Phytosteryl Macadamiate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000-02-04 - 2000-06-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The GLP-study was performed according to an internationally accepted guideline.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Test material form:

semi-solid (amorphous): gel

Remarks:

migrated information: paste

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

Twenty-one male albino Dunkin Hartley guinea pigs supplied by David Hall Limited, Burton-on-Trent, Staffordshire, UK were used. At the start of the main study the animals weighed 351 to 426g, and were approximately eight to twelve weeks old. After an acclimatisation period of at least five days, each animal was selected at random and given a number unique within the study which was written on a small area of clipped rump using a black indelible marker-pen.
The animals were housed singly or in pairs in solid-floor polypropylene cages furnished with woodflakes. Free access to mains tap water and food (Guinea Pig FD1 Diet, Special Diets Services Limited, Witham, Essex, UK) was allowed -throughout the study.
The temperature and relative humidity were set to achieve limits of 1 7 to 23 °C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was approximately fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light and twelve hours darkness.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

arachis oil

Induction 2

Concentration / amount:

Induction:
The injections were:
a) Freund's Complete Adjuvant plus distilled water in the ratio 1:1
b) a 1 % w/w formulation of the test material in arachis oil BP
c) a 1% w/w formulation of the test material in a 1:1 preparation of Freund's Complete Adjuvant plus distilled water.

One week later (Day 7), the same area on the shoulder region used previously for intradermal injections was clipped again and treated with a topical application of the undiluted test material. A filter paper patch (WHATMAN No.4: approximate size 40 mm x 20 mm), loaded with the undiluted test material as a thick, even layer was applied to the prepared skin and held in place with a strip of surgical adhesive tape covered with an overlapping length of aluminium foil.

Challenge:
A square filter paper patch (WHATMAN No.4: approximate size 20 mm x 20 mm), loaded with the undiluted test material as a thick, even layer was applied to the shorn right flank of each animal and was held in place with a strip of surgical adhesive tape. To ensure that the maximum non-irritant concentration was used at challenge, the test material at a concentration of 75% w/w in arachis oil BP was similarly applied to a skin site on the left shorn flank. The patches were occluded with an overlapping length of aluminium foil and secured with a strip of elastic adhesive bandage wound in a double layer around the torso of each animal.

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

arachis oil

Challenge 2

Concentration / amount:

Induction:
The injections were:
a) Freund's Complete Adjuvant plus distilled water in the ratio 1:1
b) a 1 % w/w formulation of the test material in arachis oil BP
c) a 1% w/w formulation of the test material in a 1:1 preparation of Freund's Complete Adjuvant plus distilled water.

One week later (Day 7), the same area on the shoulder region used previously for intradermal injections was clipped again and treated with a topical application of the undiluted test material. A filter paper patch (WHATMAN No.4: approximate size 40 mm x 20 mm), loaded with the undiluted test material as a thick, even layer was applied to the prepared skin and held in place with a strip of surgical adhesive tape covered with an overlapping length of aluminium foil.

Challenge:
A square filter paper patch (WHATMAN No.4: approximate size 20 mm x 20 mm), loaded with the undiluted test material as a thick, even layer was applied to the shorn right flank of each animal and was held in place with a strip of surgical adhesive tape. To ensure that the maximum non-irritant concentration was used at challenge, the test material at a concentration of 75% w/w in arachis oil BP was similarly applied to a skin site on the left shorn flank. The patches were occluded with an overlapping length of aluminium foil and secured with a strip of elastic adhesive bandage wound in a double layer around the torso of each animal.

No. of animals per dose:

10

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

undiluted

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: undiluted. No with. + reactions: 0.0. Total no. in groups: 10.0.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

undiluted

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: undiluted. No with. + reactions: 0.0. Total no. in groups: 10.0.

Any other information on results incl. tables

CHALLENGE CONCENTRATIONS: UNDILUTED AS SUPPLIED AND 75% w/w

Skin Reactions (Hours After Removal of Dressing)
Animal			24 Hours					48 Hours
Number		75%		100%			75%		100%
	Er	Oe	Other Er	Oe	Other	Er	Oe	Other Er	Oe	Other
1	0	0	0	0	-	0	0	0	0	-
2	0	0	0	0	-	0	0	0	0	-
3	*	*	* *	*	*	*	*	* *	*	*
4	0	0	0	0	-	0	0	0	0	-
5	0	0	0	0	-	0	0	0	0	-
6	0	0	0	0	-	0	0	0	0	-
7	0	0	0	0	-	0	0	0	0	-
8	0	0	0	0	-	0	0	0	0	-
9	0	0	0	0	-	0	0	0	0	-
10	0	0	0	0	-	0	0	0	0	-

Er = erythema            Oe = oedema              - = no other reactions noted          * = no data; animal killed for humane reasons Day 17

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test material, YOFCO MAS LOT NO. 92506, produced a 0% (0/9) sensitisation rate and was classified as a non-sensitiser to guinea pig skin. The test material did not meet the criteria for classification as a sensitiser according to EU labelling regulations. No symbol and risk phrase are required.

Executive summary:

A study was performed to assess the contact sensitisation potential of the test material in the albino guinea pig. The study was performed in compliance with the OECD Guidelines for Testing of Chemicals No. 406 "Skin Sensitisation" (adopted 17 July 1992) and Method B6 of Commission Directive 96/54/EC (which constitutes Annex V of Council Directive 67/548/EEC).

Ten test and five control animals were used for the main study.

Based on the results of sighting tests, the concentrations of test material for the induction and challenge phases were selected as follows:

Intradermal Induction                               : 1% w/w in arachis oil BP

Topical Induction                                     : undiluted as supplied

Topical Challenge                                     : undiluted as supplied and 75% w/w in arachis oil BP

 

The test material produced a 0% (0/9) sensitisation rate and was classified as a non sensitiser to guinea pig skin. The test material did not meet the criteria for classification as a sensitiser according to EU labelling regulations. No symbol and risk phrase are required.