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Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2001-11-29 to 2002-04-10
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
Chlorine dioxide (ClO2) is an unstable gas which must be stabilized with water. After skin contact, the ClO2 in aqueous solution is rapidly reduced in chlorite and chloride (see § 7.1). Therefore, it is considered as relevant to use the structurally close metabolite, sodium chlorite, to evaluate the sensitising potential of ClO2 according to a read-across approach.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2002
Report date:
2002

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
and EU Method B.6 (Skin Sensitisation)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Test was performed in 2002. LLNA method (OECD 442 B) was adopted in 2010 and also the (OECD tests n°442 C , D) was adopted in 2015.

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium chlorite
EC Number:
231-836-6
EC Name:
Sodium chlorite
Cas Number:
7758-19-2
Molecular formula:
ClHO2.Na
IUPAC Name:
sodium chlorite
Constituent 2
Reference substance name:
The test material is a 31 % aqueous solution of sodium chlorite
IUPAC Name:
The test material is a 31 % aqueous solution of sodium chlorite
Details on test material:
- Test material: Sodium chlorite (31% solution)
- Lot/Batch No: 6624 (Atofina)
- Physical state: colorless liquid
- Purity: 31.12 %
- Preparation of test substance for application:
a) for induction: In 0.9% NaCl
b) for challenge: In 0.9% NaCl
- Stability under test conditions: no data
- Storage condition of test material: at room temperature and protected from light

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Saint-Aubin-les-Elbeuf, France.
- Age at study initiation: 1 – 3 months old
- Weight at study initiation: Mean body weight males: 380 ± 31 g; females: 355 ± 13 g.
- Housing: housed individually in polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days before the beginning of the study.

-other: sex: Male and female (nulliparous and non-pregnant)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22° ± 2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12h dark / 12h light

IN-LIFE DATES: no data

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Concentrations used for induction:
- intradermal injections (day 1): 0.031 % sodium chlorite (i.e. a 0.1 % solution of a 31 % solution of sodium chlorite)
- topical application (day 8): 1.55% sodium chlorite

Concentrations used for challenge:
- 0.31 % sodium chlorite (i.e. a 1 % solution of a 31 % solution of sodium chlorite)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Concentrations used for induction:
- intradermal injections (day 1): 0.031 % sodium chlorite (i.e. a 0.1 % solution of a 31 % solution of sodium chlorite)
- topical application (day 8): 1.55% sodium chlorite

Concentrations used for challenge:
- 0.31 % sodium chlorite (i.e. a 1 % solution of a 31 % solution of sodium chlorite)
No. of animals per dose:
Thirty guinea pigs were allocated to two groups: a control group of five males and five females and a treated group of ten males and ten females.
Details on study design:
RANGE-FINDING TESTS:
A preliminary test was conducted in order to determine the concentrations to be tested in the main study.
By intradermal route (tested concentrations: 5%, 1% and 0.1% (w/w), i.e. 1.55%, 0.31% and 0.031% sodium chlorite):
- 24 hours before treatment, the dorsal region of the animals was clipped,
- intradermal injections of the dosage form preparations (0.1 mL) were performed in the interscapular region,
- cutaneous reactions were evaluated approximately 24, 48 hours and 6 days after the injections.
By cutaneous route (tested concentrations: 100%, 50%, 10%, 5%, 1% and 0.5% (w/w), i.e. 31%, 15.5%, 3.1%, 1.55%, 0.31% and 0.155% sodium chlorite):
- 24 hours before treatment, both flanks of the animals were clipped and shaved,
- the filter paper of a chamber (Finn Chamber) was fully-loaded with a dosage form preparation. The chamber was then applied to the clipped area of the skin (one concentration per flank). The chamber was held in place by means of an occlusive dressing for 24 hours,
- cutaneous reactions were evaluated 24 and 48 hours after removal of the dressings.

In order to respect the criteria for the selection of concentrations (the concentration should be well-tolerated systemically and locally, intradermal injections should cause moderate irritant effect but no necrosis or ulceration of the skin), concentration chosen for the main study was 0.1% (w/w) (i.e. 0.031% sodium chlorite).
In order to respect the criteria for the selection of concentrations (the concentrations should be well-tolerated systemically and locally, cutaneous application for the induction should cause at most weak or moderate skin reactions or be the maximal practicable concentration, cutaneous application for the challenge phase should be the highest concentration which does not cause irritant effect), concentration chosen for the topical application of the induction phase (day 8) was 5% (w/w) (i.e. 1.55% sodium chlorite). For the challenge application (day 22), it was 1% (w/w) (i.e. 0.31% sodium chlorite).

MAIN STUDY:

- INDUCTION SCHEDULE:
Intradermal route:
Day 1: six injections were made into the dermis of a 4 cm x 2 cm clipped interscapular area using a needle (0.50 x 16 mm diameter). Three injections of 0.1 mL were made into each side of this interscapsular region.
Cutaneous route:
As the test item was shown to be irritant during the preliminary test, a topical application with sodium lauryl sulphate was not necessary on day 7.
Day 8: a pad of filter paper (~8 cm2) was fully loaded with the test item at the concentration of 5% (w/w) (i.e. 1.55% sodium chlorite) and was then applied to the interscapular region of the animal of the treated group. The pad was held in place for 48 h.

- CHALLENGE SCHEDULE
On day 22, the animals received an application of the test item and vehicle. The filter paper of a chamber (Finn Chamber®) was fully loaded with the test item at the concentration of 1% (w/w) (i.e. 0.31% sodium chlorite) and was then applied to the clipped area of the skin of the posterior right flank. The vehicle was applied to the posterior left flank. The chambers were held in contact with the skin for 24 h by means of an adhesive allergenic waterproof plaster.
Challenge controls:
5 animals/sex were used as control animals and were treated with water.
Positive control substance(s):
yes
Remarks:
Mercaptobenzothiazole (as a separate study)

Results and discussion

Positive control results:
No data

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
1 % (0.31% of sodium chlorite)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No cutaneous reactions were observed.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No cutaneous reactions were observed..
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
1 % (0.31% of sodium chlorite)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No cutaneous reactions were observed.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No cutaneous reactions were observed..
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100% vehicle
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100% vehicle
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
5%
No. with + reactions:
10
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. No with. + reactions: 10.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
5%
No. with + reactions:
10
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. No with. + reactions: 10.0. Total no. in groups: 10.0.

Any other information on results incl. tables

Table 7.4.1/1: Detailed information including induction/challenge/scoring schedule for skin sensitisation test

Guinea pig maximisation test (GPMT)

Inductions

GPMT

Observations/Remarks

 

day of treatment

application

 

Induction A

0

intradermal

 

Induction B

0

topical

 

challenge

22

topical

 

scoring 1

23

No reactions were observed

 

scoring 2

24

No reactions were observed

 

Table 7.4.1/2: Result of skin sensitisation test

Number of animals with signs of allergic reactions /
number of animals in group

Negative control

Test group

Positive control

scored after 24h

0/10

0/20

10/10

scored after 48h

0/10

0/20

10/10

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
No delayed contact hypersensitivity in guinea pigs was observed.
Executive summary:

In a skin sensitisation study, Sodium chlorite 31% solution at the concentration of 1% was administered to guinea pigs by intradermal injections. This study was performed according to OECD 406 and in compliance with the Good Laboratory Practice.

It is considered as relevant to use the structurally close metabolite, sodium chlorite, to evaluate the sensitising potential of the chlorine dioxide according to a read-across approach. In fact, the chlorine dioxide (ClO2) is an unstable gas which must be stabilized with water. After skin contact, the ClO2 in solution is rapidly reduced in chlorite and chloride (see § 7.1).

Intradermal injections of Sodium chlorite were performed on day 1 at the concentration of 0.1% (w/w) in 0.9% NaCl, concentration which causes moderate irritant effect but no necrosis or ulceration of the skin. Topical application of Sodium chlorite was performed on day 8 at the concentration of 5% (w/w) in 0.9% NaCl, concentration which causes weak skin reactions.

For all groups of animals (negative and positive control groups and test substance group, challenge was performed on day 22 by a topical application of Sodium chlorite at the concentration of 1% (w/w) in 0.9% NaCl, i.e. the highest concentration which does not cause irritant effect.

The animals were observed at least once a day during the study in order to check for clinical signs and mortality

No clinical signs and no death were noted during the study. No cutaneous reactions (erythema or edema) were observed after the challenge application.

The Sodium chlorite 31% solution at the concentration of 1% (i.e. 0.31% sodium chlorite) does not induce delayed contact hypersensitivity in guinea pigs. Therefore, at concentrations where skin irritation/corrosion is not observed, the chlorine dioxide is not considered as a skin sensitiser according to a read-across approach with the sodium chlorite.

Under the test conditions, the chlorine dioxide is not classified as skin sensitiser according to the criteria of the Annex VI to the Regulation (EC) n° 1272/2008.

This skin sensitisation study is classified as acceptable. It does satisfy with the guidelines requirements for a skin sensitisation study (OECD 406) in the guinea pigs.

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