Registration Dossier
Registration Dossier
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EC number: 947-787-5 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data

Endpoint summary
Administrative data
Description of key information
No information was available on the test substance, therefore read across was used to fill the endpoint. Four key studies (Klimisch 1) were available for oleic acid and linoleic acid, using the LLNA and GMPT methods. Oleic acid and linoleic acid gave clear positive results in the LLNA with Stimulation Index (SI) values in the range between 2.6 and 14.9 for oleic acid and 1.5 and 9.1 for linoleic acid. No signs of systemic toxicity were observed for any of the test substances in any of the animals in a GPMT test. A Klimisch 4 study was also available from the Cosmetics Ingredient Review conducted with undiluted glyceryl citrate/lactate/linoleate/oleate in guinea pigs and found to be non-sensitising. Human data for glyceryl linoleate and glyceryl oleate are also available and presented in the "Exposure related observations in humans" section.
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- 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:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- 1992
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- Version / remarks:
- 1996
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Done in conjunction with the LLNA method
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source/batch No.of test material:
Sigma-Aldrich, Munich, Germany / Aldrich No. L1376
- Purity: 99% - Species:
- guinea pig
- Strain:
- other: SPF-Hsd Poc: DH
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source:
Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: yes
- Microbiological status of animals, when known:
SPF
- Weight at study initiation:
300 - 500 grams
- Housing:
Terluran cages on Altromin saw fiber bedding or Lignocel bedding
- Diet (e.g. ad libitum):
Ad libitum
- Water (e.g. ad libitum):
Ad libitum
- Acclimation period:
At least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Route:
- intradermal
- Vehicle:
- other: 1:1 (v/v) mixture of FCA/physiological saline
- Concentration / amount:
- 5%
- Day(s)/duration:
- Day 0
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 100%
- Day(s)/duration:
- Day 7 / 48 hours
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- Day 20 / 24 hours
- Adequacy of challenge:
- not specified
- No.:
- #2
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- Day 28 / 24 hours
- Adequacy of challenge:
- not specified
- No. of animals per dose:
- 10 test animals and 5 control animals
- Details on study design:
- RANGE FINDING TESTS: Performed in a preliminary experiment assessing the irritant effects of the test compound after intradermal and topical application
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: Day 0 and Day 7 (48 hours)
- Test groups: One test group exposed to linoleic acid
- Control group: One control group exposed to vehicle only
- Frequency of applications: Once
- Duration: Day 0 to Day 20
- Concentrations: Intradermal induction - 5%; Topical induction - 100%
B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: 20 and 28
- Exposure period: 24 hours
- Test groups: One test group exposed to linoleic acid
- Control group: One control group exposed to vehicle only
- Concentrations: 50%
- Evaluation (hr after challenge): 24, 48 and 72 hours
- Challenge controls:
- Vehicle only
- Positive control substance(s):
- not specified
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 2
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- other: 3rd reading
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 4
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- rechallenge
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 3
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- rechallenge
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 2
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported.
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- 100%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported.
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- other: Data inconclusive
- Conclusions:
- Due to non-specific irritative skin reactions observed, it was concluded that the challenge test concentration was too high, and no conclusion in regard to skin sensitization could be made.
- Executive summary:
Female SPF-Hsd Poc: DH guinea pigs, weighing between 300 and 500 grams, were obtained for this guinea pig maximisation test (GPMT). The guinea pigs were divided into 2 groups, a negative control group containing 5 guinea pigs, and a test group containing 10 guinea pigs. The animals recieved two induction treatments, an intradermal injection (day 0) and a topical application (day 7). The test concentrations used for the two induction tests were 5% linoleic acid for the intradermal induction and 100% linoleic acid for the topical induction. The animals also recieved two topical challenge treatments, one on day 20 and one on day 28. The test concentration used for the two challenge tests was 50% linoleic acid. Isotonic saline was used as the vehicle for the intradermal injection and vaseline was used as the vehicle for the topical exposures, and control animals were dosed with the vehicle alone. After challenge and re-challenge with linoleic acid, skin reactions were observed in the control group as frequently or even at a higher incidence than the test group. Therefore, it was concluded that the test concentration was too high and included irritative reactions and therefore no conclusion regarding skin sensitization could be drawn.
- 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:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- 1992
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- Version / remarks:
- 1996
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Done in conjunction with the LLNA method
- Species:
- guinea pig
- Strain:
- other: SPF-Hsd Poc: DH
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: yes
- Microbiological status of animals, when known: SPF
- Weight at study initiation: 300 - 500 grams
- Housing: Terluran cages on Altromin saw fiber bedding or Lignocel bedding
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Route:
- intradermal
- Vehicle:
- other: 1:1 (v/v) mixture of FCA/physiological saline
- Concentration / amount:
- 5%
- Day(s)/duration:
- Day 0
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- Day 7 / 48 hours
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 25%
- Day(s)/duration:
- Day 20 / 24 hours
- Adequacy of challenge:
- not specified
- No.:
- #2
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 25%
- Day(s)/duration:
- Day 28 / 24 hours
- Adequacy of challenge:
- not specified
- No. of animals per dose:
- 10 test animals and 5 control animals
- Details on study design:
- RANGE FINDING TESTS: Performed in a preliminary experiment assessing the irritant effects of the test compound after intradermal and topical application
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: Day 0 and Day 7 (48 hours)
- Test groups: One test group exposed to oleic acid
- Control group: One control group exposed to vehicle only
- Frequency of applications: Once
- Duration: Day 0 to Day 20
- Concentrations: Intradermal induction - 5%; Topical induction - 50%
B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: 20 and 28
- Exposure period: 24 hours
- Test groups: One test group exposed to oleic acid
- Control group: One control group exposed to vehicle only
- Concentrations: 25%
- Evaluation (hr after challenge): 24, 48 and 72 hours
- Challenge controls:
- Vehicle only
- Positive control substance(s):
- not specified
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Reading:
- other: 3rd reading
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 4
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 2
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- rechallenge
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 3
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 3
- Total no. in group:
- 10
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- 100%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported.
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- study cannot be used for classification
- Conclusions:
- For oleic acid, the skin sensitization potential was confirmed in two animals out of ten.
- Executive summary:
Female SPF-Hsd Poc: DH guinea pigs, weighing between 300 and 500 grams, were obtained for this guinea pig maximisation test (GPMT). The guinea pigs were divided into 2 groups, a negative control group containing 5 guinea pigs, and a test group containing 10 guinea pigs. The animals recieved two induction treatments, an intradermal injection (day 0) and a topical application (day 7). The test concentrations used for the two induction tests were 5% oleic acid for the intradermal induction and 50% oleic acid for the topical induction. The animals also recieved two topical challenge treatments, one on day 20 and one on day 28. The test concentration used for the two challenge tests was 25% oleic acid. Isotonic saline was used as the vehicle for the intradermal injection and vaseline was used as the vehicle for the topical exposures, and control animals were dosed with the vehicle alone. For oleic acid, one animal showed a skin reaction at 48 and 72 hours and an additional 3 animals showed skin reactions only at the 72 hours reading. Upon re-challenge, a skin reaction was reproduced only in two of the four animals. The skin sensitization was confirmed in two animals.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- secondary literature
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- no guideline followed
- GLP compliance:
- not specified
- Type of study:
- Buehler test
- Species:
- guinea pig
- Strain:
- not specified
- Sex:
- not specified
- Details on test animals and environmental conditions:
- Not specified.
- Route:
- epicutaneous, open
- Vehicle:
- unchanged (no vehicle)
- Day(s)/duration:
- 54 hours
- Adequacy of induction:
- not specified
- Route:
- epicutaneous, open
- Vehicle:
- unchanged (no vehicle)
- Day(s)/duration:
- 54 hours
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- unchanged (no vehicle)
- Day(s)/duration:
- 54 hours
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 20 test animals and 10 control animals
- Positive control substance(s):
- not specified
- Reading:
- 1st reading
- Hours after challenge:
- 30
- Group:
- test chemical
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- 2nd reading
- Hours after challenge:
- 54
- Group:
- test chemical
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Remarks on result:
- no indication of skin sensitisation
- Conclusions:
- Glyceryl citrate/lactate/linoleate/oleate did not induce sensitisation in guinea pigs.
- Executive summary:
The skin sensitisation potential of glyceryl citrate/lactate/linoleate/oleate was assessed using guinea pigs. Thirty guinea pigs were used for this study, 20 serving as test guinea pigs and 10 serving as controls. The test item was undiluted as the undiluted test item did not induce skin irritation in a preliminary test. The test item was applied via dermal application during induction phases I, II and III and the reactions were assessed 30 and 54 hours after initiation of treatment. A trigger concentration was also determined in week 4, as the highest non-irritating concentration. No skin reactions (erythema or oedema) were observed in induction phases I, II or III, or with the "trigger" concentration. Therefore, it was concluded that glyceryl citrate/lactate/linoleate/oleate did not induce sensitisation in guinea pigs.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2002
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2004
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- other: SPF-CBA/Ca01aHsd
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: SPF
- Age at study initiation: 6 to 12 weeks
- Housing: Macrolon cages on Altromin saw fiber bedding
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days
- Indication of any skin lesions: Not specified
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Vehicle:
- other: 3 + 1 (v/v) acetone/olive oil
- Concentration:
- 0, 10%, 25% and 50%
- No. of animals per dose:
- 5
- Details on study design:
- PRE-SCREEN TESTS:
- Systemic toxicity: Animals observed for systemic effects and local irritant effects
- Irritancy: Local irritant effects expressed as ear swelling were assessed by measuring ear thickness.
- Ear thickness measurements: Ear thickness measured in all groups using a mechanical calliper on day 1 and day 5
- Other measurements: Lymph node weight index (LNWI) were calculated as the arithmetic mean lymph node weight values of the test group and the negative control group
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: SI > 3
TREATMENT PREPARATION AND ADMINISTRATION: Treated by topical applications of test or control solution to the entire dorsal surface of each ear. Treatments performed once daily over three consecutive days. - Positive control substance(s):
- other: p-phenylenediamine
- Statistics:
- A one-sided t-test was used to identify a statistically significant increase in lymph node weights in treated groups vs. the vehicle control group
- Positive control results:
- Not reported
- Parameter:
- SI
- Value:
- 1.5
- Variability:
- 0.7
- Test group / Remarks:
- 10%
- Parameter:
- SI
- Value:
- 7
- Variability:
- 1.2
- Test group / Remarks:
- 25%
- Parameter:
- SI
- Value:
- 9.1
- Variability:
- 2.4
- Test group / Remarks:
- 50%
- Cellular proliferation data / Observations:
- DETAILS ON STIMULATION INDEX CALCULATION
: For each dose group, the stimulation index was calculated as the ratio of the arithmetic mean DPM (disintegrations per minute)/node values of the test group and the negative control group.
CLINICAL OBSERVATIONS: No systemic effects were observed for any of the treatments.
- Interpretation of results:
- Category 1B (indication of skin sensitising potential) based on GHS criteria
- Conclusions:
- Linoleic acid resulted in SI values above 3 and so is considered a skin sensitizer according to OECD TG 429.
- Executive summary:
Female SPF-CBA/Ca01aHsd mice, between 6 and 12 weeks old, were obtained for this local lymph node assay (LLNA). The mice were divided into 5 groups, a negative control, a vehicle control and three treatment groups, each containing 5 mice. The test substance, linoleic acid, was administered via topical application to the entire dorsal surface of each ear at concentrations of 0 (controls), 10, 25 and 50% once daily for three consecutive days. The vehicle used was 3 + 1 (v/v) acetone/olive oil. For each treatment group a stimulation index (SI) was calculated. The stimulation indexes for linoleic acid were 1.5, 7.0 and 9.1 for the 10%, 25% and 50% groups, respectively. Increased lymph node weight indexes were also observed at 25% (p<0.01) and 50% (p<0.01). An SI value > 3 indicated a positive result, therefore concentrations of 25% and 50% were considered positive as skin sensitizing. Linoleic acid was therefore considered a skin sensitizer according to the OECD TG 429.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across: supporting information
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2002
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2004
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- other: SPF-CBA/Ca01aHsd
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: SPF
- Age at study initiation: 6 to 12 weeks
- Housing: Macrolon cages on Altromin saw fiber bedding
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days
- Indication of any skin lesions: Not specified
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Vehicle:
- other: 3 + 1 (v/v) acetone/olive oil
- Concentration:
- 0, 10%, 25% and 50%
- No. of animals per dose:
- 5
- Details on study design:
- PRE-SCREEN TESTS:
- Systemic toxicity: Animals observed for systemic effects and local irritant effects
- Irritancy: Local irritant effects expressed as ear swelling were assessed by measuring ear thickness.
- Ear thickness measurements: Ear thickness measured in all groups using a mechanical calliper on day 1 and day 5
- Other measurements: Lymph node weight index (LNWI) were calculated as the arithmetic mean lymph node weight values of the test group and the negative control group
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: SI > 3
TREATMENT PREPARATION AND ADMINISTRATION: Treated by topical applications of test or control solution to the entire dorsal surface of each ear. Treatments performed once daily over three consecutive days. - Positive control substance(s):
- other: p-phenylenediamine
- Statistics:
- A one-sided t-test was used to identify a statistically significant increase in lymph node weights in treated groups vs. the vehicle control group
- Positive control results:
- Not reported
- Parameter:
- SI
- Value:
- 2.6
- Variability:
- 1.6
- Test group / Remarks:
- 10%
- Parameter:
- SI
- Value:
- 14.9
- Variability:
- 6.8
- Test group / Remarks:
- 25%
- Parameter:
- SI
- Value:
- 6.9
- Variability:
- 3.6
- Test group / Remarks:
- 50%
- Cellular proliferation data / Observations:
- DETAILS ON STIMULATION INDEX CALCULATION
: For each dose group, the stimulation index was calculated as the ratio of the arithmetic mean DPM (disintegrations per minute)/node values of the test group and the negative control group.
CLINICAL OBSERVATIONS: No systemic effects were observed for any of the treatments.
- Interpretation of results:
- Category 1B (indication of skin sensitising potential) based on GHS criteria
- Conclusions:
- Oleic acid resulted in SI values above 3 and so is considered a skin sensitizer according to OECD TG 429.
- Executive summary:
Female SPF-CBA/Ca01aHsd mice, between 6 and 12 weeks old, were obtained for this local lymph node assay (LLNA). The mice were divided into 5 groups, a negative control, a vehicle control and three treatment groups, each containing 5 mice. The test substance, oleic acid, was administered via topical application to the entire dorsal surface of each ear at concentrations of 0 (controls), 10, 25 and 50% once daily for three consecutive days. The vehicle used was 3 + 1 (v/v) acetone/olive oil. For each treatment group a stimulation index (SI) was calculated. The stimulation indexes for oleic acid were 2.6, 14.9 and 6.9 for the 10%, 25% and 50% groups, respectively. Increased lymph node weight indexes were also observed at 25% (p<0.01) and 50% (p<0.01). An SI value > 3 indicated a positive result, therefore concentrations of 25% and 50% were considered positive as skin sensitizing. The results showed a bell-shaped dose response curve as the SI value was lower at 50% than at 25%. Oleic acid was therefore considered a skin sensitizer according to the OECD TG 429.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- 1992
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- Version / remarks:
- 1996
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Done in conjunction with the LLNA method
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source/batch No.of test material:
Sigma-Aldrich, Munich, Germany / Aldrich No. L1376
- Purity: 99% - Species:
- guinea pig
- Strain:
- other: SPF-Hsd Poc: DH
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source:
Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: yes
- Microbiological status of animals, when known:
SPF
- Weight at study initiation:
300 - 500 grams
- Housing:
Terluran cages on Altromin saw fiber bedding or Lignocel bedding
- Diet (e.g. ad libitum):
Ad libitum
- Water (e.g. ad libitum):
Ad libitum
- Acclimation period:
At least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Route:
- intradermal
- Vehicle:
- other: 1:1 (v/v) mixture of FCA/physiological saline
- Concentration / amount:
- 5%
- Day(s)/duration:
- Day 0
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 100%
- Day(s)/duration:
- Day 7 / 48 hours
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- Day 20 / 24 hours
- Adequacy of challenge:
- not specified
- No.:
- #2
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- Day 28 / 24 hours
- Adequacy of challenge:
- not specified
- No. of animals per dose:
- 10 test animals and 5 control animals
- Details on study design:
- RANGE FINDING TESTS: Performed in a preliminary experiment assessing the irritant effects of the test compound after intradermal and topical application
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: Day 0 and Day 7 (48 hours)
- Test groups: One test group exposed to linoleic acid
- Control group: One control group exposed to vehicle only
- Frequency of applications: Once
- Duration: Day 0 to Day 20
- Concentrations: Intradermal induction - 5%; Topical induction - 100%
B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: 20 and 28
- Exposure period: 24 hours
- Test groups: One test group exposed to linoleic acid
- Control group: One control group exposed to vehicle only
- Concentrations: 50%
- Evaluation (hr after challenge): 24, 48 and 72 hours
- Challenge controls:
- Vehicle only
- Positive control substance(s):
- not specified
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 2
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- other: 3rd reading
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 4
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- rechallenge
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 3
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- rechallenge
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 50%
- No. with + reactions:
- 2
- Total no. in group:
- 10
- Remarks on result:
- not determinable
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported.
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- 100%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported.
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- other: Data inconclusive
- Conclusions:
- Due to non-specific irritative skin reactions observed, it was concluded that the challenge test concentration was too high, and no conclusion in regard to skin sensitization could be made.
- Executive summary:
Female SPF-Hsd Poc: DH guinea pigs, weighing between 300 and 500 grams, were obtained for this guinea pig maximisation test (GPMT). The guinea pigs were divided into 2 groups, a negative control group containing 5 guinea pigs, and a test group containing 10 guinea pigs. The animals recieved two induction treatments, an intradermal injection (day 0) and a topical application (day 7). The test concentrations used for the two induction tests were 5% linoleic acid for the intradermal induction and 100% linoleic acid for the topical induction. The animals also recieved two topical challenge treatments, one on day 20 and one on day 28. The test concentration used for the two challenge tests was 50% linoleic acid. Isotonic saline was used as the vehicle for the intradermal injection and vaseline was used as the vehicle for the topical exposures, and control animals were dosed with the vehicle alone. After challenge and re-challenge with linoleic acid, skin reactions were observed in the control group as frequently or even at a higher incidence than the test group. Therefore, it was concluded that the test concentration was too high and included irritative reactions and therefore no conclusion regarding skin sensitization could be drawn.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- 1992
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- Version / remarks:
- 1996
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Done in conjunction with the LLNA method
- Species:
- guinea pig
- Strain:
- other: SPF-Hsd Poc: DH
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: yes
- Microbiological status of animals, when known: SPF
- Weight at study initiation: 300 - 500 grams
- Housing: Terluran cages on Altromin saw fiber bedding or Lignocel bedding
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Route:
- intradermal
- Vehicle:
- other: 1:1 (v/v) mixture of FCA/physiological saline
- Concentration / amount:
- 5%
- Day(s)/duration:
- Day 0
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- Day 7 / 48 hours
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 25%
- Day(s)/duration:
- Day 20 / 24 hours
- Adequacy of challenge:
- not specified
- No.:
- #2
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 25%
- Day(s)/duration:
- Day 28 / 24 hours
- Adequacy of challenge:
- not specified
- No. of animals per dose:
- 10 test animals and 5 control animals
- Details on study design:
- RANGE FINDING TESTS: Performed in a preliminary experiment assessing the irritant effects of the test compound after intradermal and topical application
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: Day 0 and Day 7 (48 hours)
- Test groups: One test group exposed to oleic acid
- Control group: One control group exposed to vehicle only
- Frequency of applications: Once
- Duration: Day 0 to Day 20
- Concentrations: Intradermal induction - 5%; Topical induction - 50%
B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: 20 and 28
- Exposure period: 24 hours
- Test groups: One test group exposed to oleic acid
- Control group: One control group exposed to vehicle only
- Concentrations: 25%
- Evaluation (hr after challenge): 24, 48 and 72 hours
- Challenge controls:
- Vehicle only
- Positive control substance(s):
- not specified
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Reading:
- other: 3rd reading
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 4
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 2
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- rechallenge
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 3
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 25%
- No. with + reactions:
- 3
- Total no. in group:
- 10
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- 100%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- Remarks on result:
- other: Not reported.
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- study cannot be used for classification
- Conclusions:
- For oleic acid, the skin sensitization potential was confirmed in two animals out of ten.
- Executive summary:
Female SPF-Hsd Poc: DH guinea pigs, weighing between 300 and 500 grams, were obtained for this guinea pig maximisation test (GPMT). The guinea pigs were divided into 2 groups, a negative control group containing 5 guinea pigs, and a test group containing 10 guinea pigs. The animals recieved two induction treatments, an intradermal injection (day 0) and a topical application (day 7). The test concentrations used for the two induction tests were 5% oleic acid for the intradermal induction and 50% oleic acid for the topical induction. The animals also recieved two topical challenge treatments, one on day 20 and one on day 28. The test concentration used for the two challenge tests was 25% oleic acid. Isotonic saline was used as the vehicle for the intradermal injection and vaseline was used as the vehicle for the topical exposures, and control animals were dosed with the vehicle alone. For oleic acid, one animal showed a skin reaction at 48 and 72 hours and an additional 3 animals showed skin reactions only at the 72 hours reading. Upon re-challenge, a skin reaction was reproduced only in two of the four animals. The skin sensitization was confirmed in two animals.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- supporting study
- Reliability:
- 4 (not assignable)
- Rationale for reliability incl. deficiencies:
- secondary literature
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- no guideline followed
- GLP compliance:
- not specified
- Type of study:
- Buehler test
- Species:
- guinea pig
- Strain:
- not specified
- Sex:
- not specified
- Details on test animals and environmental conditions:
- Not specified.
- Route:
- epicutaneous, open
- Vehicle:
- unchanged (no vehicle)
- Day(s)/duration:
- 54 hours
- Adequacy of induction:
- not specified
- Route:
- epicutaneous, open
- Vehicle:
- unchanged (no vehicle)
- Day(s)/duration:
- 54 hours
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- unchanged (no vehicle)
- Day(s)/duration:
- 54 hours
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 20 test animals and 10 control animals
- Positive control substance(s):
- not specified
- Reading:
- 1st reading
- Hours after challenge:
- 30
- Group:
- test chemical
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- 2nd reading
- Hours after challenge:
- 54
- Group:
- test chemical
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Remarks on result:
- no indication of skin sensitisation
- Conclusions:
- Glyceryl citrate/lactate/linoleate/oleate did not induce sensitisation in guinea pigs.
- Executive summary:
The skin sensitisation potential of glyceryl citrate/lactate/linoleate/oleate was assessed using guinea pigs. Thirty guinea pigs were used for this study, 20 serving as test guinea pigs and 10 serving as controls. The test item was undiluted as the undiluted test item did not induce skin irritation in a preliminary test. The test item was applied via dermal application during induction phases I, II and III and the reactions were assessed 30 and 54 hours after initiation of treatment. A trigger concentration was also determined in week 4, as the highest non-irritating concentration. No skin reactions (erythema or oedema) were observed in induction phases I, II or III, or with the "trigger" concentration. Therefore, it was concluded that glyceryl citrate/lactate/linoleate/oleate did not induce sensitisation in guinea pigs.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2002
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2004
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- other: SPF-CBA/Ca01aHsd
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: SPF
- Age at study initiation: 6 to 12 weeks
- Housing: Macrolon cages on Altromin saw fiber bedding
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days
- Indication of any skin lesions: Not specified
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Vehicle:
- other: 3 + 1 (v/v) acetone/olive oil
- Concentration:
- 0, 10%, 25% and 50%
- No. of animals per dose:
- 5
- Details on study design:
- PRE-SCREEN TESTS:
- Systemic toxicity: Animals observed for systemic effects and local irritant effects
- Irritancy: Local irritant effects expressed as ear swelling were assessed by measuring ear thickness.
- Ear thickness measurements: Ear thickness measured in all groups using a mechanical calliper on day 1 and day 5
- Other measurements: Lymph node weight index (LNWI) were calculated as the arithmetic mean lymph node weight values of the test group and the negative control group
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: SI > 3
TREATMENT PREPARATION AND ADMINISTRATION: Treated by topical applications of test or control solution to the entire dorsal surface of each ear. Treatments performed once daily over three consecutive days. - Positive control substance(s):
- other: p-phenylenediamine
- Statistics:
- A one-sided t-test was used to identify a statistically significant increase in lymph node weights in treated groups vs. the vehicle control group
- Positive control results:
- Not reported
- Parameter:
- SI
- Value:
- 1.5
- Variability:
- 0.7
- Test group / Remarks:
- 10%
- Parameter:
- SI
- Value:
- 7
- Variability:
- 1.2
- Test group / Remarks:
- 25%
- Parameter:
- SI
- Value:
- 9.1
- Variability:
- 2.4
- Test group / Remarks:
- 50%
- Cellular proliferation data / Observations:
- DETAILS ON STIMULATION INDEX CALCULATION
: For each dose group, the stimulation index was calculated as the ratio of the arithmetic mean DPM (disintegrations per minute)/node values of the test group and the negative control group.
CLINICAL OBSERVATIONS: No systemic effects were observed for any of the treatments.
- Interpretation of results:
- Category 1B (indication of skin sensitising potential) based on GHS criteria
- Conclusions:
- Linoleic acid resulted in SI values above 3 and so is considered a skin sensitizer according to OECD TG 429.
- Executive summary:
Female SPF-CBA/Ca01aHsd mice, between 6 and 12 weeks old, were obtained for this local lymph node assay (LLNA). The mice were divided into 5 groups, a negative control, a vehicle control and three treatment groups, each containing 5 mice. The test substance, linoleic acid, was administered via topical application to the entire dorsal surface of each ear at concentrations of 0 (controls), 10, 25 and 50% once daily for three consecutive days. The vehicle used was 3 + 1 (v/v) acetone/olive oil. For each treatment group a stimulation index (SI) was calculated. The stimulation indexes for linoleic acid were 1.5, 7.0 and 9.1 for the 10%, 25% and 50% groups, respectively. Increased lymph node weight indexes were also observed at 25% (p<0.01) and 50% (p<0.01). An SI value > 3 indicated a positive result, therefore concentrations of 25% and 50% were considered positive as skin sensitizing. Linoleic acid was therefore considered a skin sensitizer according to the OECD TG 429.
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
Further information is included under 'Attached justification' in IUCLID section 13 and 'Cross-reference'. - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2002
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
- Version / remarks:
- 2004
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Version / remarks:
- 2003
- GLP compliance:
- yes
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- other: SPF-CBA/Ca01aHsd
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Females nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: SPF
- Age at study initiation: 6 to 12 weeks
- Housing: Macrolon cages on Altromin saw fiber bedding
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days
- Indication of any skin lesions: Not specified
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12 - Vehicle:
- other: 3 + 1 (v/v) acetone/olive oil
- Concentration:
- 0, 10%, 25% and 50%
- No. of animals per dose:
- 5
- Details on study design:
- PRE-SCREEN TESTS:
- Systemic toxicity: Animals observed for systemic effects and local irritant effects
- Irritancy: Local irritant effects expressed as ear swelling were assessed by measuring ear thickness.
- Ear thickness measurements: Ear thickness measured in all groups using a mechanical calliper on day 1 and day 5
- Other measurements: Lymph node weight index (LNWI) were calculated as the arithmetic mean lymph node weight values of the test group and the negative control group
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: LLNA
- Criteria used to consider a positive response: SI > 3
TREATMENT PREPARATION AND ADMINISTRATION: Treated by topical applications of test or control solution to the entire dorsal surface of each ear. Treatments performed once daily over three consecutive days. - Positive control substance(s):
- other: p-phenylenediamine
- Statistics:
- A one-sided t-test was used to identify a statistically significant increase in lymph node weights in treated groups vs. the vehicle control group
- Positive control results:
- Not reported
- Parameter:
- SI
- Value:
- 2.6
- Variability:
- 1.6
- Test group / Remarks:
- 10%
- Parameter:
- SI
- Value:
- 14.9
- Variability:
- 6.8
- Test group / Remarks:
- 25%
- Parameter:
- SI
- Value:
- 6.9
- Variability:
- 3.6
- Test group / Remarks:
- 50%
- Cellular proliferation data / Observations:
- DETAILS ON STIMULATION INDEX CALCULATION
: For each dose group, the stimulation index was calculated as the ratio of the arithmetic mean DPM (disintegrations per minute)/node values of the test group and the negative control group.
CLINICAL OBSERVATIONS: No systemic effects were observed for any of the treatments.
- Interpretation of results:
- Category 1B (indication of skin sensitising potential) based on GHS criteria
- Conclusions:
- Oleic acid resulted in SI values above 3 and so is considered a skin sensitizer according to OECD TG 429.
- Executive summary:
Female SPF-CBA/Ca01aHsd mice, between 6 and 12 weeks old, were obtained for this local lymph node assay (LLNA). The mice were divided into 5 groups, a negative control, a vehicle control and three treatment groups, each containing 5 mice. The test substance, oleic acid, was administered via topical application to the entire dorsal surface of each ear at concentrations of 0 (controls), 10, 25 and 50% once daily for three consecutive days. The vehicle used was 3 + 1 (v/v) acetone/olive oil. For each treatment group a stimulation index (SI) was calculated. The stimulation indexes for oleic acid were 2.6, 14.9 and 6.9 for the 10%, 25% and 50% groups, respectively. Increased lymph node weight indexes were also observed at 25% (p<0.01) and 50% (p<0.01). An SI value > 3 indicated a positive result, therefore concentrations of 25% and 50% were considered positive as skin sensitizing. The results showed a bell-shaped dose response curve as the SI value was lower at 50% than at 25%. Oleic acid was therefore considered a skin sensitizer according to the OECD TG 429.
- 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
Referenceopen allclose all
After challenge and re-challenge with linoleic acid, skin reactions were observed in the control group as frequently or even at a higher incidence than the test group. Therefore, it was concluded that the test concentration was too high and included irritative reactions and no conclusion regarding skin sensitization could be drawn.
For oleic acid, one animal showed a skin reaction at 48 and 72 hours and an additional 3 animals showed skin reactions only at the 72 hours reading. Upon re-challenge, a skin reaction was reproduced only in two of the four animals. The skin sensitization was confirmed in two animals.
Increased lymph node weight indices (LNWI) were observed at 25% (p<0.01) and 50% (p<0.01).
Increased lymph node weight indices (LNWI) were observed at 25% (p<0.01) and 50% (p<0.01).
After challenge and re-challenge with linoleic acid, skin reactions were observed in the control group as frequently or even at a higher incidence than the test group. Therefore, it was concluded that the test concentration was too high and included irritative reactions and no conclusion regarding skin sensitization could be drawn.
For oleic acid, one animal showed a skin reaction at 48 and 72 hours and an additional 3 animals showed skin reactions only at the 72 hours reading. Upon re-challenge, a skin reaction was reproduced only in two of the four animals. The skin sensitization was confirmed in two animals.
Increased lymph node weight indices (LNWI) were observed at 25% (p<0.01) and 50% (p<0.01).
Increased lymph node weight indices (LNWI) were observed at 25% (p<0.01) and 50% (p<0.01).
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Four key studies (Klimisch 1) were available with oleic acid and linoleic acid giving clear positive results in the LLNA test, but not in the GMPT test. An additional Klimisch 4 GMPT study was also available in which glyceryl citrate/lactate/linoleate/oleate was found to be non-sensitising. The review of LLNA and GMPT tests on unsaturated compounds by Kreiling (2008) discussed various scenarios which could lead to potential false positives in the LLNAs and suggests that formal classification and labelling for such compounds based on LLNA alone would be inappropriate. Also, although oleic acid and linoleic acid gave clear positive results in the LLNA test, there is human data available on glyceryl linoleate and oleate indicating that these substances are not skin sensitizers (see section on Exposure-related observations in humans). Based on the human data, triglycerides C16 and 18 (unsaturated) are not expected to be a skin sensitizing substances. Therefore, according to CLP Regulation (EC) No 1272/2008, Guidance on the Application of the CLP Criteria" (ECHA 2017) and the 8th Adaptation to Technical Progress to CLP Regulation, the test item is not classified as a skin sensitiser.
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