Reaction mass of copper and iron and zinc

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 06 Dec 2017 to 22 Feb 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

GLP compliance:

yes

Type of study:

Buehler test

Justification for non-LLNA method:

The guinea pig Buehler test was selected since based on the properties of the test item, it was technically not possible to prepare suitable dose formulations for performance of the LLNA study or Guinea Pig Maximization Test.

Test material

Reference

Name:

Unnamed

Type:

Constituent

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

-Animals
Species: Guinea pig
Strain: Dunkin Hartley
Condition: SPF-Quality
Source: Charles River France, L’Arbresle, France
Number of Animals:
Experimental group: 20 females
Control group : 10 females
(nulliparous and non-pregnant).
Age at the Initiation of Dosing: Young adult animals (approximately 4 weeks old) were selected.
Weight at the Initiation of Dosing: 242 to 278 g.

The Dunkin Hartley guinea pig was chosen as the animal model for this study as recognized by international guidelines as a recommended test system (e.g. OECD, FDA, MHW). The test method and number of animals are based on the test guidelines.This type of study plan was reviewed and agreed by the Laboratory Animal Welfare Officer and the Ethical Committee of Charles River Den Bosch as required by the Dutch Act on Animal Experimentation (February 1997).

-Animal Identification
At study assignment, each animal was identified using an ear tattoo.

-Environmental Acclimation
The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.

-Selection, Assignment, Replacement, and Disposition of Animals
Animals were assigned to the study at the discretion of the coordinating biotechnician according to body weights, with all animals within ± 20% of the sex mean. Animals in poor health or at extremes of body weight range were not assigned to the study. Before the initiation of dosing, a health inspection was performed and any assigned animal considered unsuitable for use in the study were replaced by alternate animals obtained from the same shipment and maintained under the same environmental conditions. The disposition of all animals was documented in the study records.

-Housing
On arrival and following assignment to the study, animals were group housed (up to 5 animals of the same sex and same dosing group together) in labeled Noryl cages (Tecniplast; 74 cm x 54 cm x 25 cm height) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room(s) in which the animals were kept were documented in the study records. Animals were separated during designated procedures/activities. Each cage was clearly labeled.

-Environmental Conditions
Target temperatures of 18 to 24°C with a relative target humidity of 40 to 70% were maintained. The actual daily mean temperature during the study period was 20 to 22°C with an actual daily mean relative humidity of 42 to 70%. A 12 hour light/12 hour dark cycle was maintained. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

-Food
Complete maintenance diet for guinea pigs (MS-H, SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum, except during designated procedures. In addition, hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least twice a week. The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.

-Water
Municipal tap-water was freely available to each animal via water bottles. Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.

-Animal Enrichment
For psychological/environmental enrichment, animals were provided with shelters (CS3B02A Play tunnels (90 mm x 5 mm x 125 mm), Datesand, Manchester, UK), except when interrupted by study procedures/activities.

-Veterinary Care
Veterinary care was available throughout the course of the study; however, no examinations or treatments were required.

Study design: in vivo (non-LLNA)

Induction

Route:

epicutaneous, occlusive

Vehicle:

other: Vaseline (BUFA, IJsselstein, The Netherlands)

Concentration / amount:

an amount equivalent to 0.5 mL of a 50% test item concentration

Day(s)/duration:

At days 1, 8 and 15 for 6 hours

Adequacy of induction:

highest technically applicable concentration used

Challenge

Route:

epicutaneous, occlusive

Vehicle:

other: Vaseline (BUFA, IJsselstein, The Netherlands)

Concentration / amount:

50% test item concentration and the vehicle (an amount equivalent to 0.1 mL each)

Day(s)/duration:

At day 29 for 6 hours

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Experimental group: 20 females, Control group: 10 females.

Details on study design:

-Main Study
The concentrations and induction method were selected based on the results of the preliminary irritation study.

INDUCTION - Experimental animals
Days 1, 8 and 15:
The left side of the scapular region was clipped and epidermally treated with an amount equivalent to 0.5 mL of a 50% test item concentration, using a Metalline patch (2x3 cm) mounted on Medical tape, which was held in place with Micropore tape and subsequently Coban elastic bandage. After 6 hours, the dressings were removed and the skin cleaned of residual test item using water. Immediately after removal of the last induction application on Day 15, the treated area was assessed for irritation.

INDUCTION - Control animals
The control animals were treated as described for the experimental animals, except that, instead of the test item, the vehicle was administered.

CHALLENGE - All animals
Day 29:
Epidermal application of a 50% test item concentration and the vehicle (an amount equivalent to 0.1 mL each) to the clipped, right flank of all animals, using Patch Test Plasters (Curatest®, Lohmann, Almere, The Netherlands). The patches were held in place with Micropore tape and subsequently Coban elastic bandage. After 6 hours, the dressings were removed and the skin cleaned of residual test item using water. The treated sites were assessed for challenge reactions 24 hours and 48 hours after removal of the last dressing.

-Mortality/Moribundity Checks
Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals were not removed from cage during observation, unless necessary for identification or confirmation of possible findings.

-Toxicity
Observations for toxicity were performed once daily throughout the study.

-Body Weights
Animals were weighed individually on Day 1 (predose) and at termination of the study.

-Terminal Procedures
No necropsy for gross macroscopic examination was performed according to study plan.

-ANALYSIS
All results presented in the tables of the report are calculated using values as per the raw data rounding procedure and may not be exactly reproduced from the individual data presented. The results for the experimental animals at the challenge application(s) were compared with the results for the control animals. Positive skin reactions (grade 1 or more) were considered signs of sensitization, provided that such reactions were not observed or were less persistent in the control group. Other local effects were also taken into account. A sensitization rate (%) was calculated for each concentration as follows: the number of sensitized animals at one concentration as a proportion of the total number of animals of the experimental group. The results were evaluated according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (including all amendments) and the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Challenge controls:

The vehicle (an amount equivalent to 0.1 mL) was applied.

Positive control substance(s):

yes

Results and discussion

Positive control results:

A reliability check is carried out at regular intervals to check the sensitivity of the test system and the reliability of the experimental techniques as used by Charles River Den Bosch. In this study, performed in Dec 2017, females of the Dunkin Hartley guinea pig (from Charles River France, L’Arbresle, France) were checked for the sensitivity to Alpha- Hexylcinnamaldehyde, technical grade.

The skin reactions observed in eight experimental animals in response to the 50% test item concentration in the challenge phase were considered indicative of sensitisation, based on the absence of any response of a similar grade in the control animals. These results lead to a sensitisation rate of 80 percent to the 50% concentration.

From these results, it was concluded that the female guinea pig of the Dunkin Hartley strain is an appropriate animal model for the performance of studies designed to evaluate the sensitizing potential of a test item in a Buehler type of test.

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

50%

No. with + reactions:

0

Total no. in group:

20

Results 2

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

50%

No. with + reactions:

0

Total no. in group:

20

Results 3

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

50%

No. with + reactions:

0

Total no. in group:

10

Results 4

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

50%

No. with + reactions:

0

Total no. in group:

10

Any other information on results incl. tables

-Toxicity / Mortality

No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study.

-Body weights

Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

There was no evidence that Carrier FK-800 had caused skin hypersensitivity in the guinea pig, since no responses were observed in the experimental animals in response to a 50% test item concentration in the challenge phase.

This result indicates a sensitization rate of 0 per cent.

Based on these results Carrier FK-800 does not have to be classified and has no obligatory labelling requirement for sensitization by skin contact according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Executive summary:

The objective of this study was to evaluate whether Carrier FK-800 induces contact hypersensitivity in guinea pigs after three epidermal exposures under the conditions described in this report.

The study was carried out in compliance with the guidelines described in:

·        OECD Guideline 406. Skin Sensitization,1992.

·        EC No 440/2008 Part B. Skin Sensitization: Buehler,May 2008.

·        EPA Health Effects Test Guideline OPPTS 870.2600. Skin Sensitization,March 2003.

·        Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan (JMAFF), November 2000, including the most recent revisions.

Test item concentrations selected for the main study were based on the results of a preliminary study. Based on the results, the test item concentration selected for the main study was a 50% concentration for the epidermal induction exposures. A 50% test item concentration was selected for the challenge phase. In the Main study, twenty experimental animals were epidermally treated on three occasions (Days 1, 8 and 15) with a 50% concentration of the test item and ten control animals were similarly treated, but with the vehicle alone (Vaseline). Two weeks after the last induction exposure, all animals were challenged with a 50% concentration and Vaseline.

No skin reactions were evident after the third induction exposure in the experimental and control animals.

No skin reactions were evident after the challenge exposure in the experimental and control animals.

No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study.

Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

 

There was no evidence that Carrier FK-800 had caused skin hypersensitivity in the guinea pig, since no responses were observed in the experimental animals in response to a 50% test item concentration in the challenge phase.

This result indicates a sensitization rate of 0 per cent.

Based on these results Carrier FK-800 does not have to be classified and has no obligatory labelling requirement for sensitization by skin contact according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2015) (including all amendments) andRegulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).