Oils, fish, oxidized

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

end on 26-APR-2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study performed according to OECD guideline and GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

modified as IMDS test

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Oils, fish, oxidized

EC Number:

269-122-1

EC Name:

Oils, fish, oxidized

Cas Number:

68187-75-7

Molecular formula:

Not necessary

IUPAC Name:

Triglycerides C12-24, even, saturated and unsaturated, oxidized

Details on test material:

- Name of test material (as cited in study report): triglyceride, C12-24, saturated and unsaturated, oxidized- Substance type: UVCB- Physical state: liquid- Stability under test conditions: data not available- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:

mouse

Strain:

NMRI

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS- Source: from the company Charles River Deutschland GmbH, D-97633 Sulzfeld- Age at study initiation: 9-10 weeks- Weight at study initiation: 31 to 34 g- Housing: transparent macrolone cages (type 150, floor area 810 cm2) with 6 animals in each cages- Diet: a pelleted complete rodent diet "Altromin 1324", ad libitum- Water: free access to bottles of drinking water of domestic quality, which was acidified with hydrochloric acid to pH 2.5 in order to prevent microbial growth- Acclimation period: at least 5 daysENVIRONMENTAL CONDITIONS- Temperature: 21 +/- 3°C- Humidity: at least 30% and preferably not exceed 70%- Air changes: 10 per hr- Photoperiod: 12 hrs dark / 12 hrs lightIN-LIFE DATES: the experimental phase was carried out between November 9th, 2009 and November 12th, 2009

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

25, 50 and 100% (v/v)

No. of animals per dose:

6 animals per dose

Details on study design:

RANGE FINDING TESTS: data not availableMAIN STUDYANIMAL ASSIGNMENT AND TREATMENT- Name of test method: LLNA / IMDS- Criteria used to consider a positive response:a differentiation index (DI) is calculated from the different parameters of investigation which describes the relation between the activation of the local skin-draining lymph nodes and the skin inflammation at the site of topical treatment as well as it contributes to distinguish between inflammatory (non-specific) and allergic (specific) reaction:- 0 < DI < 1 : inflammation- DI > 1 : allergic reactionDI = (% max. LN cell count index increase) / (% max. ear thickness increase)TREATMENT PREPARATION AND ADMINISTRATION:Twenty-four albino mice of the strain Crl:NMRI in groups consisting of six animals each were treated with three concentrations of the test item (100%, 50% and 25% v/v) or only with the vehicle acetone/olive oil (4/1 v/v) on three consecutive days. An amount of 25µL of the test substances was applied on the dorsal side of each ear.

Positive control substance(s):

not specified

Statistics:

data not available

Results and discussion

Positive control results:

the last positive test results was registered in August 2009 with differentiation index (DI) of 1.63 and 1.64

In vivo (LLNA)

Results

Parameter:

SI

Remarks on result:

other: see table

Any other information on results incl. tables

Table 1: calculation of means, standard deviation and indices of the ear thickness, for the weight of ear tissues and lymph nodes as well as number of lymph node cells

Parameter of investigation		Negative control	Triglyceride, C12-24, saturated and unsaturated, oxidized
		Acetone/olive oil (4:1 v/v)	100%	50%	25%
Increase in ear thickness	Mean of ear thickness on day 4 (mm)	0.19	0.20	0.20	0.19
	SD of ear thickness on day 4 (% of mean)	1	5	3	3
	Index of ear thickness	1.00	1.06	1.04	1.01
Weight of ear (diameter 7mm)	Mean value (g)	0.0098	0.0109	0.0109	0.0107
	SD (% of mean)	4	10	7	7
	Index of ear thickness	1.00	1.10	1.11	1.09
Weight of lymph nodes (2 pieces)	Mean value (g)	0.0099	0.0104	0.0087	0.0100
	SD (% of mean)	12	15	27	15
	Index of LN weight	1.00	1.05	0.88	1.02
Number of lymph node cells	Mean value	9,286,667	11,113,333	11,293, 333	11,656,000
	SD (% of mean)	28	25	35	16
	Stimulation index (index of number of LN cells)	1.00	1.20	1.22	1.26

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

Based on the result of the study described neither a specific (sensitizing) nor a non-specific (irritant) stimulation potential shall be attributed to the test item in the tested concentrations of 100%; 50% and 25% (v/v).

Executive summary:

The test item Triglycerides, C12-24, even, saturated and unsaturated, oxidized was investigated for the existence of a specific (sensitizing) or non-specific (irritant) stimulation potential by using the modified Local Lymph Node Assay (IMDS).

Twenty-four albino mice of the strain Crl:NMRI in groups consisting of six animals each were treated with three concentrations of the test item (100%, 50% and 25% v/v) or only with the vehicle acetone/olive oil (4/1 v/v) on three consecutive days. An amount of 25µL of the test substances was applied on the dorsal side of each ear. The determination of ear thickness, ear weight, lymph node weight and lymph node cell count compared to the negative control (vehicle group) should provide the information whether the test item has a specific (sensitizing) or non-specific (irritant) stimulation potential.

In the test group treated with 25% (v/v) test item one animal (No. 21) was excluded from the study evaluation because of abnormally enlarged lymph nodes. But in any case at least four animals per group were available.

During the study, a positive increase of the thickness of ears was not recorded on day 4 in any animal of the test groups in comparison with the negative control. The determination of the ear weights showed a slight increase in the test groups compared to the negative control. The weight of the lymph nodes in the test groups was not appreciable increased in comparison with the negative control. Counting the cells of the lymph nodes showed increases of proliferation in the animals of the three test groups compared to the control, but the positive threshold value was not transgressed.

Since the positive threshold values of the indices for the lymph node cell count and the increase of the ear thickness were not transgressed a calculation of the differentiation indices was not necessary.

Based on the result of the study described neither a specific (sensitizing) nor a non-specific (irritant) stimulation potential shall be attributed to the test item in the tested concentrations of 100%; 50% and 25% (v/v).