Reaction mass of 3-[(2-aminoethyl)ammonio]propane-1-sulfonate and 3,3'-(ethane-1,2-diyldiammonio)dipropane-1-sulfonate and ethane-1,2-diamine

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

May 12, 1981

Deviations:

no

Principles of method if other than guideline:

according to the OECD guideline 406 with particular reference to the following publications by B. Magnusson and A.M. Kligman: "The identification of contact allergens by animal assay. The quinea pig maximization test" (J. Invest. Dermatol., 52, 268-276, 1969) and "Allergic Contact Dermatitis in the Guinea Pig" (Thomas Publisher, Springfield, Illinois, 1970)

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

At the time of study completion, the LLNA method was not adopted.

Test material

Test material information

Constituent 1

Reference substance name:

3-[(2-aminoethyl)amino]propane-1-sulfonic acid

Cas Number:

14235-54-2

Molecular formula:

C5H14N2O3S

IUPAC Name:

3-[(2-aminoethyl)amino]propane-1-sulfonic acid

Details on test material:

- Name of test material (as cited in study report): EPS
- Physical state: yellowish-brown, viscous liquid
- Analytical purity: > 99 %
- Lot/batch No.: LA 92018 SC
- Stability under test conditions: at least 2 years
- Storage condition of test material: ambient

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Pirbright white

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Firma Winkelmann, Versuchstierzucht, Gartenstr. 27, 4799 Borchen, Germany
- Weight at study initiation: male: 305-498 g; female: 304-378 g
- Housing: collective housing up to a maximum of 5 animals per cage (Makrolon type IV)
- Diet: ad libitum; Ssniff-G, pellets, 1.0 cm long, 0.5 cm diameter
- Water: ad libitum; Makrolon drinking bottles, drinking water as for human consumption
- Acclimation period: 13 days (range finding), 8 days (main test, females), 22 days (main test, males)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 °C
- Humidity (%): 30- 70 %
- Photoperiod (hrs dark / hrs light): artificial lightning (120 lux) from 7.00 a.m. - 7.00 p.m.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

water

Induction 2

Concentration / amount:

induction exposure: 1 % in aqua ad iniectabilia
challenge exposure: 100 %

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

water

Challenge 2

Concentration / amount:

induction exposure: 1 % in aqua ad iniectabilia
challenge exposure: 100 %

No. of animals per dose:

20 test and 20 control animals

Details on study design:

RANGE FINDING TESTS:
Intradermal Injection: final concentration of the test article was 5% in aqua ad iniect. and Freund's complete adjuvant (FCA). Since the highest permissible concentration produced redness, lower concentration (2.5 and 1 %) were tested. Six animals were employed, skin reactions being recorded 48 h after treatment.
Dermal Appliction: The test article was used undiluted (100%). Occlusive bandage was used. Two animals were employed and skin reactions were recorded 48 h post applicationem.

MAIN STUDY
A. INDUCTION EXPOSURE
First stage:
Three pairs of intradermal injections of 0.1 ml volume were made symmetrically in two rows on either side (clipped short with electric clippers and cleaned with 70 % (v/v) ethanol) of the spine
Test group:
Injection 1: Freund's complete adjuvant (FCA) 50 % (w/w) diluted in aqua ad iniect.
Injection 2: the test article diluted in aqua ad iniect. (final concentration: 1 %)
Injection 3: the test article diluted in FCA/aqua ad iniect. (final concentration: 1 %)
Control group:
Injection 1: FCA 50 % (w/w) diluted in aqua ad iniect.
Injection 2: aqua ad iniect.
Injection 3: aqua ad iniect. diluted 50 % (w/w) in FCA
Second stage: 7 days after the intradermal injections dermal application was initiated.
The area was reclipped and pretreated with 10% sodium lauryl sulphate in vaseline, 24 hours before application of the test article at a concentration of 100 %. The test article was spread in a thick layer (to saturation) over a 4 x 5 cm patch (filter paper). The latter was firmly secured over the previous injection sites by an occlusive dressing for 48 h. Control animals received a patch loaded with the vehicle alone.

B. CHALLENGE EXPOSURE
- No. of exposures: one
- Day(s) of challenge: 14 days after the second stage of induction
- Exposure period: 24 h
- Test animals: 100 % of the test article on the left flank
- Control animals: vehicle on the right flank
- Site: 5 x 5 cm clipped and shaved area on each flank
- Concentrations: maximal non-irritating concentration (100 %)
- Evaluation (hr after challenge): 24 and 48 h after patch removal

Positive control substance(s):

yes

Remarks:

2,4 dinitrochlorobenzene and benzocaine

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

100 %

No. with + reactions:

3

Total no. in group:

20

Clinical observations:

erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100 %. No with. + reactions: 3.0. Total no. in groups: 20.0. Clinical observations: erythema.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100 %

No. with + reactions:

1

Total no. in group:

20

Clinical observations:

erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100 %. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: erythema.

Results 3

Reading:

other: total

Group:

test chemical

Dose level:

100 %

No. with + reactions:

3

Total no. in group:

20

Clinical observations:

erythema

Remarks on result:

other: Reading: other: total. Group: test group. Dose level: 100 %. No with. + reactions: 3.0. Total no. in groups: 20.0. Clinical observations: erythema.

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information mild

Conclusions:

Sensitization rate (%) at 24 h: 15
Sensitization rate (%) at 48 h: 5

Executive summary:

The potential skin sensitizing properties of "EPS" were assessed in the guinea pig maximization test using 20 test and 20 control animals. Following induction exposure to the test article or the vehicle, the animals were subjected two weeks later to challenge exposure with the test article. Allergic responses to the challenge procedure were evaluated 24 and 48 h after the end of the exposure period.

According to Magnusson and Klingman (J. Invest. Dermatol., 52, 268 -276, 1969), the test article "EPS" may be classified as a "mild sensitizer".