Amyris balsamifera, ext.

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test was performed according to valid method (human maximisation test; Kligman 1966), but not according to OECD guideline or GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1972

Report date:

1972

Materials and methods

Test guideline

Qualifier:

no guideline followed

Version / remarks:

Test was performed according to valid method (human maximisation test; Kligman 1966)

Principles of method if other than guideline:

- Principle of test: Test was performed according to valid method (human maximisation test; Kligman J.I.D.; Vol 47; No 5; 393-409; 1966)

GLP compliance:

no

Remarks:

GLP was not yet enforced at the time

Type of study:

patch test

Justification for non-LLNA method:

As the results of a previously performed human maximisation test are already available, this data will be used for the WoE evaluation.

Test material

Test material information

Constituent 1

Reference substance name:

Amyris balsamifera, ext.

EC Number:

291-076-6

EC Name:

Amyris balsamifera, ext.

Cas Number:

90320-49-3

Molecular formula:

Not applicable due to the UVCB nature of the substance.

IUPAC Name:

Essential oil of Amyris obtained from the wood of Amyris balsamifera (Rutaceae) trees by steam distillation.

Test material form:

liquid: viscous

Details on test material:

For specific details on test material if available - See in RSS
Name of test material (as cited in study report or in reference): Amyris oil

Specific details on test material used for the study:

Not specified further

In vivo test system

Test animals

Species:

other: Human

Strain:

not specified

Sex:

male

Details on test animals and environmental conditions:

Inmate volunteers

Study design: in vivo (non-LLNA)

Induction

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

Not Specified

Day(s)/duration:

5 alternate-day 48 hour periods

Adequacy of induction:

other: Non-irritant substance, but skin pre-treated with 5% aqueous sodium lauryl sulphate

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

Not Specified

Day(s)/duration:

48 hours

Adequacy of challenge:

other: Skin pretreated with 1h application of 10% aqeous sodium Lauryl Suplhate under occlusion, before challenge.

No. of animals per dose:

25 volunteers

Details on study design:

RANGE FINDING TESTS: The materials were pre-tested on five subjects. Amyris Oil was applied to the (untreated) backs of 5 healthy male volunteers for 48 hours under occlusion. After this period the site was inspected, and no sign of irritation was observed. Based on this result the maximisation test was performed using an SLS pre-treatment.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: five (alternate-days)
- Exposure period: 5 alternate-day 48-hour periods
- Test groups: 25 male healthy inmate volunteers
- Control group: n/a
- Site: volar forearms
- Frequency of applications: once per exposure
- Duration: 48-hour periods
- Pre-treatment: patch sites were pre-treated for 34 hours with 5% aqueous sodium lauryl sulphate under occlusion

B. CHALLENGE EXPOSURE
- No. of exposures: once
- Day(s) of challenge: 10 days after last application (10 day rest period)
- Exposure period: 48 hours
- Test groups: 25 male healthy inmate volunteers
- Control group: n/a
- Site: volar forearms (fresh site)
- Evaluation (hr after challenge): on removal of the patch and 24 hours thereafter
- Pre-treatment: Challenge applications were preceded by one-hour applications of 10% aqueous sodium lauryl sulphate under occlusion.

Challenge controls:

not specified

Positive control substance(s):

not specified

Results and discussion

Positive control results:

not specified

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

pure

No. with + reactions:

2

Total no. in group:

25

Clinical observations:

Contact sensitisation observed, clinical effects are not further specified

Remarks on result:

positive indication of skin sensitisation

Results 2

Key result

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

Pure

No. with + reactions:

2

Total no. in group:

25

Clinical observations:

Contact sensitisation observed, clinical effects are not further specified

Remarks on result:

positive indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:

other: Sub-category 1B: potential to produce sensitisation

Remarks:

based on CLP criteria

Conclusions:

Under the conditions of the test, the substance produced 2 cases of contact-sensitisation and can be considered a weak sensitizer. Based on the results of this study the test substance should therefore be classified as a skin sensitizer (Sub-category 1B) in accordance with the CLP Regulation (1272/2008/EC).

Executive summary:

Pre-Testing: The materials were pre-tested on five subjects. Amyris Oil was applied to the (untreated) backs of 5 healthy male volunteers for 48 hours under occlusion. After this period the site was inspected, and no sign of irritation was observed. Based on this result the maximisation test was performed using an SLS pre-treatment.

Maximization test: (Kligman; J.I.D.; Vol 47; No 5; 393-409; 1966) the material was applied under occlusion to the volar forearms of 25 male healthy inmate volunteers, for five alternate-day 48-hour periods. The patch sites were pre-treated for 24 hours with 5% aqueous sodium lauryl sulphate under occlusion. Following a ten-day rest-period, challenge patches of the material was applied under occlusion to fresh sites for 48 hours. Challenge applications were preceded by one-hour applications of 10% aqueous sodium lauryl sulphate under occlusion. The challenge sites were read on removal of the patch and 24 hours thereafter.

Under the conditions of the test, the substance produced 2 cases of contact-sensitisation and can be considered a weak sensitizer. Based on the results of this study the test substance should therefore be classified as a skin sensitizer (Sub-category  1B) in accordance with the CLP Regulation (1272/2008/EC).