Reaction products of fatty acid chlorides, C8-12 (even numbered) with glycine and sodium hydroxide

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

The study was performed prior to the establishment of LLNA.

Test material

Test material information

Constituent 1

Reference substance name:

Sodium cocoyl glycinate (SCG) [INCI]

IUPAC Name:

Sodium cocoyl glycinate (SCG) [INCI]

Details on test material:

- Physical state: liquid
- Stability under test conditions: stable at room temperature
- Storage condition of test material: At room temperature (range of 20 ± 5 °C), light protected.

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland GmbH
- Age at study initiation: 5-6 weeks
- Weight at study initiation: 282 – 388 g
- Housing: Individually in Makrolon type-4 cages with standard softwood bedding
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 guinea pig breeding / maintenance diet, ad libidum
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 2008-09-18 to 2008-10-13


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 / 12

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

water

Concentration / amount:

5 % / 0.1 ml

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

50% /0.3 ml

Day(s)/duration:

24h

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Challenge

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

5% /0.2 mL

Day(s)/duration:

24h

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

5 control animals (males)
10 treated animals (males)

Details on study design:

RANGE FINDING TESTS:
A: Intradermal injections:
Four intradermal injections (0.1 mL/site) of a 1:1 (v/v) mixture of Freund's Complete Adjuvant/physiological saline (day 1) (shaved neck of one guinea pig)
Five days later: Four intradermal injections (0.1 mL/site) at concentrations of H = 15 %, I = 10 % and J = 5 % of the test item in purified water (clipped flank of the same guinea pig)
Dermal reactions assessed 24 hrs later.

B: Epidermal applications:
Four intradermal injections (0.1 mL/site) of a 1:1 (v/v) mixture of Freund's Complete Adjuvant/physiological saline (day 1) (shaved necks of two guinea pigs)
Five days later: Epidermal application with the test item at K = 15 %, L = 10 %, M = 5 % and N = 3 % in purified water (shaved flanks of the same two guinea pigs)
Dermal reactions assessed 24 and 48 hrs after removal of the bandage.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal / epicutaneously)
- Exposure period: injected / 48 h occlusive
- Site: scapular region (clipped free of hair)
- Frequency of applications: 1 / 1
- Duration: 0 - 8 days
- Concentrations:
1st application: 5% intradermal; 2nd application: 50% epicutaneous

B.
CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
-Exposure period: 24 hrs
- Site: left flank (test item); right flank: vehicle (clipped free of hair)
- Concentrations: 5% occlusive epicutaneous
- Evaluation (hr after challenge): 24 and 48 hrs

Positive control substance(s):

yes

Remarks:

ALPHA-HEXYLCINNAMALDEHYDE

Results and discussion

Positive control results:

Discrete/patchy to moderate/confluent erythema with or without scaling was observed in nine out of ten test animals at the 24- and 48-hour reading after the challenge treatment with ALPHA-HEXYLCINNAMALDEHYDE at 1 % in PEG 300 (left shoulder). Five test animals showed discrete/patchy erythema at the 24-hour reading after treatment with ALPHAHEXYLCINNAMALDEHYDE at 0.1 % in PEG 300 (left flank). No skin effect was observed in the control group.
Based on the above mentioned findings in an adjuvant sensitization test (M&K-test) in guinea pigs and in accordance to Commission Directive 2001/59/EC, ALPHAHEXYLCINNAMALDEHYDE has to be classified and labelled as a skin sensitizer.

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

5%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

no indication of skin sensitisation

Results 2

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

5 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

no indication of skin sensitisation

Results 3

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Results 4

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: none.

Any other information on results incl. tables

 

Skin reactions in the intradermal pretest
Animal No.	Concentration (%)	Reactions
671, male	75	Moderate and confluent erythema Oedema and necrosis
	50	Moderate and confluent erythema Oedema and necrosis
	25	Moderate and confluent erythema Oedema and necrosis
689, male	15	Moderate and confluent erythema Necrosis
	10	Moderate and confluent erythema Necrosis
	5	Discrete/patch erythema

 

Skin reactions in the intradermal pretest
Animal No.	Concentration (%)	Reactions after removal of badange
		24h	28h
672, male	100	Intense erythema and swelling; Fissures	Moderate and confluent erythema; Scaling
	75	Intense erythema and swelling; Fissure	Moderate and confluent erythema; Scaling
	50	Moderate and confluent erythema	Moderate and confluent erythema; Scaling
	25	Discrete/patchy erythema	No visible change
673, male	100	Intense erythema and swelling; Fissures	Moderate and confluent erythema; Scaling
	75	Intense erythema and swelling; Fissures	Moderate and confluent erythema; Scaling
	50	Discrete/patchy erythema	No visible change
	25	Discrete/patchy erythema	Discrete/patchy erythema
690, male	15	Discrete/patchy erythema	No visible change
	10	No visible change	No visible change
	5	No visible change	No visible change
	3	No visible change	No visible change
691, male	15	Discrete/patchy erythema	No visible change
	10	Discrete/patchy erythema	No visible change
	5	No visible change	No visible change
	3	No visible change	No visible change

 

 

Skin reactions after epidermal induction of with test item treated animals
Animal No.	Reactions after removal of badange
	24h	48h
679	Discrete/patchy erythema	Discrete/patchy erythema
680	Discrete/patchy erythema	Discrete/patchy erythema
681	Discrete/patchy erythema	No visible change
682	Discrete/patchy erythema	Discrete/patchy erythema
683	Discrete/patchy erythema	Discrete/patchy erythema
684	Discrete/patchy erythema	Discrete/patchy erythema
685	Discrete/patchy erythema	No visible change
686	Discrete/patchy erythema	Discrete/patchy erythema
687	Moderate and confluent erythema	Discrete/patchy erythema
688	Discrete/patchy erythema	Discrete/patchy erythema

 

 

Skin reactions after epidermal challenge of with test item treated animals
Animal No.	Reactions after removal of badange
	24h	48h
679	No visible change	No visible change
680	No visible change	No visible change
681	No visible change	No visible change
682	No visible change	No visible change
683	No visible change	No visible change
684	No visible change	No visible change
685	No visible change	No visible change
686	No visible change	No visible change
687	No visible change	No visible change
688	No visible change	No visible change

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Hostapon SG was investigated for its skin isensitization property according to the Guideline OECD 406 (M&K). It is not skin sensitizing.

Executive summary:

Hostapon SG was investigated for its skin sensitization property according to the OECD 406 (M&K). As for induction, ten male guinea pigs were treated intradermally at concentration of 5% in water and one week later epidermally at concentration of 50 % in water. Five control animals were treated with vehicle only. Two weeks after the epidermal induction, all animals were epidermally challenged at concentration of 5%. None of the animals responded. Hostapon SG is not skin sensitizing.