Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-08-29 to 1992-03-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable without restriction because it was conducted according to EPA OTS 798.4100.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EPA OTS 798.4100 (Skin Sensitisation)
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
Buehler test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: F-133, thermocracked kerosene, CAS No. 68333-23-3
- Substance type:
- Physical state: liquid
- Analytical purity: 1005 petroleum hydrocarbons
- Storage condition of test material: materials were stored at temperature less than 27 degrees celsius
- Specific gravity: approximately 0.74
- Flash point: -42 degrees celsius, extremely flammable

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Sasco, Inc., Omaha, Nebraska
- Age at study initiation: young adult
- Weight at study initiation: 300 to 500 grams
- Housing: individually housed in stainless steel, wire mesh bottom cages
- Diet: fresh Agway certified guinea pig feed provided ad libitum, throughout both acclimation and study periods
- Water: provided ad libitum via automatic watering system
- Acclimation period: 14 days for induction phase animals; 7 days for dose selections phase animals


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 26 degrees celsius
- Humidity (%): 40 to 70%
- Air changes (per hr): no less than ten changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light/ 12 hours dark


IN-LIFE DATES: From: 1990-08-29 To: 1990-10-12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Mineral oil
Concentration / amount:
Of test substance at:

Dose selection (irritation) phase: undiluted F-133, 1:2 v/v, 1:4 v/v, and 1:8 v/v F-133

Induction phase: undiluted

Challenge phase: 1:4 v/v
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Mineral oil
Concentration / amount:
Of test substance at:

Dose selection (irritation) phase: undiluted F-133, 1:2 v/v, 1:4 v/v, and 1:8 v/v F-133

Induction phase: undiluted

Challenge phase: 1:4 v/v
No. of animals per dose:
Of test substance at:

Dose selection phase: six animals

Induction phase and challenge phase: ten animals

Details on study design:
RANGE FINDING TESTS: Six guinea pigs were used for the dose selection phase. Each animal was dosed with either undiluted, or 1:2 v/v, 1:4 v/v, or 1:8 v/v mineral oil dilutions. The respective dilutions were applied to four pads, placed on the shaved areas, and occlusively wrapped. the wrappings were removed after six hours and the skin was scored using the Draize method after 24 hours.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: three
- Exposure period: 6 hours
- Test groups: one group of ten guinea pigs dosed with 0.5 ml of undiluted F-133
- Control group: one group of 10 guinea pigs were dosed with 0.3% DNCB in 80% ethanol/water and served as the positive control; one group of 4 guinea pigs were dosed with 0.5 mL of undiluted test substance to serve as the challenge control; one group of 4 guinea pigs were dosed with mineral oil to serve as the vehicle control group
- Site: shaved strip of hair along one side of the dorsal midline
- Frequency of applications: one 6 hour exposure per week
- Duration: 3 weeks
- Concentrations: undiluted test article


B. CHALLENGE EXPOSURE
- No. of exposures: one
- Day(s) of challenge: the challenge phase took place 14 days following the last induction application
- Exposure period: 6 hours
- Test groups: the induction/challenge group consists of 10 guinea pigs dosed with a 1:4 F-133 dilution of the test article
- Control group: four guinea pigs treated with 1:4 dilution of F-133 in mineral oil as challenge control, ten guinea pigs treated with 0.2% DNCB in 80% ethanol/water as induction/ challenge positive control, four guinea pigs treated with 0.2% DNCB in 80% ethanol/water as challenge control, four guinea pigs in the mineral oil as vehicle control
- Site: shaved strip of hair along one side of the dorsal midline on the opposite side from the induction test site.
- Concentrations: 1:4 dilution of F-133
- Evaluation (hr after challenge): 24 and 48 hours after exposure


Challenge controls:
1:4 dilution of F-133 challenge control; 0.2% DNCB challenge control
Positive control substance(s):
yes
Remarks:
1,4 dinitrochlorobenzene (2,4 DNCB)

Study design: in vivo (LLNA)

Statistics:
Response grade, severity index, and incidence were generated for each treatment period. The response grade is the sum of the erythema and edema scores based on the standard Draize scoring system. The severity index is the sum of all response grades at 24 and 48 hours, multiplied by the number of animals with that given response grade, divided by the number of animals in the group. The incidence is the number of animals exhibiting a response grade of one or more at either 24 or 48 hours.

Results and discussion

Positive control results:
DNCB induced an appropriate positive response in the induction and challenge phases and is considered a delayed contact sensitizer, under the conditions of this test.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
1:4 dilution
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Severity Index (erythema + oedema) = 0.0
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1:4 dilution. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Severity Index (erythema + oedema) = 0.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
1:4 dilution
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
Severity Index (erythema + oedema) = 0.0
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 1:4 dilution. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: Severity Index (erythema + oedema) = 0.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.2%
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
Severity Index (erythema + oedema) = 3.8
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.2%. No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: Severity Index (erythema + oedema) = 3.8.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
1:4 dilution
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Severity Index (erythema + oedema) = 0.0
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1:4 dilution. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Severity Index (erythema + oedema) = 0.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
1:4 dilution
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
Severity Index (erythema + oedema) = 0.0
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 1:4 dilution. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: Severity Index (erythema + oedema) = 0.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.2%
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
Severity Index (erythema + oedema) = 3.0
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.2%. No with. + reactions: 10.0. Total no. in groups: 10.0. Clinical observations: Severity Index (erythema + oedema) = 3.0.

Any other information on results incl. tables

The scores after the challenge treatment are summarised below


Group

Time

Response Grade Range

Incidence

F-133 Induction/Challenge group

24 h

0

0/10

 

48 h

0

0/10

F-133 Challenge control

24 h

0

0/4

 

48 h

0

0/4

DNCB Induction/Challenge positive control

24 h

3-5

10/10

 

48 h

2-4

10/10

DNCB Challenge control

24 h

0-1

3/4

 

48 h

0-1

3/4

Vehicle control

24 h

0

0/4

 

48 h

0

0/4


On the basis of the above response, the test material was not sensitizing.

No. with positive reactions:

Induction Phase:

Treatment 1 Scores: 9 out of 10 (test group); dose: 0.5 ml of undiluted F-133

Treatment 1 Scores: 9 out of 10 (positive control); dose: 0.5 ml of 0.3% DNCB

Treatment 2 Scores: 10 out of 10 (test group); dose: 0.5 ml of undiluted F-133

Treatment 2 Scores: 10 out of 10 (positive control); dose: 0.5 ml of 0.3% DNCB

Treatment 3 Scores: 10 out of 10 (test group); dose: 0.5 ml of undiluted F-133

Treatment 3 Scores: 10 out of 10 (positive control); dose: 0.5 ml of 0.3% DNCB

Challenge Scores:

0 out of 10 (Induction/Challenge test group); dose: 0.5 ml of F-133 1:4 dilution in mineral oil

0 out of 4 (F-133 Challenge Control); dose: 0.5 ml of F-133 1:4 dilution in mineral oil

10 out of 10 (DNCB Induction/Challenge Group); dose: 0.5 ml of 0.3% DNCB in 80% ethanol

3 out of 4 (DNCB Challenge Control); dose: 0.5 ml of 0.2% DNCB in 80% ethanol

0 out of 4 (Vehicle Challenge Control); dose: 0.5 ml of mineral oil

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
During the challenge phase, exposure of a 1:4 dilution of thermocracked kerosine to induction phase test animals did not yield higher response grades, severity, or incidence than those associated with the naive challenge control group exposed to thermocracked kerosine.

During the challenge phase, exposure of 0.2% DNCB to induction phase positive control animals elicited significantly higher response grades, severity indices, and incidence over the naive DNCB challenge control group. This indicates an appropriate positive response.

The vehicle irritation control group was free of dermal irritation during the challenge phase therefore, mineral oil used as a dilutant is not an irritant.

Therefore, under the conditions of this study, F-133 is not considered a delayed contact sensitizer.
Executive summary:

In a dermal sensitisation study using thermocracked kerosine in mineral oil, male young adult Pig/Hartley guinea pigs were tested using a modified Buehler technique.

During the challenge phase, a second exposure of a 1:4 dilution of thermocracked kerosine to induction test animals did not yield higher response grades, severity, or incidence than those associated with the naive challenge control group exposed to thermocracked kerosine. During the challenge phase, exposure of 0.2% DNCB to induction positive control animals elicited significantly higher response grades, severity indices, and incidence over the naive DNCB challenge control group. The vehicle irritation control group was free of dermal irritation during the challenge phase. Therefore, under the conditions of this study, thermocracked kerosine is not considered a delayed contact sensitiser and DNCB induced an appropriate positive response.

This study received a Klimisch score of 1 and is classified as reliable without restriction because it was conducted according to EPA OTS 798.4100.