Renewable hydrocarbons (kerosene type fraction)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study on supporting substance (with limited range, C8-C12) conducted in compliance with GLP; acceptable, well-documented study report which meets basic scientific principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

This test method was selected because the first-choice method according to REACH Annex VII §8.3, the Murine Local Lymph Node Assay, is known to give false positive results with hydrocarbon substances.

Test material

Test material information

Constituent 1

Reference substance name:

Kerosine (Fischer-Tropsch), limited range, C8-12 - branched and linear

IUPAC Name:

Kerosine (Fischer-Tropsch), limited range, C8-12 - branched and linear

Details on test material:

- Substance type: Organic
- Physical state: Liquid
- Analytical purity: 100%
- Impurities (identity and concentrations): Information not available
- Isomers composition: Information not available
- Purity test date: December 06, 1995
- Expiration date of the lot/batch: Information not available
- Stability under test conditions: Stable under normal temperature and pressure
- Storage condition of test material: Under cool conditions (approximately 4°C)

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Age at study initiation: Not reported
- Weight at study initiation: Not reported
- Housing: Stainless-steel cages with grid floors and tops
- Diet (e.g. ad libitum): commercially available pelleted guinea pig diet (Guinea pig F.D.1), was fed without restriction
- Water (e.g. ad libitum): free access
- Acclimation period: at least 6 days, up to max. 16 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 °C (range 15 to 23 °C)
- Humidity (%): 55% R.H. (range 50 to 70% R.H.)
- Air changes (per hr): at least 10 complete air changes per hour (without re-circulation)
- Photoperiod (hrs dark / hrs light): 12 hours artifical light per day, no natural light

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

paraffin oil

Induction 2

Concentration / amount:

Concentrations of test material and vehicle used at each stage of induction:
- First induction: 50% w/v in paraffin oil
50% w/v in FCA
- Topical induction: undiluted as supplied

Concentrations of test material and vehicle used at each stage of challenge:
- Topical challenge: 50% v/v in paraffin oil
10% v/v in paraffin oil

Challenge

Challenge 1

Route:

epicutaneous, semiocclusive

Vehicle:

paraffin oil

Challenge 2

Concentration / amount:

Concentrations of test material and vehicle used at each stage of induction:
- First induction: 50% w/v in paraffin oil
50% w/v in FCA
- Topical induction: undiluted as supplied

Concentrations of test material and vehicle used at each stage of challenge:
- Topical challenge: 50% v/v in paraffin oil
10% v/v in paraffin oil

No. of animals per dose:

TEST GROUP: 20 (10m/10f)
CONTROL GROUP: 20 (10m/10f)

Details on study design:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period:
after intradermal injection: 7 days;
after topical application: 15 days.
- Test groups: 20 animals (ten male and 10 female animals)
- Control group: 20 animals
- Site: area of skin 8 cm x 6 cm, overlying the scapulae
- Frequency of applications:
after intradermal injection: 7 days;
after topical application: 15 days.
- Duration:
after topical application the test side was covered with occlusive dressing for 48 hours
- Concentrations:
intradermal injection: 50% v/v Shell MDS (M) Kerosine in paraffin oil and 50% v/v Shell MDS (M) Kerosine in adjuvant;
topical application: Shell MDS (M) Kerosine.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: on day 22 of test
- Exposure period: 24 and 48 hours
- Test groups: 20 animals (ten male and 10 female animals)
- Control group: 20 animals
- Site: area of skin 8 cm x 6 cm, overlying the scapulae
- Concentrations: 10 and 50% v/v Shell MDS (M) Kerosine in paraffin oil;
- Evaluation (hr after challenge): 24 and 48 hours

Challenge controls:

Challenge application of paraffin oil alone

Positive control substance(s):

yes

Remarks:

Hexylcinnamic aldehyde (HCA)

Results and discussion

Positive control results:

Challenge application of paraffin oil alone caused a positive response in two control animals.

Challenge application of 50% v/v HCH in propylene glycol to the positive control study gave rise to a positive response in nine positive control and one control animals.

A positive response was evident in nine positive control and two control animals following challenge application of 30% v/v HCH in propylene glycol.

No dermal response was evident following challenge application of propylene glycol alone.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

50% (v/v)

No. with + reactions:

9

Total no. in group:

20

Clinical observations:

Slight erythema, exfoliation

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50% (v/v). No with. + reactions: 9.0. Total no. in groups: 20.0. Clinical observations: Slight erythema, exfoliation.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

50% (v/v)

No. with + reactions:

7

Total no. in group:

20

Clinical observations:

Slight erythema, eschar formation, exfoliation

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% (v/v). No with. + reactions: 7.0. Total no. in groups: 20.0. Clinical observations: Slight erythema, eschar formation, exfoliation.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

50% (v/v)

No. with + reactions:

13

Total no. in group:

19

Clinical observations:

Slight erythema, eschar formation, exfoliation, oedema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50% (v/v). No with. + reactions: 13.0. Total no. in groups: 19.0. Clinical observations: Slight erythema, eschar formation, exfoliation, oedema.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

50% (v/v)

No. with + reactions:

14

Total no. in group:

19

Clinical observations:

Slight erythema, eschar formation, exfoliation, oedema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50% (v/v). No with. + reactions: 14.0. Total no. in groups: 19.0. Clinical observations: Slight erythema, eschar formation, exfoliation, oedema.

Results 5

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

10% (v/v)

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

no response (exfoliation of 1 animal)

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10% (v/v). No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no response (exfoliation of 1 animal).

Results 6

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

10 % (v/v)

No. with + reactions:

2

Total no. in group:

20

Clinical observations:

slight erythema, exfoliation

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10 % (v/v). No with. + reactions: 2.0. Total no. in groups: 20.0. Clinical observations: slight erythema, exfoliation.

Results 7

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

10% (v/v)

No. with + reactions:

5

Total no. in group:

20

Clinical observations:

slight erythema, exfoliation

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10% (v/v). No with. + reactions: 5.0. Total no. in groups: 20.0. Clinical observations: slight erythema, exfoliation.

Results 8

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

10% (v/v)

No. with + reactions:

7

Total no. in group:

20

Clinical observations:

slight erythema, eschar formation, exfoliation

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10% (v/v). No with. + reactions: 7.0. Total no. in groups: 20.0. Clinical observations: slight erythema, eschar formation, exfoliation.

Any other information on results incl. tables

Signs of irritation during induction: 50% v/v - moderate erythema and oedema; no dermal response was evident following a similar administration of 50% v/v of test material in paraffin oil.

Occluded topical induction of test material as supplied gave rise to slight erythema, eschar formation, exfoliation, yellow staining, loss of flexibility and oedema.

Evidence of sensitization at each challenge: Challenge application of 50% v/v test material in paraffin oil gave rise to eschar formation or oedema in six test and five control animals. Exfoliation was evident in seventeen test and nineteen control animals. Challenge application of 10% v/v test material in paraffin oil caused eschar formation in two test and no control animals; a further seven test and two control animals showed slight erythema. Exfoliation was evident in twelve test and sixteen control animals. Challenge application of paraffin oil alone gave rise to slight erythema in two control animals. Exfoliation was evident in thirteen test and fourteen control animals. On this basis it is considered that a significant dermal response was observed in no test animal following challenge with 50% v/v test material in paraffin oil and in two test animals following challenge application of 10% v/v test material in paraffin oil.

Other observations: Abrasions were evident in the majority of test animals after second (topical) induction; the control animals remained in overt good health. One test animal was found dead on Day 2; necropsy revealed incomplete collapse of the lungs. All surviving animals achieved anticipated overall bodyweight gains.

Primary irritation screen

Phase 1 - Intradermal administration 

Animal number and sex	Concentration of Test material	Dermal responses at
		24 hours	time after injection 48 hours	7 days
311M	50% v/v in vehicle+	1	1	0
	30% v/v in vehicle	1	I	0
	10% v/v in vehicle	1	0	0
	50% v/v inFCA	2d	2d	0
	30% v/v in FCA	2	2	0
	10% v/v in FCA	2	1	0
317F	50% v/v in vehicle+	1	0	0
	30%o v/v in vehicle	1	0	0
	10% v/v in vehicle	1	0	0
	50% v/v in FCA	2	1	0
	30% v/v in FCA	2	1	0
	10% v/v in FCA	1	0	0
312M	5% v/v in vehicle	0	0	0
	3% v/v in vehicle	0	0	0
	1%) v/v in vehicle	1p	0p	0
	5% v/v in FCA	1	0	0
	3% v/v in FCA	1	0	0
	1% v/v in FCA	1	0	0
318F	5% v/v in vehicle	0	0	0
	3% v/v in vehicle	0	0	0
	1 % v/v in vehicle	1	0	0
	5% v/v in FCA	1	1	0
	3% v/v in FCA	0	0	0
	1% v/v in FCA	0	0	0

0      No response

1      Slight erythema

2      Moderate erythema

d      Test site discoloured

p      pallor

FCA Freunds Complete Adjuvant

+      Maximum practicable concentration (Section 3.5)

 

Primary irritation screen

Phase 2 - Topical induction administration 

Animal number and sex	Concentration of Test material in vehicle	Dermal responses at time after removal of dressings
		24 hours	48 hours	7 days
313M	100%+	0	0	0
	50% v/v	0	0	0
	30% v/v	0	0	0
	10% v/v	0	0	0
319F	100%+	0	0f	0
	50% v/v	0	0	0
	30% v/v	0	0	0
	10% v/v	0	0	0

 

0      No response

f       Exfoliation

+      Maximum practicable concentration (Section 3.5)

 

Primary irritation screen

Phase 3 - Topical challenge administration 

Animal number and sex	Concentration of Test material in vehicle	Dermal responses at time after removal of dressings
		24 hours	48 hours
314M	100%+	0	0f
	50% v/v	0	0
	30% v/v	0f	0f
	10% v/v	0	0
315M	100%+	0f	0f
	50% v/v	0	0
	30% v/v	0	0
	10% v/v	0	0
320F	100%+	3'e	3'e
	50% v/v	0	0
	30% v/v	1	0f
	10% v/v	0	0

0      No response

1      Slight erythema

3'e   Eschar formation

f       Exfoliation

+      Maximum practicable concentration (Section 3.5)

First phase of induction - dermal responses after intradermal injection

Control Group 

Response	Number of animals showing response+
	A		B		C
	Male	Female	Male	Female	Male	Female
No response			10	10	2
Slight erythema	1				1	1
Moderate erythema	9	10			7	9
Pallor					1

+   Ten animals in each sex-group

A:  FCA

B:  Vehicle

C:  Vehicle in FCA

  

Test Group 

Response	Number of animals showing response+
	A		B		C
	Male	Female	Male	Female	Male	Female
No response			10	9
Moderate erythema	10	9			10	9
Oedema					9	9

+      Ten animals in each sex-group - one female animal was found dead on Day 2.

A:     FCA

B:     Test material in vehicle

C:     Test material in FCA

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

It was concluded that, under the conditions of this study, repeated administration of 'Kerosine (Fischer-Tropsch), limited range, C8-12, branched and linear', an analogue to 'Kerosine (Fischer-tropsch), C8-16 - branched and linear' had a weak potential to cause delayed contact hypersensitivity in guinea pigs. The incident of significant response was below the EEC trigger threshold for classification as a dermal sensitizer (according to EC No. 1272/2008).

Executive summary:

In a guinea pig maximisation test, conducted in accordance with OECD 406 and GLP, 'Kerosine (Fischer-Tropsch), limited range, C8-12, branched and linear', was tested at challenge concentrations of 50% v/v and 10% v/v in paraffin oil. Intradermal induction was performed at a concentration of 50% v/v, and topical induction used undiluted test substance.

Challenge application of 50% v/v gave rise to eschar formation or oedema in six test and five control animals, moderate erythema in one test animal and slight erythema in test and six control animals. Exfoliation was evident in seventeen test and nineteen control animals.

Challenge application of 10% v/v caused eschar formation in two test and no control animals; a further seven test and two control animals showed slight erythema. Exfoliation was evident in twelve test and sixteen control animals.

Challenge application of paraffin oil alone gave rise to slight erythema in two control animals. Exfoliation was evident in 13 test and 14 control animals. 

Abrasions were evident in the majority of test animals after the second (topical) induction; the control animals remained in overt good health. One test group animal was found dead on Day 2; necropsy revealed incomplete collapse of the lungs. There were no clinical observations and no effects on overall bodyweight gains.

A significant dermal response (a reaction more marked than the most severe evident amongst the control animals) was observed in no test animal following challenge with 50% v/v 'Kerosine (Fischer-Tropsch), limited range, C8-12, branched and linear', in paraffin oil, and in 2/20 test animals following challenge application of 10% v/v 'Kerosine (Fischer-Tropsch), limited range, C8-12, branched and linear', in paraffin oil.

It was concluded that, under the conditions of this study, repeated administration of 'Kerosine (Fischer-Tropsch), limited range, C8-12, branched and linear', an analogue to 'Kerosine (Fischer-tropsch), C8-16 - branched and linear' had a weak potential to cause delayed contact hypersensitivity in guinea pigs. The incident of significant response was below the EEC trigger threshold for classification as a dermal sensitizer (EC No 1272/2008).