Fatty acids, rape-oil, hydrogenated, reaction products with diethylenetriamine, di-Me sulfate-quaternized

Endpoint summary

Description of key information

Based on the results of the read across GPMT and the confirmatory HRIPT, the test substance, 'di-C18-22 AAEMIM-MS' is considered to be non-sensitising to the skin.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

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

Adequacy of study:

key study

Study period:

From April 01, 2009 to June 16, 2009

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Refer to section 13 of IUCLID for details on the read-across justification. The study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

as layed down in Commission Regulation (EC) No 440/2008 of 30 May 2008

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

GPMT study was already available with the read across substance

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
Albino Dunkin Hartley Guinea Pig, CRL:(HA)BR, SPF
- Age at study initiation: 4-6 weeks
- Weight at study initiation: pretest groups: 332 - 349 g, test and control group: 330 - 366 g
- Housing: individually in Makrolon type-4 cages with standard softwood bedding ("Lignocel", Schill AG, 4132 Muttenz, Switzerland).
- Diet: ad libitum, pelleted standard Provimi Kliba 3418 guinea pig breeding / maintenance diet batch no. 82/08, Provimi Kliba AG, 4303 Kaiseraugst, Switzerland
- Water: ad libitum, tap water as for human consumption
- Acclimation period: animals of test and control group were acclimatised, no data on acclimation length


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30 - 70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): automatically controlled light cycle of 12 h light and 12 h dark

Route:

intradermal and epicutaneous

Vehicle:

polyethylene glycol

Concentration / amount:

Induction: 0.5 % (V/V) in intradermal, 10 % (V/V) epicutaneous challenge: 1 % (V/V)
Concentration selection based on a small scale dose range studies

Route:

epicutaneous, occlusive

Vehicle:

polyethylene glycol

Concentration / amount:

Induction: 0.5 % (V/V) in intradermal, 10 % (V/V) epicutaneous challenge: 1 % (V/V)
Concentration selection based on a small scale dose range studies

No. of animals per dose:

treatment group: 10
control group: 5

Details on study design:

RANGE FINDING STUDIES
Intradermal injection:
For intradermal injections (0.1 mL/site) of a 1:1 (v/v) mixture of Freund's Complete Adjuvant/physiological saline were made into the shaved neck of three guinea pig. Six days later intradermal injections (0.1 mL/site) were made into the clipped flank . One animal received spatially divided injections of 25, 15, and 10 % , one of 5, 2.5, and 1 % and one of 0.5 and 0.1 %. Concentrations were formulated in polyethylene glycol 300 (PEG 300). Dermal reactions were assessed 24 hours later.

Dermal application:
Four intradermal injections (0.1 mL/site) of a 1:1 (v/v) mixture of Freund's Complete Adjuvant/physiological saline were made into the shaved neck of two guinea pigs. Six days later, four patches of filter paper (3 x 3 cm) were saturated with the test item at 100% (technically the highest possible concentration to be applied sufficiently), 75%, 50% and 25% in PEG 300 and applied to the clipped and shaved flanks of the same guinea pigs. The amount or volume of test item preparation applied was approximately 0.2 g at 100%, 75% and 50% or 0.2 mL at 25%. The concentration causing mild (to moderate) local skin reaction and the concentration being the highest tested non-irritating concentration could not be determined after the epidermal pretest described above. Therefore, a second pretest was performed with two additional naive guinea pigs, treated in the same way as those described previously, with the concentrations of 15%, 10%, 5% and 1% in PEG 300, using an application volume of approximately 0.2 mL.

MAIN STUDY
A. INDUCTION EXPOSURE
- First stage (test day 1)
An area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair. Three pairs of intradermal injections (0.1 mL/site) were made symmetrically in two rows on either side of the sine just within the boundaries of a 4 x 6 cm area in the clipped region.
-- Test groups:
1. 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline
2. The test item at 0.5% in PEG 300
3. The test item at 0.5% in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline
- Control group:
1. 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline
2. PEG 300
3. 1:1 (w/w) mixture of PEG 300 in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline

- Second stage - One week after the intradermal injections (test day 8), the scapular area (approximately 6 x 8 cm) was again clipped and shaved free of hair prior to the application. A 2 x 4 cm patch of filter paper was saturated with the test item at 10% in PEG 300 and placed over the injection sites of the test animals. The volume of test item preparation applied was approximately 0.3 mL. The patch was covered with aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The occlusive dressings were left in place
for 48 hours. The epidermal application procedure described ensured intensive contact of the test item.

The guinea pigs of the control group were treated as described above with PEG 300 only, applied at a volume of approximately 0.3 mL.

The reaction sites were assessed 24 and 48 hours after removal of the bandage for erythema and oedema according to the method of Magnusson and Kligman.

B. CHALLENGE EXPOSURE (test day 22)
The test and control guinea pigs were challenged two weeks after the epidermal induction application and were treated in the same way.

Two patches (3 x 3 cm) of filter paper were saturated with the test item at the highest tested nonirritating concentration of 1% (applied to the left flank) and the vehicle only (PEG 300 applied to the right flank) using the same method as for the epidermal application. The volume of test item preparation and vehicle alone applied was approximately 0.2 mL. The dressings were left in place for 24 hours.

C. OBSERVATION and SCORING
The test item skin area of the animals used for the epidermal pretest and challenge was depilated approximately 21 hours after the patches have been removed, using an approved depilatory cream (VEET Cream, Reckitt & Colman AG, 4123 Allschwil, Switzerland). The depilation was performed to clean the stratum corneum from the possible staining produced by the test item and to facilitate the reading of the possible skin reaction. The depilatory cream was placed on the patch sites and surrounding areas, and left on for up to 3-5 minutes. It was then thoroughly washed off with a stream of warm, running water. The animals were then dried with a disposable towel, and returned to their cages.

The scoring system was performed by visual assessment of erythema and oedema.
0 = no visible change
1 = discrete or patchy erythema
2 = moderate and confluent erythema
3 = intense erythema and swelling

Any other gross lesions not covered by this scoring system were described.

The grading method used for the pretest, induction and challenge was identical. It was performed 24 ± 2 hours after removal of the patches for the intradermal and epidermal pretest and induction and for challenge and repeated 24 ± 2 hours later (48-hour grades) for the epidermal pretest,
epidermal induction and challenge.

OTHER OBSERVATIONS
- Viability / Mortality: Daily from delivery of the animals to the termination of the test.
- Clinical Signs / Grading of Skin Response: Daily from delivery of the animals to the termination of test. Skin responses were graded during the pretest, induction and challenge periods.
- Body Weights: At delivery/acclimatization start, at the end of the pretest, at test day 1 (day of treatment) and at the termination of the study.

POSITIVE CONTROL (RELIABILITY CHECK):
- The sensitivity and reliability of the experimental technique employed was assessed by use of ALPHA-HEXYLCINNAMALDEHYDE which is recommended by the OECD 406 Guidelines and is known to have moderate skin sensitisation properties in the guinea pig strain. The results from the most recent test run (Harlan Laboratories Study C16261, performed from 08-Oct-2008 to 14-Nov-2008) were included in the study report.

STATISTICAL ANALYSIS
Descriptive statistics (means and standard deviations) were calculated for body weights. No inferential statistics were used.

Positive control substance(s):

yes

Remarks:

Alpha-hexylcinnamaldehyde, periodically tested, most recent test run performed from 08-Oct-2008 to 14-Nov-2008

Positive control results:

All test animals showed skin reactions after the challenge treatment with Alpha-hexylcinnamaldehyde at 3% in PEG 300. No skin effect was observed in the animals treated with Alpha-hexylcinnamaldehyde at 1% in PEG 300. No skin effect was observed in the control group.

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

1%

No. with + reactions:

1

Total no. in group:

10

Clinical observations:

discrete patchy erythema

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

1%

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

1%

No. with + reactions:

0

Total no. in group:

5

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

1%

No. with + reactions:

0

Total no. in group:

5

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

other: Vehicle (PEG) control in initiated animals

Dose level:

0%

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

other: Vehicle 9PEG) control in initiated animals

Dose level:

0%

No. with + reactions:

0

Total no. in group:

10

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

3% in PEG 300

No. with + reactions:

5

Total no. in group:

5

Remarks on result:

positive indication of skin sensitisation

RESULTS

Range finding studies:

Intradermal injection: All tested concentrations caused skin reactions:

- 25 and 10 %: moderate and confluent erythema (score 2) with swelling

- 10, 5, 2.5 and 1 %: moderate and confluent erythema (score 2)

- 0.5 %: discrete or patchy erythema (score 1)

- 0.1 %: discrete or patchy erythema (score 1) with blanching

Dermal application: The test item at a concentration of 1 % caused no skin reactions. Concentration depended skin reactions after application of 5 to 100 %:

- 100 and 75 %: at 24 and 48 h scoring intense erythema and swelling (score 3), at 48 h with scaling and necrosis

- 50 %: at 24 and 48 h scoring moderate and confluent erythema (score 2), at 24 with swelling, at 48 h with scaling and necrosis

- 25 %: at 24 h scoring in both animals moderate and confluent erythema (score 2), at 48 h scoring one animals with discrete or patchy erythema (score 1) with scaling and one animals with moderate and confluent erythema (score 2) with scaling and necrosis

- 15, 10 and 5 %: at 24 and 48 h scoring discrete or patchy erythema (score 1), at 48 h with scaling

- 1 %: at 24 and 48 h scoring no visible changes

Main study:

- Induction:

-- intradermal: The expected and common findings were observed in the control and test group after the different applications using FCA intradermally. These findings consisted of erythema, oedema, necrotizing dermatitis, encrustation and exfoliation of encrustation.

-- epicutaneous: No erythematous or oedematous reaction was observed in the animals treated with PEG 300 only. Discrete or patchy (score 1) to moderate and confluent (score 2) erythema was observed in all animals at the 24- and 48 -hour scorings after treatment with the test item at 10% in PEG 300; in detail at 24 h scoring 8/10 animals with discrete or patchy erythema (score 1), 2/10 animals with moderate and confluent erythema (score 2), at 48 h scoring 9/10 animals with discrete or patchy erythema (score 1), 1/10 animals with moderate and confluent erythema (score 2). Scaling was observed in 70% of the test animals at the 48-hour reading.

- Challenge

-- test group: Discrete or patchy erythema (score 1) was observed in 1/10 of the test animals at the 24-hour scoring after treated with the test item at 1% in PEG 300. No local skin reaction was observed at the 48-hour scoring.

No skin reactions were observed in the test animals when treated with PEG 300 only.

-- control group: No skin reactions were observed in the animals when treated with PEG 300 alone or when treated with the test item at 1% in PEG 300.

OTHER OBSERVATIONS

- Viability / Mortality: There were no deaths during the course of the study, hence no necropsies were performed.

- Clinical Signs: No signs of systemic toxicity were observed in the animals.

- Body Weights: The body weight of the animals was within the range commonly recorded for animals of this strain and age.

Interpretation of results:

other: CLP criteria not met

Conclusions:

Based on the results of the read across study, a similar non-sensitising behaviour can be expected for the test substance, 'di-C18-22 AAEMIM-MS'.

Executive summary:

A study was conducted to determine the skin sensitisation potential of the read across substance, 'di-C16-18 and C18-unsatd. AAEMIM-MS' (active: 100%) using Guinea Pig Maximisation test (GPMT) method, according to OECD Guideline 406, in compliance with GLP.A pre-test was conducted to determine non-irritating concentrations to be used in the main study.The sensitivity of the test system was assessed by use of the positive control substance Alpha-exylcinnamaldehyde.In the main test, one treated group of ten maleDunkin Hartley guinea pigsreceived 0.5% of read across substance in a PEG 300 and in an emulsion of Freund’s Complete Adjuvant (FCA)/physiological saline (50/50, w/w), on Day 1 by intradermal injections in the nuchal region and48 h topical applications of 10% in PEG 300 on Day 8,under occlusive conditions. A control group of five male guinea pigs were intradermally induced with PEG 300 and FCA/physiological saline on Day 1 and epidermally induced with PEG 300 under occlusion on Day 8. Two weeks after epidermal induction the control and test animals were challenged by epidermal application of the test substance at 1% in PEG 300 and PEG 300 alone under occlusive dressing. Cutaneous reactions were evaluated at 24 and 48 h after removal of the dressing. No toxic signs were evident in the guinea pigs of the control or test group. No deaths occurred. Ten percent of the test animals showed discrete or patchy erythema at the 24 h scoring after the challenge treatment with test substance at 1% (w/v) in PEG 300; at the 48 h scoring the 10 animals were free of skin reactions. None of the 5 control animals showed skin reactions at challenge. Under the study conditions, the read across substance was considered to be non-sensitising to the skin (Arcelin, 2009). Based on the results of the read across study, a similar non-sensitising behaviour can be expected for the test substance, 'di-C18-22 AAEMIM-MS'.

Reference 2

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

From December 2, 1999 to January 6, 2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

according to guideline

Guideline:

other: Repeat Insult Patch Test (RIPT) Sensitive (or Standard) Panel

Principles of method if other than guideline:

A Human Repeated Insult Patch Test (HRIPT) was conducted to determine the skin sensitivity potential of the test substance, following epicutaneous induction with 5% test substance concentrations to 65 healthy volunteers for 24 h under a semi-occlusive conditions. After a rest period of ca. 2 weeks, the substance was applied under the same conditions to a new site as a challenge. A the end of 24 h, the semi-occluded patch is removed and the site is read for immediate response. Follow-up readings are made 24 and 72 h later. The skin effects are then scored for irritation/sensitisation reactions.

GLP compliance:

not specified

Type of study:

patch test

Justification for non-LLNA method:

Human study

Species:

other: Human

Sex:

male/female

Details on test animals and environmental conditions:

The subjects were informed of the nature of the test, including possible adverse reactions. Written informed consent was obtained. Additionally, the subjects were considered dependable and able to read and understand instructions. The subjects did not exhibit any physical or dennatological condition that would have precluded application of the test article.

Route:

epicutaneous, semiocclusive

Vehicle:

water

Concentration / amount:

5% aqeous solution approximately 0.2 mL

Day(s)/duration:

24 h application followed by 24 h rest after 2 cycles 48 h rest period upto nine applicaitons

Adequacy of induction:

other: If a subject developed a positive reaction of 2 level erythema or greater during teh induction phase

Route:

epicutaneous, semiocclusive

Vehicle:

water

Concentration / amount:

0.2 mL of 5% solution

Day(s)/duration:

24 h and 72 h

No. of animals per dose:

Total: 65 subjects, 57 females and 8 males, ranging in age Horn 20 to 69 years were empanelled for this test

Details on study design:

Procedure:
Induction phase: Approximately 0.2 mL of the test substance was placed onto 2-cm square semi-occlusive patches (scanned surgical tape/webril non-woven cotton) which was applied to the back of each subject between the scapulae and waist, adjacent to the spinal mid-line. The subjects were instructed to remove the patch 24-h aster application. 24 h post periods follows the Tuesday and Thursday removals and 48 h rest periods followed each Saturday removal. The site was scored by trained examiner just prior to the next patch application. In general this procedure was repeated every Monday, Wednesday and Friday until 9 applications of the test substance had been made.
Procedurally, if a subject developed a positive reaction of 2-level erythema or greater during the induction phase or, at the discretion of the principal investigator, if the skin response warranted a change in site, the patch would be applied to a previously unpatched, adjacent site for the next application. If a 2-level reaction (or greater) occurred at the new site, no further application would be made. However, any reactive subjects would be subsequently challenged patch tested.

Challenge phase:
After rest period of approximately two weeks (no application of the test substance) the challenge patch was applied to a previously unpatched (virgin) test site. The site was scored 24 and 72 h after application. All subjects were instructed to report any delayed skin reactivity that might have occured after the final challenge patch reading. When warranted, selected test subjects were called back to the clinic for additional patch reading. When warranted, selected test subjects were called back to the clinic for additional examinations and scoring to determine possible increased or decreases in challenge patch reactivity. Skin responses for both the induction and challenge phases of the study were scored according to the following 6-point scale:
0=No evidance of any effect
+=Barely perceptible (minimal, faint, uniform or spotty erythea)
1=Mild (Pink, uniform erythema covering most of the contact site)
2=Moderate (Pink-red erythema with/without petchiae or paules)
4=Severe (Deep-red erythema with/without vesiculation or weeping)

All other observed dermal sequelae (i.e edema, dryness, hypo-or hyper-pigmentation, etc) were approximately recorded and described as mild, moderate or severe.

Positive control substance(s):

not specified

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

5% aqueous solution

No. with + reactions:

0

Total no. in group:

65

Clinical observations:

none

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

5% aqueous solution

No. with + reactions:

0

Total no. in group:

65

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

test chemical

Dose level:

5% aqueous solution

No. with + reactions:

0

Total no. in group:

65

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Remarks on result:

not measured/tested

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

5% aqueous solution

No. with + reactions:

0

Total no. in group:

65

Clinical observations:

None

Remarks on result:

no indication of skin sensitisation

61 subjects satisfactorily completed the test procedure. Four (4) test panelists discontinued for reasons unrelated to the conduct of the study. Discontinued panelist data are shown, up to the point of discontinuation, but are not used in the Results, Discussion or Conclusions sections of this final report. A single transient, barely-perceptible (+) non-specific patch test response was observed on one (1/61) test panelist (Subj. No.:56) during the Induction and/or Challenge phases of the study. This low-grade, nonspecific response was not considered irritant nor allergic in nature.

Results:

Subject nuber	Subject initial	age	sex	Induction exposure No.									Challenge reading (Hrs)
				1	2	3	4	5	6	7	8	9	24	72
1	DA	69	M	0	0	0	0	0	0	0	0	0	0	0
2	IP	62	F	0	0	0	0	0	0	0	0	0	0	0
3	SR	22	F	0	0	0	0	0	0	0	0	0	0	0
4	LG	20	F	0	0	0	0	0	0	0	0	0	0	0
5	MG	24	M	0	0	0	0	0	0	0	0	0	0	0
6	BB	41	F	0	0	0	0	0	0	0	0	0	0	0
7	TS	29	F	0	0	0	0	0	0	0	0	0	0	0
8	SP	50	F	0	0	0	0	0	0	0	0	0	0	0
9	RF	45	F	0	0	0	0	0	0	0	0	0	0	0
10	SJ	38	F	0	0	0	0	0	0	0	0	0	0	0
11	SM	51	F	0	0	0	0	0	0	0	0	0	0	0
12	SM	47	F	0	0	0	0	0	0	0	0	0	0	0
13	TB	46	F	0	0	0	0	0	0	0	0	0	0	0
14	EK	51	F	0	0	0	0	0	0	0	0	0	0	0
15	CS	57	F	0	0	0	0	0	0	0	0	0	0	0
16	FW	63	M	0	0	0	0	0	0	0	0	0	0	0
17	CW	66	F	0	0	0	0	0	0	0	0	0	0	0
18	CL	36	F	0	0	0	0	0	0	0	0	0	0	0
19	GM	42	F	0	0	0	0	0	0	0	0	0	0	0
20	LM	51	M	0	0	0	0	0	0	0	Disc
21	EH	65	F	0	0	0	0	0	0	0	0	0	0	0
22	DM	36	F	0	0	0	0	0	0	0	0	0	0	0
23	GB	49	M	0	0	0	0	0	0	0	0	0	0	0
24	SC	56	F	0	0	0	0	0	0	0	0	0	0	0
25	DS	42	F	0	0	0	0	0	0	0	0	0	0	0
26	VJ	48	F	0	0	0	0	0	0	0	0	0	0	0
27	MM	46	F	0	0	0	0	0	0	0	0	0	0	0
28	MR	43	F	0	0	0	0	0	0	0	0	0	0	0
29	DP	32	F	0	0	0	0	0	0	0	0	0	0	0
30	MS	54	F	0	0	0	0	0	0	0	0	0	0	0
31	DT	49	F	0	0	0	0	0	0	0	0	0	0	0
32	WG	34	F	0	0	0	0	0	0	0	0	0	0	0
33	WK	68	M	0	0	0	0	0	0	0	0	0	0	0
34	PK	67	F	0	0	0	0	0	0	0	0	0	0	0
35	HB	28	F	0	0	0	0	0	0	0	0	Disc
36	CJ	32	F	0	0	0	0	0	0	0	0	0	0	0
37	PS	39	F	0	0	0	0	0	0	0	0	0	0	0
38	GS	60	F	0	0	0	0	0	0	0	0	0	0	0
39	SB	42	F	0	0	0	0	0	0	0	0	0	0	0
40	TF	29	F	0	0	0	0	0	0	0	0	0	0	0
41	NL	67	F	0	0	0	0	0	0	0	0	0	0	0
42	PS	49	F	0	0	0	0	0	0	0	0	0	0	0
43	JM	32	F	0	0	0	0	0	0	0	0	0	0	0
44	YD	37	F	0	0	0	0	0	0	0	0	0	0	0
45	LR	33	F	0	0	0	0	0	0	0	0	0	0	0
46	PA	35	M	0	0	0	0	0	0	0	0	0	0	0
47	DG	44	F	0	0	0	0	0	0	0	0	0	0	0
48	VS	42	F	0	0	Disc
49	LA	28	F	0	0	0	0	0	0	0	0	0	0	0
50	RB	67	F	0	0	0	0	0	0	0	0	0	0	0
51	KG	36	F	Disc
52	NL	37	F	0	0	0	0	0	0	0	0	0	0	0
53	TL	37	F	0	0	0	0	0	0	0	0	0	0	0
54	KG	42	F	0	0	0	0	0	0	0	0	0	0	0
55	TB	22	F	0	0	0	0	0	0	0	0	0	0	0
56	BS	69	F	0	0	0	0	0	0	0	0	0	0	0
57	HF	39	F	0	0	0	0	0	0	0	0	0	0	0
58	KS	51	F	0	0	0	0	0	0	0	0	0	0	0
59	SR	37	F	0	0	0	0	0	0	0	0	0	0	0
60	CS	69	F	0	0	0	0	0	0	0	0	0	0	0
61	GM	53	F	0	0	0	0	0	0	0	0	0	0	0
62	MP	68	F	0	0	0	0	0	0	0	0	0	0	0
63	DY	41	F	0	0	0	0	0	0	0	0	0	0	0
64	JF	36	M	0	0	0	0	0	0	0	0	0	0	0
65	LE	44	F	0	0	0	0	0	0	0	0	0	0	0

0 = No reaction

+ = Barely Perceptible (minimal, faint, unifonn or spotty erytllema)

I = Mild (pink, unifonn erytllema covering most of the contact site)

2 = Moderate (pink-red erytllema visibly unifonn in entire contact site)

3 = Marked (bright-red erytllema with/without petechiae or papules)

4 = Severe (deep-red erytllema with/without vesiculation or weeping)

Interpretation of results:

other: CLP criteria not met

Conclusions:

Under the study conditions, the test substance was not considered to be sensitising to the human skin at 5% test concentration (ca. 2% active ingredient)

Executive summary:

A study was conducted to determine the skin sensitisation potential of a formulation containing 39% of the test substance, ‘di-C18-22 AAEMIM-MS’ according to human repeated insult patch test (HRIPT) method, in compliance with GLP. In the induction phase, a 5% test substance preparation was applied epicutaneously 3 times weekly to 65 (57 female and 8 male) healthy volunteers for 24 h under semi-occlusive conditions. After a rest period of ca. 2 weeks, the substance was applied under the same conditions to a new site as a challenge. At the end of 24 h, the semi-occluded patch was removed and the site was read for immediate response. Follow-up readings were made 24 and 72 h later. The skin effects were then scored for irritation and allergic responses (such as erythema with or without vesiculation, weeping, petechiae or papules, edema, dryness and hypo/hyper-pigmentation). No visible irritation or allergic contact dermatitis was observed in any of the 65 individuals following challenge. Under the study conditions, the test substance was not considered to be sensitising to the human skin at 5% test concentration (ca. 2% active ingredient) (RCTS, 2002).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information:

Study 1:

A study was conducted to determine the skin sensitisation potential of the read across substance, 'di-C16-18 and C18-unsatd. AAEMIM-MS' using Guinea Pig Maximisation test (GPMT) method, according to OECD Guideline 406, in compliance with GLP.A pre-test was conducted to determine non-irritating concentrations to be used in the main study.The sensitivity of the test system was assessed by use of the positive control substance Alpha-exylcinnamaldehyde.In the main test, one treated group of ten maleDunkin Hartley guinea pigsreceived 0.5% of read across substance in a PEG 300 and in an emulsion of Freund’s Complete Adjuvant (FCA)/physiological saline (50/50, w/w), on Day 1 by intradermal injections in the nuchal region and48 h topical applications of 10% in PEG 300 on Day 8,under occlusive conditions. A control group of five male guinea pigs were intradermally induced with PEG 300 and FCA/physiological saline on Day 1 and epidermally induced with PEG 300 under occlusion on Day 8. Two weeks after epidermal induction the control and test animals were challenged by epidermal application of the test substance at 1% in PEG 300 and PEG 300 alone under occlusive dressing. Cutaneous reactions were evaluated at 24 and 48 h after removal of the dressing. No toxic signs were evident in the guinea pigs of the control or test group. No deaths occurred. Ten percent of the test animals showed discrete or patchy erythema at the 24 h scoring after the challenge treatment with test substance at 1% (w/v) in PEG 300; at the 48 h scoring the 10 animals were free of skin reactions. None of the 5 control animals showed skin reactions at challenge. Under the study conditions, the read across substance was considered to be non-sensitising to the skin (Arcelin, 2009).Based on the results of the read across study, a similar non-sensitising behaviour can be expected for the test substance, 'di-C18-22 AAEMIM-MS'.

Study 2:

A study was conducted to determine the skin sensitisation potential of a formulation containing 39% of the test substance, ‘di-C18 -22 AAEMIM-MS’ according to human repeated insult patch test (HRIPT) method, in compliance with GLP. In the induction phase, a 5% test substance preparation (i.e., equivalent to ca. 2% active ingredient) was applied epicutaneously 3 times weekly to 65 (57 female and 8 male) healthy volunteers for 24 h under semi-occlusive conditions. After a rest period of ca. 2 weeks, the substance was applied under the same conditions to a new site as a challenge. At the end of 24 h, the semi-occluded patch was removed and the site was read for immediate response. Follow-up readings were made 24 and 72 h later. The skin effects were then scored for irritation and allergic responses (such as erythema with or without vesiculation, weeping, petechiae or papules, edema, dryness and hypo/hyper-pigmentation). No visible irritation or allergic contact dermatitis was observed in any of the 65 individuals following challenge. Under the study conditions, the test substance was not considered to be sensitising to the human skin at 2% a.i. test concentration (RCTS, 2002).

Overall, based on the available results from read across GPMT and confirmatory HRIPT, the test substance, 'di-C18-22 AAEMIM-MS' is considered to be non-sensitising to the skin.

Justification for classification or non-classification

Based on the available study results from read across GPMT and confirmatory HRIPT, the test substance, 'di-C18-22 AAEMIM-MS', does not warrant classification for skin sensitisation, according to the EU CLP criteria (Regulation 1272/2008/EC).