Acetate 'mono' and Longifolene Coeur both tested in a study similar to OECD TG 401.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1964

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

This information is used for read across to Cedryl Acetate EOA

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Version / remarks:

1987

Deviations:

yes

Remarks:

Not all animals were of the same sex, no data on whether animals that died during the test were necropsied.

GLP compliance:

not specified

Remarks:

The study is a publication

Test type:

other: not reported

Limit test:

no

Species:

rat

Strain:

Osborne-Mendel

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: young adults
- Fasting period before study: 18 hrs
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

No data

Doses:

No data

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: two weeks
- Frequency of observations and weighing: close observation, no frequency reported
- Necropsy of survivors performed: no

Statistics:

The LD50's were computed by the method of Litchfield & Wilcoxon (1949), slope function and confidence limits were also reported.

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

44 750 mg/kg bw

Based on:

test mat.

95% CL:

>= 33 650 - < 59 520

Mortality:

One animal died after 4 hours and one after 11 days.

Clinical signs:

Depression, scrawny appearance, rough, wet fur.

Body weight:

No data

Gross pathology:

No data

Interpretation of results:

other: Not acute harmful

Remarks:

according to Regulation (EC) No. 1272/2008 and its amendments.

Conclusions:

In this acute oral toxicity test an LD50 of 44,750 mg/kg bw was determined. Based on these results, Cedryl acetate does not need to be classified for acute oral toxicity in accordance with the criteria outlined in Annex I of the CLP Regulation (1272/2008/EC).

Executive summary:

In this study, 10 rats (5 males and 5 females) were treated with Cedryl acetate via oral gavage. Observation period was 14 days after treatment and toxic signs and mortality were reported. An LD50 is calculated based on the results. Two animals died during the study, one after 4 hours and one after 11 days. The rats showed depression, scrawny appearance, rough and wet fur during the observation period. The acute oral LD50 for the substance in male and female rats was calculated to be 44,750 mg/kg bw, with a 95% Confidence Interval of 33,650-59,520 mg/kg bw. Based on the results of this study, Cedryl acetate does not need to be classified for acute oral toxicity in accordance with the criteria outlined in Annex I of the CLP Regulation (1272/2008/EC).

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2018

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Read-across information.

Justification for type of information:

The read across justification is presented in the Endpoint summary Acute toxicity. The accompanying files are also attached there.

Reason / purpose for cross-reference:

read-across source

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Remarks on result:

other: read-across from Longifolene Coeur

Interpretation of results:

other: Not acute harmful

Remarks:

according to Regulation (EC) No. 1272/2008 and its amendments.

Conclusions:

The LD50 value for the substance is >5000 mg/kg bw, based on the results of the source substance.

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 December 1983 - 02 January 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

This information is used for read across to Cedryl Acetate EOA

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Labs, Wilmington, Massachusetts
- Age at study initiation: no data
- Weight at study initiation: 180 - 200 g
- Fasting period before study: 18 hours
- Housing: Housed in groups, according to sex, or individually in stainless steel 4" wire mesh cages.
- Diet: Free access to Wayne Lab Blox.
- Water: Free access to fresh tap water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 22 ± 3
- Humidity (%): 30 to 70
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: Dose levels were based on a dose range finding study.

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: The rats were observed immediately and at 1, 4 and 24 hours after dosing and twice daily for 14 days for pharmacotoxic, CNS effects and mortality. On day 14 body weights were recorded.
- Necropsy of survivors performed: The rats were sacrificed by CO2 inhalation and a gross necropsy was performed.

Statistics:

Not performed.

Preliminary study:

In a dose range finding study, four fasted animals, two per sex, were administered to the test substance at 500, 1600 and 5000 mg/kg bw, orally, by gavage. Signs observed included diarrhea, decreased activity and body tone, hypersensitivity and poor grooming. None of the animals died.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred.

Clinical signs:

Clinical signs observed included decreased activity, abnormal gait, poor grooming, diarrhea, abnormal stance and piloerection.

Body weight:

No data.

Gross pathology:

Terminal necropsy revealed no test substance related lesions in none of the animals.

Interpretation of results:

other: Not acute harmful.

Remarks:

according to Regulation (EC) No. 1272/2008 and its amendments.

Conclusions:

The acute oral toxicity test showed an LD50 of > 5000 mg/kg bw.

Executive summary:

In this study performed equivalent to OECD TG 401 guideline and GLP principles, 10 rats (5 males and 5 females) were administered to the substance at a dose level of 5000 mg/kg bw. No mortality was seen. Clinical signs observed included decreased activity, abnormal gait, poor grooming, diarrhea, abnormal stance and piloerection. Terminal necropsy revealed no test substance related lesions in none of the animals.

Based on these results, the substance is not acute harmful and the acute oral LD50 for the substance in male and female rats was determined to be >5000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The acute oral toxicity result is of sufficient quality and adequate for this dossier.

Additional information

Acute oral toxicity is assessed based on read-across from Cedryl acetate ('mono') and Longifolene Coeur to Cedryl Acetate EOA. The executive summaries of the source information is presented below followed by the read-across rationale.

Cedryl Acetate 'mono':

In a study equivalent to OECD TG 401, 10 rats (5 males and 5 females) were treated with Cedryl acetate via oral gavage. Observation period was 14 days after treatment and toxic signs and mortality were reported. An LD50 is calculated based on the results. Two animals died during the study, one after 4 hours and one after 11 days. The rats showed depression, scrawny appearance, rough and wet fur during the observation period. The acute oral LD50 for the substance in male and female rats was calculated to be 44,750 mg/kg bw, with a 95% Confidence Interval of 33,650-59,520 mg/kg bw.

Longifolene Coeur:

In a study equivalent to OECD TG 401 and GLP principles, 10 rats (5 males and 5 females) were administered to the substance at a dose level of 5000 mg/kg bw. No mortality was seen. Clinical signs observed included decreased activity, abnormal gait, poor grooming, diarrhea, abnormal stance and piloerection. Terminal necropsy revealed no test substance related lesions in none of the animals. Based on these results, the substance is not acute harmful and the acute oral LD50 for the substance in male and female rats was determined to be >5000 mg/kg bw.

The acute oral toxicity of Cedryl Acetate EOA based on read across from data available for Cedryl Acetate ‘mono’ (CAS #77-54-3) and Longifolene Coeur (CAS #475-20-7)

 

Introduction and hypothesis for the analogue approach

Cedryl acetate EOA consists of one major, two minor constituents and a number of impurities, all containing a hydrocarbon fused-ring system. Half of the constituents have an acetate attached to this system the other half does not have this ester group. The constituents are sub-grouped into two main types: Cedryl acetate-type and Longifolene Coeur (hydrocarbon-fused-ring-type).

For Cedryl Acetate EOA there are no acute oral toxicity data available.In accordance with Article 13 of REACH, lacking information should be generated whenever possible by means other than vertebrate animal tests, i.e. applying alternative methods such as in vitro tests, QSARs, grouping and read-across.For assessing the acute oral toxicity of Cedryl Acetate EOA, the analogue approach is selected because for one of its constituents, Cedryl Acetate ‘mono’ representing the acetate group data is available. For the hydrocarbon-fused-ring type the analogue, Longifolene Coeur has acute oral toxicity data. These data can be used for read across.

Hypothesis:Cedryl Acetate EOA as a whole has similar acute oral toxicity as Cedryl Acetate ‘mono’ considering the acetate type and as Longifolene Coeur considering the hydrocarbon-fused-ring-type.

Available information:For Cedryl Acetate ‘mono’ the acute oral LD50 is > 5000 mg/kg bw (similar to OECD TG 401, Rel. 2). For Longifolene Coeur the LD50 is > 5000 mg/kg bw (OECD TG 401, Rel. 2).

Target chemical and source chemical(s)

Chemical structures of the target chemical and the source chemical(s) are shown in the data matrix, including physico-chemical properties and available toxicologicalinformation.

Purity / Impurities

The unidentified impurities of Cedryl Acetate EOA are not considered to have a significant influence on the acute oral toxicity.

Analogue approach justification

According to Annex XI 1.5 read across can be used to replace testing when the similarity can be based on a common backbone and a common functional group. When using read across the result derived should be applicable for C&L and/or risk assessment and it should be presented with adequate and reliable documentation.

Analogue selection: Cedryl Acetate EOA has a major constituent which isCedryl Acetate ‘mono’. This ‘mono’ has a similar fused ring system as all constituents and has an acetate group, which half of the constituents of Cedryl Acetate EOA have too. The other half without the acetate group can be represented by Longifolene Coeur, which has a similar fused ring system but not this acetate group.

Structural similarities and differences: Cedryl Acetate EOA components all have a hydrocarbon fused-ring system similar to Cedryl Acetate ‘mono’ and Longifolene Coeur. All constituents have some double bonds. The key difference is that Cedryl Acetate ‘mono’ (representing) the subgroup of Cedryl Acetate EOA) has an acetate group, which Longifolene type’ does not have and this type consists of solely hydrocarbons. That this ester bond does not influence the acute oral toxicity is presented below.

Toxico-kinetic, Absorption: Cedryl Acetate EOA will be similar absorbed compared to both source chemicals (Cedryl Acetate ‘mono’ and Longifolene Coeur based on the similarities in chemical structure and physico-chemical properties. All constituents have low water solubility and high (estimated) log Kow values( >5).Metabolism: The constituents of Cedral Acetate EOA with the acetate group will be easily cleaved by carboxyl esterases (Toxicological handbooks and e.g. Saghir et al. 1997) which will follow the same pattern as Cedryl Acetate ‘mono’. The (methylated) double bonds in other constituents of Cedral Acetate EOA will be prone for oxidation and give the corresponding alcohols. This is similar to the metabolic pathway of Longifolene Coeur and its double bonds.

Toxico-dynamics: In view of the similarities in backbones and functional groups between target and both sources the acute oral toxicity will be similar too.

Uncertainty of the prediction:In view of the reasoning above no other uncertainties are identified.

Data matrix

The relevant information on physico-chemical properties and toxicological characteristics are presented in the data matrix below.

Conclusions on hazard and risk assessment

For Cedryl Acetate EOA no acute oral toxicity is available and read across is used to fill this gap. When using read across the result derived should be applicable for C&L and/or risk assessment and be presented with adequate and reliable documentation.The text presented here fulfils the documentation. Cedryl Acetate ‘mono’ and Longifolene Coeur can be used as analogues. These analogues both have LD50s of > 5000 mg/kg bw and therefore Cedryl Acetate EOA has an LD50 > 5000 mg/kg bw.

Final conclusion on hazard and risk assessment: Cedryl Acetate EOA has an LD50 > 5000 mg/kg bw.

 

References

Saghir, M., Werner, J., Laposata, M., 1997, Rapid in vivo hydrolysis of fatty acid ethyl esters, toxic nonoxidative ethanol metabolites. Am. J. Physiol. - Gastrointestinal and Liver, 273, G184-G190

 

Data matrix to support the read across to Cedryl acetate EOA from Cedryl acetate and Longifolene Coeur for acute oral toxicity

Common names	Cedryl Acetate EOA							Cedryl acetate	Longifolene Coeur
	Target	Target	Target	Target	Target	Target	Target	Source	Supporting source
	Cedryl Acetate type			Longifolene type
Chemical structures
Typical concentration (%)	30-45 (major)	<10	<10	15-30 (minor)	8-16 (minor)	<10	<10
CAS no				32435-95-3	22567-43-7			77-54-3	475-20-7
Einecs	944-520-4							201-036-1	207-491-2
REACH		2018						2018	2018
Molecular weight	264	264	264	204	204	204	202	264	204
Phys-Chem data*
WS, mg/l	0.55	0.55	0.17	0.07	0.15	0.13	0.22	0.55	0.25
Log Kow	5.33	5.33	5.94	6.12	5.74	5.82	6.19	5.33 (6 exp.)	5.48 (5 exp.)
Human health
Acute oral toxicity LD50 in mg/kg bw	>5000 (OECD TG 401)	>5000 (Read across)	>5000 (Read across)	>5000 (Read across)	>5000 (Read across)	>5000 (RA)	>5000 (RA)	>5000 (OECD TG 401)	>5000 (OECD TG 401)

* Episuite v4.11 ; RA = Read across

Justification for classification or non-classification

Based on the results, the substance does not need to be classified for acute oral toxicity according to EU CLP (EC No. 1272/2008 and its amendments).