Disodium 2-hydroxyethyliminodi(acetate)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD TG 406, EPA TG OPPTS 870.2600, EEC Method B.6, JMAFF Agchem Test Guidelines and in accordance with the Principles of Good Laboratory prcatice (GLP)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF Agchem Test Guidelines

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes

Type of study:

Buehler test

Justification for non-LLNA method:

The study was done before in 2002 before the LLNA became the required test method

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 2-hydroxyethyliminodi(acetate)

EC Number:

205-187-4

EC Name:

Disodium 2-hydroxyethyliminodi(acetate)

Cas Number:

135-37-5

Molecular formula:

C6H9NO5.Na2

IUPAC Name:

disodium 2-hydroxyethyliminodi(acetate)

Constituent 2

Reference substance name:

Na2HEIDA

IUPAC Name:

Na2HEIDA

Details on test material:

- Name of test material (as cited in study report): XUS 40855.01
- Physical state: clear, light yellow liquid
- Composition of test material, percentage of components: active ingredient as Na2HEIDA 27.5%
- Lot/batch No.: PK2001D602
- Storage condition of test material: at room temperature

In vivo test system

Test animals

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Hilltop Lan Animals Inc., Scottdale, PA
- Age at study initiation: young adults approximately 8 -9 weeks of age
- Weight at study initiation: males: 469-506 grams and females: 451-493 grams
- Housing: The animals were housed indivdually in suspended stainless steel cages.
- Diet (e.g. ad libitum): PMI certified guinea pig chow #5026 (Purina Mills, Inc.) was provided adlibitum to the animals throughout the study.
- Water (e.g. ad libitum): Municipal tap water treated by reverse osmosis was available ad libitum throughout the study.
- Acclimation period: minimum of 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-23 °C
- Humidity (%): 32-57%
- Air changes (per hr): 10-15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Induction 2

Concentration / amount:

100% undiluted test material

Challenge

Challenge 1

Route:

epicutaneous, open

Vehicle:

unchanged (no vehicle)

Challenge 2

Concentration / amount:

100% undiluted test material

No. of animals per dose:

10 male + 10 female

Details on study design:

RANGE FINDING TESTS: The range finding test was conducted at 100, 75, 50 and 25% w/v in deionised water administered topically to 4 guinea pigs (one animal/concentration). The four conentrations were applied to clipped area of each animal. Following chamber application, the trunk of each animal was wrapped with elatic wrap and secured with adhesive tape to prevent removal of the chambers and the animal returned to its cage. Approximately 6 hours after chamber application, the binding materials were removed. The test sites were then wiped with gauze moistened with deionized water followed by dry gauze to remove test material residue and animals returned to their cages. The test sites of these animals were graded for irritation at approximately 24 and 48 hours following chamber application.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 (days 0, 7 and 13)
- Exposure period: 3 consecutive induction exposures
- Test groups: 1
- Control group: no control group
- Site: left back side of each animal
- Frequency of applications: once on days 0, 7 and 13
- Duration: once on days 0, 7 and 13
- Concentrations: A dose of 0.3 ml of the test material was placed on a 25 mm Hilltop chamber backed by adhesive tape (occlusive patch) and placed to the clipped surface.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: day 27
- Exposure period: 6 hours
- Test groups: 1
- Control group: 1
- Site: right back side of each animal
- Concentrations: 100% undiluted test material was used
- Evaluation (hr after challenge): 24 and 48 hours following chamber removal

Challenge controls:

A group of 5 male + 5 female guinea pigs served as challenge control

Positive control substance(s):

yes

Remarks:

1-Chloro-2,4-dinitrobenzene (DNCB) and alpha-Hexylcinnamaldehyde (HCA)

Results and discussion

Positive control results:

Following induction at 0.1% w/v DNCB in acetone/ethanol and challenge at levels of 0.1% and 0.05% w/v DNCB in acetone/ethanol, a contact sensitization response was observed, thereby demonstrating the susceptibility of the test system to this sensitizing agent.
Following induction at 5% w/v HCA in ethanol and challenge at levels of 2.5% and 1% w/v HCA in acetone, a contact sensitization response was observed, thereby demonstrating the susceptibility of the test system to this sensitizing agent.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

100 % undiluted

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100% undiluted

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

none

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

100% undiluted

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

100% undiluted

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

none

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the study, XUS 40855.001 is not considered to be a contact skin sensitizer in guinea pigs.

Executive summary:

The dermal sensitization potential of XUS 40855.01, which contained 27% Na2HEIDA as the active ingredient, was evaluated in Hartley-derived albino guinea pigs. Ten male and ten female guinea pigs were topically treated with 100% XUS 40855.01, once per week, for three consecutive weeks.

Following a two-week rest period, a challenge was performed whereby the twenty test and ten previously untreated (naive) challenge control guinea pigs were topically treated with 100% XUS 40855.01. Challenge responses in the test animals were compared with those of the challenge control animals.

Following challenge with 100% XUS 40855.01, dermal scores of 0 were noted in all of the test and challenge control animals. Group mean dermal scores were noted to be 0.0 in the test and challenge control animals.

Following induction at 0.1% w/v DNCB (1-Chloro-2,4-dinitrobenzene) in acetone/ethanol and challenge at levels of 0.1% and 0.05% w/v DNCB in acetone/ethanol, a contact sensitization response was observed and following induction at 5% w/v HCA (á-Hexylcinnamaldehyde ) in ethanol and challenge at levels of 2.5% and 1% w/v HCA in acetone, a contact sensitization response was observed, thereby demonstrating the susceptibility of the test system to this sensitizing agent.

Based on the results of this study, XUS 40855.01 is not considered to be a contact sensitizer in guinea pigs.