Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: - | 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
Skin sensitisation in vivo: guinea pig maximisation test, Magnusson-Kligman, Pirbright White guinea pig m/f, 10 (test item), 5 (control), 25% intradermal, 100% epicutaneous induction, 100% epicutaneous challenge (OECD 406, GLP): sensitising
Skin sensitisation in vivo: guinea pig maximisation test, Magnusson-Kligman, Dunkin-Hartley guinea pig male, 20 (test item), 10 (control), 2.5% intradermal, 100% epicutaneous induction, 100% / 50% (1st) and 30% / 15% (2nd) epicutaneous challenge (OECD 406, GLP): not sensitising
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Remarks:
- in vivo study was conducted prior to REACH implementation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 1995-01-10 - 1995-02-17 (experimental phase)
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- OECD-Guideline for Testing of Chemicals Section 4, 406 "Skin Sensitisation", adopted July 17, 1992
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Study was performed prior to REACH implementation and adoption of LLNA OECD TG.
- Species:
- guinea pig
- Strain:
- other: Pirbright White
- Remarks:
- Albino
- Sex:
- male/female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Interfauna, Süddeutsche Versuchstierfarm, Oberer Bann 37, D-78532 Tuttlingen
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: younger than one year
- Housing: The animals housed individually in Macrolon cages (area 780 cm²).
- Diet (e.g. ad libitum): Raiffeisen Ringfutter, Trockenfutter 52 P, Raiffeisen Kraftfutterwerk Kehl
- Water (e.g. ad libitum): tap water of Karlsruhe, additive:20 mg ascorbic acid and 100 mg citric acid per 100 ml water, free access for the animals by daily changing of the watering-bottles
- Acclimation period: five days
- Indication of any skin lesions: none stated; animals were without morbid signs
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23°C
- Humidity (%):30-70%
- Photoperiod (hrs dark / hrs light): 12:12
- IN-LIFE DATES:
From: 1995-01-20 (start of acclimatization)
To: 1995-02-17 (reading of challenge reactions) - Route:
- intradermal and epicutaneous
- Vehicle:
- other: Freund's complete adjuvant & water or sesame oil
- Concentration / amount:
- - Induction, day 0, intradermal: Three intradermal injections were made in a row:
1) Freund's Complete Adjuvant (FCA) (No. F-5881, Sigma Chemical Company, P.O. Box 14508, St. Louis, USA) diluted with aqua ad iniectabilia in the ratio 1+1 and shaken up. The volume of each injection was 0.1 ml.
2) Test item, diluted with sesame oil in the ratio 1 + 3. The volume of each injection was 0.1 ml. The solutions administered were not older than 1 hour.
3) First, the test item is emulsified in FCA in the ratio 2 + 3. This emulsion is further diluted with H2O in the ratio 5 + 3. The resulting mixture containing 25 % of the test item is then injected. The volume of each injection was 0.1 ml
- Induction, dermal (day 8): A filter paper (2x4 cm²) was fully-loaded with approx. 1 ml undiluted test item. It was then applied with the loaded side down to the test area - Day(s)/duration:
- intradermal: single application / epicutaneous: 48 h
- Adequacy of induction:
- highest technically applicable concentration used
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: sesame oil
- Concentration / amount:
- a linen patch loaded with the test substance was applied
- Day(s)/duration:
- for 24 h, on day 21, 2 weeks after topical induction; evaluation 24&48h after removal of the bandage
- Adequacy of challenge:
- other: undiluted; highest concentration possible
- No. of animals per dose:
- 10 (test item)
5 (control)
group I: (test substance) (animals No. 220-229)
group II: (control) (animals No. 230-234)
male animals: No. 225-229; 232-234
female animals: No. 220-224; 230-231 - Details on study design:
- RANGE FINDING TESTS:
The concentration of the test substance for each induction exposure should be the highest to cause mild irritation.
In order to check the animal tolerability of the test concentration applied, a small-scale pilot study with two albino guinea pigs was performed. After clipping away their hair the following intradermal injections were made in each dorsal side:
above: 100 µl test item diluted with sesame oil in the ratio 1:3
below: 100 µl test item diluted with sesame oil in the ratio 1:7
During the next 3 days the skin area around each prick spot was checked for skin irritation effects. No erythema or oedema could be observed. Further testing with more concentrated test item solution was omitted in consequence of the high viscosity of the test substance.
MAIN STUDY
After randomization and assignment to both groups the animals were acclimatized to the test conditions for 5 days prior to the administration.
The study is chronologically subdivided into the acclimatization period (day -5 to -1), the induction phase (day 0 to 8) with an intradermal administration of the test substance on day 0 and a dermal one on day 8, followed by a phase free from treatment (day 9 to day 19) during which a hypersensitive state is developed in both phases. The challenge phase (dermal administration on day 20) follows with subsequent valuation of potential sensitisation reactions on the days 22 and 23.
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: day 0, day 8
- Test groups:
- Induction, day 0, intradermal: An area of 4 x 6 cm² on the dorsum was clipped with an electric clipper. Three intradermal injections were made in a row:
1) Freund's Complete Adjuvant (FCA) (No. F-5881, Sigma Chemical Company, P.O. Box 14508, St. Louis, USA) diluted with aqua ad iniectabilia in the ratio 1+1 and shaken up. The volume of each injection was 0.1 ml.
2) Test item, diluted with sesame oil in the ratio 1 + 3. The volume of each injection was 0.1 ml. The solutions administered were not older than 1 hour.
3) First, the test item is emulsified in FCA in the ratio 2 + 3. This emulsion is further diluted with H2O in the ratio 5 + 3. The resulting mixture containing 25 % of the test item is then injected. The volume of each injection was 0.1 ml
- Causing of a local irritation (day 7): As expected no dermal irritations were noticed in both groups. Therefore, the fur of all guinea pigs was clipped in the dorsal test area with an electric clipper once again. Twenty-four hours before topical induction application the test area was painted with 0.5 ml of a mixture of 10 % sodium lauryl sulfate (No. 20763, anal. grade, Serva Feinbiochemica GmbH & Co. KG, Heidelberg, Germany) in vaseline (DAB 10, Carl Roth GmbH + Co., Karlsruhe, Germany) in order to create a local dermal irritation.
- Induction, dermal (day 8): Eight days after intradermal induction the topical dermal induction was performed. A filter paper (2x4 cm2) was fully-loaded with approx. 1 ml undiluted Additin M 10306. It was then applied with the loaded side down to the test area marked with a rectangular outline and held in contact by an occlusive dressing for a period of 48 hours. In detail, the filter paper was covered with the impermeable non-stripped, non-irritating plastic material of the reverse of a patch (width 6 cm) (Werovil, No. 15 00 20, WERO-MEDICAL, D-65232 Taunusstein, Germany). It was held in place by an occlusive bandage made from a dressing (width 8 cm, with selvages, coated completely with cellophane) (No. 09 00 22, WERO-MEDICAL). The bandage was fixed with 1.25 cm broad strips of Adhesive Tape ST (WERO-MEDICAL).
- Control group:
- Induction, day 0, intradermal: An area of 4 x 6 cm² on the dorsum was clipped with an electric clipper. Three intradermal injections were made in a row: Volume of each injection: 0.1 ml.
1) Freund's Complete Adjuvant (FCA) (No. F-5881, Sigma Chemical Company, P.O. Box 14508, St. Louis, USA) diluted with aqua ad iniectabilia in the ratio 1+1 and shaken up. The volume of each injection was 0.1 ml.
2) Sesame oil; the volume of each injection was 0.1ml.
3) Sesame oil emulsified in FCA m the ratio 2+1. This emulsion is further diluted with H2O in the ratio 3 + 1. The resulting mixture containing 50 % sesame oil is the injected. The volume of each injection was 0.1 ml.
- Causing of a local irritation (day 7): As expected no dermal irritations were noticed in both groups. Therefore, the fur of all guinea pigs was clipped in the dorsal test area with an electric clipper once again. Twenty-four hours before topical induction application the test area was painted with 0.5 ml of a mixture of 10 % sodium lauryl sulfate (No. 20763, anal. grade, Serva Feinbiochemica GmbH & Co. KG, Heidelberg, Germany) in vaseline (DAB 10, Carl Roth GmbH + Co., Karlsruhe, Germany) in order to create a local dermal irritation.
- Induction, dermal (day 8): The animals of the control group were treated in the same way as the test item group. However, instead of the test substance the pure vehicle sesame oil was applied dermally under the filter paper. The exposure area was protected with an occlusive bandage, too.
- Site: dorsum
- Frequency of applications: single application per day
- Duration: single application (day 0), 48h (day 8)
B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: The challenge phase (dermal administration on day 20, 2 weeks after last induction) follows with subsequent valuation of potential sensitisation reactions on the days 22 and 23.
- Exposure period: 24h
- Test groups / Control group:
Challenge (day 21): The animals were challenged two weeks after the topical dermal induction.
The hair was removed from two 3x5 cm2 areas on both flanks by clipping with an electric clipper. To both groups a linen patch loaded with the test substance was applied to the left flank and a linen patch loaded with the vehicle (sesame oil) was applied to the right flank.
An occlusive dressing made from the materials specified above protected the exposure areas during a period of 24 hours.
- Site: flanks
- Concentrations: undiluted
- Evaluation (hr after challenge): Reading of challenge reactions (days 23 + 24): The challenge sites were evaluated 24 and 48 hours after removal of the bandage. - Challenge controls:
- none
- Positive control substance(s):
- yes
- Remarks:
- The sensitivity of the guinea pig strain used and the reliability of the experimental technique were assessed by use of p-phenylenediamine which is known to have mild-to-moderate skin sensitization properties in a separate study.
- Positive control results:
- Mortality / toxicological symptoms
Group III (reference substance): Guinea pig No. 187 was found dead on day 21 of the reference substance study. None of the surviving animals of group III showed signs of toxicological symptoms.
Group IV (control): Guinea pig No. 192 was found dead on day 20 of the reference substance study. Guinea pig No. 194 was found dead on day 21 of the reference substance study. None of the surviving animals of group IV showed signs of toxicological symptoms.
Body weight
At the end of the experiment all surviving animals had an increase in weight compared to the beginning of the study.
Classification of the findings caused by the reference substance p-phenylenediamine
As stated in "Criteria of judgement" the relevance of the observations is based upon the irritation score and the number of animals reacting as compared with the negative control group. Four of the surviving 9 animals showed symptoms of allergic reactions. Therefore, the potency of p-phenylenediamine to cause allergic symptoms is shown as well as the sensitivity of the guinea pig strain used.
allergic reactions animal challenge reaction
No. day 23 day 24
180 1 1
181 0 0
182 0 0
183 1 0
erythemata noted
184 1 0
185 1 1
186 0 0
187 + a)
188 0 0
189 0 0 - Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 4
- Total no. in group:
- 10
- Clinical observations:
- None of the guinea pigs of group I died during the experiment or showed signs of toxicological symptoms.
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- Four of the animals of the test group showed symptoms of allergic reactions. Therefore, the test substance shows skin sensitisation potential under the experimental conditions.
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 100%
- No. with + reactions:
- 2
- Total no. in group:
- 10
- Clinical observations:
- None of the guinea pigs of group I died during the experiment or showed signs of toxicological symptoms.
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- 100%
- No. with + reactions:
- 0
- Total no. in group:
- 5
- Clinical observations:
- None of the guinea pig of group II died during the experiment or showed signs of toxicological symptoms.
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- 100%
- No. with + reactions:
- 0
- Total no. in group:
- 5
- Clinical observations:
- None of the guinea pig of group II died during the experiment or showed signs of toxicological symptoms.
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Dose level:
- 0.5% (intradermal, epicutaneus induction & challenge)
- No. with + reactions:
- 4
- Total no. in group:
- 9
- Clinical observations:
- death of the 10th animal
- Remarks on result:
- positive indication of skin sensitisation
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- positive control
- Dose level:
- 0.5% (intradermal, epicutaneus induction & challenge)
- No. with + reactions:
- 2
- Total no. in group:
- 9
- Clinical observations:
- death of the 10th animal
- Interpretation of results:
- Category 1 (skin sensitising) based on GHS criteria
- Remarks:
- EU implementation
- Conclusions:
- This study was performed in compliance with OECD-Guideline 406 under GLP, is well-documented and with sufficient information for classification, the results can hence be considered as reliable. Positive and negative controls gave the appropriate results. Under the conditions of this experiment the test substance produced skin reactions in 4 out of 10 animals of the treatment group.
According to Regulation 1272/2008, when an adjuvant type guinea pig test method for skin sensitisation is used, a response of at least 30 % of the animals is considered as positive.
According to the guideline, a minimum of 10 animals is used in the treatment group and at least 5 animals in the control group. When fewer than 20 test and 10 control guinea pigs have been used, and it is not possible to conclude that the test substance is a sensitiser, testing in additional animals to give a total of at least 20 test and 10 control animals is strongly recommended.
The present test is an adjuvant test, and 40% of the animals showed reactions in the form of discrete or patchy erythema. Hence, the substance gave a positive reaction acc. Regulation 1272/2008, and so also the number of animals is sufficient. According to Commission Regulation (EU) No 286/2011 of 10 March 2011, Table 3.4.4, Animal test results for sub-category 1B, a substance must be classified as skin sensitizer Cat. 1B, if in a GPMT ≥ 30 % of the animals are responding at > 1 % intradermal induction dose. Intradermal induction dose was 25%, and 40% responded, so a subcategorisation into Skin Sens. Cat. 1B ist indicated. - Executive summary:
The skin sensitisation study was performed according to the method of Magnusson & Kligman in compliance with OECD-Guideline 406. Two groups (test substance and control) with 10 guinea pigs (test substance) and 5 guinea pigs (control) were employed, both male and female animals contained.
Under the conditions of this experiment, i.e. after the intradermal and dermal applications of the test item with subsequent challenge four animals showed allergic reactions. As expected, the animals of the control group showed no allergic reactions.
During the experiment the body weight of all animals had a normal physiological growth. Resulting from this study the test substance has skin sensitisation potential. A histopathological examination was not performed.
The substance must be regarded as a potential skin sensitizer.
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Remarks:
- in vivo study was conducted prior to REACH implementation
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 1995-05-31 - 1995-07-15 (experimental phase)
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- Adopted: 17 July 1992
- Deviations:
- no
- GLP compliance:
- yes
- Remarks:
- self-declaration
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Study was performed prior to REACH implementation and adoption of LLNA OECD TG.
- Specific details on test material used for the study:
- STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature - Species:
- guinea pig
- Strain:
- Dunkin-Hartley
- Sex:
- male
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: D. Hall, Newchurch, Staffordshire, England
- Microbiological status of animals, when known:
- Age at study initiation: 4-5 weeks
- Weight at study initiation: 307 to 367 g
- Housing: in groups of five in suspended metal cages with wire mesh floors
- Diet (e.g. ad libitum): vitamin C enriched guinea pig diet FD2 ad libitum
- Water (e.g. ad libitum): drinking water ad libitum
- Acclimation period: 5 days
- Indication of any skin lesions: none stated, animals were healthy
ENVIRONMENTAL CONDITIONS
- Temperature (°C): approx. 21°C
- Humidity (%): 30-70%
- Air changes (per hr): approx. 15
- Photoperiod (hrs dark / hrs light): 12:12 - Route:
- intradermal
- Vehicle:
- other: Alembicol D
- Remarks:
- a product of coconut oil
- Concentration / amount:
- 2.5%
- Day(s)/duration:
- single application on day 1
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- unchanged (no vehicle)
- Concentration / amount:
- 100% as supplied
- Day(s)/duration:
- starting on day 7, for 48h
- Adequacy of induction:
- other: undiluted, non-irritating substance
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Alembicol D
- Remarks:
- a product of coconut oil
- Concentration / amount:
- 50%
- Day(s)/duration:
- on day 21, for 24h
- No.:
- #2
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Alembicol D
- Remarks:
- a product of coconut oil
- Concentration / amount:
- 30% and 15%
- Day(s)/duration:
- on day 28, for 24h
- No. of animals per dose:
- 6 (pre-study)
10 (control)
20 (test group) - Details on study design:
- RANGE FINDING TESTS:
Animals for these investigations were pre-treated with an intradermal injection of Freund's complete adjuvant, 50:50 with water for irrigation, approx. 1 week prior to the start of the preliminary investigations.
The intradermal and topical irritancy of a range of dilutions of the test substance was investigated to identify where possible (a) concentrations of the test substance that would produce irritation suitable for the induction phase of the main study and (b) a maximum non-irritant concentration by the topical route of administration for the challenge phase.
Based on the reults of the preliminary investigations, the following concentrations of the test item were selected:
Induction intradermal injection: 2.5% v/v in Alembicol D: This was the highest concentration intradermally that caused some irritation but did not adversely affect the animals.
Induction topical application: as supplied
Topical challenge: as supplied and 50% v/v in Alembicol D
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: day 1 and day 7 for 48h
- Test groups:
Induction intradermal injections: A 40*60 mm area of the dorsal skin on the scapular region of the guinea pig was clipped free of hair with electric clippers. Three pairs of intradermal injections were made into a 20*40 mm area within the clipped area in a row.
Injectables for the test animals were prepared as follows:
1. Freund’s complete adjuvant was diluted with an equal volume of water for irrigation.
2. Test item, 2.5% v/v in Alembicol D
3. Test item, 2.5% v/v in Alembicol D in a 50:50 mixture of Freund’s complete adjuvant and Alembicol D
Induction topical applications: Six days after the injections, the same 40*60 mm interscapular area was clipped and shaved free of hair and the site was pre-treated by gentle rubbing with 0.5 ml per site of 10% v/v sodium lauryl sulphate in petrolatum. 24 h later a 20*40 mm patch of Whatman No. 3 paper was saturated with approx. 0.4 ml of the test item as supplied. The patch was placed on the skin of the test animals and covered by a length of impermeable plastic adhesive tape. This was firmly secured by elastic adhesive bandage and fixed with plastic adhesive tape. The dressing was left in place for 48h.
The dermal reactions were observed after each induction phase in both control and test animals by group.
- Control group:
During the induction phase, 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. The dermal reactions were observed after each induction phase in both control and test animals by group.
- Site: dorsal skin
- Frequency of applications: Day 1 and 6
- Duration: single injection / 48h
- Concentrations: 2.5% / undiluted
B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: 2 weeks after induction, and 1 week thereafter
- Exposure period: each for 24h
- Test groups / Control group:
The control and test animals were challenged topically two weeks after the topical induction application using the test item as supplied and 50% v/v in Alembicol D. Hair was removed by clipping and then shaving from an area on the left flank of each guinea pig. A 20*20 patch of Whatman No. 3 paper was saturated with approx. 0.2 ml of the test item as supplied and applied to the flank. The test item 50% v/v in Alembicol D was applied in a similar manner to a posterior site. The patches were sealed to the flank for 24h under strips of “Blenderm” covered by “Elatoplast” wound round the trunk and secured with “Sleek”.
A second challenge application was made one week later. The method employed was similar to that described above, with the exception that on this occasion the test item, 30% and 15%, was applied to the right flank of the control and test animals.
- Site: flank
- Concentrations: as supplied and 50% v/v in Alembicol D / 30% and 15%
- Evaluation (hr after challenge): The challenge sites were evaluated 24, 48, and 72 h after removal of the pathes.
OTHER:
Observations:
Clinical signs: All animals were observed daily for signs of ill health or toxicity
Bodyweight: The body weight of each guinea pig on the main study was recorded on day 1 (day of intradermal injections) and on the last day observations were made of dermal responses to the challenge applications.
Dermal responses: The dermal reactions resulting from intradermal injection and topical application on the preliminary study, and topical application at the challenge were assessed using the following numerical system:
Erythema and Eschar Formation
No erythema 0
Very slight erythema (barely perceptible) 1
Well defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema 4
Oedema Formation
No oedema 0
Very slight oedema (barely perceptible) 1
Slight 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 area of exposure) 4
The diameter (mm) of the dermal response at the intradermal injection sites was recorded in the preliminary study only to assist in the choice of concentrations for the main study.
Any other lesion not covered in this scoring system was described.
The challenge sites were evaluated 24, 48, 72 h after removal of the patches. - Positive control substance(s):
- yes
- Remarks:
- hexyl cinnamic aldehyde
- Positive control results:
- Results of 7 independent trials:
Positive Inconclusive Negative
10/10 0/10 0/10
9/10 0/10 1/10
10/10 0/10 0/10
9/10 0/10 1/10
10/10 0/10 0/10
10/10 0/10 0/10
8/10 1/10 1/10 - Reading:
- 1st reading
- Group:
- other: negative control and test group
- Dose level:
- undiluted / 50%
- Clinical observations:
- No signs of ill health or toxicity were recorded. Bodyweight increases were recorded for all guinea pigs over the period of the study.
- Remarks on result:
- not determinable
- Remarks:
- Following the first challenge application, dermal responses (generally slight erythema and oedema) were seen for the majority of the test and control animals. The degree of irritation seen precluded the precise evaluation of the test animals response. Therefore a second challenge was carried out, one week later, using lower concentrations of the test substance.
- Key result
- Reading:
- 2nd reading
- Group:
- test chemical
- Dose level:
- 30% / 15%
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Clinical observations:
- No signs of ill health or toxicity were recorded. Bodyweight increases were recorded for all guinea pigs over the period of the study.
- Remarks on result:
- no indication of skin sensitisation
- Remarks:
- No dermal reactions were seen in either test or control animals following the second challenge application.
- Reading:
- 2nd reading
- Group:
- negative control
- Dose level:
- 30% / 15%
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Clinical observations:
- No signs of ill health or toxicity were recorded. Bodyweight increases were recorded for all guinea pigs over the period of the study.
- Reading:
- other: not stated
- Group:
- positive control
- Dose level:
- 10% intradermal induction, undiluted epicutaneous induction, 50% and undiluted challenge
- No. with + reactions:
- 10
- Total no. in group:
- 10
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- in 4 of 7 studies
- Reading:
- other: not stated
- Group:
- positive control
- Dose level:
- 10% intradermal induction, undiluted epicutaneous induction, 50% and undiluted challenge
- No. with + reactions:
- 9
- Total no. in group:
- 10
- Remarks on result:
- positive indication of skin sensitisation
- Remarks:
- in 3 of 7 studies
- Interpretation of results:
- GHS criteria not met
- Conclusions:
- This study was performed in compliance with OECD-Guideline 406 under GLP, is well-documented and with sufficient information for classification, the results can hence be considered as reliable. Positive and negative controls gave the appropriate results.
Following the first application challenge application, dermal responses (generally slight erythema and oedema) were seen for the majority of the test and control animals. The degree of irritation seen precluded the precise evaluation of the test animals response. Therefore a second challenge was carried out, one week later, using lower concentrations of the test substance. No dermal reactions were seen in either test or control animals following the second challenge application. Based on the results of the second challenge application there was no evidence of skin sensitization in any of the twenty test animals.
According to Regulation 1272/2008, when an adjuvant type guinea pig test method for skin sensitisation is used, a response of at least 30 % of the animals is considered as positive.
According to the guideline, a minimum of 10 animals is used in the treatment group and at least 5 animals in the control group. When fewer than 20 test and 10 control guinea pigs have been used, and it is not possible to conclude that the test substance is a sensitizer, testing in additional animals to give a total of at least 20 test and 10 control animals is strongly recommended.
The present test is an adjuvant test, and none of the animals showed reactions over control. Hence, the substance gave a negative reaction acc. Regulation 1272/2008, and does not need to be regarded as skin sensitizer. Also, the number of animals is sufficient. - Executive summary:
The skin sensitisation study was performed according to the method of Magnusson & Kligman in compliance with OECD-Guideline 406. Two groups (test substance and control) with 20 male guinea pigs (test substance) and 10 male guinea pigs (control) were employed.
Based on the results of a preliminary study and in compliance with the guideline, the following dose levels were selected:
Intradermal injection: 2.5% v/v in Alembicol D
Topical application: as supplied
First challenge application: as supplied and 50% v/v in Alembicol D
Second challenge application: 30% and 15% v/v in Alembicol D
Based on the results of the second challenge the test item did not produce evidence of skin sensitization (delayed contact hypersensitivity) in any of the twenty test animals. The test item does not need to be regarded as skin sensitizer acc. Regulation 1272/2008.
Referenceopen allclose all
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (sensitising)
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
There are two independent OECD 406 guideline studies available, both are considered as reliable and valid. They were both performed according to the method of Magnusson and Kligman. However, they gave contradicting results.
In the first study, under the conditions of this experiment the test substance produced skin reactions in 4 out of 10 animals of the treatment group. In the control group, no indication of irritation upon challenge was observed.
In the second study, the second challenge the test item did not produce evidence of skin sensitization (delayed contact hypersensitivity) in any of the twenty test animals
In both tests, substances were tested in concentrations leading to the required grade of irritation. The fact that two different strains (Pirbright White resp. Dunkin-Hartley) were used, may also not lead to an explanation why the results are contradicting. There are rare cases in which a strain difference is noted (Anderson, Maybach, 1985, Curr. Probl. Derm., vol. 14, pp. 263-290), but it is highly unlikely that this is the case here.
They are both adjuvant tests, and according to Regulation 1272/2008, when an adjuvant type guinea pig test method for skin sensitisation is used, a response of at least 30 % of the animals is considered as positive.
In the first test, 40% of the animals showed reactions in the form of discrete or patchy erythema. Hence, the substance gave a positive reaction acc. Regulation 1272/2008, and should be classified as skin sensitizer.
In the second study, in the second challenge the test item did not produce evidence of skin sensitisation (delayed contact hypersensitivity) in any of the twenty test animals. Hence, the substance does not need to be classified as skin sensitiser acc. Regulation 1272/2008, taking only in account the results of the second study.
However, out of precautionary reasons, as it is unknown why the two tests gave different results, the substance should be classified as skin sensitizer acc. Regulation 1272/2008. According to Commission Regulation (EU) No 286/2011 of 10 March 2011, Table 3.4.4, Animal test results for sub-category 1B, a substance must be classified as skin sensitizer Cat. 1B, if in a GPMT ≥ 30 % of the animals are responding at > 1 % intradermal induction dose. Intradermal induction dose was 25%, and 40% responded, so a subcategorisation into Skin Sens. Cat. 1B is indicated.
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.