Eldew APS-307

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 9 November 2006 and 6 June 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: 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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): Phytosteryl/decyltetradecyl MyristoyImethylaminopropionate
- Substance type: Organic UVCB
- Physical state: Wax
- Analytical purity: 100%
- Lot/batch No.: 609074
- Expiration date of the lot/batch: 6 September 2010
- Stability under test conditions: Stable under the following storage conditions
- Storage condition of test material: Room temperature, dark place in well-closed containers (Storage temperature: 18 to 23 °C)

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source:
Japan Laboratory Animals Inc.

- Age at study initiation:
At the start of the study the animals were seven weeks old.

- Weight at study initiation:
At the start of the study the animals were in the weight range of 384 to 492 g.

- Housing:
The animals were housed individually or in groups of 2 animals in stainless-steel wire-mesh cages (W 266 x D 266 x H 200 mm: Riko Denki Co., Ltd.)

- Diet (e.g. ad libitum):
Allowed free access to RC4 pelleted diet (Oriental Yeast Co., Ltd., Lot Nos.: 060921 and 061018) using stainless-steel feeders

- Water (e.g. ad libitum):
Tap water (Fujimi Water Union) via an automatic water supplying system.

- Acclimation period:
Twenty one days


ENVIRONMENTAL CONDITIONS

- Temperature (°C):
The temperature was controlled to remain within the target ranges of 17 to 32 °C

- Humidity (%):
The humidity was controlled to remain within the target ranges of 25 to 81%.

- Air changes (per hr):
Rate of air ventilation was approximately 11 to 14 times per hour

- Photoperiod (hrs dark / hrs light):
The lighting was controlled by a time switch to give twelve hours continuous light (07:00 to 19:00) and twelve hours darknness.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: Vehicle 1: Liquid Paraffin Vehicle 2: Polyethylene glycol 300

Induction 2

Concentration / amount:

PRELIMINARY STUDY
Preliminary Study 1 (to select dose concentrations for intradennal induction): 25, 10, 5 and 2 w/w %
Preliminary Study 2 (to select dose concentrations for topical induction and challenge exposure): 100%, 50, 25, 10, 5 and 2.5 w/w %

MAIN STUDY
Intradermal Induction
Test article Sensitization group: 1:1 Emulsion of physiological saline and FCA; 10 w/w % Test formulation; 1:1 emulsion of 20 w/w % test formulation (test article mixed with FCA) and physiological saline.

Control Group: 1:1 Emulsion of physiological saline and FCA; Liquid paraffin.

Topical Induction
Test article Sensitization group: 100 % Test article.

Control group: Polyethylene glycol 300.

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: Vehicle 1: Liquid Paraffin Vehicle 2: Polyethylene glycol 300

Challenge 2

Concentration / amount:

PRELIMINARY STUDY
Preliminary Study 1 (to select dose concentrations for intradennal induction): 25, 10, 5 and 2 w/w %
Preliminary Study 2 (to select dose concentrations for topical induction and challenge exposure): 100%, 50, 25, 10, 5 and 2.5 w/w %

MAIN STUDY
Intradermal Induction
Test article Sensitization group: 1:1 Emulsion of physiological saline and FCA; 10 w/w % Test formulation; 1:1 emulsion of 20 w/w % test formulation (test article mixed with FCA) and physiological saline.

Control Group: 1:1 Emulsion of physiological saline and FCA; Liquid paraffin.

Topical Induction
Test article Sensitization group: 100 % Test article.

Control group: Polyethylene glycol 300.

No. of animals per dose:

10 animals - test group
5 animals - Control group
4 animals - Preliminary study group

Details on study design:

PREPARATION OF TEST ARTICLE

Preliminary study
1) Preliminary Study 1 (to select dose concentrations for intradermal induction)
After the test article (1.25,0.5,0.25 and 0.1 g) was weighed, liquid padin was added to make 5 g each, and the mixture was melted, stining in a double boiler at about 40°C (prepared concentrations: 25,10,5 and 2 w/w %).

2) Preliminary Study 2 (to select dose concentrations for topical induction and challenge exposure)
On the starting day of the preliminary study, 2.5 mL of physiological saline was added to 2.5 mL of Freund's Complete Adjuvant (FCA) to prepare a 1 :1 emulsion.
Seven days later, the undiluted test article (100%) was melted, stining in a double boiler at about 40°C, and 2 mL was measured and used for application. After the test article (2.5, 1.25, 0.5 and 0.25 g) was weighed, polyethylene glycol 300 was added to make 5 g each (prepared
concentrations: 50,25, 10 and 5 w/w %). Additionally, 0.25 g of the test article was weighed, and then polyethylene glycol 300 was added to make 10 g (prepared concentration: 2.5 w/w %). In preparation, the test article was melted, stining in a double boiler at about 40 °C, after adding polyethylene glycol 300.

Preparations at Intradermal Induction, Topical Induction and Challenge Exposure
1) Intradermal Induction
After 0.5 g of the test article was weighed, liquid paraffin was added to make 5 g, and the mixture was melted, stirring in a double boiler at about 40°C (prepared concentration: 10 w/w %). After 1.0 g of the test article was weighed, FCA was added to make 5 g, and the mixture was melted, stining in a double boiler at about 40°C (pqared concentration: 20 w/w %). 2.5 mL of physiological saline was added to 2.5 mL of the 20 w/w % test formulation
to prepare a 1 : 1 emulsion (final concentration of the test formulation: 10 %).

2) Topical Induction
After melting the undiluted test article (100%) in a double boiler at about 40°C, 5 mL was measured and used for application.

3) Challenge Exposure
After melting the undiluted test article (100%) in a double boiler at about 40°C, 5 mL was measured and used for application. Additionally, 2.5 g of the test article was weighed, polyethylene glycol 300 was added to make 5 g, and the mixture was melted, stirring in a double boiler at about 40°C (prepared concentration: 50 w/w %).

Preparation of 1:1 Emulsion of Physiological Saline and Freund's Complete Adjuvant (FCA)
At indermal induction, 10 rnL of FCA was added to 10 mL of physiological saline to prepare a 1 :1 emulsion.

Preparation of Petrolatum Containing 10 (w/w)% Sodium lauryl sulfate (SLS)
On the day before the topical induction, 1 g of SLS was weighed and white petrolatum was added to make 10 g. The mixture was mixed well using a mortar, melted in a warm water bath, and collected with a syringe.

PRELIMINARY STUDY TESTS:
A preliminary study was conducted to select dose concentrations for intradermal induction, topical induction and challenge exposure.

Preliminary study 1 (to select dose concentrations for intradermal induction):
Two animals without abnormalities (Animal numbers 1 and 2) were selected, fur of the dorsal skin was clipped and shaved, and 0.1 mL of each test formulation at various concentrations (25, 10,5 and 2 w/w %) was injected intradermaly to the application areas on the back closer to the neck region.

Preliminary study 2 (to select dose concentrations for topical induction and challenge exposure):
Another two animals without abnormalities (Animal numbers: 3 and 4) were selected, fur on the scapula (2 x 4 cm area) was clipped and shaved, and 0.1 mL of 1:1 emulsion of physiological saline and FCA was injected intradermaly to each of the 4 comers of the area. After 7 days, fur of the left and right flanks (3 x 7 cm areas) was clipped and shaved, and the test formulations at 6 concentrations (100 %, 50, 25, 10, 5 and 2.5 w/w %) were applied. For
application, 0.1 mL of each test formulation was put on a patch of 1.5 cm in diameter and the patch was applied to the application areas on the right side and left side of the back. The patch was removed at 24 hours after application, and the application area was cleaned with absorbent cotton soaked with polyethylene glycol 300.

Clinical Observation and Measurement of Body weight
In the preliminary study, animals were observed for clinical signs once daily from the starting day of study to the end of observation and weighed on the day of administration. The animals in the preliminary study to select dose concentrations for challenge exposure were weighed twice:once on the day of administration of 1:1 emulsion of physiological saline with FCA and once on the day of administration of the test article.

Observation and scoring of Skin Reactions
Observation of skin reactions was performed at 24 and 48 hours after intradermal injection and 24 and 48 hours after removal of topical application, and skin reactions were scored according to the following criteria (Magnusson & Kligman's criteria: 1969, 1970):

Degree of skin Reactions Score
No visible change 0
Discrete or patchy erythema 1
Moderate and confluent erythema 2
Intense erythema and swelling 3

RESULTS FOR PRELIMINARY STUDIES
Results of Preliminary Study and selection of Dose Concentrations for Induction and Challenge
The results of the preliminary study are shown in Table 1 (Attachment 1).

Preliminary study 1(to select dose concentrations for intradermal induction):
The treatment at 25 w/w % resulted in erythema and edema (score 3) with necrosis in 2/2 animals each at 24 and 48 hours after application. The treatment at 10 and 5 w/w % produced erythema of score 2 or 1 and the treatment at 2 w/w % resulted in erythema of score 1 in 2/2 or 1/2 animals at 24 and 48 hours after application, although no necrosis was noted in any animals. on the basis of te results, the dose concentration for intradermal induction was set at 10 w/w %, the maximum concetration without necrosis.

Preliminary study 2 (to select dose concentrations for topical induction and challenge exposure):
Application at each concentration of 100, 50, 25,10,5 or 2.5 w/w % was associated with no skin reactions at 24 or 48 hours after removal of application. Therefore, 100 %, the maximum concentration was selected for topical induction. Two concentrations, 100%, the maximum concentration inducing no initation, and 50 w/w %, half of the maximum concentration, were selected for challenge exposure.


MAIN STUDY
A. INDUCTION EXPOSURE

INTRADERMAL INDUCTION: On the day before intradermal induction, fur of a 4 x 6 cm area above the scapula was clipped with an electric clipper and further shaved with an electric shaver as closey as possible. On the next day, 0.1 mL each of the intrademal sensitizers listed in Text Table 4 (Attachment 1). Substances for Induction was intrademally injected to the induction areas A, B and C within the 2 x 4 cm area on the back closer to the neck region (see Attachment 1, Fig 2 - Application areas for induction and challenge).

TOPICAL INDUCTION: On Day 6 after intrademal induction, the same area was clipped and shaved, and 0.2 mL of petrolatum containing 100 w/w% SLS was applied topically to area D (Fig. 2, attachment 1) between the intradermal induction areas (2 x 4 cm). On the next day, this area was cleaned with absorbent cotton soaked with ether 0.2 mL each of topical sensitizers listed in Text Table 4 (attachment 1) was applied to a patch of 2 x 4 cm and the patch was applied to the induction area D4 and covered occlusively with a polyethylene film tape for topical induction for 48 hours.


B. CHALLENGE EXPOSURE

Twenty days after the intradermal induction, fur of the left flank (approximately 3 x 7 crn) of each animal in the test article sensitization group and the control group was clipped with an electric clipper and after shaved with an electric shaver as closely as possible. Next day, 3 areas (see attachment 1 for E to G for challenge areas) were provided, 0.1 mL each of the test formulations at two concentrations (100% and 50 w/w %) and polyethylene glycol 300 were applied to a patch of 1.5 cm in diameter, and applied to the animals, changing the areas for each animal according to Text Table 5 (attachment 1) showing location of application areas. Each patch was fixed in place with a polyethylene film tape for 24 hours for occhsive application of challenge. After the end of the 24-hour application, the patch was removed and the application area was cleaned with absorbent cotton soaked with polyethylene glycol 300.

Clinical Observation:
Animals were observed for clinical signs once daily from the starting day of induction (Day 0) to the end of skin observation after challenge (Day 24).

Measurement of Body Weight;
All animals were weighed on the starting day of induction (Day 0), day of topical induction (Day 7), day of challenge exposure (Day 21), and the final day of observation (Day 24). Mean body weight and standard deviation were calculated per group on each day.

Observation and Scoring of Skin Reactions:
Observation of skin reactions was performed at 24 hours after intradennal induction (for each area), immediately after removal of application for topical induction, at 24 and 48 hours after removal of challenge application, and erythema and edema were scored and recorded according to Magnusson & Kligman's criteria for Evaluation of Skin Reactions. Changes other than erythema and edema observed in the skin were also recorded. Representative animals (2 animals per group) were photographed at 24 hours afier removal of challenge application.

Evaluation:
The severity of skin reactions of individual animals was scored and the mean score for each test group at each observation after challenge was calculated, and the positive reaction rate [{(the number of animals with skin reactions of score 1 or greater) / (the total number of animals)} x 100] was determined. The sensitization potential of the test article was assessed by comparison of the mean score and positive reaction rate between the sensitization group and the control group, and the test article was judged to have sensitization potential if they were higher in the sensitization group than in the control group. The results obtained at 24 hours after intradermal induction and immediately after removal of patch for topical induction were regarded as reference data. For the data, group mean score was calculated in the same manner.

Statistical analysis was not performed in the evaluation of this study.

RESULTS FOR MAIN STUDY

Skin Reactions at lntradermal Induction:

Results are shown in Table 2 (Attachment 1).
Observation was done at 24 hours after intradermal injection.

1) Test Article Sensitization Group (see application areas A to C in Fig. 2 in Attachment 1).
At the injection area (A) of 1 :1 emulsion of physiological saline and FCA, erythema and edema (score 3) were observed at right and left areas in all animals (10/10 animals). The mean score was 3.0 each for right and left areas. For the injection area of 10 wlw% test formulation (area B), erythema of score 1 or 2 was observed in all animals in right and left areas. The mean score was 1.4 on left side and 1.3 on right side. At the injection area (C) of 1 :1 emulsion of 20 w/w % test formulation (test article mixed with FCA) and physiological saline, erythema and edema (score 3) were observed at right and left areas in all animals. The mean score was 3.0 at every injection area. Given the above, since there was no necrosis at injection areas of the test article (areas B and C), the set concentrations were considered to be appropriate.

2) Control Group
At the injection areas of 1:1 emulsion of physiological saline and FCA (Freund's Complete Adjuvant), all animals (5/5 animals) developed erythema and edema (score 3) at both areas A and C on right and left sides. The mean score was 3.0 each at areas A and C on right and left sides. For the injection area of liquid paraffin (area B), no skin reaction was observed at right or left injection area in any animals. The mean score was 0 at every injection area of both sides.

Skin Reactions at Topical Induction

Results are shown in Attachment 1, Table 3.
Observation was done immediately after removal of patches for topical induction.

1) Test Article Sensitization Group (substance for induction: 100% test article)
Erythema of score 1 or 2 was observed in all (10/10) animals, and the mean score was 1.8. The skin reaction was considered to involve some effects of the petrolatum containing 10% SLS which was applied on the day before topical induction. The procedure for induction was judged to be appropriate since the changes observed at the topical induction areas were mild to moderate erythema and no severe initation reaction was observed in any animals.

2) Control Group (substance for induction: polyethylene glycol 300)
Erythema of score 1 or 2 was observed in all (5/5) animals, and the mean score was 1.4. Eschar formation was observed only in 1/5 animals. The cause of eschar formation was unclear, but it suggested a specifically intense effect caused by application of the petrolatum containing 10% Sodium lauryl sulfate (SLS).

Skin Sensitization

Results are shown in Table 4 and Appendices 1 and 2 (Attachment 1)

1) Test Article Sensitization Group
At the area of challenge exposure to test article at 100% and 50 w/w %, no skin reactions were observed at 24 or 48 hours after removal of challenge application in any of the 10 animals.
Mean score was 0 at all observation times and the positive reaction rate was 0% at concentrations of 100% and 50 w/w%.
At the area of challenge exposure to polyethylene glycol 300 (vehicle), no skin reactions were observed at 24 or 48 hours after removal of challenge application. Mean score was 0 at all observation times and the positive reaction rate was 0%.

2) Control Group
Similarly to the test article sensitization group, at the area of challenge exposure to the test article, no skin reactions were observed at 24 or 48 hours after removal of challenge application at concentrations of 100% or 50 w/w% in any of the 5 animals. Mean score was 0 at all observation times and the positive reaction rate was 0% at concentrations of 100% and 50 w/w %.
At the area of challenge exposure to polyethylene glycol 300 (vehicle), no skin reactions were observed at 24 or 48 hours after removal of challenge application. Mean score was 0 at all observation times and the positive reaction rate was 0%.

Clinical Signs
Results are shown in Table 5 and Appendix 3 (Attachment 1).
There were no abnormal clinical signs in any animal in any group during the observation period.

Body Weight
Results are shown in Table 6 and Appendix 4 (Attachment 1).
On Day 24, the final day of observation, 8/10 animals in the test article sensitization group and 3/5 animals in the control group showed decreases in body weight in comparison with the previous values (Day 21). The degree of the decrease was 2 to 13 g, but there were no abnormalities in the clinical observation in any animal. The decreases in body weight were judged to be caused mainly by the procedure for occlusive application for challenge, and at least not to be test article-related.


ENVIRONMENTAL FACTORS THAT MIGHT HAVE AFFECTED THE RELIABILITY OF STUDY RESULTS

1) The acceptable ranges of temperature and humidity in the animal room where this study was conducted were set at 23 ± 5 °C (temperature) and 55 ± 25% (humidity) in the Protocol; however, the temperature and humidity deviated from each range on many days as shown in (i) and (ii) below.

(i) The humidity deviated from the upper limit of the acceptable range temporarily (81% at maximum) on one day (November 11,2006) due to cleaning in the animal room.

(ii) The temperature deviated from the upper limit of the acceptable range 5 times in total (32°C at maximum), from 10:00 to 11:00 (intermittently), &from 15:00 to 16:00 (intermittently) and at 20:00 (temporarily) on November 13 and at 5:00 (temporarily) and 9:00 (temporarily) on November 14,2006. The humidity deviated h m the lower limit of the acceptable range
one time (25% at minimum), from 10:00 to 11:00 (intermittently) on November 13,2006. The temperature deviated from the lower limit of the acceptable range 4 times in total (17 °C at minimum), at 17:00 (temporarily) on November 14, 10:00 (temporarily) on December 13,
& from 11:30 to 14:00 (intermittently) on December 21 and from 11:00 to 15:00 (intermittently) on December 22,2006. The cause of each deviation was unclear. As shown above, the temperature and humidity deviated from each range on many days during the animal experiment. However, each deviation occurred intermittently or temporarily and the deviation time did not extend over a long, continuous period. Additionally, no abnormal clinical signs were observed in any animals, and thus it was judged that they had no effect on the skin sensitization evaluation.

2) The Protocol stated that the preliminary study was to be conducted at 4 prepared concentrations of the test article, 25, 10, 5 and 1 w/w % to select dose concentrations for intradermal induction. However, it turned out afterwards that the actual concentration prepared for the lowest concentration (1 w/w %) was 2 w/w %. This was a deviation from the Protocol due to an error in preparation. Since no necrosis was observed at 10 w/w % or below, there was no problem on selecting dose concentrations for intrademal induction and it was judged to have no effect on the skin sensitization evaluation.

Otherwise there were no environmental factors that might have affected the reliability of the study results.

Challenge controls:

Twenty days after the intradermal induction, fur of the left flank (approximately 3 x 7 cm) of each animal in the test article sensitization group and the control group was clipped with an electric clipper and after shaved with an electric shaver as closely as possible. Next day, 3 areas (see attachment 1 for E to G for challenge areas) were provided, 0.1 mL each of the test formulations at two concentrations (100% and 50 w/w %) and polyethylene glycol 300 were applied to a patch of 1.5 cm in diameter, and applied to the animals, changing the areas for each animal according to Text Table 5 (attachment 1). Location of Application Areas. Each patch was fixed in place with a polyethylene film tape for 24 hours for occhsive application of challenge. After the end of the 24-hour application, the patch was removed and the application area was cleaned with absorbent cotton soaked with polyethylene glycol 300.

Measurement of Body Weight;
All animals were weighed on the starting day of induction (Day 0), day of topical induction (Day 7), day of challenge exposure (Day 21), and the final day of observation (Day 24). Mean body weight and standard deviation were calculated per group on each day.

Observation and Scoring of Skin Reactions:
Observation of skin reactions was performed at 24 hours after intradennal induction (for each area), immediately afier removal of application for topical induction, at 24 and 48 hours after removal of challenge application, and erythema and edema were scored and recorded according to Magnusson & Kligman's criteria for Evaluation of Skin Reactions. Changes other than erythema and edema observed in the skin were also recorded. Representative animals (2 animals per group) were photographed at 24 hours afier removal of challenge application.

Positive control substance(s):

yes

Remarks:

In this study, no positive control group was provided, however, a study using the positive control article, MBT (2-Mercaptobenzothiazole) was conducted and the results are attached.

Study design: in vivo (LLNA)

Concentration:

Not applicable

No. of animals per dose:

Not applicable

Details on study design:

Not applicable

Statistics:

Not Applicable

Results and discussion

Positive control results:

In this study, no positive control group was provided, however, a study using the positive control article, MBT (2-Mercaptobenzothiazole) was conducted and the results are attached.
In the study, the sensitization potential of MBT was clearly demonstrated and thus it was confirmed that the experimental procedure is appropriate and there was no problem in the sensitivity of guinea pigs used in the study.

In vivo (non-LLNA)

Results

Results 1

Reading:

other: Challenge Exposure

Hours after challenge:

24

Group:

test chemical

Dose level:

Test Article (100%)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 24.0. Group: test group. Dose level: Test Article (100%). No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Results 2

Reading:

other: Challenge Exposure

Hours after challenge:

48

Group:

test chemical

Dose level:

Test article (100 %)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 48.0. Group: test group. Dose level: Test article (100 %). No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: None.

Results 3

Reading:

other: Challenge Exposure

Hours after challenge:

24

Group:

test chemical

Dose level:

Test article in PEG300 (50 w/w %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

None

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 24.0. Group: test group. Dose level: Test article in PEG300 (50 w/w %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: None.

Results 4

Reading:

other: Challenge Exposure

Hours after challenge:

48

Group:

test chemical

Dose level:

Test article in PEG300 (50 w/w %)

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 48.0. Group: test group. Dose level: Test article in PEG300 (50 w/w %). No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Results 5

Reading:

other: Challenge Exposure

Hours after challenge:

24

Group:

test chemical

Dose level:

PEG300

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 24.0. Group: test group. Dose level: PEG300. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Results 6

Reading:

other: Challenge Exposure

Hours after challenge:

48

Group:

test chemical

Dose level:

PEG300

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 48.0. Group: test group. Dose level: PEG300. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Results 7

Reading:

other: Challenge Exposure

Hours after challenge:

24

Group:

other: Control

Dose level:

Test article (100 %)

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 24.0. Group: other: Control. Dose level: Test article (100 %). No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 8

Reading:

other: Challenge Exposure

Hours after challenge:

48

Group:

other: Control

Dose level:

Test article (100%)

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 48.0. Group: other: Control. Dose level: Test article (100%). No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 9

Reading:

other: Challenge Exposure

Hours after challenge:

24

Group:

other: Control

Dose level:

Test article in PEG300 (50 w/w %)

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 24.0. Group: other: Control. Dose level: Test article in PEG300 (50 w/w %). No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 10

Reading:

other: Challenge Exposure

Hours after challenge:

48

Group:

other: Control

Dose level:

Test article in PEG300 (50 w/w %)

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 48.0. Group: other: Control. Dose level: Test article in PEG300 (50 w/w %). No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 11

Reading:

other: Challenge Exposure

Hours after challenge:

24

Group:

other: Control

Dose level:

PEG300

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 24.0. Group: other: Control. Dose level: PEG300. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 12

Reading:

other: Challenge Exposure

Hours after challenge:

48

Group:

other: Control

Dose level:

PEG300

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: other: Challenge Exposure. . Hours after challenge: 48.0. Group: other: Control. Dose level: PEG300. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: Not applicable.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Not applicable

Any other information on results incl. tables

Not Applicable

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

On the basis of the results described above, it was concluded that Phytosteryl/decyltetradecyl Myristoylmethylaminopropionate had no skin sensitization potential.

Executive summary:

Introduction

As a part of safety evaluation of Phytosterly/decyltetradecy Myristoylmethylaminopropionate, a skin sensitization study in guinea pigs was carried out by the Maximization Test. This study was conducted in compliance with "OECD Guidelines for Testing of Chemicals 406" (OECD Council: July 17,1992).

A skin sensitization study was conducted in Hartley albino guinea pigs by the Maximization Test, in order to evaluate the skin sensitization potential of Phytosteryl/decyltetradecyl Myristoyhethylaminopropionate. Two test groups were provided: a test article sensitization group (10 animals) and a control group (5 animals). In the test article sensitization group, intradermal induction was done at 10 w/w%, the maximum concentration inducing no necrosis, topical induction was done at 100 w/w%, the maximum concentration, and then challenge exposure was done at 100%, the maximum concentration inducing no irritation and 50 w/w%, half of the maximum concentration In the control group, intradermal induction was done by liquid paraffin, topical induction was done by polyethylene glycol 300, and then challenge exposure was done in the same manner as for the test article sensitization group. The animals in each test group were also treated with polyethylene glycol 300 at challenge exposure. Observation of skin reactions was performed at 24 and 48 hours after removal of challenge application to determine any skin sensitization potential.

Results

The results are described in the following:

1) Skin Sensitization:

In the test article sensitization group, no skin reactions were observed at any application areas of Phytosteryl/decyltetradecyl M yristoylmethylaminopropionate at 24 or 48 hours after removal of challenge application at concentrations of 100% or 50 w/w%. The mean score was 0 at each time point at both concentrations, and the positive reaction rate was 0 %. In the control group, no skin reactions were observed at the area of challenge application at concentrations of 100% or 50 w/w%, and the positive reaction rate was 0% for both concentrations. At the area treated with polyethylene glycol 300, no skin reactions were observed in any group.

2) Clinical Signs and Body Weight:

There were no test article-related abnormalities in clinical observation or body weight during the observation period in any group.

Conclusion

On the basis of the results described above, it was concluded that Phytosteryl/decyltetradecyl Myristoylmethylaminopropionate had no skin sensitization potential.