Oils, fish, oxidized, bisulfited, sodium salts

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 01-29, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study (OECD 429, EU Method B.42)

Justification for data waiving:

other:

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Oils, fish, oxidized, bisulfited, sodium salts

EC Number:

307-037-4

EC Name:

Oils, fish, oxidized, bisulfited, sodium salts

Cas Number:

97488-98-7

Molecular formula:

Not applicable

IUPAC Name:

not available

Details on test material:

- Name of test material (as cited in study report): FLL sample 3
- Chemical name: Oils, fish, oxidized, bisulfited, sodium salts (triglycerides, C12 to C24, even, saturated and unsaturated, bisulfited, sodium salt)
- Physical state: viscous liquid
- Purity: 90%
- Impurities: 10% water content by Karl Fischer
- Lot/batch No.: LS06921
- Expiration date of the lot/batch: 13 October 2010
- Storage condition of test material: room temperature in the dark
- Other: amber color
- All other template details: Not reported

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/Ca (CBA/CaOlaHsd)

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd, Oxon, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 15-23 g
- Housing: individually housed in suspended solid-floor polypropylene cages furnished with softwood woodflakes.
- Diet (e.g. ad libitum): ad libitum access to 2014 Teklad Global Rodent diet
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 degrees C
- Humidity (%): 30-70%
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs light / 12 hours dark

IN-LIFE DATES: not reported

Study design: in vivo (LLNA)

Vehicle:

other: butanone

Concentration:

100%, 50% or 25% v/v in butanone

No. of animals per dose:

4

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: undiluted test substance used
- Irritation: no signs of systemic toxicity or skin irritation were noted
- Lymph node proliferation response: not measured

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: at least a threefold increase in 3HTdR incorporation compared to control values

TREATMENT PREPARATION AND ADMINISTRATION: test material was diluted in butanone and applied to the dorsal surface of each ear of the mice for three consecutive days with a three day observation period. The test material formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette.

Positive control substance(s):

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

Statistics:

The EC3 value (concentration of test material expected to cause a 3-fold increase in 3HTdR incorporation) was calculated using the following equation: EC3 = c + [[(3-d)/(b-d)] x (a-c)], where a = lowest concentration giving stimulation index >3, b = actual stimulation index caused by 'a', c = highest concentration failing to produce a stimulation index of 3, and d = actual stimulation index caused by 'c'.

Results and discussion

Positive control results:

A group of 5 animals treated with 25 µl per ear of alpha-Hexylcinnamaldehyde as a solution in butanone as a concentration of 15% v/v produced a stimulation index of 6.63.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: See Table 2

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: See Table 2

Any other information on results incl. tables

There were no deaths, signs of systemic toxicity or skin irritation noted in the test or control animals during the test.

EC3 = 34.8 % test material v/v in butanone

Table 2. Disintegrations per Minute (dpm), Disintegrations per Minute/Node and Stimulation Index

Concentration (%v/v) in butanone	Dpm	Dpm/Node	Stimulation Index	Result
Vehicle	4709.30	588.66	Not applicable	Not applicable
25	10685.05	1335.63	2.27	Negative
50	19510.38	2438.80	4.14	Positive
100	22634.04	2829.26	4.81	Positive

 

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

Given additional information provided in the "Overall Remarks, Attachments" section, the weakly positive responses in the LLNA should not result in a label of skin sensitization.

Executive summary:

Study summary:

Introduction. A study was performed to assess the skin sensitisation potential of the test material in the CBA/Ca strain mouse following topical application to the dorsal surface of the ear. The method was designed to meet the requirements of the following:

- OECD Guideline for the Testing of Chemicals No. 429 “Skin Sensitisation: Local Lymph Node Assay” (adopted 24 April 2002)

- Method B.42 Skin Sensitisation (Local Lymph Node Assay) of Commission Regulation (EC) No. 440/2008

 

Methods. Following a preliminary screening test in which no clinical signs of toxicity were noted at a concentration of 100%, this concentration was selected as the highest dose investigated in the main test of the Local Lymph Node Assay. Three groups, each of four animals, were treated with 50µl (25µl per ear) of the undiluted test material or the test material as a solution in butanone at concentrations of 50% or 25% v/v. A further group of four animals was treated with butanone alone.

 

Results. The stimulation index expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle control group are as follows:

Concentration (% v/v) in butanone	Stimulation Index	Result
25	2.27	Negative
50	4.14	Positive
100	4.81	Positive

 

The concentration of test material expected to cause a 3-fold increase in³HTdR incorporation (EC₃) value) was calculated to be 34.8% v/v in butanone.

 

Conclusion. The test material was considered to be a sensitiser under the conditions of the test.

However, given additional information provided in the Attachment, the weakly positive responses in the LLNA should not result in a label of skin sensitization.