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Description of key information

The substance is not considered to be a skin sensitiser.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Study period:
June 1982
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: well conducted and reported. No information about GLP.
Justification for type of information:
See attached (in chapter 13 of IUCLID) document with the justification for the category/read-across approach.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
not specified
Type of study:
Buehler test
Justification for non-LLNA method:
Exiisting study conducted in 1982.
Specific details on test material used for the study:
- AEPD Lot No. 6G28DF18
Species:
guinea pig
Strain:
Hartley
Sex:
male
Details on test animals and environmental conditions:
Animals purchased at least 1 week prior to study and allowed to acclimatise. Healthy animals free of disease were selected for the study. 24 hours prior to test, animals examined for skin injury. Those with no injury were used in test. Animals were fed Purina Certified Rabbit Chow, ad libitum, certified free of contaminants by supplier. Animals were given tap water ad libitum. Water was tested to ensure levels of contaminants are equivalent to or less than recommended levels as per the primary drinking water regulations.
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
0.5 ml of a 0.5% aqueous solution of AEPD (first 5 applications); 0.5 ml of 0.05% aqueous solution of AEPD for the final 5 applications
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
0.5 ml of a 0.5% aqueous solution of AEPD (first 5 applications); 0.5 ml of 0.05% aqueous solution of AEPD for the final 5 applications
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
10
Details on study design:
Fifty male guinea pigs weighing 250-300 g were divided into five groups of 10 each. The animals ' backs and flanks were shaved free of hair. Group I (treatment group) was topically treated with 0.5 ml of 0.5% P-1050 aqueous solution and covered with a gauze under an occlusive patch. Group IV (positive control) was similarly treated with 0.5 ml of 0.3% dinitrochlorobenzene solution (DNCB, solubilized in a minimum volume of alcohol and made to volume with saline). Groups V, VII, and VIII (negative controls) were similarly treated with 0.5 ml of saline. After 24 hours exposure the patches were removed, the treated skin sites were cleaned and scored at 24 and 48 hours for erythema and edema according to Draize (Draize, J.H., "Appraisal of the Safety' of Chemicals in Foods, Drugs, and cosmetic. Assoc. of Food & Drug Officials of the United States, p. 48, 1957).

At 48 hours the topical application procedure was repeated with each group of animals two to three times a week until a total of 10 applications have been made. After the last application, the animals were allowed to rest for two weeks. On the first day of the third week (or the 36th day after the first application), the animals in each group were challenged as follows: Group I and Group V animals were challenged only with 1% and 0.05% of P-1050. Group IV and Group VIII animals were challenged with 0.3% DNCB solution solubilized in acetone instead of alcohol. Group VII (a negative control) was challenged with saline. After 24 h exposure, the patches were removed and the sites cleaned. At this time the challenge sites were depilated with "Nair" .Three hours after removal of the hair the challenge sites were scored for inflammatory skin reactions (erythema and edema). These sites are scored again at 48 h.
If the test material at challenge induces skin reactions in a large number of treatment group animals compared to the negative control, then the material is considered a sensitizer. The positive control group serves as an internal control for the test.
Challenge controls:
Negative control:
Saline treated (group VII) - saline treated throughout induction and challenged with saline
Irritation controls
Group V: treated with saline during induction, challenged with AEPD
Group VIII: treated with saline during induction, challenged with DNCB
Positive control substance(s):
yes
Remarks:
dinitrochlorobenzene (DNCB), 0.3% solution
Positive control results:
Positive control responded as expected witha clear sensitising response at 24 and 48 hours (8 out of 10 animals)
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
0.05%
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: 0.05%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
1%
No. with + reactions:
4
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1%. No with. + reactions: 4.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
0.05%
No. with + reactions:
1
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.05%. No with. + reactions: 1.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
1%
No. with + reactions:
1
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1%. No with. + reactions: 1.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.3%
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.3%. No with. + reactions: 8.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.3%
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.3%. No with. + reactions: 8.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: Irritation Control for AEPD
Dose level:
0.05%
No. with + reactions:
6
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: Irritation Control for AEPD. Dose level: 0.05%. No with. + reactions: 6.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Irritation Control for AEPD
Dose level:
0.05%
No. with + reactions:
6
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: Irritation Control for AEPD. Dose level: 0.05%. No with. + reactions: 6.0. Total no. in groups: 10.0.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
other: Irritation Control for AEPD
Dose level:
1%
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: Irritation Control for AEPD. Dose level: 1%. No with. + reactions: 8.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Irritation Control for AEPD
Dose level:
15%
No. with + reactions:
6
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: Irritation Control for AEPD. Dose level: 15%. No with. + reactions: 6.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: Irritation control for DNCB
Dose level:
0.3%
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: Irritation control for DNCB. Dose level: 0.3%. No with. + reactions: 2.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Irritation Control for DNCB
Dose level:
0.3%
No. with + reactions:
5
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: Irritation Control for DNCB. Dose level: 0.3%. No with. + reactions: 5.0. Total no. in groups: 10.0.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
saline
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: saline. No with. + reactions: 0.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
saline
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: saline. No with. + reactions: 0.0. Total no. in groups: 10.0.

Based on the results of the study, AEPD was irritating but not a sensitiser

Interpretation of results:
not sensitising
Conclusions:
AEPD was not sensitising, but a skin irritant (under the conditions of the study)
Executive summary:

A topical contact sensitization test was conducted in the m a l e guinea pigs according to Buehler's procedure (Buehler, E. V. "Delayed Contact Hypersensitivity in the Guinea Pig", It Arch. Dermat. -91: 171-175, 1965). Fifty male guinea pigs weighing 250-300 g were divided into five groups of 10 each. The animals ' backs and flanks were shaved free of hair. Group I (treatment group) was topically treated with 0.5 ml of 0.5% P-1050 aqueous solution an8 covered with a gauze under an occlusive patch. Group IV (positive control) was similarly treated with 0.5 rnl of 0.3% dinitrochlorcbenzene solution (DNCB, solubilizcd in a minimum volume of alcohol and made to volume with saline). Groups V, VfI, and VIII (negative controls) were similarly treated with 0.5 ml of saline. After 24 hours exposure the patches were removed, the treated skin sites were cleaned and scored at 24 and 48 hours for erythema and edema according to Draize. At 48 hours the topical application procedure was repeated with each group of animals two to three times a week until a total of 10 applications have been made.

In the initial test all the animals in the test and the negative controls developed skin rashes and the skin sensitization reactions could not be evaluated. The topical sensitization test was repeated with a new batch of animals. During the induction period (initial ten applications) some of the animals in Group 1 showed mild erythema when treated with 0.5% solution of P-1050, so the last five applications were made with 0.05% solution, The animals in Group IV (DNCB) showed mild skin reactions during the entire induction period, When challenged with 0.05% solution of P-1050, one animal in Group I (treatment) showed skin reactions at 48 h and six animals in Group V (negative control) showed skin reactions at 24 and 48 h. A rechallenge with 1% solution of P-1050, elicited skin reactions in more animals of the negative control group (eight) than of the treatment group (four). A t challenge with 0.3% DNCB solution, more of the animals in the positive control (Group IV) showed skin reactions than in the negative,control (Group VIII) group of animals at 24 and 48 hous. The negative control (Group VII) did not show any skin reactions with saline.

Endpoint:
skin sensitisation: in vitro
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail
Principles of method if other than guideline:
To test direct peptide reactivity which is a key pathway leading to skin sensitisation, the test substance was investigated for peptide depletion by chemical reaction. The assay method established by Natsch and Gfeller (2008) was validated and improved in the testing facility and utilised in this study.
GLP compliance:
yes
Type of study:
other: peptide binding assay
Key result
Parameter:
other: AMPD was tested for peptide reactivity by monitoring peptide depletion. Under the conditions used in this study, there is no evidence of that AMPD contains direct protein reactivity that can result in skin sensitization.
Remarks on result:
no indication of skin sensitisation

The test substance was completely soluble in acetonitrile and was not precipitated by mixing with peptide solutions. After 24 h incubation, there was no colour change or a precipitate observed from the test substance. The test article did not have any UV absorbance at 220 nm through entire HPLC chromatography and therefore there was no interference with HPLC-UV analyses for peptides. Furthermore, the test substance did not interfere with the MS detections used in the test system that were monitoring higher than 700.0 m/z. Using the established calibration curve, the concentrations of free peptide were calculated for each sample (Table 1). Average peptide depletion by the test substance was 4.22 ± 1.84%. Negative and positive controls resulted in 4.83 ± 1.66% and 96.13 ± 0.21% peptide depletion, respectively. These results confirmed the assay was valid. Because there was no peptide depletion by the test substance, no further analysis was performed to measure dimerized- or oxidized-peptide by the test substance.

 

Table 1: Individual data from free peptide quantitation and average peptide depletion

Group

 Replicate#

Analyte Peak Area (counts)

 Peptide conc. (mM)

Peptide depletion (%)

 Average depletion (%)

Test substance

1

15300000

98.03

1.97

4.22 ± 1.84

2

15200000

97.37

2.63

3

14700000

94.07

5.93

4

14700000

94.07

5.93

5

14900000

95.39

4.61

Negative control

1

14600000

93.40

6.60

4.83 ± 1.66

2

14900000

95.39

4.61

3

15100000

96.71

3.29

Positive control

1

1030000

3.78

96.22

96.13 ± 0.21

2

1080000

4.11

95.89

3

1020000

3.71

96.29
Interpretation of results:
not sensitising
Conclusions:
Under the test conditions, there is no evidence that the test substance contains direct protein reactivity which would cause skin sensitisation.
Executive summary:

Direct peptide reactivity is a key component of the pathway leading to skin sensitisation, XU-12398.00 (AMPD) was tested for peptide reactivity by monitoring peptide depletion by chemical reactions using the assay method established by Natsch and Gfeller (2008) with minor modifications. After a 24-hour incubation with one tenth molar ratio of the standard peptide, XU-12398.00 (AMPD) resulted in minimal peptide depletion, which was within the range of the negative control. The positive control substance depleted most of free peptides. Under the conditions used in this study, there is no evidence of that XU-12398.00 (AMPD) contains direct protein reactivity that can result in skin sensitisation.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Burnett et al. (2009) reviewed the sensitisation potential of cosmetic formulations containing AMPD in human test subjects using a repeated insult patch test (RIPT). 0.073% AMPD was tested in a group of 30 human test subjects and was applied to the arm daily, 4 days per week for 2 weeks, alternating arms daily. In additon, an occlusive patch was applied on the first day of the test. After the 2 -week application period, there was a 2 -week nontreatment period. After this period, the test subjects were challenged with a reapplication of the formulation to the test site along with an occlusive patch at an adjacent site. The original patch, challenge patch, and open challenge test sites were read at 24, 48 and 96 h. No reaction were observed in any of the test subjects. The formulation containing 0.073% AMPD was neither a primary irritant nor a senistiser, and the fomulation was safe under the conditions of the study. A modified RIPT of a cosmetic formulation containing 0.5% AMPD was performed in a group of 39 women and 20 men. The test material (0.5 mL) was applied to a semiopen patch on the arm of each test subject ever Monday, Tuesday, Wednesday, and Thursday for 2 weeks. The patch sites were graded approx. 24 h after application. In addition, a closed patch was applied to each test subject on the first day of the study and on the day of challenge. No patches were applied for 2 weeks after the induction phase. On the Monday following the nontreatment period, challenge patches were applied to the original test site and an adjacent site; the second closed patch was also applied at this time. The challenge sites were graded 1, 2, and 4 days after application. Slight erythema was noted at 1 adjacent application site at each of the grading times, but it was not clear whether these reaction occurred in the same test subject. Under the test conditions, the cosmetic formulation containing 0.5% AMPD was not a sensitiser.

The irritation and sensitisation potential of a mascara containing 1.92% AMPD using113 test subjects. A semioccluded patch was used to apply 0.2 mL of the test material to the interscapular region of the subjects, and the patch was affixed 24 h before removal. The induction phase consisted of patch applications 3 times a week for a total of 9 applications. Following a 2 -week nontreatment period, a challenge patch was applied to a virgin site adjacent to the induction patch site. The challenge patch was removed after 24 h, and the site was scored for reaction at 24 and 72 h post application. No indication of potential dermal irritation or allergic contact sensitisation by the cosmetic product containing AMPD was observed.

Reference (not cited in IUCLID)

Burnett, C.L. et al. (2009) Final Amended Report on Safety Assessment on Aminomethyl Propanol and Aminomethyl Propanediol. Internation Journal of Toxicology. 28, 141S-161S

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification