Reaction mass of 1,2-Diacetoxy-3-butene and 1,2-Dihydroxy-3-butene, monoacetate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003-04-16 - 2003-06-30

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

GLP compliance:

yes (incl. QA statement)

Remarks:

Experimental Toxicology and Ecology, BASF Aktiengesellschaft, 67056 Ludwigshafen/Rhein, Germany

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

445-630-0

EC Name:

-

Molecular formula:

Unspecified

IUPAC Name:

1-(acetyloxy)but-3-en-2-yl acetate; 2-hydroxybut-3-en-1-yl acetate

Details on test material:

- Name of test material (as cited in study report): Vinylglykoldiacetat ca. 75 %ig
- Test substance No.: 02/0526-1
- Physical state: Liquid / colorless, clear
- Analytical purity: 74.8 area% (73.3 g/100 g)
- Other constituents:
4-Acetoxy-3-hydroxy-1-butene and 2-Acetoxy-1-hydroxy-1-butene: 17.3 area-%.(17.0 g/100 g)
3,4-Diacetoxy butane: 2.0 area-% (2.0 g/100 g)
1-Acetoxy-2-hydroxy butane: 0.8 area-% (0.8 g/100 g)
2-Acetoxy-1-hydroxy butane : 0.4 area-% (0,3 g/100 g)
4-Acetoxy crotonaldehyd: 1.8 area-% (1.8 g/100 g)
- Lot/batch No.: Kanister 1, 2, 4, 5
- Date of production: Oct. 21, 2001
- Stability under test conditions: The stability under storage conditions was confirmed by reanalysis
- Storage condition of test material: Room temperature, exclusion of oxygen (storage under nitrogen)

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Hsd Poc: DH (SPF)

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen, FRG
- Identification: ear tattoo
- Age at study initiation: about 5 weeks
- Weight at study initiation: 255 g – 321 g
- Housing: 5 per cage in stainless steel wire mesh cages with plastic-coated grating, minimum floor area: 2,000 cm2
- Diet (e.g. ad libitum): Kliba Labordiät (Kaninchen/ Meerschweinchen-Haltungsdiät) Provimi Kliba SA, Kaiseraugst, Switzerland; ad libitum
- Water (e.g. ad libitum): Tap water; ad libitum
- Acclimation period: 6 days before the first test substance application.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12 / 12

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

olive oil

Induction 2

Concentration / amount:

Intradermal induction: 5%
Epicutanous induction: 100%
Challenge: 100%

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

olive oil

Challenge 2

Concentration / amount:

Intradermal induction: 5%
Epicutanous induction: 100%
Challenge: 100%

No. of animals per dose:

10 (test group), 5 (control group)

Details on study design:

RANGE FINDING TESTS:
Intradermal Pretest:
- Amount applied: 6 intradermal injections in groups of two per animal were applied to each animal.
A) front row: 2 injections each of 0.1 ml Freund's adjuvant without test substance emulsified with 0.9% aqueous NaCI-solution in a ratio of 1 : 1
B) middle row: 2 injections each of 0.1 ml of a test substance formulation in an appropriate vehicle at the selected concentration
C) back row: 2 injections each of 0.1 ml Freund's adjuvant / 0.9% aqueous NaCI-solution (1 : 1) with test substance at the selected concentration.
- Site of application: neck region
- No. of animals: 2 per test substance concentration
- Reading: 24 h after the beginning of application

Epicutaneous Pretest:
- Amount applied: 2 x 2 cm gauze patches (6 Iayers surgical gauze Ph. Eur. from Lohmann GmbH & Co. KG) containing 0.5 ml of the test substance were applied to the skin of the flanks under an occlusive dressing. The dressing consisted of rubberized linen patches (4 x 4 cm from Russka), patches of Idealbinde (5 x 5 cm from Pfälzische Verbandstoff-Fabrik) and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG.
- Duration of exposure: 24 h
- Site of application: flank
- Number of test animals: 3
- Readings:1 h, 24 h and 48 h after removal of the patches


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 6 intradermal injections in groups of two per animal were given:
A) front row: 2 injections each of 0.1 ml Freund's adjuvant without test substance emulsified with 0.9% aqueous NaCI-solution in a ratio of 1 : 1
B) middle row: 2 injections each of 0.1 ml of a test substance formulation in an appropriate vehicle at the selected concentration
C) back row: 2 injections each of 0.1 ml Freund's adjuvant / 0.9% aqueous NaCI-solution (1 : 1) with test substance at the selected concentration.
- Epicutaneous induction was carried out one week after intradermal induction:
2 x 4 cm gauze patches (6 Iayers surgical gauze Ph. Eur. from Lohmann GmbH & Co. KG) containing 1 ml of test substance were applied to the skin of the neck region under an occlusive dressing for 48 h. The dressing consisted of rubberized linen patches (4 x 6 cm from Russka) and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG.
- Test group: 10 animals
- Control group: 5 animals



B. CHALLENGE EXPOSURE
- No. of exposures: 1, 2 x 2 cm gauze patches (6 layers surgical gauze Ph. Eur. from Lohmann GmbH & Co. KG) containing the test substance were applied to the skin of the flank under an occlusive dressing. The dressing consisted of rubberized linen patches (4 x 4 cm from Russka), patches of Idealbinde (5 x 5 cm from Pfälzische Verbandstoff-Fabrik) and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG.
- Day(s) of challenge: 14 days after the epicutaneous induction.
- Exposure period: 24 h
- Test group: 10 animals
- Control group: 5 animals
- Site: flank
- Concentrations: 0.5 ml, 100%
- Evaluation (hr after challenge): 24 and 48 h after the removal of the patch

EVALUATION
The number of animals with skin findings at 24 and/or 48 hours after the removal of the patch was taken into account for the determination of the sensitization rate. The evaluation "sensitizing" results if at least 30 per cent of the test animals exhibit skin reactions in this adjuvant test.
The evaluation is based an the criteria of Annex VI of the Commission Directive 67/548/EEC that were in place an the date of report signature.

Positive control substance(s):

no

Remarks:

A positive control (reliability check) was performed as separate study twice a year in the laboratory using alpha-hexylcinnamaldehyde techn. 85%. The test system was shown to be able to detect sensitizing compounds under the test conditions chosen.

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:

5

Total no. in group:

10

Clinical observations:

discrete or patchy erythema in 2 test group animals; moderate and confluent erythema in 2 test group animals; moderate and confluent erythema and dry skin in 1 test group animal

Remarks on result:

other: see Remark

Remarks:

Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100%. No with. + reactions: 5.0. Total no. in groups: 10.0. Clinical observations: discrete or patchy erythema in 2 test group animals; moderate and confluent erythema in 2 test group animals; moderate and confluent erythema and dry skin in 1 test group animal.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100%

No. with + reactions:

4

Total no. in group:

10

Clinical observations:

discrete or patchy erythema and scaling in 3 test group animals; moderate and confluent erythema and scaling in 1 test group animal

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100%. No with. + reactions: 4.0. Total no. in groups: 10.0. Clinical observations: discrete or patchy erythema and scaling in 3 test group animals; moderate and confluent erythema and scaling in 1 test group animal.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

100%

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 5.0.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

100%

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 5.0.

Any other information on results incl. tables

Additional results:

- the test substance caused a yellow-brown discoloration of the application area, however this did not impair evaluation of erythema formation.

- body weights: The expected body weight gain was generally observed in the course of the study.