Reaction mass of 2-methylpropan-2-ol and acetone and water

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, OECD Guideline study.

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)

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Type:

Constituent

Details on test material:

Batch No. S-122009-311853
Purity: 100%

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

Age: 8 - 12 weeks
Body weight: 15 - 23 g
Temperature: 19 - 25 degrees C
Humidity: 30 - 70 %

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

Undiluted, 50%, or 25%

No. of animals per dose:

4

Details on study design:

Three groups, each of four animals, were treated with 50 uI (25 uI per ear) of the undiluted test material or the test material as a solution in dimethyl formamide at concentrations of 50% or 25% v/v. A further group of four animals was treated with dimethyl formamide alone.

Five days following the first topical application of the test material or vehicle (Day 6) all mice were injected via the tail vein with 250 IJI of phosphate buffered saline (PBS) containing 3H-methyl thymidine (3HTdR: 80 IJCi/ml, specific activity 2.0 Ci/mmol, ARC UK Ltd) giving a total of 20 IJCi to each mouse.

Clinical Observations: All animals were observed twice daily on Days 1, 2 and 3 and on a daily basis on Days 4, 5 and 6. Any signs of toxicity or signs of ill health during the test were recorded.

Bodyweights: The bodyweight of each mouse was recorded on Day 1 (prior to dosing)
and Day 6 (prior to termination).

Termination: Five hours following the administration of 3HTdR all mice were killed by carbon dioxide asphyxiation. The draining auricular lymph nodes from the four mice were excised and pooled for each experimental group. For each group 1 ml of PBS was added to the pooled lymph nodes.

Preparation of Single Cell Suspension: A single cell suspension of pooled lymph node cells was prepared by gentle mechanical disaggregation through a 200-mesh stainless steel gauze. The lymph node cells were rinsed through the gauze with 4 ml of PBS into a petri dish labelled with the project number and dose concentration. The lymph node cell suspension was transferred to a centrifuge tube. The petri dish was washed with an additional 5 ml of PBS to remove all remaining lymph node cells and these were added to the centrifuge tube. The pooled lymph node cells were pelleted at
1400 rpm (approximately 190 g) for ten minutes. The pellet was resuspended in 10 ml of PBS and re-pelleted. To precipitate out the radioactive material, the pellet was resuspended in 3 ml of 50/0 Trichloroacetic acid (TCA).

Positive control substance(s):

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

Results and discussion

Positive control results:

At a concentration of 15% v/v in DMF, the stimulation index was 5.16 (positive)

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: 25% Fuel-A: 0.54 50% Fuel-A: 0.58 100%: Fuel-A: 0.34

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Vehicle: 7238 25% Fuel-A: 3892 50% Fuel-A: 4216 100% Fuel-A: 2459

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

Fuel-A was non-sentisiting in the local lymph node assay.