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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
supporting study
Study period:
no data
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Old study (1947), no guideline. Protocol different from standard method even if few method details were  reported. HMD tested as aqueous solution at 2% and 1%
Cross-reference
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1947

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
The aim of this study was to analysed the skin sensitisation potential of Hexamethylene Diamine (HMD) at 2% and 1%. Experiment was realised with 10 Guinea Pig.
A 2% HMD aqueous solution was applicated (Epicutaneous route) of clipped Guinea Pig.Three successive applications were performed the first day on the same area . Then, six successive subcutaneous injections of solution of 1% HMD were performed on the left side through 13 days followed by a 8-day rest period.Then a final patch application was performed as the beginning with 1% and 2% HMD aqueous solution. Skin lesions were recorded after patch removal. Finally, a seventh final test substance subcutaneous injection was administered to the animals.
Finally Guinea Pigs were sacrified with Illum gas two days after final injection and a gross-autopsy was performed for some animals.
Skin reactions were analysed and weight measured for approximatively one month through both treatment period.
GLP compliance:
no
Remarks:
Study performed before GLP establishment
Type of study:
intracutaneous test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Hexamethylene Diamine at 2% and 1%

In vivo test system

Test animals

Species:
guinea pig
Strain:
not specified
Sex:
not specified
Details on test animals and environmental conditions:
No data

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Induction: aqueous solution of 2% hexamethylene diamine (epicutaneous route) and 1% (intradermal route)
Challenge: aqueous solution of 2% hexamethylene diamine (epicutaneous route) and 1% (intradermal route)
Challengeopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Induction: aqueous solution of 2% hexamethylene diamine (epicutaneous route) and 1% (intradermal route)
Challenge: aqueous solution of 2% hexamethylene diamine (epicutaneous route) and 1% (intradermal route)
No. of animals per dose:
10
Details on study design:
No additional data
Challenge controls:
No data
Positive control substance(s):
no

Study design: in vivo (LLNA)

Statistics:
No data

Results and discussion

Positive control results:
No data

Any other information on results incl. tables

Table 7.2.1 : Weight evolution of Guinea Pigs after both treatments

Time after treatment started

#G.Pig

# treatment

% gain or loss in weight

0 hours

10

Initial Patch

-

Wed ¿ 16-29-47 Subcu. Inj. started

72 hours

10

1

+4.77%

6 days

10

3

+5.20%

10 days

10

5

+6.73%

13 days

10

6

+4.05%

11-11 to 11-19 inc. Rest Period

17 days

10

Rest period

+6.63%

20 days

10

Rest period

+3.70%

Thurs 11-20 Final Patch

24 days

10

Final Patch

+0.55%

27 days

10

Final sub-inj.

+6.83%

28 days

10

Final sub-inj.

-2.29%

29 days

10

Final sub-inj.

+0.39%

Applicant's summary and conclusion

Interpretation of results:
other: no sensitisation after final patch application
Conclusions:
Under the test conditions, no skin sensitisation was observed in any animals after final patch application. Necrosis was observed 24-hour after the last injections following the final patch application.
Executive summary:

The skin sensitisation potential of Hexamethylene Diamine (HMD) at 2% and 1% was tested (Hardy, 1947b). Experiment was realized with 10 Guinea Pig.

A 2% HMD aqueous solution was applicated (epicutaneous route) on back (near shoulder) of clipped Guinea Pig.Three successive applications were performed the first day on the same area . Then, six successive subcutaneous injections of solution of 1% HMD were performed on the left side through 13 days followed by a 8-day rest period. Then the challenge (epicutaneous and intradermal routes) was performed with 2% and 1% HMD aqueous solution. Skin reactions were analysed. Finally Guinea Pigs were sacrified with Illum gas two days after final injection (challenge) and a gross-autopsy was performed for some animals. The body weight gain was measured.

Animals showed necrotic area at site of injection of HMD 3 days after treatment. Swelling occured at site of injection at 6 days post treatment. Gross-autopsy was performed for three Guinea Pig.

Skin between shoulders appear normal. Site of last injection marked by area of hemmorhage and necrosis, first and second red area on overlying skin surface. Internal organs grossly normal.

After initial patch application, within ten minutes, G. Pigs began hopping around cage definite erythema on left side. One hour after application, HMD showed signs of becoming necrotic + dark brown. Three hours post application, application site was definitely necrotic dark brown. At the end of the experiment, necrotic areas beginning to slough off slightly.

Under the test conditions, necrosis was observed 24-hour after the last injections following the final patch application. Based on these results, no conclusion can be made.