Ceraphyl 55

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): Ceraphyl 55
- Data received: 03/30/04
- Physical state: Clear liquid
- Sample prepartion: The test article was mixed with the vehicle, AOO (Acetone:Olive Oil) 4:1 to appropriate concentrations
- Lot/batch No.: P1743
- Storage condition of test material: Room temperature and humidity

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/J

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Jackson Laboratories
- Received on: 05/18/04
- Age at study initiation: Approximately 7 weeks old
- Weight at study initiation: 20-24 g
- Housing: Animals were housed 1/cage in suspended wire cages. Bedding was placed beneath the cages and changed at least three times/week
- Diet (e.g. ad libitum): Fresh PMI (Diet #5001) freely available
- Water (e.g. ad libitum): Freely available
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature and Humidity: Controlled
- Photoperiod (hrs dark / hrs light): 12 hr light/12 hr dark

Study design: in vivo (non-LLNA)

Positive control substance(s):

yes

Remarks:

hexylcinnamic aldhehyde (CAS No 101-86-0) 1.471 ml 25% HCA was added to 3.529 ml of AOO (Acetone: Olive Oil)

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

Group 1: Naive
Group 2: AOO -Vehicle
Group 3: 25% HCA (Control)
Group 4: 5% Ceraphyl 55
Group 5: 10% Ceraphyl 55
Group 6: 25% Ceraphyl 55
Group 7: 50% Ceraphyl 55

No. of animals per dose:

Six groups of 5 CBA/J female mice

Details on study design:

The test material, vehicle or control, was applied and spread to the dorsum of each ear of the mice by topical application one time per day for three consecutive days. The naive group was not treated. A volume of 25 ul/ear of was the test material, vehicle or control was applied using a micro pipette. Animals were observed once daily for pharmacological effects and mortality. Bodyweights were recorded at the beginning of the study and prior to termination. Ears were observed at Day 1, 3 and 6.
Five days following the first dose mice were sacrificed. Five hours before the termination, mice were treated with 5-bromo-2'-deoxy-uridine at dose of 100 mg/kg by peritoneal injection using a 27 gauge needle. This is a thymidine analogue that becomes incorporated into DNA of proliferating cells including proliferating nodal lymphocytes. The auricular lymphocytes were isolated and single cell suspensions were generated.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

The Stimulation Index (SI) was calculated for each group as follow:
For each animal, the number of BrdU+cell in the node was determined by cytometry and the mean ± SD was then calculated for each group. The mean BrdU+LNC in each test article treated group was then divided by that of the mean number of BrdU+LNC in the vehicle control group.

Results and discussion

In vivo (LLNA)

Results

Parameter:

SI

Remarks on result:

other: see Remark

Remarks:

Group 1: Naive SI=159 S.D=±0.79 Group 2: AOO -Vehicle SI=1.00 S.D=±0.71 Group 3: 25% HCA (Control) SI=16.85 S.D=±11.77 Group 4: 5% Ceraphyl 55 SI=2.14 S.D=±0.44 Group 5: 10% Ceraphyl 55 SI=1.69 S.D=±0.81 Group 6: 25% Ceraphyl 55 SI=6.96 S.D=±7.71 Group 7: 50% Ceraphyl 55 SI=23.77 S.D=±3.25

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test article Ceraphyl 55 is a potential dermal sensitiser as indicated by a positive result (SI>3).

Executive summary:

A Local Lymph Node Assay (LLNA) study was conducted to determine the sensitising potential of the test material Ceraphyl 55. The mice were divided in 7 groups and treated as follows: Group 1: Naive; Group2: AOO -Vehicle; Group 3: 25% HCA (positive control); Group 4: 5% Ceraphyl 55; Group 5: 10% Ceraphyl 55; Group 6: 25% Ceraphyl 55; Group 7: 50% Ceraphyl 55. A volume of 25 ul was applied topically to the dorsum of each ear one time per day for three consecutive days. Animals were observed once daily through the study for pharmacological effect and mortality. Body weights were recorded before the test and prior to termination. Ears measurements were taken on Day 1, 3, and 6. Five day following the initial dose and 5 hours before the sacrifice, a dose of 100 mg/kg 5-bromo-2'-deoxy-uridine was administrated by peritoneal injection using a 27 gauge needle. This is a thymidine analogue that becomes incorporated into DNA of proliferating cells including proliferating nodal lymphocytes. The auricular lymphocytes were isolated and single cell suspensions were generated. The mean Stimulation Index (SI) was calculated for each group with the following results:

Group 1: Naive SI=159 S.D=±0.79

Group 2: AOO -Vehicle SI=1.00 S.D=±0.71

Group 3: 25% HCA (Control) SI=16.85 S.D=±11.77

Group 4: 5% Ceraphyl 55 SI=2.14 S.D=±0.44

Group 5: 10% Ceraphyl 55 SI=1.69 S.D=±0.81

Group 6: 25% Ceraphyl 55 SI=6.96 S.D=±7.71

Group 7: 50% Ceraphyl 55 SI=23.77 S.D=±3.25

Based on the criteria of this test, if the SI is≥3, the test article is considered to be a skin sensitiser. The result of the test shows that the SI of 25% Ceraphyl 55 (v/v) (Acetone/Olive oil 4:1) and 50% Ceraphyl 55 are 6.96 and 23.77, respectively. Therefore Ceraphyl 55 is considered to be a skin sensitiser.