Registration Dossier
Registration Dossier
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EC number: 263-038-9 | CAS number: 61789-18-2
Endpoint summary
Administrative data
Description of key information
Based on the results of the read across studies, the test substance is considered to be non-sensitising to the skin.
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Study period:
- June 1, 1978
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study
- Justification for type of information:
- Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Deviations:
- yes
- Remarks:
- No information on reliability check. Induction concentration could have been higher: 0.2% caused slight erythema in 1 of the two animals in the range finding test. However, the study is still considered sufficient for the hazard identification.
- GLP compliance:
- no
- Remarks:
- GLP was not yet compulsory at date of study
- Type of study:
- Buehler test
- Justification for non-LLNA method:
- A LLNA was not conducted with the test substance as a Buehler test was already available allowing assessment of the skin sensitisation potential of the substance. Moreover, the test substance is corrosive hence any additional testing is not required as per Annex VII column 2 of REACH Regulation (1907/2006).
- Species:
- guinea pig
- Strain:
- Dunkin-Hartley
- Sex:
- male/female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Redfern Animal Breeders, Jasons Farm, Old Hay, Brenchley, Kent, UK
- Weight at study initiation: Young, withing range 300 – 400 g
- Sex: Male and female - Route:
- epicutaneous, occlusive
- Vehicle:
- water
- Concentration / amount:
- For the determination of a non-irritating concentration: 0.1, 0.2, 0.5, 1.0, 5.0, 10 and 20.0 % w/v (0.4 mL was applied over 6 h)
Dermal induction: 1000 µg test substance / ml (0.4 ml, 0.1 % w/v) for 6 h under closed patches
Challenge: 400 µg test substance/ml (0.4 ml, 0.1 % w/v) for 6 h under closed patches on untreated other flank. - Route:
- epicutaneous, occlusive
- Vehicle:
- water
- Concentration / amount:
- For the determination of a non-irritating concentration: 0.1, 0.2, 0.5, 1.0, 5.0, 10 and 20.0 % w/v (0.4 ml was applied over 6 h)
Dermal induction: 1000 µg test substance / ml (0.4 ml, 0.1 % w/v) for 6 h under closed patches
Challenge: 400 µg test substance/ml (0.4 ml, 0.1 % w/v) for 6 h under closed patches on untreated other flank. - No. of animals per dose:
- Dose range finding: 16 animals (8 groups of 2 animals)
Treatment group: 20 animals (10 male, 10 female)
Controlgroup: 10 animals (5 male, 5 female) - Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 0.1% w/v in acetone (challenge dose)
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Remarks on result:
- other: 3/20 showed a grade of 0.5; in control 2/10 showed a grade 0.5
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 0.1% w/v in acetone (challenge dose)
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Remarks on result:
- other: no erythema was observed in any of the animals
- Key result
- Reading:
- other: Incidence
- Hours after challenge:
- 48
- Group:
- negative control
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Remarks:
- (acetone)
- Key result
- Reading:
- other: Incidence
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- other: CLP criteria not met
- Remarks:
- (not sensitising)
- Conclusions:
- Under the conditions of the Buehler test, the test substance is considered to be non-sensitizing.
- Executive summary:
A study was conducted to determine the sensitising potential of the read across substance, C12 -14 TMAC (purity not specified), according to a method equivalent to the Buehler test protocol (OECD Guideline 406). A pre-test was conducted to determine non-irritating concentrations to be used in the main study. For the main study the induction was carried out at: topical 0.1% w/v in aqueous ethanol for 6 h, repeated after 7 and 14 d. Challenge was done two weeks after the last induction treatment (Day 28): control and test animals received 0.1% w/v in acetone for 6 h on previously untreated site under closed patches. After 18 h the sites were treated with depilatory cream, rinsed and dried. After 3 h, challenge sites were evaluated for erythema on a scale of 0-3. Evaluation was repeated 24 h later. Results of the first grading were: 0/20 (3/20 showed a grade of 0.5; in control 2/10 showed a grade 0.5). Second grading: 0/20 (no erythema was observed in any of the animals). Under the conditions of the Buehler test, the read across substance was not sensitizing. (Jones, 1978). Based on the results of the read across study, the test substance can also be considered to be non-sensitising to the skin.
.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Remarks:
- GPMT test
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- From 6 February, 1984 to 5 March, 1984
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- A LLNA was not conducted with the test substance as a GPMT was already available allowing assessment of the skin sensitisation potential of the substance. Moreover, the test substance is corrosive hence any additional testing is not required as per Annex VII column 2 of REACH Regulation (1907/2006).
- Species:
- guinea pig
- Strain:
- Dunkin-Hartley
- Sex:
- female
- Details on test animals and environmental conditions:
- Forty female albino guinea-pigs were obtained and acclimated to the laboratory environment. Animal identifications were made by ear tattoo unique number method. The animals were randomly allocated to test and control groups (20 animals to each group). The animals were housed in cages with wire mesh floors and were given free access to tap water and a vitamin C-enriched guinea pig diet. Hay was given once weekly. Animal room temperature was 21ºC and relative humidity 30-70%. 15 air changes per hour air exchange was maintained and lighting was controlled by means of a time switch to give 12 hours of light in each 24 hour period. All animals were observed daily for signs of ill health or toxic signs.
- Route:
- intradermal
- Vehicle:
- other: Water for injection
- Concentration / amount:
- 0.1% v/v test substance in water for injection and 0.1% v/v test substance in a 50:50 mixture of Freund's complete adjuvant and water for injection
- Day(s)/duration:
- Day 0
- Adequacy of induction:
- not specified
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Water for injection
- Concentration / amount:
- 2×4 cm patch of whatman paper saturated with 5% v/v test substance in distilled water
- Day(s)/duration:
- 48 h
- Adequacy of induction:
- not specified
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Water for injection
- Concentration / amount:
- 2×4 cm patch of whatman paper saturated with 0.2 ml of 1% v/v test substance in distilled water, applied to an anterior site on the flank
- Day(s)/duration:
- 24 h
- Adequacy of challenge:
- not specified
- No.:
- #2
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Water for injection
- Concentration / amount:
- 2×4 cm patch of whatman paper saturated with 0.2 ml of 0.5% v/v test substance in distilled water, applied to an posterior site on the flank
- Day(s)/duration:
- 24 h
- Adequacy of challenge:
- not specified
- No. of animals per dose:
- 20 animals each for test substance and Freud's treated control
- Positive control substance(s):
- not specified
- Key result
- Reading:
- other: Combined results over 24-72 h
- Group:
- test chemical
- Dose level:
- 1% v/v (0.33% a.i.) (challenge conc)
- No. with + reactions:
- 11
- Total no. in group:
- 20
- Clinical observations:
- showed grade 2 erythema with grade 1 or 2 edema at all time points
- Remarks on result:
- other: test concentrations relatively too high; likely to be fasle positive reaction
- Key result
- Reading:
- other: Combined results over 24-72 h
- Group:
- test chemical
- Dose level:
- 1% v/v (0.33% a.i.) (challenge conc)
- No. with + reactions:
- 7
- Total no. in group:
- 20
- Clinical observations:
- showed grade 1 erythema
- Remarks on result:
- other: not considered sufficient to conclude as a positive skin sensitisation
- Key result
- Reading:
- other: Combined results over 24-72 h
- Group:
- test chemical
- Dose level:
- 0.5 % v/v (0.17% a.i.) (challenge conc)
- No. with + reactions:
- 2
- Total no. in group:
- 20
- Clinical observations:
- showed grade 2 erythema
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- i.e., equivalen to 10% incidence
- Key result
- Reading:
- other: Combined resuls 24-72 h
- Group:
- test chemical
- Dose level:
- 0.5% v/v (0.17% a.i.) (challenge conc)
- No. with + reactions:
- 9
- Total no. in group:
- 15
- Clinical observations:
- showed grade 1 erythema response
- Remarks on result:
- other: not considered sufficient to conclude as a positive skin sensitisation
- Key result
- Reading:
- other: Combined resuls 24-72 h
- Group:
- negative control
- Dose level:
- Negative control
- No. with + reactions:
- 1
- Total no. in group:
- 20
- Remarks on result:
- no indication of skin sensitisation
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- other: CLP criteria not met
- Remarks:
- (not sensitising)
- Conclusions:
- Based on the results, the test substance can be considered to be non-sensitising
- Executive summary:
A study was conducted to determine the skin sensitisation potential of the test substance, Coco TMAC (33% active), according to OECD Guideline 406. A pre-test was conducted to determine non-irritating concentrations to be used in the main study. In the main study, female albino guinea pigs received on Day 1 an intracutaneous injection of 0.1 mL of 0.1% (i.e., 0.033% a.i.) test substance in water for injection with Freund’s complete adjuvant (1:1) on the clipped part of the flank (approximately 6 x 4 cm). On Day 7, animals were applied an epicutaneous occlusive patch saturated with 5% v/v (i.e., 1.7% a.i.) in distilled water for 48 h. During the induction period the control animals were treated similarly to the test animals with the exception that the test compound was omitted from the intradermal injections and topical application. Finally, on Day 21, guinea pigs were challenged epicutaneously (occlusive application) with 0.2 mL of 1% (0.33% a.i.) and 0.5% (i.e., 0.17% a.i.) of test substance in distilled water at the clipped anterior and posterior sites of the flank respectively. The challenged sites were evaluated 24, 48 and 72 h after removal of patches. The numerical scores were awarded to the dermal reactions by the challenge application.As per the study report, the dermal reactions observed in test animals were concluded to be positive in 12/20 test animals, slightly reacting in 6/20 test animals and similar to control (or negative) in 2/20 test animals. Therefore, the test substance was concluded to be a skin sensitizer (Seaber, 1984).
However, based on below arguments it can be concluded that the test substance can be considered to be ‘non-sensitising’ despite the conclusion of the study report:
- The two key indicators that a dermal response has the characteristics associated with skin sensitisation are: (1) an increasing intensity with time, and (2) the ability to be reproduced upon rechallenge (Kligman and Basketter, 1995; Frankild et al., 1997). A grade 1 reaction (typically barely perceptible erythema) is not a sufficient basis, in the absence of other evidence, to arrive at any firm conclusion on the intrinsic toxicological hazard of substance.
- The results table of the above Seaber, 1984 study shows:6 to 7 control animals that reacted positive to 1% (i.e., 0.33% a.i.) indicating that this concentration was possibly too high for challenge.
- The results of challenge with 0.5% (i.e., 0.17% a.i.) shows: Onlyone animal at 24 h that reacted positive in the control group. In thetest group: 14/20 positive, of which 12 did not get a reaction above 1; and in some cases even did not last up to the last examination day, which is indicative for a more pronounced irritation reaction than actual sensitization. Only 2 animals showed a grade 2 erythema of which one accompanied by a grade 1 oedema. This would indicate more a 2/20 positive results, concluding to an overall negative.
- No reactions to intra dermal injections have been reported to verify the possible influence of corrosive/necrosis reactions due to the test substance, which could lead to ‘angry skin syndrome’, which could cause an increased reaction to irritants leading to false sensitisation reactions.
- On comparing to skin sensitization studies available with other structurally similar substances (i.e., C12 TMAB, C14 TMAB or C12-14 TMAC), it can be concluded that the concentrations for challenge in the study with Coco TMAC, were almost 7 times higher for the high concentration, and 16.6 times higher for the low concentration, which was still over 3 times higher than the high concentration in the other studies. Despite these relative high challenge concentrations, the level of reported reactions was rather mild, with only 2/20 a reaction above slight erythema.
- Results with read across substance, C12-14 TMAC (summarised above), which varies from Coco TMAC in lacking the presence of C16 chains at >10% concentrations, was found to be non-sensitising in a Buehler test (Jones, 1978).
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- supporting study
- Study period:
- From August 10, 1982 to September 04, 1982
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Justification for type of information:
- Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- GLP compliance:
- not specified
- Type of study:
- guinea pig maximisation test
- Species:
- guinea pig
- Strain:
- Dunkin-Hartley
- Sex:
- male
- Details on test animals and environmental conditions:
- Test animals
- Source: D. Hall, Newchurch, Staffordshire
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 357-450 g
- Diet: Vitamin C enriched Guinea pig diet F.D.I. (Special Diets Services Limited); ad libitum
- Water: Tap water ad libitum
Environmental conditions
- Temperature (°C): 21°C
- Humidity (%): 30-70%
- Air changes (per h): 15
- Photoperiod (dark / light): 12 h/12 h - Route:
- intradermal
- Vehicle:
- water
- Concentration / amount:
- 0.01% w/w in water for injection
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- other: Topical, occlusive
- Vehicle:
- water
- Concentration / amount:
- 0.5% w/w in distilled water
- Day(s)/duration:
- 48 h
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- other: Topical, occlusive
- Vehicle:
- water
- Concentration / amount:
- 0.05% and 0.01% w/w in distilled water
- Day(s)/duration:
- 24 h
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 15
- Details on study design:
- Range finding study:
The intradermal and topical irritancy of a range of aqueous dilutions of test substance was investigated to identify the following:
- irritant concentrations of the test substance suitable for the induction phase of the main study
- non-irritant concentration by the topical route of administration for the challenge phase
Main study
A. Induction exposure
- No. of exposures: Intradermal injections - three, topical application - once
- Exposure period: 48 h (topical application)
- Test groups: Intradermal injections - 4 X 6 cm2 area of dorsal skin on the scapular region of the animals was clipped free of hair with electric clippers. Three pairs of intradermal injections were made simultaneously into this area. Injectables were prepared as follows:
1. Freund's complete adjuvant was diluted with an equal volume of water for injection
2. Test substance, 0.01% w/w in water for injection
3. Test substance, 0.01% w/w in a 50:50 mixture of Freund's complete adjuvant and water for injection.
Topical application - One week after the injections, the same 4 X 6 cm2 interscapular area was clipped and shaved free of hair. A 2 X 4 cm2 patch of Whatman No. 3 paper was saturated with test substance, 0.5% w/w in distilled water. The patch was placed on the skin and covered by a length of impermeable plastic adhesive tape (5 cm width "Blenderm"). This, in turn, was firmly secured by elastic adhesive bandage ("Elastoplast" 5 cm width) wound round the torso of the animal and fixed with "Sleek" impervious plastic adhesive tape. The dressing was left in place for 48 h.
- Control group: During the induction period the control animals were treated similarly to the test animals with the exception that the test substance was omitted from the intradermal injections and topical application.
B. Challenge exposure
- No. of exposures: Once
- Exposure period: 24 h
- Test groups: The animals were challenged topically two weeks after the induction period using test substance, 0.05% and 0.01% w/w in distilled water. Hair was removed by clipping and then shaving from the left flank of each animal. A 2 X 2 cm2 Whatman No. 3 paper was saturated with approximately 0.2 mL of the test substance in a similar fashion to that used for the topical induction application. The patch was sealed to the flank for 24 h under a 5 cm strip of "Blenderm" covered by "Elastoplast" wound round the trunk and secured with "Sleek".
- Control group: The animals of the control group were similarly challenged.
- Site: Left flank
- Concentrations: 0.05% and 0.01% w/w in distilled water
- Evaluation (h after challenge): 24, 48 and 72 h
Other: Scoring system:
Erythema and eschar formation:
No erythema: 0
Slight erythema (barely perceptible): 1
Well-defined erythema: 2
Moderate erythema: 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth): 4
Oedema formation:
No oedema: 0
Slight oedema (barely perceptible): 1
Well-defined oedema (edges of area well-defined by definite raising): 2
Moderate oedema (raised approximately 1 mm): 3
Severe oedema (raised more than 1 mm and extending beyond the area of exposure): 4 - Challenge controls:
- The animals were challenged 2 weeks after the induction period.
- Positive control substance(s):
- no
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 0.05% w/w
- No. with + reactions:
- 0
- Total no. in group:
- 15
- Clinical observations:
- None
- Reading:
- 1st reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 0.05% w/w
- No. with + reactions:
- 0
- Total no. in group:
- 15
- Clinical observations:
- None
- Reading:
- 1st reading
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 0.05% w/w
- No. with + reactions:
- 0
- Total no. in group:
- 15
- Clinical observations:
- None
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 0.01% w/w
- No. with + reactions:
- 0
- Total no. in group:
- 15
- Clinical observations:
- None
- Reading:
- 1st reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 0.01% w/w
- No. with + reactions:
- 0
- Total no. in group:
- 15
- Clinical observations:
- None
- Reading:
- 1st reading
- Hours after challenge:
- 72
- Group:
- test chemical
- Dose level:
- 0.01% w/w
- No. with + reactions:
- 0
- Total no. in group:
- 15
- Clinical observations:
- None
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 0.05% w/w in distilled water
- No. with + reactions:
- 3
- Total no. in group:
- 15
- Clinical observations:
- Localised dermal reaction restricted to a small area of the challenge site
- Reading:
- 1st reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- 0.05% w/w in distilled water
- No. with + reactions:
- 1
- Total no. in group:
- 15
- Clinical observations:
- Localised dermal reaction restricted to a small area of the challenge site
- Reading:
- 1st reading
- Hours after challenge:
- 72
- Group:
- negative control
- Dose level:
- 0.05% w/w in distilled water
- No. with + reactions:
- 0
- Total no. in group:
- 15
- Clinical observations:
- None
- Interpretation of results:
- other: CLP criteria not met
- Remarks:
- (not sensitising)
- Conclusions:
- Based on the results of the read across study, the test substance, Coco TMAC, was considered to be non-sensitiser to the skin.
- Executive summary:
A study was conducted to determine the skin sensitisation potential of the read across substance, lauryl trimethyl ammonium bromide (C12 TMAB; Purity not specified), according to the method similar to OECD Guideline 406, using a guinea pig maximisation test. Read across substance concentrations for the main study were determined on the basis of preliminary investigations. In the induction phase, three pairs of intradermal injections (Freund's complete adjuvant diluted with an equal volume of water for injection, 0.01% w/w read across substance in water for injection, and 0.01% w/w read across substance in a 50:50 mixture of Freund's complete adjuvant and water for injection) were made simultaneously in the 4 x 6 cm area of dorsal skin on the scapular region of the animals. One week after the intradermal injections, a 2 x 4 cm occlusive patch (0.5% w/w read across substance in distilled water) was placed topically on the skin and left for 48 h. During induction phase, the control animals were treated similarly to the test animals with the exception that the read across substance was omitted from the intradermal injections and topical application. Challenge exposure was made two weeks after the induction phase. A 2 x 2 cm occlusive patches with approximately 0.2 mL each of 0.05% and 0.01% read across substance in distilled water were applied to anterior and posterior site, respectively. The patches were kept to the place for 24 h. The challenge sites were evaluated after 24, 48 and 72 h of patch removal. No dermal reactions were seen in any of the test animals during the observation period. Under the study conditions, the read across substance was considered to be non-sensitiser to the skin (Liggett and Seaber, 1982). Based on the results of the read across study, the test substance can also be considered to be non-sensitising to the skin.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- read-across based on grouping of substances (category approach)
- Adequacy of study:
- key study
- Study period:
- From September 26, 1994 to November 24, 1994
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- KL2 due to RA
- Justification for type of information:
- Refer to the Quaternary ammonium salts (QAS) category or section 13 of IUCLID for details on the category justification.
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- GLP compliance:
- yes
- Type of study:
- Buehler test
- Justification for non-LLNA method:
- Before LLNA method implementation
- Species:
- guinea pig
- Strain:
- other: Pirbright-White
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Hoechst AG, Kastengrund, SPF breeding
- Weight at study initiation: 227 - 299 g (average 269 g)
- Housing: in groups of 5 in Type 4 macrolon cages
- Diet (e.g. ad libitum): Altromin diet for guinea pigs, Altromin GmbH, Lage/Lippe, Germany
- Water (e.g. ad libitum): tap water ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 23°C
- Humidity (%): 40 - 70%
- Photoperiod (hrs dark / hrs light): 12/12 - Route:
- epicutaneous, occlusive
- Vehicle:
- water
- Concentration / amount:
- For the determination of a non-irritating concentration: 0.1, 1.0, 4.0, 20.0 and 100.0% w/v
Dermal induction: 4% w/v
Challenge: 1% w/v - Route:
- epicutaneous, occlusive
- Vehicle:
- water
- Concentration / amount:
- For the determination of a non-irritating concentration: 0.1, 1.0, 4.0, 20.0 and 100.0% w/v
Dermal induction: 4% w/v
Challenge: 1% w/v - No. of animals per dose:
- 20 for treated group
10 for controls - Details on study design:
- For details, kindly refer to the attached background material section of the IUCLID.
- Positive control substance(s):
- no
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 1% in water
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Clinical observations:
- None
- Remarks on result:
- no indication of skin sensitisation
- Remarks:
- ((evaluation of erythema and edema on Day 30)
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 1% in water
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Clinical observations:
- None
- Remarks on result:
- no indication of skin sensitisation
- Remarks:
- (evaluation of erythema and edema on Day 31)
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Remarks:
- (evaluation of erythema and edema on Day 30)
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Remarks:
- (evaluation of erythema and edema on Day 31)
- Key result
- Reading:
- other:
- Group:
- positive control
- Remarks on result:
- not measured/tested
- Interpretation of results:
- other: CLP criteria not met
- Remarks:
- (not sensitising)
- Conclusions:
- Under the study conditions, the read across substance was considered to be non-sensitizing.
- Executive summary:
A study was conducted to determine the sensitising potential of read across substance, C16 TMAC (30% active in water) in guinea-pigs according to OECD Guideline 406 (Buehler method) and EU Method B6, in compliance with GLP. A pre-test was conducted to determine the non-irritating concentrations to use in the main study. During the induction phase (Days 1-15), the test animals were exposed to 0.5 mL of the read across substance at 4% w/v via an occlusive bandage placed on the shaved skin of the left flank. After 6 hours, the bandage was removed and the skin was washed with warm tap water. Observations of the treated skin were made approximately 24 hours later. On Day 29, the test and control animals were exposed to 0.5 mL of the read across substance at 1% w/v via an occlusive bandage placed on the shaved skin of the right flank. On Days 30 and 31, a macroscopic evaluation of the treated skin was made and animal bodyweights were recorded. During the dermal induction phase (Days 1-5), animals presented light to clearly defined erythema and very light edema. In the control group, no effects were seen on the treated skin. In the challenge phase, 24 and 48 hours after the occlusive bandage was removed, no effects were observed in any animals of the treated or control groups. During the main test, there were no signs of toxicity and the bodyweight gain of the test animals was comparable to that of the controls. Under the study conditions, the read across substance was considered to be non-sensitising (Bury D, 1994). Based on the results of the read across study, the test substance can also be considered to be non-sensitising to the skin.
- Endpoint:
- skin sensitisation: in vitro
- Data waiving:
- study scientifically not necessary / other information available
- Justification for data waiving:
- the study does not need to be conducted because the substance is classified as skin corrosion (Category 1, 1A, 1B or 1C)
- other:
- Reason / purpose for cross-reference:
- data waiving: supporting information
- Reason / purpose for cross-reference:
- data waiving: supporting information
Referenceopen allclose all
Results for dose range finding study:
Skin
reactions were scored after 24 h. Results:
20%: severe erythema 2/2;
10%: severe erythema 2/2;
5%:
severe- moderate 2/2;
1%: severe-
moderate 4/4;
0.5%: moderate-slight 2/2;
0.2%: slight erythema 1/2;
0.1%: No erythema 2/2.
Results for main study:
First grading: 0/20 (3/20 showed a grade of 0.5; in control 2/10 showed a grade 0.5). Second grading: 0/20 (no erythema was observed in any of the animals)
The test substance tested has a narrower chain length distribution compared to full coco. The results from this substance however are fully valid for evaluation of TMAC as:
- The tested substance constitutes already for 70% TMAC, without some additional shorter and longer chain lengths present..
Principally, aspects of sensitisation are related to possible reactivity and protein binding, which are properties that are independent to chain length.
The challenge was performed with exposure to 1 and 0.5% concentrations under closed conditions for 24 h.
The results table shows 6 to 7 control animals that reacted positive to 1% (i.e., 0.33% a.i.) indicating that this concentration was possibly too high for challenge.
Results to challenge with 0.5% (i.e., 0.17% a.i.):
- In control only one animal at 24 h only.
- Treated: 14/20 positive, of which 12 do not get a reaction above 1 which in some cases even did not last up to the last examination day, which is indicative for a more pronounced irritation reaction than actual sensitization.Only 2 animals showed a grade 2 erythema of which one accompanied by a grade 1 oedema. This would indicate more a 2/20 positive results, concluding to an overall negative.
No reactions to intra dermal injections have been reported to verify the possible influence of corrosive/necrosis reactions due to the test substance, which could lead to ‘angry skin syndrome’, which could cause an increased reaction to irritants leading to false sensitisation reactions.
Further, on comparing to skin sensitization studies available with other structurally similar substances, it can be concluded that the concentrations for challenge in the study with Coco TMAC, were almost 7 times higher for the high concentration, and 16.6 times higher for the low concentration, which was still over 3 times higher than the high concentration in the other studies (see below table comparing the test conditions of read across studies). Despite these relative high challenge concentrations, the level of reported reactions were rather mild, with only 2/20 a reaction above slight erythema.
CRO |
Hazleton |
Huntingdon |
Huntingdon |
Huntingdon |
Huntingdon |
Report |
1378-110/113 |
82477D/CCO 13/SS |
82475D/CCO 13/SS |
82476D/CCO 13/SS |
84233D/KKM 7/SS |
year |
1978 |
07 October 1982 |
1982 |
07 October 1982 |
1984 |
Substance |
(C12-14-TMAC) |
C14 TMAB |
C12 TMAB |
C14 TMAB |
Coco TMAC (active ingredient33%) |
Guideline / GLP |
P&G procedure / QA |
M&K / QA - pre-GLP |
M&K / QA - pre-GLP |
M&K / QA - pre-GLP |
OECD 406 / QA - pre-GLP |
Design |
30 animals: 15 control/ 15 test |
30 animals: 15 control/ 15 test |
30 animals: 15 control/ 15 test |
30 animals: 15 control/ 15 test |
40 animals: 20 control/ 20 test |
Induction |
Preliminary irritation testing: 0.2% was slight irritating; 0.1% was non-irritating |
Preliminary irritation testing |
Preliminary irritation testing |
Preliminary irritation testing |
Preliminary irritation testing |
Intra-dermal |
none |
- Freund's complete adjuvant |
- Freund's complete adjuvant |
- Freund's complete adjuvant |
- Freund's complete adjuvant |
topical |
0.1% in ethanol (Highest non-irritating concentration), occlusive 6 hr on day 1, 8 and 15. (Control animals not treated at all) |
0.5% v/v in distilled water, occlusive 48 hr |
0.5% v/v in distilled water, occlusive 48 hr |
0.5% v/v in distilled water, occlusive 48 hr |
5%(1.7%)v/v in distilled water, occlusive 48 hr |
observations |
|
not reported |
not reported |
not reported |
not reported |
Challenge |
0.1% in acetone, occlusive 6 hr. |
0.05% and 0.01% v/v in distilled water, 24 hr occlusive |
0.05% and 0.01% v/v in distilled water, 24 hr occlusive |
0.05% and 0.01% v/v in distilled water, 24 hr occlusive |
1% (0.33%) and 0.5% (0.17%) in distilled water 24 hr occlusive |
Results: |
Control: 2/10 showed erythema 0.5 after 24 hours only |
Control: 0.05%: no reactions; 0.01%: 1/15 showed localised erythema ast some time points (1L) |
Control: 0.05%: 1/15 localised erythema (1L) at 24h; 0.01%: 2/15 showed 1L at 24 , of which 1 also at 48 hr |
Control: 0.05%: 3/15 showed localised erythema ast some time points (1L); 0.01%: no reactions |
Control: 1%(0.33%): 6/20 showed localised erythema (1L) at all time points, plus 1 at 24 hr; 0.5%(0.17%): 1/20 showed 1L at 24 hr. |
|
0.1%: 3/20 showed erythema 0.5 after 24 hours only |
0.05%: 2/15 showed 1L at some time points; additional 1 animals incidental 2L reaction at 24 h. |
0.05%: No reactions |
0.05%: 3/15 showed 1L at some time points; additional 2 animals incidental 2L reaction at 24 h. |
1%(0.33%): 11/20 showed grade 2 erythema with grade 1 or 2 edema at all time points, and additional 7/20 showed grade 1 erythema. |
|
0.01%: 1/15 showed 1L at at 24 hr. |
0.01%: No reactions |
0.01%: 3/15 showed 1L at some time points; additional 1/15 showed 2L at 24 and 48 hr. |
0.5%(0.17%): 2/20 showed grade 2 erythema; additional 9/15 showed grade 1. |
|
Conclusion |
Negative: 0/20 |
Negative: 0/15 positive |
Negative: 0/15 positive |
Negative: 0/15 positive |
Positive: 18/20 (own evaluation negative for 0.17%: 2/20 ) |
Considering the above information along with the results of the read across studies, it can be concluded that the test substance, Coco TMAC is not sensitizing.
a) Dermal reactions elicited by challenge application in the test animals:
Guinea-pig number |
E = Erythema O = Oedema |
Score |
Results Positive (+) Negative (-) |
|||||
24 h |
48 h |
72 h |
||||||
A |
P |
A |
P |
A |
P |
|||
620 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
621 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
622 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
623 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
624 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
625 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
626 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
627 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
628 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
629 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
730 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
731 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
732 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
733 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
734 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
b) Dermal reactions elicited by challenge application in the Freund's treated controls:
Guinea-pig number |
E = Erythema O = Oedema |
Score |
Results Positive (+) Negative (-) |
|||||
24 h |
48 h |
72 h |
||||||
A |
P |
A |
P |
A |
P |
|||
630 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
631 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
632 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
633 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
634 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
635 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
636 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
637 |
E O |
0 0 |
1L 0 |
0 0 |
1L 0 |
0 0 |
0 0 |
- |
638 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
639 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
735 |
E O |
1L 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
736 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
737 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
738 |
E O |
0 0 |
0 0 |
1L 0 |
0 0 |
0 0 |
0 0 |
- |
739 |
E O |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
0 0 |
- |
L = Localised dermal reaction (restricted to a small area of the challenge site)
A = Anterior site, exposed to test substance, 0.05% w/w in distilled water
P = Posterior site, exposed to test substance, 0.01% w/w in distilled water
Determination of a non-irritating concentration
Exposure of guinea pig skin to 100 or 20% w/v test substance resulted in moderate erythema and very light to light edema. At 4% w/v, the animals showed light / clearly defined erythema, and in one animal very light edema. There were no signs of irritation at 1 or 0.2% w/v.
The doses of 4 and 1% w/v were therefore selected for the induction and challenge phases, respectively.
Dermal induction phase
During the induction phase (Days 1 - 15), animals presented light to clearly defined erythema and very light edema. In the control group, no effects were seen on the treated skin.
Challenge phase
24 and 48 h after the occlusive bandage was removed, no effects were observed in any animals of the treated or control groups.
Clinical signs and bodyweight
During the main test, there were no signs of toxicity and bodyweight gain of the test animals was comparable to that of controls.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
Study 1: A study was conducted to determine the sensitising potential of the test substance, C12-14 TMAC (purity not specified), according to a method equivalent to the Buehler test protocol (OECD Guideline 406). A pre-test was conducted to determine non-irritating concentrations to be used in the main study. For the main study the induction was carried out at: topical 0.1% w/v in aqueous ethanol for 6 h, repeated after 7 and 14 d. Challenge was done two weeks after the last induction treatment (Day 28): control and test animals received 0.1% w/v in acetone for 6 h on previously untreated site under closed patches. After 18 h the sites were treated with depilatory cream, rinsed and dried. After 3 h, challenge sites were evaluated for erythema on a scale of 0-3. Evaluation was repeated 24 h later. Results of the first grading were: 0/20 (3/20 showed a grade of 0.5; in control 2/10 showed a grade 0.5). Second grading: 0/20 (no erythema was observed in any of the animals). Under the conditions of the Buehler test, the test substance was not sensitizing (Jones, 1978).
Study 2:A study was conducted to determine the skin sensitisation potential of the test substance, TMAC C, according to OECD Guideline 406. A pre-test was conducted to determine non-irritating concentrations to be used in the main study. In the main study, female albino guinea pigs received on Day 1 an intracutaneous injection of 0.1 mL of 0.1% (i.e., 0.033% a.i.) test substance in water for injection with Freund’s complete adjuvant (1:1) on the clipped part of the flank (approximately 6 x 4 cm). On Day 7, animals were applied an epicutaneous occlusive patch saturated with 5% v/v (i.e., 1.7% a.i.) in distilled water for 48 h. During the induction period the control animals were treated similarly to the test animals with the exception that the test compound was omitted from the intradermal injections and topical application. Finally, on Day 21, guinea pigs were challenged epicutaneously (occlusive application) with 0.2 mL of 1% (0.33% a.i.) and 0.5% (i.e., 0.17% a.i.) of test substance in distilled water at the clipped anterior and posterior sites of the flank respectively. The challenged sites were evaluated 24, 48 and 72 h after removal of patches. The numerical scores were awarded to the dermal reactions by the challenge application.As per the study report, the dermal reactions observed in test animals were concluded to be positive in 12/20 test animals, slightly reacting in 6/20 test animals and similar to control (or negative) in 2/20 test animals. Therefore, the test substance was concluded to be a skin sensitizer (Seaber, 1984).
Study 3: A study was conducted to determine the skin sensitisation potential of the read across substance, lauryl trimethyl ammonium bromide (Purity not specified), according to the method similar to OECD Guideline 406, using a guinea pig maximisation test. Read across substance concentrations for the main study were determined on the basis of preliminary investigations. In the induction phase, three pairs of intradermal injections (Freund’s complete adjuvant diluted with an equal volume of water for injection, 0.01% w/w read across substance in water for injection, and 0.01% w/w read across substance in a 50:50 mixture of Freund’s complete adjuvant and water for injection) were made simultaneously in the 4 x 6 cm area of dorsal skin on the scapular region of the animals. One week after the intradermal injections, a 2 x 4 cm occlusive patch (0.5% w/w read across substance in distilled water) was placed topically on the skin and left for 48 h. During induction phase, the control animals were treated similarly to the test animals with the exception that the read across substance was omitted from the intradermal injections and topical application. Challenge exposure was made two weeks after the induction phase. A 2 x 2 cm occlusive patches with approximately 0.2 mL each of 0.05% and 0.01% read across substance in distilled water were applied to anterior and posterior site, respectively. The patches were kept to the place for 24 h. The challenge sites were evaluated after 24, 48 and 72 h of patch removal. No dermal reactions were seen in any of the test animals during the observation period. Under the study conditions, the read across substance was considered to be non-sensitiser to the skin (Liggett and Seaber, 1982).
Study 4: A study was conducted to determine the sensitising potential of read across substance, C16 TMAC (30% active in water) in guinea-pigs according to OECD Guideline 406 (Buehler method) and EU Method B6, in compliance with GLP. A pre-test was conducted to determine the non-irritating concentrations to use in the main study. During the induction phase (Days 1-15), the test animals were exposed to 0.5 mL of the read across substance at 4% w/v via an occlusive bandage placed on the shaved skin of the left flank. After 6 hours, the bandage was removed, and the skin was washed with warm tap water. Observations of the treated skin were made approximately 24 hours later. On Day 29, the test and control animals were exposed to 0.5 mL of the read across substance at 1% w/v via an occlusive bandage placed on the shaved skin of the right flank. On Days 30 and 31, a macroscopic evaluation of the treated skin was made, and animal bodyweights were recorded. During the dermal induction phase (Days 1-5), animals presented light to clearly defined erythema and very light edema. In the control group, no effects were seen on the treated skin. In the challenge phase, 24 and 48 hours after the occlusive bandage was removed, no effects were observed in any animals of the treated or control groups. During the main test, there were no signs of toxicity and the bodyweight gain of the test animals was comparable to that of the controls. Under the study conditions, the read across substance was considered to be non-sensitising (Bury D, 1994).
Overall assessment
Based on below arguments, the test substance should be considered as not sensitising despite the conclusion of the Seaber (1984) study report:
• The two key indicators that a dermal response has the characteristics associated with skin sensitisation are: (1) an increasing intensity with time, and (2) the ability to be reproduced upon rechallenge (Kligman and Basketter, 1995; Frankildet al., 1997). A grade 1 reaction (typically barely perceptible erythema) is not a sufficient basis, in the absence of other evidence, to arrive at any firm conclusion on the intrinsic toxicological hazard of substance.
• The results table of the Seaber (1984) study shows: 6 to 7 control animals that reacted positive to 1% (i.e., 0.33% a.i.) indicating that this concentration was possibly too high for challenge.
• The results of challenge with 0.5% (i.e., 0.17% a.i.) shows: Only one animal at 24 h that reacted positive in the control group. In the test group: 14/20 positive, of which 12 did not get a reaction above 1; and in some cases even did not last up to the last examination day, which is indicative for a more pronounced irritation reaction than actual sensitization. Only 2 animals showed a grade 2 erythema of which one accompanied by a grade 1 oedema. This would indicate more a 2/20 positive results, concluding to an overall negative.
• No reactions to intra dermal injections have been reported to verify the possible influence of corrosive/necrosis reactions due to the test substance which could lead to ‘angry skin syndrome’ and cause an increased reaction to irritants leading to false sensitisation reactions.
• Compared to skin sensitization studies with other structurally similar substances (i.e., C12 TMAB, C14 TMAB or C12-14 TMAC), the concentrations for challenge in the study with TMAC C were almost 7 times higher for the high concentration, and 16.6 times higher for the low concentration, which was still over 3 times higher than the high concentration in the other studies. Despite these relative high challenge concentrations, the level of reported reactions was rather mild, with only 2/20 a reaction above slight erythema.
• Results with read across substance, C12-14 TMAC (summarised above), which varies from TMAC C in lacking the presence of C16 chains at >10%, was found to be non-sensitising in a Buehler test (Jones, 1978). Similar results were also observed with C16 TMAC in a Buehler test (summarised above) as well as C12 TMAB in a GPMT (summarised above).
The biocide assessment report available from RMS Italy on TMAC C also concluded the active substance to be not a skin sensitiser, according to the available data (ECHA biocides assessment report, 2016). Therefore, based on the available information and in line with the biocides assessment report, the test substance is considered to be non-sensitising to the skin.
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Based on the results of the read across studies, the test substance does not warrant a classification for the skin sensitisation endpoint, according to the EU CLP criteria (Regulation EC 1272/2008).
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