Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2-5-2006 to 2-6-2006
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to GLP and valid methods, therefore it is considered adequate, reliable and relevant for classification.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
other company data
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Principles of method if other than guideline:
Magnusson and Kligman maximization procedure
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Chloramin B; sodium N-chlorobenzenesulphonamide trihydrate
- Substance type: Biocide
- Physical state: White to light yellow powder
- Analytical purity: 77% (CAS 127-52-6).
- Impurities (identity and concentrations): 0.6% NaOH.
- Composition of test material, percentage of components: 77% CAS 127-52-6 (98% pure), 22.4% Water, 0.6% NaOH.
- Isomers composition: Not applicable.
- Purity test date: No data.
- Lot/batch No.: 54/MA/20/06
- Expiration date of the lot/batch: 03/2011
- Stability under test conditions: Stable, c.f. Chapter 4.18.
- Storage condition of test material: Stable, c.f. Chapter 4.18.

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: BFA
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Breeding farm BioTest s.r.o., Konárovice, 281 25, CZ, RČH CZ 21760152
- Age at study initiation: 6-7 weeks at receiving
- Weight at study initiation: 356-506 g
- Fasting period before study: No data
- Housing: Monitored conditions, microbiologically defined background, according to SOP No. 40 ; sterilized shavings of soft wood as bedding
- Diet (e.g. ad libitum): MAK 1 pelleted standard diet for experimental animals ad libitum (producer: Bergman, Kocanda Mill)
- Water (e.g. ad libitum): Drinking tap water with ascorbic acid ad libitum (quality corresponding to Regulation No. 252/2004 Czech Coll. Of Law)
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C (permanently monitored)
- Humidity (%): 30-70% (permanently monitored)
- Air changes (per hr): Not provided
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 25-4-2006 To: 2-6-2006

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: topical applications: Vaselinum album ; intradermal injections: water for injections
Concentration / amount:
Induction – intradermal injections: 1% test substance in water for injections
Induction – topical application: 20% test substance in vaseline
Challenge – topical application: 1% test substance in vaseline

Total amount of the test substance used for main test was 2.27 g:
Induction – intradermal injections: 0.12 g
Induction – topical application: 2 g
Challenge – topical application: 0.15 g
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: topical applications: Vaselinum album ; intradermal injections: water for injections
Concentration / amount:
Induction – intradermal injections: 1% test substance in water for injections
Induction – topical application: 20% test substance in vaseline
Challenge – topical application: 1% test substance in vaseline

Total amount of the test substance used for main test was 2.27 g:
Induction – intradermal injections: 0.12 g
Induction – topical application: 2 g
Challenge – topical application: 0.15 g
No. of animals per dose:
Pilot experiment: 5 animals (2M + 3F)
Test groups: 20 animals (10M + 10F) in the treatment group and 10 animals (6M +4F) in the control group
Details on study design:
RANGE FINDING TESTS: 5 animals were used in the pilot experiment.
Induction – intradermal injections: 1% test substance in water for injections (discrete erythema of the skin in pilot experiment) was chosen for the main study.
Induction – topical application: 20% test substance in Vaseline ( confluent erythema in pilot experiment) was chosen for the main study.
Challenge – topical application: 1% test substance in Vaseline ( fully absorbed and no visible changes in pilot experiment) was chosen for the main study.



MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Day 0: three pairs of intradermal injections: Day 6: occlusive topical application
- Exposure period: Topical application under occlusive dressing for 48 hours
- Test groups: 20 animals (10M + 10F)
- Control group: 10 animals (6M +4F)
- Site: Injections: in the shoulder region, Topical : 2 x 4 cm area (not specified) cleared of hair
- Concentrations: Induction – intradermal injections: 1% test substance in water for injections
Induction – topical application: 20% test substance in Vaseline

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 20 was day of challenge
- Exposure period: 24 hours under occlusion
- Test groups: In treated and control animals 1% test substance in vaseline was applied to the left flank and vaseline only was applied to the right flank of the animals
- Site: Topical: flanks cleared of hair (treatment 2 x 2 cm)
- Concentrations: Challenge – topical application: 1% test substance in Vaseline
- Evaluation (hr after challenge): Approximately 21 hours after removing the patch, the challenge area were cleaned and shaved; approximately 3 hours later (approx. 48 hours from the start of application of the challenge dose) the skin reaction was observed and recorded; approximately 24 hours after this observation a second observation was made and skin reaction was again recorded.
Challenge controls:
yes: topical application: 1% test substance in Vaseline applied to the left flank of the animals (idem as in the induced animals).
Positive control substance(s):
yes
Remarks:
benzocaine

Results and discussion

Positive control results:
Reliability of the experimental technique is checked periodically in about six-month interval by the experiment with known sensitizer bezocaine. For intradermal application the 5% concentration in olive oil was used and the challenge exposure was performed by the same concentration in vaselina alba.
The result of last experiment (March 2006): positive skin reaction – 4 animals (total number of animals = 10) i.e; 40%. According to the guideline the minimal number of animals with positive skin reaction is expected to be 30%. The result is satisfactory.
The results of reference study test are described in VUOS-CETA internal report No. 0643.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
1% in vaseline
No. with + reactions:
9
Total no. in group:
20
Clinical observations:
discrete erythema on the left flank
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1% in vaseline. No with. + reactions: 9.0. Total no. in groups: 20.0. Clinical observations: discrete erythema on the left flank.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
1% in vaseline
No. with + reactions:
6
Total no. in group:
20
Clinical observations:
discrete erythema of left flank
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 1% in vaseline. No with. + reactions: 6.0. Total no. in groups: 20.0. Clinical observations: discrete erythema of left flank.
Reading:
other: first and second reading
Group:
negative control
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading:

Any other information on results incl. tables

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

Inductions

 

Concentration of test substance 

Day of treatment

Application

Post-challenge observations

24 h

48 h

Induction 1

1 %

0

intradermal

-

-

Induction 2

20 %

6

topical

-

-

Challenge

1 %

20

topical

9/20*

6/20*

Table 2. Average body weight of animals throughout the study

 

Number of animals

Average body weight of animals at study start

(g)

Average body weight of animals at study end        (g)

Average weight increment            

(g)

Exposed group

20

425.04

570.69

145.65

Control group

10

399.20

555.11

155.91

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance, Chloramine B trihydrate, caused a positive reaction in 9 of 20 guinea pigs, e.g. in 45% of animals which were exposed to the test substance.
Executive summary:

The test substance, Chloramine B trihydrate, was tested for the assessment of skin allergic effects using albino guinea pigs (strain BFA).

The test was performed according to EU Method B.6, Skin sensitisation, which is analogous to OECD Test Guideline No. 406, Skin sensitisation. The Magnusson and Kligman maximization procedure was followed. The experiment proceeded in three phases: intradermal induction - topical induction – topical challenge. Potential skin reactions were evaluated at the end of experiment.
The test substance caused a positive reaction in 9 of 20 guinea pigs, e.g. in 45% of animals which were exposed to the test substance.