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EC number: 260-892-4 | CAS number: 57674-14-3
- 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
Not sensitising
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Remarks:
- an in vitro or in chemico skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available
- Type of information:
- other: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- From June 27 to August 8, 1994
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- Justification for Read Across is detailed in the endpoint summary.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- adopted July 17, 1992
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- an in vitro or in chemico skin sensitisation study does not need to be conducted because adequate data from an in vivo skin sensitisation study are available
- Species:
- guinea pig
- Strain:
- Himalayan
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd. Wolferstrasse 4, CH-4414 Fullinsdorf/Switzerland.
- Animals life-stage: females nulliparous and non-pregnant.
- Age at study initiation: 5-7 weeks.
- Weight at study initiation: 226 - 362 g.
- Housing: individually in Makrolon type-3 cages with autoclaved standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet: ad libitum. Pelleted standard Kliba 342, Batch nos. 63/94 (at delivery of the animals to 03-August-94) and 64/94 (from 04-August-1994 to termination of test) guinea pig breeding/ maintenance diet ("Kliba", Klingentalmuhle AG, CH-4303 Kaiseraugst).
- Water: ad libitum. Community tap water. Once weekly additional supply of ascorbic acid (1 g/l) via the drinking water -Identification: By unique cage number and corresponding ear tags.
- Acclimation period: one week for one control and the test group under test conditions after health examination. Ten days for the second control group (which was delivered on the test day 24). No acclimatization for the animals of the pretest groups, except for the one epidermal pretest group. Only animals without any visual signs of illness were used for the study.
ENVIRONMENTAL CONDITIONS
- Temperature: 21 - 25 °C.
- Humidity: 52-70 %.
- Air changes: 10-15 air changes/hour.
- Photoperiod: 12 hours artificial fluorescent light (approx. 100 Lux) and 12 hours dark.
- Other: music during the light period. - Route:
- intradermal and epicutaneous
- Vehicle:
- other: ethanol was used for the intradermal applications and vaselinum album for the epidermal applications. The Freund's Complete Adjuvant/physiological saline (1:1) was used as a vehicle for the intradermal induction in the main study.
- Concentration / amount:
- Induction:
Concentration for intradermal injection: 5 % *
Concentration for epicutaneous application: 50 % *
*concentration selected based on pretest I
1st Challenge:
Concentration for epicutaneous application: 50 %*
* concentration selected based on pretest I
2nd Challenge:
Concentration for epicutaneous application: 50 and 25 %**
**concentration selected based on pretest I - Route:
- epicutaneous, occlusive
- Vehicle:
- other: ethanol was used for the intradermal applications and vaselinum album for the epidermal applications. The Freund's Complete Adjuvant/physiological saline (1:1) was used as a vehicle for the intradermal induction in the main study.
- Concentration / amount:
- Induction:
Concentration for intradermal injection: 5 % *
Concentration for epicutaneous application: 50 % *
*concentration selected based on pretest I
1st Challenge:
Concentration for epicutaneous application: 50 %*
* concentration selected based on pretest I
2nd Challenge:
Concentration for epicutaneous application: 50 and 25 %**
**concentration selected based on pretest I - No. of animals per dose:
- Preliminary study: intradermal injections group: 2 animals; epicutaneous applications group: 4 animals.
Main study: control group: 10 animals; test group: 20 treated animals. - Details on study design:
- TEST ARTICLE PREPARATION
The test article and the vehicle were placed into a glass beaker on a tared Mettler PM 480 balance and weight/weight dilutions were prepared. Homogeneity of test article in ethanol and FCA/ physiological saline (1/1) was reached using a magnetic stirrer and a spatula was used for the test article in vaselinum album.
The preparations were made immediately prior to each dosing.
PRETEST
INTRADERMAL INJECTIONS
Intradermal injections (0.1 ml/site) were made into the clipped flank of two guinea-pigs at concentrations of 5, 3 and 1 % of the test substance in ethanol. The resulting dermal reactions were assessed 24 h later. For intradermal induction application a 5 % test substance dilution was selected.
EPIDERMAL PRETEST I
Both flanks of each of 4 guinea pigs were clipped and shaved just prior to the application. Thereafter 4 patches of filter paper ( 2 x 2 cm) were saturated with the test substance at 50 % (this concentration used was found to be the most qualified to assure an optimum technical application procedure), 25 %, 15% and 10 % of the test substance in vaselinum album and applied to the clipped and shaved flanks. The patches were covered by a strip of aluminum foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. This procedure ensured the intensive contact of the test substance. The dressings were removed after an exposure period of 24 hours. 21 hour after removing of the dressing the application site was depilated with an approved depilatory cream to clean the application site from staining produced by the test substance, so that possible erythema reactions were clearly visible at that time. The depilatory was placed on the patch sites and surrounding areas, and left on for 3-5 minutes. It was then thoroughly washed off with a stream of warm, running water. The animals were then dried with a disposable towel, and returned to their cages. The reaction sites were assessed 24 and 48 hours after removal of the bandage for erythema and oedema on a numerical basis according to Draize score. The allocation of the different test dilutions to the sites on the animals was alternated in order to minimize site to site variation in responsiveness. For the epidermal induction and first challenge procedure the test substance at 50 % was selected.
MAIN STUDY
INDUCTION
Intradermal injections / performed on test Day 1: an area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair. Three pairs of intradermal injections (0.1 ml/site) were made at the border of a 4 x 6 cm area in the clipped region as follows:
Test Group:
1 - 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2 - the test substance, diluted to 5 % with ethanol.
3 - the test substance diluted to 5 % by emulsion in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
Control Group:
1 - 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2 - ethanol
3 - 1:1 (w/w) mixture of ethanol in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
Epidermal applications / performed on test Day 8
On 7 d and 23.5 h prior to the epidermal application the scapular area (approx. 6 x 8 cm) was clipped, shaved free of hair and the test area was pretreated with 10 % Sodium-Lauryl-Sulfate (SLS) in paraffinum perliquidum as no primary irritation had been observed in the pretest. The SLS was massaged into the skin with a glass rod without bandaging. This 10 % concentration of SLS enhances sensitization by provoking a mild inflammatory reaction.
On 8 d, a 2 x 4 cm patch of filter paper was saturated with the test substance (50 % in vaselinum album) and placed over the injection sites of the test animals. The patch was covered with aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The dressings were left in place for 48 h. The epidermal application procedure described ensured intensive contact of the test substance. The guinea pigs of the control group I were treated as described above with vaselinum album only. Reaction sites were assessed for erythema and oedema 24 and 48 h after removal of the dressing, using the numerical grading system according to Draize.
FIRST CHALLENGE / performed on test Day 22
The test group and control group I were challenged two weeks after the epidermal induction application. The test and control guinea-pigs were treated in the same way. Hair was clipped and shaved from a 5 x 5 cm area on the left and right flank of each guinea-pig just prior to the application. Two patches ( 2 x 2 cm) of filter paper were saturated with the highest non-irritating concentration of 50 % (left flank) and the vehicle only (vaselinum album, applied to the right flank) using the same method as for the epidermal application. The dressing were left in place for 24 h. 21 h after removing of the dressing the test sites were depilated with an approved depilatory cream (VEET Cream, Reckitt & Colman AG, CH-4123 Allschwil). The cream was placed on the patch sites for 3-5 minutes and then washed off with a stream of warm running water. When the application sites were clean and any stains from the test substance removed the animals were dried with a disposable paper towel and returned to their cages. 25 and 48.5 h after the removal of the dressing the application sites were assessed for erythema and oedema using the numerical scoring system according to Draize.
EPIDERMAL PRETEST II / performed on test Day 26
Very slight to well-defined erythematous reactions were observed in the control group I after the first challenge when treated with the test substance at 50 % in vaselinum album. Therefore, a second epidermal pretest was performed prior to the second challenge to confirm the highest non-irritating concentration of 50 % in vaselinum album determined in the pretest I. The second epidermal pretest was performed in the same way and in the same concentrations as the first pretest. Four additional animals were used. The test substance at 50 % in vaselinum album was considered to be the highest nonirritating concentration used for the second challenge procedure. In addition, the test substance at 25 % in vaselinum album was applied in the second challenge.
SECOND CHALLENGE / performed on test Day 31
A second challenge was performed nine days after the first challenge. The treatment procedure used for the animals in the test group was the same as that described for the first challenge except the applications were made to the opposite flanks of the guinea pigs. Ten additional untreated control animals were used and treated in the same conditions as the test group. The test substance at 50 % and 25 % in vaselinum album was applied on the right cranial and right caudal flank.
OBSERVATIONS
In addition to the sensitizing reactions the following observations and data were recorded during the test and observation period:
- viability/Mortality: daily during the observation period.
- clinical Signs (local/systemic): daily during the observation period.
- skin reactions: at the times specified during the induction and challenge periods.
- body weights: at the beginning of the acclimatization period, at day one and at the termination of the test. The animals of the epidermal pretest II were weighed at their delivery and at the termination of test. The animals of the control group II were only weighed at termination of the test.
- necropsy: necropsy was performed in one animal of the intradermal pretest which was found dead on test 27 day. No necropsies were performed on the animals sacrificed at termination of the observation period. The surviving animals were sacrificed with an intraperitoneal injection of Narcoren at a dose of at least 5.1 ml/kg bw (equivalent to 810 mg sodium pentobarbitone/kg bw) and discarded. - Challenge controls:
- First challenge: test substance (50 % (left flank)) and Vaseline album (right flank).
Second challenge: test substance (50 % (right cranial flank) 25 % (right caudal flank) (ten additional untreated control animals were used). - Positive control substance(s):
- yes
- Remarks:
- 2-mercaptobenzothiazol and 4-aminobenzoic acid ethyl ester
- Positive control results:
- Positive response was observed in 60 % of treated animals after the epidermal challenge application.
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 50 % epidermal application (left flank)
- No. with + reactions:
- 1
- Total no. in group:
- 20
- Clinical observations:
- very slight erythematous reaction
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 50 % epidermal application (left flank)
- No. with + reactions:
- 11
- Total no. in group:
- 20
- Clinical observations:
- very slight or welldefined erythematous reactions
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 50 % epidermal application (right cranial flank)
- No. with + reactions:
- 1
- Total no. in group:
- 20
- Clinical observations:
- very slight erythematous reaction
- Reading:
- rechallenge
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- 25 % epidermal application (right caudal flank)
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Clinical observations:
- No positive reactions were observed
- Reading:
- rechallenge
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 50 % epidermal application (right cranial flank)
- No. with + reactions:
- 3
- Total no. in group:
- 20
- Clinical observations:
- very slight erythematous reaction
- Reading:
- rechallenge
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 25 % epidermal application (right caudal flank)
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Clinical observations:
- No positive reactions were observed
- Interpretation of results:
- not sensitising
- Remarks:
- Migrated information according to the CLP Regulation Criteria used for interpretation of results: EU
- Conclusions:
- Not sensitising.
- Executive summary:
Method
To assess the allergenic potential of the test item in albino guinea pigs the Maximization-Test of B. Magnusson and A.M. Kligman (1969) was used. Ten females were used as control group and 20 females were used as test group.
The highest non-irritating test article concentration used for challenge application was 50 % in vaselinum album.
Due to positive skin reaction in the control group, a second challenge was performed with 10 additional control animals. Prior to the second challenge, a second epidermal pretest was performed to confirm the first epidermal pretest in which the test article at 50 % was determined as the highest non-irritating concentration.
Results
No toxic symptoms were evident in the guinea pigs of the control or test group.
After the first challenge, positive skin reactions were observed in the control group. Therefore a second challenge was performed with an additional control group. The test article at 50 % in vaselinum album was determined twice as the highest non-irritating concentration. However by comparison, the second challenge did not confirm the first challenge with the test article at 50 % and the interpretation is therefore very ambiguous. Only the results with the test article at 25 % could be taken into account in which no skin reactions were observed in the control and test group.
The test article applied at 25 % in vaselinum album should be considered to be a non-sensitizer when described under the test conditions.
Conclusion
According to the CLP Regulation (EC 1272/2008) a substance in considered a skin sensitizer when an adjuvant type test method for skin sensitisation is used, a response of at least 30 % of the animals is considered as positive. For a non-adjuvant Guinea pig test method a response of at least 15 % of the animals is considered positive.
The test item applied at 50 % caused a 10 % of positive results at 24 hours and a 15 % at 48 hours; at 25 %, the substance did not cause positive results.
Reference
MAIN STUDY
SKIN EFFECTS AFTER INTRADERMAL INDUCTION PERFORMED ON TEST DAY 1
Control group I
- Injection site 1 (1:1 (v/v) mixture of FCA and physiological saline): the area around the injection site was oedematous and erythematous from test Day 2 to 5, and became necrotic from test Day 6 to 10, followed by encrustation and exfoliation of encrustation up to the termination of test.
- Injection site 2 (ethanol): the reactions observed were identical to those obtained at injection site 1 with the mixture of FCA and physiological saline
- Injection site 3 (1:1 (v/v) mixture of FCA and physiological saline): the reactions observed were identical to those obtained at injection site 1 with the mixture of FCA and physiological saline. As the animals were bandaged with the semi-occlusive dressing no observations of the skin were possible on test Day 9.
Test group
- Injection site 1 (1:1 (v/v) mixture of FCA and physiological saline): the reactions observed were identical to those obtained in the control group with the mixture of FCA and physiological saline, at injection site 1.
- Injection site 2 (5 % solution of test article in ethanol): the area around the injection site was oedematous and red discoloured from test day 2 to 5 and 2 to 6 respectively, necrotic from test day 6 to 10, followed by encrustation and exfoliation of encrustation up to the termination of test.
- Injection site 3 (5 % solution of test article in a 1:1 (v/v) mixture of FCA and physiological saline): the reactions observed were identical to those obtained at injection site 2 with the 5 % solution of test article in ethanol.
SKIN EFFECTS AFTER EPIDERMAL INDUCTION PERFORMED ON TEST DAY 8
Control group I
No erythematous or oedematous reaction was observed in the animals treated with vaselinum album only.
Test group
As the test article stained the skin red, it was not possible to determine whether erythema was present. However, no oedema was observed. Red discoloration was noted from test day 10 to 24. All animals of the control and test group were pretreated with a 10 % SLS in paraffinum perliquidum.
SKIN EFFECTS AFTER THE FIRST AND SECOND CHALLENGE PERFORMED ON TEST DAY 22 AND 31 RESPECTIVELY
Control group I
First Challenge
No positive reactions were observed in the animals treated with vaselinum album alone.
Red discoloration was noted from test day 23 (after removal of the dressing) to 24 (prior to the depilation).
One animal was observed with a very slight erythematous reaction at the 24-hour reading when treated with the test article at 50 % in vaselinum album. At the 48 -hour reading, 6 animals (out of 10) were observed with very slight or well-defined erythematous reactions.
Control group II
Second Challenge
No positive reactions were observed in the animals either when treated with vaselinum album alone or when treated with the test article at 25 % in vaselinum album.
One animal was observed with a very slight erythematous reaction at the 24-hour reading when treated with the test article at 50 % in vaselinum album. At the 48 -hour reading 3 animals (out of 20) were observed with the same reactions.
Red discolouration was noted from test day 32 (after removal of the dressing) to 33 (prior to the depilation).
Test Group
First Challenge
No positive reactions were observed in the animals treated with vaselinum album alone.
One animal was observed with a very slight erythematous reaction at the 24-hour reading when treated with the test article at 50 % in vaselinum album. At the 48 -hour reading, 11 animals (out of 20) were observed with very slight or well-defined erythematous reactions.
Red discolouration was noted from test day 23 (after removal of the dressing) to 24 (prior to the depilation).
Second Challenge
No positive reactions were observed in the animals either when treated with vaselinum album alone or when treated with the test article at 25 % in vaselinum album.
One animal was observed with a very slight erythematous reaction at the 24-hour reading when treated with the test article at 50 % in vaselinum album. At the 48 -hour reading 3 animals (out of 20) were observed with the same reactions.
Red discolouration was noted from test day 32 (after removal of the dressing) to 33 (prior to the depilation).
VIABILITY / MORTALITY / MACROSCOPIC FINDINGS
One animal (no. 468) of the intradermal pretest was found dead on test day 27.
At necropsy many dark red foci (D = 10 mm) were observed in the lungs.
CLINICAL SIGNS, SYSTEMIC
No symptoms of systemic toxicity were observed in the animals.
BODY WEIGHTS
One animal of the control group lost weight during the acclimatization period. The loss of weight should be considered to be of incidental nature.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
In order to assess the skin sensitisation potential of the Acid Red 357 (AR357), the available data on the structural analogue Acid Red 315 (AR315 - Similar Substance 01) were considered.
AR357 and AR315 have a comparable molecular weight, steric hindrance, polarisation of the whole molecule and both are very water soluble (418.11 g/l at 20 °C and pH 6.84 and 135 g/l at 20 °C, respectively). The respective partition coefficients (log Pow: -1.49 in the case of AR357 and log Pow: 1.15 in the case of AR315) show that both substances have a greater affinity with the water phase, than the octanol phase.
AR357 and AR315 are chromate pyrazol sodium salts: AR357 is a trisodium, while AR315 is a disodium salt. They share the same structure, except for the fact that AR357 has two sulphonated groups, instead one as AR315. These differences are expected to not impact the skin sensitisation potential, thus the read across can be considered as reliable.
To assess the allergenic potential of the test item in albino guinea pigs the Maximization-Test of B. Magnusson and A.M. Kligman (1969) was used. The highest non-irritating test article concentration used for challenge application was 50 % in vaselinum album.
Due to positive skin reaction in the control group, a second challenge was performed with 10 additional control animals. Prior to the second challenge, a second epidermal pretest was performed to confirm the first epidermal pretest in which the test article at 50 % was determined as the highest non-irritating concentration.
No toxic symptoms were evident in the guinea pigs of the control or test group.
After the first challenge, positive skin reactions were observed in the control group. Therefore a second challenge was performed with an additional control group. The test article at 50 % in vaselinum album was determined twice as the highest non-irritating concentration. However by comparison, the second challenge did not confirm the first challenge with the test article at 50 % and the interpretation is therefore very ambiguous. Only the results with the test article at 25 % could be taken into account in which no skin reactions were observed in the control and test group.
The test article applied at 25 % in vaslinum album should be considered to be a non-sensitizer when described under the test conditions.
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
According to CLP Regulation (EC 1272/2008), 3.4 Respiratory or skin sensitisation section, skin sensitizer means a substance that will lead to an allergic response following skin contact.
The criteria to classify a substance as skin sensitizer, on the basis of results from test animals, are reported into the second adaptation to technical progress*. A substance in considered a skin sensitizer when:
- an adjuvant type test method for skin sensitisation is used and a response of at least 30 % of the animals is considered as positive;
- for a non-adjuvant Guinea pig test method a response of at least 15 % of the animals is considered positive;
- a stimulation index of three or more is considered a positive response in the local lymph node assay.
In the Guinea Pig Maximization Test, the test item applied at 50 % caused a 10 % of positive results at 24 hours and a 15 % at 48 hours; at 25 %, the substance did not cause positive results.
In conclusion, the substance does not meet the criteria for classification as skin sensitizer, according to the CLP Regulation (EC 1272/2008).
*Commission Regulation (EU) No 286/2011 of 10 March 2011, amending, for the purposes of its adaptation to technical and scientific progress, Regulation (EC) No 1272/2008 of the European Parliament and of the Council on classification, labelling and packaging of substances and mixtures.
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