Grapefruit, ext.

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

June 20, 1995 - July 28, 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was performed in accordance with internal standard operating procedures, and internally audited by a Quality Assurance Unit.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

other: method of Ritz and Buehler 1980

Deviations:

yes

Principles of method if other than guideline:

The potential of Grapefruit oil to produce delayed contact hypersensitivity in guinea pigs was evaluated using an adaptation of the method of Ritz and Buehler (Ritz HL and Buehler EV. Current Concepts in Cutaneous Toxicity, ed. Drill VA and Lazar T. (Academic Press, 1980) pp 25-40).

GLP compliance:

yes

Type of study:

Buehler test

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague Dawley, inc.
- Weight at study initiation: 405 - 478 grams at start of the primary irritation phase; 352 - 487 grams at start of the induction phase.
- Housing: individually housed in wire mesh suspension cages
- Diet: Teklad Guinea Pig Diet ad libitum
- Water: ad libitum
- Acclimation period: at least four days

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12-hour light / 12-hour dark cycle

Route:

epicutaneous, occlusive

Vehicle:

other: diethyl phthalate

Concentration / amount:

primary irritation phase: undiluted, 50%, 25%, 10%, 5%, 2.5%, 1%, 0.5%
induction phase: 25%
challenge phase: 5%, 1.5%, 0.5%

Route:

epicutaneous, occlusive

Vehicle:

other: diethyl phthalate

Concentration / amount:

primary irritation phase: undiluted, 50%, 25%, 10%, 5%, 2.5%, 1%, 0.5%
induction phase: 25%
challenge phase: 5%, 1.5%, 0.5%

No. of animals per dose:

primary irritation phase: 8 animals
main test: 20 test animals, 10 vehicle control animals

Details on study design:

RANGE FINDING TESTS: Irritation screening
- Concentrations: undiluted, 50%, 25%, 10%, 5%, 2.5%, 1%, 0.5%
- Site: back

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: three weeks
- Test groups: 25% w/v formulation in diethyl phthalate
- Control group: undiluted diethyl phthalate
- Site: left shoulder
- Frequency of applications: interval between exposures varied from 6 to 9 days
- Duration: 6 hours (approximately)
- Concentrations: 25% test substance

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: two weeks after last induction exposure
- Exposure period: 6 hours
- Test groups: same 20 test animals as during induction period
- Control group: same 10 control animals as during induction period
- Site: previously untreated test sites
- Concentrations: 5%, 1.5%, and 0.5% test substance in diethyl phthalate for both the test animals and the control animals; and undiluted diethyl phthalate for the control animals
- Evaluation (hr after challenge): the day following primary challenge exposure, evaluation was repeated the following day.

Challenge controls:

Animals receiving undiluted diethyl phthalate during the induction period and different concentrations of test substance during the challenge period.

Positive control substance(s):

no

Positive control results:

not applicable
Historical positive control data were added to the study report.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

5 %

No. with + reactions:

9

Total no. in group:

10

Clinical observations:

9 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 5 %. No with. + reactions: 9.0. Total no. in groups: 10.0. Clinical observations: 9 animals showed slight, patchy erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

1.5 %

No. with + reactions:

9

Total no. in group:

10

Clinical observations:

9 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 1.5 %. No with. + reactions: 9.0. Total no. in groups: 10.0. Clinical observations: 9 animals showed slight, patchy erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0.5 %

No. with + reactions:

9

Total no. in group:

10

Clinical observations:

9 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0.5 %. No with. + reactions: 9.0. Total no. in groups: 10.0. Clinical observations: 9 animals showed slight, patchy erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0

No. with + reactions:

8

Total no. in group:

10

Clinical observations:

8 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0. No with. + reactions: 8.0. Total no. in groups: 10.0. Clinical observations: 8 animals showed slight, patchy erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

5 %

No. with + reactions:

17

Total no. in group:

19

Clinical observations:

17 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 5 %. No with. + reactions: 17.0. Total no. in groups: 19.0. Clinical observations: 17 animals showed slight, patchy erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

1.5 %

No. with + reactions:

18

Total no. in group:

19

Clinical observations:

18 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 1.5 %. No with. + reactions: 18.0. Total no. in groups: 19.0. Clinical observations: 18 animals showed slight, patchy erythema.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.5 %

No. with + reactions:

11

Total no. in group:

19

Clinical observations:

11 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0.5 %. No with. + reactions: 11.0. Total no. in groups: 19.0. Clinical observations: 11 animals showed slight, patchy erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

5.0 %

No. with + reactions:

8

Total no. in group:

10

Clinical observations:

8 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 5.0 %. No with. + reactions: 8.0. Total no. in groups: 10.0. Clinical observations: 8 animals showed slight, patchy erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

1.5 %

No. with + reactions:

6

Total no. in group:

10

Clinical observations:

6 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 1.5 %. No with. + reactions: 6.0. Total no. in groups: 10.0. Clinical observations: 6 animals showed slight, patchy erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0.5 %

No. with + reactions:

6

Total no. in group:

10

Clinical observations:

6 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0.5 %. No with. + reactions: 6.0. Total no. in groups: 10.0. Clinical observations: 6 animals showed slight, patchy erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0

No. with + reactions:

5

Total no. in group:

10

Clinical observations:

5 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0. No with. + reactions: 5.0. Total no. in groups: 10.0. Clinical observations: 5 animals showed slight, patchy erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

5 %

No. with + reactions:

10

Total no. in group:

19

Clinical observations:

10 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5 %. No with. + reactions: 10.0. Total no. in groups: 19.0. Clinical observations: 10 animals showed slight, patchy erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

1.5 %

No. with + reactions:

10

Total no. in group:

19

Clinical observations:

10 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1.5 %. No with. + reactions: 10.0. Total no. in groups: 19.0. Clinical observations: 10 animals showed slight, patchy erythema.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

0.5 %

No. with + reactions:

8

Total no. in group:

19

Clinical observations:

8 animals showed slight, patchy erythema

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0.5 %. No with. + reactions: 8.0. Total no. in groups: 19.0. Clinical observations: 8 animals showed slight, patchy erythema.

No control animals exhibited grades greater than 0.5 (slight, patchy erythema). A test animal must therefore at least exhibit a grade 1 at 48 hours to be considered positive (According to the selection criteria defined in the 'interpretation of the results').

Interpretation of results:

other: not sensitising

Remarks:

Based on CLP criteria

Conclusions:

The test substance grapefruit oil showed no sensitising properties in guinea pigs under the conditions of this test.

Executive summary:

The potential of grapefruit oil (25% w/v in diethyl phtalate), to produce delayed contact hypersensitiviy in guinea pigs was evaluated using an adaptation of the method of Ritz and Buehler. Following primary challenge using grapefruit oil as 5%, 1.5%, and 0.5% w/v formulations in diethyl phtalate, responses in the test group were compared to those of the vehicle control group. The incidence of test animals exhibiting a positive response to levels of 5%, 1.5%, and 0.5% test substance was 0 of 19 for all concentrations. These data indicate that sensitisation to grapefruit oil had not been induced. Based on the EU criteria outlined in 1272/2008/EC and 67/548/EEC, grapefruit oil 25% is not a sensitiser.

Reference 2

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 07 to 18, 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP study conducted in compliance with OECD guideline 429 with minor deviations: no certificate of analysis of the test substance. Due to the read-across purpose it was given a Klimisch 2 rating, in accordance with the ECHA Practical guide #6 on the reporting of read-across in IUCLID.

Justification for type of information:

The justification for read across is provided in the attached background material of the chapter summary.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

no certificate of analysis of the test substance

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Species:

mouse

Strain:

other: CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Ltd, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 17.7-20 g
- Housing: Housed in a group of 4 mice (maximum) per cage
- Diet (e.g. ad libitum): Diet (RM1) (Special Diet Services Limited, Witham, UK), ad libitum
- Water (e.g. ad libitum): Mains water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30-70%
- Air changes (per hour): 15/hour (minimum)
- Photoperiod (hours dark / hours light): 12 hours dark / 12 hours light

Vehicle:

other: ethanol/diethyl phthalate (3: 1 v/v)

Concentration:

0, 10, 25, 50, 75 or 100% v/v in ethanol/diethyl phthalate (3: 1 v/v)

No. of animals per dose:

Four females

Details on study design:

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: Local Lymph Node Assay
- Criteria used to consider a positive response: One or more concentrations of the test substance should elicit a 3-fold or greater increase in isotope incorporation relative to the vehicle control group.

TREATMENT PREPARATION AND ADMINISTRATION: 25 µL of test material at concentrations of 0 (vehicle control), 10, 25, 50, 75 or 100% v/v in ethanol/diethyl phthalate (3: 1 v/v) were applied to the dorsal surface of each ear on Days 1, 2 and 3. On Day 6, 250 μL phosphate buffered saline (PBS) containing 20 μCi of 3H-methyl thymidine (specific gravity: 2.0 Ci/mmol) was injected into the tail vein of each experimental mouse. Five hours later, all mice were killed by inhalation of halothane vapour followed by cervical dislocation and the draining auricular lymph node of each ear was excised into PBS. A single cell suspension of lymph node cells (LNC) was prepared by gentle separation through 200-mesh stainless steel gauze. LNC were washed thrice with 10 mL of PBS and precipitated with 5% trichloroacetic acid (TCA) at 4 °C. Pellets were re-suspended in 1 mL TCA and transferred to scintillation vials containing 10 mL of scintillation fluid (Optiphase) for 3H-counting. The number of radioactive disintegrations per minute (DPM) was measured using a Packard Tri-Carb Liquid Scintillation Counter.

Positive control substance(s):

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

Statistics:

No data

Positive control results:

Mean DPM
- Vehicle (acetone/olive oil 4:1 v/v) = 2263
- Test material (5 % w/v) = 5496
- Test material (10 % w/v) = 7571
- Test material (25 % w/v) = 24752

Stimulation Index
- Test material (5 % w/v) = 2.4
- Test material (10 % w/v) = 3.3
- Test material (25 % w/v) = 10.9

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: - Vehicle = 2511 - Test material (10% v/v) = 3319 - Test material (25% v/v) = 8554 - Test material (50% v/v) = 9916 - Test material (75% v/v) = 22063 - Test material (100% v/v) = 16259

Key result

Parameter:

SI

Value:

1.3

Test group / Remarks:

10% v/v

Key result

Parameter:

SI

Value:

3.4

Test group / Remarks:

25% v/v

Key result

Parameter:

SI

Value:

4

Test group / Remarks:

50% v/v

- Dermal reactions: Application of the test substance did not result in an increase in visual levels of irritancy to the skin on or around the ear area for the duration of the study.

Table 1 - Skin sensitisation potential of d-limonene in ethanol/diethyl phthalate (3:1 v/v)

 

Concentration of test substance (% v/v)	Number of lymph nodes assayed	Disintegrations per min (dpm)	dpm per lymph node	Test:control ratio
0 (vehicle only)	8	2511	314	Not applicable
10	8	3319	415	1.3
25	8	8554	1069	3.4
50	8	9916	1240	4.0
75	8	22063	2758	8.8
100	8	16259	2032	6.5
EC3	Estimated to be 22% (5500 µg/cm2)

 

Table 2 - Skin sensitisation potential of the positive control substance (hexylcinnamaldehyde)

 

Concentration of hexylcinnamaldehyde (% w/v)	Number of lymph nodes assayed	Disintegrations per min (dpm)	dpm per lymph node	Test:control ratio
0 (vehicle only)	8	2263	283	Not applicable
5	8	5496	687	2.4
10	8	7571	946	3.3
25	8	24752	3094	10.9

Interpretation of results:

other: Sensitising category 1

Remarks:

Based on CLP criteria

Conclusions:

Based on the results of the study for read-across substance D-limonene, Grapefruit oil "May cause sensitisation by skin contact", according to the criteria of Annex VI to the Directive 67/548/EEC and ‘Category 1’ according to the CLP Regulation (EC) N° (1272-2008).

Executive summary:

In a local lymph node assay performed in CBA/Ca strain mice according to OECD guideline 429 and in compliance with GLP, groups of mice (4 females/dose) were applied with 25 µL of d-limonene at concentrations of 0 (vehicle control), 10, 25, 50, 75 or 100% v/v in ethanol/diethyl phthalate (3: 1 v/v) to the dorsal surface of each ear for three consecutive days. On Day 6, all animals were injected with 3H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised and measured for radioactivity expressed as number of disintegrations per minute (DPM). Historic data of hexylcinnamaldehyde (5, 10 and 25 % w/v) in acetone/olive oil (4:1 v/v) was used as the data for positive control group.

 

Mean DPM for 0, 10, 25, 50, 75 or 100% d-limonene were observed to be 2511, 3319, 8554, 9916, 22063 or 16259 dpm, respectively. Stimulation index for 10, 25, 50, 75 or 100% d-limonene were calculated to be 1.3, 3.4, 4.0, 8.8 or 6.5, respectively. The estimated concentration giving rise to a 3 fold increase in lymphocyte proliferation (EC3) was 22% v/v (5500 µg/cm2). No increase in visual levels of irritancy to the ear skin was observed during the study.

 

Under the test conditions, d-limonene is classified as ‘R43 May cause sensitisation by skin contact’, according to the criteria of Annex VI to the Directive 67/548/EEC and ‘Category 1’ according to the CLP Regulation (EC) N° (1272-2008).

Reference 3

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP study conducted in compliance with OECD guideline 429 with minor deviations: no certificate of analysis of the test substance. Due to the read-across purpose it was given a Klimisch 2 rating, in accordance with the ECHA Practical guide #6 on the reporting of read-across in IUCLID.

Justification for type of information:

The read across justification is presented in the document attached to this record.

Reason / purpose for cross-reference:

read-across source

Statistics:

No data

Positive control results:

Mean DPM
- Vehicle (acetone/olive oil 4:1 v/v) = 2263
- Test material (5 % w/v) = 5496
- Test material (10 % w/v) = 7571
- Test material (25 % w/v) = 24752

Stimulation Index
- Test material (5 % w/v) = 2.4
- Test material (10 % w/v) = 3.3
- Test material (25 % w/v) = 10.9

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: - Vehicle = 2511 - Test material (10% v/v) = 3319 - Test material (25% v/v) = 8554 - Test material (50% v/v) = 9916 - Test material (75% v/v) = 22063 - Test material (100% v/v) = 16259

Key result

Parameter:

SI

Value:

1.3

Test group / Remarks:

10% v/v

Key result

Parameter:

SI

Value:

3.4

Test group / Remarks:

25% v/v

Key result

Parameter:

SI

Value:

4

Test group / Remarks:

50% v/v

- Dermal reactions: Application of the test substance did not result in an increase in visual levels of irritancy to the skin on or around the ear area for the duration of the study.

Table 1 - Skin sensitisation potential of d-limonene in ethanol/diethyl phthalate (3:1 v/v)

 

Concentration of test substance (% v/v)	Number of lymph nodes assayed	Disintegrations per min (dpm)	dpm per lymph node	Test:control ratio
0 (vehicle only)	8	2511	314	Not applicable
10	8	3319	415	1.3
25	8	8554	1069	3.4
50	8	9916	1240	4.0
75	8	22063	2758	8.8
100	8	16259	2032	6.5
EC3	Estimated to be 22% (5500 µg/cm2)

 

Table 2 - Skin sensitisation potential of the positive control substance (hexylcinnamaldehyde)

 

Concentration of hexylcinnamaldehyde (% w/v)	Number of lymph nodes assayed	Disintegrations per min (dpm)	dpm per lymph node	Test:control ratio
0 (vehicle only)	8	2263	283	Not applicable
5	8	5496	687	2.4
10	8	7571	946	3.3
25	8	24752	3094	10.9

Interpretation of results:

other: Sensitising Category 1

Remarks:

Based on CLP criteria

Conclusions:

Under the test conditions, d-limonene is classified as ‘R43 May cause sensitisation by skin contact’, according to the criteria of Annex VI to the Directive 67/548/EEC and ‘Category 1’ according to the CLP Regulation (EC) N° (1272-2008).

Executive summary:

In a local lymph node assay performed in CBA/Ca strain mice according to OECD guideline 429 and in compliance with GLP, groups of mice (4 females/dose) were applied with 25 µL of d-limonene at concentrations of 0 (vehicle control), 10, 25, 50, 75 or 100% v/v in ethanol/diethyl phthalate (3: 1 v/v) to the dorsal surface of each ear for three consecutive days. On Day 6, all animals were injected with 3H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised and measured for radioactivity expressed as number of disintegrations per minute (DPM). Historic data of hexylcinnamaldehyde (5, 10 and 25 % w/v) in acetone/olive oil (4:1 v/v) was used as the data for positive control group.

 

Mean DPM for 0, 10, 25, 50, 75 or 100% d-limonene were observed to be 2511, 3319, 8554, 9916, 22063 or 16259 dpm, respectively. Stimulation index for 10, 25, 50, 75 or 100% d-limonene were calculated to be 1.3, 3.4, 4.0, 8.8 or 6.5, respectively. The estimated concentration giving rise to a 3 fold increase in lymphocyte proliferation (EC3) was 22% v/v (5500 µg/cm2). No increase in visual levels of irritancy to the ear skin was observed during the study.

 

Under the test conditions, d-limonene is classified as ‘R43 May cause sensitisation by skin contact’, according to the criteria of Annex VI to the Directive 67/548/EEC and ‘Category 1’ according to the CLP Regulation (EC) N° (1272-2008).

Reference 4

Endpoint:

skin sensitisation: in vitro

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

an in vitro skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (sensitising)

Additional information:

The key study is a local lymph node assay performed with major constituent limonene which was read across to grapefruit oil. The study was performed in CBA/Ca strain mice according to OECD guideline 429 and in compliance with GLP.

Mean DPM for 0, 10, 25, 50, 75 or 100% d-limonene were observed to be 2511, 3319, 8554, 9916, 22063 or 16259 dpm, respectively. Stimulation index for 10, 25, 50, 75 or 100% d-limonene were calculated to be 1.3, 3.4, 4.0, 8.8 or 6.5, respectively. The estimated concentration giving rise to a 3 fold increase in lymphocyte proliferation (EC3) was 22% v/v (5500 µg/cm2). No increase in visual levels of irritancy to the ear skin was observed during the study.

Under the test conditions, d-limonene is classified as ‘R43 May cause sensitisation by skin contact’, according to the criteria of Annex VI to the Directive 67/548/EEC and ‘Category 1’ according to the CLP Regulation (EC) N° (1272-2008).

 

Three supporting studies on the skin sensitising potential of grapefruit oil were available for the dossier, a human repeated insult patch test, a human maximisation study and a delayed contact hypersensitivity study in guinea pigs.

In the repeated insult patch test (included in 7.10.4), the sensitising properties of grapefruit oil were investigated in humans. 106 subjects completed the study, none of them showed sensitisation.

In the maximisation test (included in 7.10.4), exposure to grapefruit oil also did not induce sensitisation in 24 healthy adult males.

In the delayed contact hypersensitivity study (Buehler Technique), the potential of grapefruit oil to induce skin sensitisation was investigated. After the challenge with several concentrations of test substance, none of the guinea pigs showed sensitisation compared to the controls.

Based on the results of the supporting studies, it can be concluded that the substance graperfruit oil is not a sensitiser. However, as the major constituent of grapefruit oil is limonene, which is classified for skin sensitisation, grapefruit oil will be classified for skin sensitisation.

Migrated from Short description of key information:
Skin sensitisation::
Read across based on d-limonene content: sensitising

Justification for classification or non-classification

Based on the skin sensitising properties of the major constituent of grapefruit oil, d-limonene, grapefruit oil has to be labelled as skin sensitiser in accordance with the EU criteria outlined in Annex VI of 67/548/EEC and Annex I of 1272/2008/EC.