Alkenes, C15-18-branched and linear, maleated, hydrolyzed

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-02-01 to 2010-02-1

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

Method B42 Skin Sensitisation

Deviations:

no

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:

(6E)-3,13-diethylpentadec-6-ene; (6E)-tetradec-6-ene; (7E)-hexadec-7-ene; (8E)-nonadec-8-ene; 2-[(6E)-3,13-diethylpentadec-6-en-8-yl]butanedioic acid; 2-[(8E)-nonadec-8-en-7-yl]butanedioic acid; 2-[(8E)-tetradec-8-en-7-yl]butanedioic acid; 2-[(9E)-hexadec-9-en-8-yl]butanedioic acid

EC Number:

700-363-1

Cas Number:

1335203-20-7

IUPAC Name:

(6E)-3,13-diethylpentadec-6-ene; (6E)-tetradec-6-ene; (7E)-hexadec-7-ene; (8E)-nonadec-8-ene; 2-[(6E)-3,13-diethylpentadec-6-en-8-yl]butanedioic acid; 2-[(8E)-nonadec-8-en-7-yl]butanedioic acid; 2-[(8E)-tetradec-8-en-7-yl]butanedioic acid; 2-[(9E)-hexadec-9-en-8-yl]butanedioic acid

Details on test material:

- Name of test material : Hydrolysed reaction products of furan-2,5-dione and C15-18-(linear and branched)-alkenes
- Physical state: Amber coloured extremely viscous liquid
- Lot/batch No.: AF405
- Expiration date of the lot/batch: 31 December 2010
- Storage condition of test material: room temperature in the dark

In vivo test system

Test animals

Species:

mouse

Strain:

other: CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

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


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


IN-LIFE DATES: From: February 2010 To: 17 February 2010

Study design: in vivo (LLNA)

Vehicle:

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

Concentration:

0, 10, 25 and 50% in acetone/olive oil 4:1

No. of animals per dose:

Preliminary dose: one mouse
Main test: four mice/dose

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: The test substance is expected to be soluble in acetone/olive oil
- Irritation: Clinical observations, bodyweight and mortality were recorded


MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response: Sensitiser if at least one concentration of the test material results in a threefold or greater increase in 3HTdR incorporation compared to control values. Any test material failing to produce a threefold or greater increase in 3HTdR incorporation will be classified as a "non sensitiser".
- Lymph node proliferation response: Number of radioactive disintegration per minute per lymph node and as the ratio of 3HTdR incorporation into lymph node cells of test nodes relative to that recorded for the control nodes (Stimulation Index)

TREATMENT PREPARATION AND ADMINISTRATION:
Groups of four mice were treated with the test material at concentrations of 50%, 25% or 10% w/w in acetone/olive oil 4:1. The mice were treated by daily application of 25 µl of the appropriate concentration of the test material to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3). Five days following the first topical application of the test material or vehicle (Day 6) the surviving mice were injected via the tail vein with 250 µl of phosphate buffered saline (PBS) containing 3H methyl thymidine (3HTdR: 80 µCi/ml, specific activity 2.0 Ci/mmol, ARC UK Ltd) giving a total of 20 µCi to each mouse. Five hours later, the mice were killed and the draining auricular lymph nodes from the surviving mice were excised, collected and prepared for analysis of the disintegration per minute of 3H-methyl thymidine by scintillation counting.

Positive control substance(s):

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

Statistics:

Not applicable

Results and discussion

Positive control results:

The Stimulation Index expressed as the mean radioactive incorporation for the treatment group divided by the mean radioactive incorporation of the vehicle control group is 3.12 for Hexylcinnamaldehyde, tech., 85% in acetone/olive oil (4:1). Based on this result, it is concluded that it is a sensitiser.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Value:

6.65

Test group / Remarks:

10% DCI-30.n w/w in acetone/olive oil (4:1)

Results 2

Parameter:

SI

Value:

7.2

Test group / Remarks:

25% DCI-30.n w/w in acetone/olive oil (4:1)

Results 3

Parameter:

SI

Value:

9.02

Test group / Remarks:

50% DCI-30.n w/w in acetone/olive oil (4:1)

Results 4

Parameter:

other: disintegrations per minute (DPM)

Value:

64 063.33

Test group / Remarks:

10% DCI-30.n w/w in acetone/olive oil (4:1)

Results 5

Parameter:

other: disintegrations per minute (DPM)

Value:

52 001.84

Test group / Remarks:

25% DCI-30.n w/w in acetone/olive oil (4:1)

Results 6

Parameter:

other: disintegrations per minute (DPM)

Value:

86 798.47

Test group / Remarks:

50% DCI-30.n w/w in acetone/olive oil (4:1)

Any other information on results incl. tables

During the preliminary screening test, no signs of systemic toxicity were noted. Mild redness to the ears and neck was reported on Day 3 and on Day 4. Based on this information the dose levels selected for the main test were 50%, 25% and10% w/w in acetone/olive oil 4:1.

During the main study, one animal in the 25% w/w in actone/olive oil group died on day 6. No signs of toxicity were reported in any of the remaining animals. Mild redness to the ears and neck was noted in animals treated with the test material at concentrations of 50% or 25% w/w in acetone/olive oil 4:1.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (skin sensitising) based on GHS criteria

Conclusions:

The Stimulation Index for the 10% w/w, 25% w/w and 50% w/w Hydrolysed reaction products of furan-2,5-dione and C15-18-(linear and branched)-alkenes concentration in olive oil groups were reported to be 6.65, 7.20 and 9.02. The test is considered to be a sensitiser since the Stimulation Index is greater than 3. It is classified as a skin sensitiser Category 1 and assigned the hazard statement H317 "May cause an allergic skin reaction" according to Regulation (EC) No 1272/2008.

Executive summary:

In a preliminary study, one mouse was treated with a 50% w/w Hydrolysed reaction products of furan-2,5-dione and C15-18-(linear and branched)-alkenes in acetone/olive oil (4:1). No clinical signs were reported and this dose was selected as the highest dose in the main test of the Local Lymph Node Assay. Hydrolysed reaction products of furan-2,5-dione and C15-18-(linear and branched)-alkenes was applied to the dorsal surface of each ear of three groups of four mice at the following concentration 50% w/w, 25% w/w and 10% w/w Hydrolysed reaction products of furan-2,5-dione and C15-18-(linear and branched)-alkenes in acetone/olive oil (4:1). A negative control was only treated with acetone/olive oil (4:1).

The applications were made for three consecutive days (Days 1, 2 and 3). Five days after the first topical application, the mice were injected 250 µl of phosphate buffered saline (PBS) containing³H-methyl thymidine. Five hours later, the mice were killed and the draining auricular lymph nodes from the surviving mice were excised, collected and prepared for analysis of the disintegration per minute of ³H-methyl thymidine by scintillation counting. The proliferation of lymph node cells was expressed as the number of radioactive disintegrations per minute per lymph node (disintegration perminute/node) and as the ration of 3HTdR incorporation into lymph node cells of test nodes relative to that recorded for the control nodes (Stimulation Index).

During the preliminary screening test, no signs of systemic toxicity were noted. Mild redness to the ears and neck was reported on Day 3 and on Day 4. Based on this information the dose levels selected for the main test were 50%, 25% and10% w/w in acetone/olive oil 4:1. During the main study, one animal in the 25% w/w in acetone/olive oil group died on day 6. No signs of toxicity were reported in any of the remaining animals.Mild redness to the ears and neck was noted in animals treated with the test material at concentrations of 50% or 25% w/w in acetone/olive oil 4:1. Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period.

The Stimulation Index for the 10% w/w, 25% w/w and 50% w/w Hydrolysed reaction products of furan-2,5-dione and C15-18-(linear and branched)-alkenes concentration in olive oil groups were reported to be 6.65, 7.20 and 9.02. The test is considered to be a sensitiser since the Stimulation Index is greater than 3. It is classified as a skin sensitiser Category 1 and assigned the hazard statement H317 "May cause an allergic skin reaction" according to Regulation (EC) No 1272/2008.