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EC number: 308-859-6 | CAS number: 98679-19-7
- 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
Skin sensitisation
Administrative data
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 25 Jul - 07 Sep 2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Cross-reference
- Reason / purpose for cross-reference:
- read-across source
- Remarks:
- link to target
Reference
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- supporting study
- Study period:
- 25 Jul - 07 Sep 2012
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study with acceptable restrictions
- Remarks:
- read-across
- Justification for type of information:
- REPORTING FORMAT FOR THE ANALOGUE APPROACH
1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The present analogue approach contemplates Phosphoric acid, butyl ester, branched, compd. with 2-ethyl-N-(2-ethylhexyl)-1-hexanamine (CAS 98679-19-7) as target substance for read across from the source substance Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates (CAS 96690-34-5). The read-across approach is used to evaluate the hazardous potential of the target substance with respect to ecotoxicological endpoints and human health for REACH Annex VII.
Based on similar chemical structures, read-across based on different compounds having the same type of effect(s) as described in scenario 2 of the Read-Across Assessment Framework document can be used as a basis for assessment.
The target substance (Phosphoric acid, butyl ester, branched, compd. with 2-ethyl-N-(2-ethylhexyl)-1-hexanamine) and the source substance (Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates) are both organic UVCB substances.
In consequence, read-across can be justified due to the high structural similarities as well as common properties, which will be outlined in detail below.
2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source: Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates (EC 306-227-4, CAS 96690-34-5)
The substance is manufactured from the educts phosphorus pentoxide (CAS 1314-56-3, EC 215-236-1), 2-butanol (CAS 78-92-2, EC 201-158-5), iso-butanol (CAS 78-83-1, EC 201-148-0) and Amines, C12-14-tert-alkyl (CAS 68955-53-3, EC 273-279-1).
Target: Phosphoric acid, butyl ester, branched, compd. with 2-ethyl-N-(2-ethylhexyl)-1-hexanamine (EC 308-859-6, CAS 98679-19-7)
The substance is manufactured from the educts phosphor pentoxide (CAS 1314-56-3, EC 215-236-1), 2-butanol (CAS 78-92-2, EC 201-158-5), iso-butanol (CAS 78-83-1, EC 201-148-0) and bis(2-ethylhexyl)amine (CAS 106-20-7, EC 203-372-4)
Both substances do not contain any impurities which may impact the feasibility of read-across.
3. ANALOGUE APPROACH JUSTIFICATION
Both substances are organic UVCBs and are manufactured from the educts phosphorus pentoxide (CAS 1314-56-3, EC 215-236-1), 2-butanol (CAS 78-92-2, EC 201-158-5), iso-butanol (CAS 78-83-1, EC 201-148-0) in similar ratios, the only slight difference is the amine part. In the source chemical, Amines, C12-14-tert-alkyl is used, and in the target bis(2-ethylhexyl)amine.
So, the reaction products have a similar distribution pattern with regard to varied alkyl phosphate species (mainly dialkyl phosphates). Both used amines have similar molecular weights, i.e. 185.35-213.41 g/mol (CAS 68955-53-3) and 241.46 g/mol (CAS 106-20-7), which are hence not expected to alter their toxicokinetic behaviour essentially. The fact that one amine is mono-substituted and the other di-substituted, is also not considered to impact the suitability for read-across, as amines are considered to be rather stable functional groups, e.g. hydrolytically stable, and are not considered to be altered to a relevant extent in the body. Both amines are not covalently bound to the phosphates, so they are expected to dissociate into similar dissociation products, too.
Last but not least, both substances show similar physico-chemical properties. They are both liquid at all relevant handling temperatures, both are characterized by negative glass transition temperatures they do not boil but decompose, have similar densities and low vapour pressures indicating that they are no volatile liquids. They have both logPow values and water solubilities which, despite slight variations, are not expected to alter essentially their uptake and distribution through the body.
As there is not sufficient data on both source and target substance available, and the amine is considered to be the main difference and also not covalently bound to the phosphates, data on human health relevant endpoints is retrieved from publically available data sources, i.e. ACToR (https://actor.epa.gov/actor/searchidentifier.xhtml) and RTECS (http://ccinfoweb.ccohs.ca/rtecs/search.html). Further, ecotoxicological properties were estimated via US EPA EpiSuite ECOSAR Class Program v1.11.
Data indicate that both amines produce severe irritating reactions when applied into the eye or onto the skin. Further, the acute dermal toxicity LD50 values are nearly identical in the rabbit (acute tox. Cat. 4), and when applied orally, the available LD50 values similarly indicate that a classification as acute toxic Cat. 4 is triggered. With regard to ecotoxicological properties, the estimated values slightly differ. However, they consistently trigger the same classification, as both substances are not biodegradable; the source chemical is not, so the same can be applied to the amines.
So in consequence, taking into account the similar manufacturing process, i.e. identical and similar educts, and so similar reaction products, similar physico-chemical properties of the source and target chemical and similar (eco)toxicological properties of the source and target amines, the read-across from Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates to Phosphoric acid, butyl ester, branched, compd. with 2-ethyl-N-(2-ethylhexyl)-1-hexanamine is scientifically justified
4. DATA MATRIX
Table: Data Matrix, source and target chemical
Endpoint Source: CAS 96690-34-5 Target: CAS 98679-19-7
Physical state at 20°C, 1013 hPa liquid liquid
Glass transition temperature -44.95 °C -72.1 °C (onset)
Decomposition 243.36 °C 167.1°C (onset)
Density 0.97 g/cm³ 0.968 g/cm³
Vapour pressure ≤ 54 Pa at 20°C < 6.7 Pa at 20°C
logPow ≥ 0.69 - ≤ 5.6 (estimated) - 0.61 at 23°C
Water solubility 1322 mg/L at 25 °C and pH 3.57 17.14 g/l at 20°C
Surface tension 42.9 mN/m at 20°C and 0.66g/l 37.8 mN/m ≤ST ≤ 41.2 mN/m at 20°C and 1g/l
Flash point 1 35°C at 101.325 kPa 107.5°C at 101.3 kPa
Table: Data Matrix, source and target amine
Endpoint Source amine: CAS 68955-53-3 Target: CAS 106-20-7
Acute oral toxicity LD50 = 552 mg/kg (m/f mice, 470 - 719 mg/kg) (ACToR) LD50 = 800µL/kg (mouse, intraperitoneal) (ACToR, RTECS)
LD50 = 320 mg/kg (male rats) (ACToR; RTECS) LD50 = 1640mg/kg (rat) (ACToR, RTECS)
LD50 = 300 mg/kg (rat) (RTECS)
Acute dermal toxicity LD50 = 251 mg/kg (m/f rats, 190 - 322 mg/kg) (ACToR) LD50 = 1190uL/kg (rabbit) (ACToR, RTECS)
LD50 = 1.12 g/kg (rabbits, 0.83 - 1.51) (ACToR, RTECS)
Acute toxicity inhalation LC50 = 157 ppm (female rats, 1.19 mg/L; 90 - 249 ppm) (ACToR, RTECS)
LC50 > 231 ppm (male rats, 1.75 mg/L) (ACToR, RTECS) No data
LC50 > 940 mg/m³/4h (rats) (RTECS)
Skin irritation Severe reaction (rabbit, Draize test) (RTECS) Severe reaction (rabbit, Draize test) (RTECS)
Severe reaction (rabbit, open irritation test) (RTECS)
Mild reaction (rabbit, open irritation test) (RTECS)
Eye irritation Severe reaction (rabbit, Draize test) (RTECS) Severe reaction (rabbit, Draize test) (RTECS)
Genetic Toxicity No data Negative ±S9 (Ames test) (ACToR)
Fish 96-hr LC50 0.644 mg/L* 0.047 mg/L*
0.632 mg/L** 0.016 mg/L**
Daphnid 48-hr LC50 0.104 mg/L* 0.010 mg/L*
0.458 mg/L** 0.014 mg/L**
Green Algae 96-hr EC50 0.047 mg/L* 0.003 mg/L*
0.939 mg/L** 0.059 mg/L**
Fish ChV 0.015 mg/L* 0.000546 mg/L*
0.082 mg/L** 0.003 mg/L**
Daphnid ChV 0.012 mg/L* 0.00131 mg/L*
0.088 mg/L** 0.004 mg/L**
Green Algae ChV 0.019 mg/L* 0.00134 mg/L*
0.424 mg/L** 0.039 mg/L**
* Aliphatic Amines
** Neutral Organic SAR (Baseline Toxicity) - Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- 17 July 1992
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Test was done before LLNA as first-choice method for in-vivo testing was set into force.
- Species:
- guinea pig
- Strain:
- other: Crl: HA
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 5 weeks
- Weight at study initiation: 355 - 420 g
- Housing: animals were kept in groups in Terluran-cages on Altromin saw fibre bedding (lot no. 300312).
- Diet: autoclaved hey and Altromin 3122 maintenance diet for guinea pigs (rich in crude fibre, lot no. 1725), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12 - Route:
- intradermal
- Vehicle:
- physiological saline
- Concentration / amount:
- 1.5%
- Day(s)/duration:
- single injection
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- 2
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 3%
- Day(s)/duration:
- 1
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 5 (controls), 10 (test groups)
- Details on study design:
- RANGE FINDING TESTS: a preliminary test with 7 animals was performed to determine appropriate treatment concentrations for the main assay. Cutaneous reactions (erythema and edema) were evaluated 24, 48 and 72 h after injection or topical application with the test substance.
By intradermal route:
- 1 animal: intradermal administrations of the test substance emulsified in physiological saline at concentrations of 2.5 and 5%
- 1 animal: intradermal administrations of the test substance emulsified in physiological saline at concentrations of 1 and 1.5%
By cutaneous route;
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 50 and 100% for 24 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 50 and 100% for 48 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 12.5 and 25% for 24 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 12.5 and 6% for 48 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 3 and 6% for 24 h
Scored erythema and edema are listed in Table 1 under “Any information on materials and methods incl. tables”.
Based on the results (mild-to-moderate skin irritation, but well tolerated systemically), the following concentrations were chosen for the main assay:
- intradermal induction: 1.5%; epicutaneous induction: 6%; challenge application: 3%
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl
Injection 2: test substance in physiological saline 0.9% NaCl
Injection 3: test substance in a 1:1 mixture (v/v) FCA/ physiological saline 0.9% NaCl
Epicutaneous: test substance in vaseline
- Control group:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl
Injection 2: 100% vaseline
Injection 3: 50% (v/v) formulation of vaseline in a 1:1 mixture (v/v) FCA/ physiological saline 0.9% NaCl
Epicutaneous: vaseline
- Site: shoulder region (intradermal and epicutaneous)
- Frequency of applications: twice (Day 0 and Day 7)
- Duration: 0-7
- Concentrations: intradermal 1.5%, epicutaneous 6%
B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 20
- Exposure period: 24 h
- Test groups: test substance and vaseline only
- Control group: test substance and vaseline only
- Site: left flank (test substance) and right flank (vehicle = intraspecific control)
- Concentrations: 3%
- Evaluation (hr after challenge):24 and 48 h after patch removal
OTHER:
On Day 6 (approx. 24 h before epicutaneous application of the test substance), the test area of the animals was clipped and they were treated with 0.5 g of 10% sodium lauryl sulphate in vaseline in order to induce local irritation. - Challenge controls:
- The control group is actually a challenge control.
- Positive control substance(s):
- yes
- Remarks:
- mercaptobenzothiazole (2 and 15% (w/w))
- Positive control results:
- The sensitivity and reliability of the experimental technique was routinely checked using the positive control substance mercaptobenzothiazole. Ten animals were intradermally (2%) and epicutaneously (25%) induced with the positive control substance, followed by challenge treatment at 15% concentration. The sensitisation rate after challenge application of the positive control substance was 100%, thus fulfilling the criteria for a positive result (≥ 30%) and verifying the sensitivity and reliability of the assay.
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- induction: 0%; challenge: 3%
- No. with + reactions:
- 0
- Total no. in group:
- 5
- Remarks on result:
- no indication of skin sensitisation
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- induction: 1.5 and 6%; challenge: 3%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Clinical observations:
- 1/10: erythema grade 1
- Remarks on result:
- other: slight evidence of sensitisation
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Dose level:
- induction: 2 and 25%; challenge: 15%
- No. with + reactions:
- 10
- Total no. in group:
- 10
- Clinical observations:
- 3/10: erythema grade 1; 5/10: erythema grade 2; 2/10: erythema grade 3; 1/10: edema grade 1; 1/10: edema grade 2
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- induction: 0%; challenge: 3%
- No. with + reactions:
- 0
- Total no. in group:
- 5
- Remarks on result:
- no indication of skin sensitisation
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- induction: 1.5 and 6%; challenge: 3%
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- positive control
- Dose level:
- induction: 2 and 25%; challenge: 15%
- No. with + reactions:
- 10
- Total no. in group:
- 10
- Clinical observations:
- 2/10: erythema grade 1; 5/10: erythema 2; 3/10: erythema grade 3; 1/10: edema grade 1; 1/10: edema grade 2
- Remarks on result:
- positive indication of skin sensitisation
- Interpretation of results:
- other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008
- Conclusions:
- CLP: not classified
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 012
- Report date:
- 2012
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 406 (Skin Sensitisation)
- Version / remarks:
- 17 July 1992
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.6 (Skin Sensitisation)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.2600 (Skin Sensitisation)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany
- Type of study:
- guinea pig maximisation test
- Justification for non-LLNA method:
- Test was done before LLNA as first-choice method for in-vivo testing was set into force.
Test material
- Reference substance name:
- Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates
- EC Number:
- 306-227-4
- EC Name:
- Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates
- Cas Number:
- 96690-34-5
- Molecular formula:
- C4H11O4P to C8H20O7P2 as representative molecular formula of the composition as specified in section 1.2
- IUPAC Name:
- Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates
Constituent 1
- Specific details on test material used for the study:
- Physical state at 20°C, 1013 hPa: liquid
Glass transition temperature: -44.95 °C
Decomposition: 243.36 °C
Density: 0.97 g/cm³
Vapour pressure: ≤ 54 Pa at 20°C
log Pow: ≥ 0.69 - ≤ 5.6 (estimated)
Water solubility: 1322 mg/L at 25 °C and pH 3.57
Surface tension: 42.9 mN/m at 20°C and 0.66g/l
Flash point: 135°C
In vivo test system
Test animals
- Species:
- guinea pig
- Strain:
- other: Crl: HA
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 5 weeks
- Weight at study initiation: 355 - 420 g
- Housing: animals were kept in groups in Terluran-cages on Altromin saw fibre bedding (lot no. 300312).
- Diet: autoclaved hey and Altromin 3122 maintenance diet for guinea pigs (rich in crude fibre, lot no. 1725), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Study design: in vivo (non-LLNA)
Inductionopen allclose all
- Route:
- intradermal
- Vehicle:
- physiological saline
- Concentration / amount:
- 1.5%
- Day(s)/duration:
- single injection
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 50%
- Day(s)/duration:
- 2
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- other: Vaseline
- Concentration / amount:
- 3%
- Day(s)/duration:
- 1
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- 5 (controls), 10 (test groups)
- Details on study design:
- RANGE FINDING TESTS: a preliminary test with 7 animals was performed to determine appropriate treatment concentrations for the main assay. Cutaneous reactions (erythema and edema) were evaluated 24, 48 and 72 h after injection or topical application with the test substance.
By intradermal route:
- 1 animal: intradermal administrations of the test substance emulsified in physiological saline at concentrations of 2.5 and 5%
- 1 animal: intradermal administrations of the test substance emulsified in physiological saline at concentrations of 1 and 1.5%
By cutaneous route;
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 50 and 100% for 24 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 50 and 100% for 48 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 12.5 and 25% for 24 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 12.5 and 6% for 48 h
- 1 animal: topical treatment with the test substance dissolved in vaseline at concentrations of 3 and 6% for 24 h
Scored erythema and edema are listed in Table 1 under “Any information on materials and methods incl. tables”.
Based on the results (mild-to-moderate skin irritation, but well tolerated systemically), the following concentrations were chosen for the main assay:
- intradermal induction: 1.5%; epicutaneous induction: 6%; challenge application: 3%
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl
Injection 2: test substance in physiological saline 0.9% NaCl
Injection 3: test substance in a 1:1 mixture (v/v) FCA/ physiological saline 0.9% NaCl
Epicutaneous: test substance in vaseline
- Control group:
Intradermal (3 pairs of injections):
Injection 1: a 1:1 mixture (v/v) FCA/physiological saline 0.9% NaCl
Injection 2: 100% vaseline
Injection 3: 50% (v/v) formulation of vaseline in a 1:1 mixture (v/v) FCA/ physiological saline 0.9% NaCl
Epicutaneous: vaseline
- Site: shoulder region (intradermal and epicutaneous)
- Frequency of applications: twice (Day 0 and Day 7)
- Duration: 0-7
- Concentrations: intradermal 1.5%, epicutaneous 6%
B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 20
- Exposure period: 24 h
- Test groups: test substance and vaseline only
- Control group: test substance and vaseline only
- Site: left flank (test substance) and right flank (vehicle = intraspecific control)
- Concentrations: 3%
- Evaluation (hr after challenge):24 and 48 h after patch removal
OTHER:
On Day 6 (approx. 24 h before epicutaneous application of the test substance), the test area of the animals was clipped and they were treated with 0.5 g of 10% sodium lauryl sulphate in vaseline in order to induce local irritation. - Challenge controls:
- The control group is actually a challenge control.
- Positive control substance(s):
- yes
- Remarks:
- mercaptobenzothiazole (2 and 15% (w/w))
Results and discussion
- Positive control results:
- The sensitivity and reliability of the experimental technique was routinely checked using the positive control substance mercaptobenzothiazole. Ten animals were intradermally (2%) and epicutaneously (25%) induced with the positive control substance, followed by challenge treatment at 15% concentration. The sensitisation rate after challenge application of the positive control substance was 100%, thus fulfilling the criteria for a positive result (≥ 30%) and verifying the sensitivity and reliability of the assay.
In vivo (non-LLNA)
Resultsopen allclose all
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- negative control
- Dose level:
- induction: 0%; challenge: 3%
- No. with + reactions:
- 0
- Total no. in group:
- 5
- Remarks on result:
- no indication of skin sensitisation
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- test chemical
- Dose level:
- induction: 1.5 and 6%; challenge: 3%
- No. with + reactions:
- 1
- Total no. in group:
- 10
- Clinical observations:
- 1/10: erythema grade 1
- Remarks on result:
- other: slight evidence of sensitisation
- Key result
- Reading:
- 1st reading
- Hours after challenge:
- 24
- Group:
- positive control
- Dose level:
- induction: 2 and 25%; challenge: 15%
- No. with + reactions:
- 10
- Total no. in group:
- 10
- Clinical observations:
- 3/10: erythema grade 1; 5/10: erythema grade 2; 2/10: erythema grade 3; 1/10: edema grade 1; 1/10: edema grade 2
- Remarks on result:
- positive indication of skin sensitisation
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- negative control
- Dose level:
- induction: 0%; challenge: 3%
- No. with + reactions:
- 0
- Total no. in group:
- 5
- Remarks on result:
- no indication of skin sensitisation
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- induction: 1.5 and 6%; challenge: 3%
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Remarks on result:
- no indication of skin sensitisation
- Key result
- Reading:
- 2nd reading
- Hours after challenge:
- 48
- Group:
- positive control
- Dose level:
- induction: 2 and 25%; challenge: 15%
- No. with + reactions:
- 10
- Total no. in group:
- 10
- Clinical observations:
- 2/10: erythema grade 1; 5/10: erythema 2; 3/10: erythema grade 3; 1/10: edema grade 1; 1/10: edema grade 2
- Remarks on result:
- positive indication of skin sensitisation
Applicant's summary and conclusion
- Interpretation of results:
- other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No. 1272/2008
- Conclusions:
- CLP: not classified
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