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Toxicological information

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was conducted in 2009.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name of test material (as cited in study report): Daltolac XR 159
- Lot/batch No.: RZW023
- Expiration date of the lot/batch: Twenty one months at customer
- Physical Description: Light yellow viscous liquid
- Composition/Purity: Reaction mass of 2,2'-Oxydiethanol, propoxylated and formaldehyde polymer with benzenamine and 2-methyloxirane
Stability: The test article was expected to be stable under standard laboratory conditions for the duration of testing.
Storage Conditions: Room temperature (10-30ºC)

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
Total Number: 23
8 Primary irritation (Dose Range (DR) study)
10 Test article group
5 vehicle control group

Gender: Male and female
Age Range: DR Study: 3 weeks, young adult
Main Study: 4 weeks, young adult at the start of dosing for main study. Records of dates of birth for animals used in this study are retained in the Calvert archives.
Body Weight Range: DR study: 279 to 330 grams
Main study: 330 to 390 grams at the outset (Day 1) of the study.
Animal Source: Elm Hill Breeding Laboratories, Chelmsford, MA
Experimental History: Purpose-bred and experimentally naïve at the outset of the study.
Identification: Ear tag and cage card.

ENVIRONMENTAL CONDITIONS
Housing: Animals were group housed individually in compliance with USDA Guidelines. The room in which the animals were kept was documented in the study records. No other species were kept in the same room.
Lighting: 12 hours light/12 hours dark
Room Temperature: 17.2 to 26.1°C
Relative Humidity: 17 to 73%
Food: All animals had access to Harlan Teklad Rodent Diet (certified) ad libitum. The lot number(s) and specifications of each lot used are archived at Calvert. No contaminants were known to be present in the certified diet at levels that would be expected to interfere with the results of this study. Analysis of the diet was limited to that performed by the manufacturer, records of which are maintained in the Calvert archives.
Water: Tap water was available ad libitum, to each animal via an automatic watering device. The water is routinely analyzed for contaminants as per Calvert SOP’s. No contaminants were known to be present in the water at levels that would be expected to interfere with the results of this study. Results of the water analysis are maintained in the Calvert archives.
Acclimation: Study animals were acclimated to their housing for a minimum of 7 days prior to dosing.

IN-LIFE DATES: From: 5 April 2009 To: 11 May 2009

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Mineral oil and FCA/H2O for intradermal induction. For topical induction and challenge the test article was used as received.
Concentration / amount:
The test article concentration selected for intradermal induction was 5% and topical was 100%. The challenge dose was the highest non-irritating concentration (HNIC) of the test article which was determined to be 100%.
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Mineral oil and FCA/H2O for intradermal induction. For topical induction and challenge the test article was used as received.
Concentration / amount:
The test article concentration selected for intradermal induction was 5% and topical was 100%. The challenge dose was the highest non-irritating concentration (HNIC) of the test article which was determined to be 100%.
No. of animals per dose:
8 animals DR study
10 animals main study
5 animals control group
Details on study design:
Intradermal Dose Range: Prior to initiation of the induction, the irritation potential was determined. Four naive animals were exposed to four different concentrations (0.5, 1.0, 3.0 and 5.0%) of the test article in mineral oil by intradermal techniques described in site preparation and intradermal induction stage. In this test, both sides of the animal were clipped and exposed to the various concentrations of the test article. For grading of the response, the procedure described below for primary challenge was used, except only 24-hour grades were obtained.

Topical Dose Range: Four naive guinea pigs were exposed to several different concentrations (25, 50, 75 and 100%) of the test article. The procedure described in topical induction period was employed except that the wrapping were removed at 20 hours after doing. Twenty-one hours (20 to 22 hours) after unwrapping the sites were depilated. Four hours later (3-5 hours) the sites were examined for skin reactions (24-hour grade). The location of each of the concentrations of test article differed in each of the four animals to compensate for any site-to-site variations.

Site Preparation and Intradermal Induction Stage: The hair from an area over the shoulders of the guinea pigs was removed 24 to 48 hours prior to dosing using a clipper. The test article and vehicle control groups with and without FCA were injected in the shoulder region of each animal. A row of three injections, on each side of the animals, (i.e., 6 in all), was made as follows:
(1 & 2) 0.1 mL of FCA (1:1 mixture with distilled water)
(3 & 4) 0.1 mL of the test article in the vehicle (5% v/v) or the vehicle alone
(5 & 6) 0.1 mL of the test article in FCA (1:1) at 5% v/v or the vehicle emulsified with FCA and the vehicle (1:1)
Injections 1 & 2 were given close to each other nearest to the head and injection 5 & 6 are given most caudally; injections 3 & 4 are given between injection sites 1 & 2 and 5 & 6. The injection sites were just within the boundaries of a 2 x 4 cm patch, which was applied one week later for the topical induction.

Topical Induction Stage: After six days, the test sites were reclipped free of hair. Since irritation was observed in the animals dosed with the test article during the topical range-finding study, no pretreatment was performed.

One week after intradermal injections (Day 8), 0.3 mL of the test article (100%) was spread over a 2 x 4 cm filter paper and applied to the injection site area and occluded using Blenderm® tape. The Blenderm® tape was held in place with an appropriate bandage. A minimum of forty-eight hours later the dressings were removed. This same procedure was employed with the vehicle control article (0.3 mL) in the vehicle control animals. After the 48-hour exposure period, the induction sites were unwrapped and any residual material removed with gauze and mineral oil.

Challenge Period: On Day 22, the test article group and vehicle control group animals were challenged with occluded patches of the test article (100%) and vehicle for 24 hours on the left and right flanks, respectively. The hair was removed from these areas using a clipper 24 hours before application of the patches on Day 21. A 2 x 2 cm filter paper was saturated (0.2 mL) with the test article (100%) and applied to the left flank. Another 2 x 2 cm filter paper was saturated (0.2 mL) with the vehicle (mineral oil) and applied to the right flank. The same occlusive technique as for topical induction was employed.
Approximately twenty four hours later, the wrapping was removed and the sites wiped clean with gauze and mineral oil. Twenty one hours after unwrapping, the challenge sites were depilated with Nair Lotion Hair Remover (lot # L7340, pink lotion). Three hours later the sites were graded for elicited skin reactions (24-hour grade). Approximately twenty-four hours later the sites were graded a second time (48-hour grade).
Challenge controls:
On Day 22, the vehicle control group animals were challenged with occluded patches of the test article (100%) and vehicle for 24 hours on the left and right flanks, respectively.
Positive control substance(s):
yes
Remarks:
1-chloro-2, 4-dinitrobenzene (DNCB)

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
Dermal reaction
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: Dermal reaction.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
Dermal reaction
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: Dermal reaction.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Dermal reaction
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Dermal reaction.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
Dermal reaction
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: Dermal reaction.
Reading:
other:
Hours after challenge:
0
Group:
positive control
Dose level:
0
No. with + reactions:
0
Total no. in group:
0
Remarks on result:
other: Positive control not used

Any other information on results incl. tables

 

Incidence of Dermal Irritation Scores at Challenge

 

 

Dermal Irritation Scores

 

 

Induction

Challenge

24-hour Grade

48-hour Grade

 

 

Treatment

Treatment

0

1

2

3

4

0

1

2

3

4

 

 

Vehicle Control

Daltolac XR 159

@100%

4

0

1

0

0

4

1

0

0

0

 

 

Mineral oil

5

0

0

0

0

5

0

0

0

0

 

 

Test

Article

Daltolac XR 159

@100%

2

2

3

3

0

0

1

6

0

3

 

 

Mineral oil

 

 

10

0

0

0

0

10

0

0

0

0

 

 

 

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information
Conclusions:
In conclusion, under the conditions of this study, an intradermal induction of Daltolac XR 159 at 5% with a topical induction at 100%, followed by a topical challenge at 100% to guinea pigs did elicit a dermal sensitization response at 24 and 48 hours post treatment. Therefore, Daltolac XR 159 is considered to be a contact sensitizer in Guinea Pigs.
Executive summary:

The purpose of this study was to determine if the test article elicits a delayed dermal contact hypersensitivity response in guinea pigs.

Daltolac XR 159 was evaluated in the Magnusson-Kligman guinea pig maximization model of delayed hypersensitivity. Prior to experimental initiation of the induction, the irritation potential of the test article was determined with a dose range-finding studies (intradermal and topical) utilizing 8 naïve guinea pigs. Based upon these results, the intradermal dose utilized was 5%, the topical induction dose was 100% and the challenge dose utilized was 100% in the main study.

 

The main study was conducted with 10 animals for the test article group and five animals for the vehicle control group. 

No mortality was observed in any vehicle control or test article group animals in the study. No clinical signs were observed in any vehicle control group animals in the study. Decreased body tone was observed in one test article treated female on Day 21 through 25 of the study. No other clinical signs were noted in the test article treated group during the study.

Under the conditions of this study, an intradermal induction of Daltolac XR 159 at 5% with a topical induction at 100%, followed by a topical challenge at 100% to guinea pigs did elicit a dermal sensitization response at 24 and 48 hours post treatment. Therefore, Daltolac XR 159 is considered to be a contact sensitizer in Guinea Pigs.