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EC number: 240-474-8 | CAS number: 16423-68-0
- 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
A sensitive mouse lymph node assay (SLNA) was performed on Female BALB/c strain mice, 6-8 weeks old to assess the skin sensitization potential of the test chemical. A sensitive mouse lymph node assay (SLNA) developed for the identification of contact allergens and succeeded in enhancement of the sensitivity of the LLNA using Freund’s complete adjuvant (FCA) and two application phases: intradermal injection and repeated topical application. In this method, a group of 3 mice were intra dermally injected with 50 µl of test chemical –FCA emulsion into two sites of the abdominal skin at both sides of the ventral line. After 5 days 25 µl of test chemical in vehicle was applied to both sides of each ear for three consecutive days. Control mice were treated by intradermal injection of vehicle-FCA emulsion into the abdomen and then after 5 days they were exposed to vehicle alone on the ears for three consecutive days. The next day following the final topical application, auricular lymph nodes were excised and a single-cell suspension of LNC was prepared by mechanical disaggregation through a sterile 200-mesh gauge and washed once with Hank's balanced salt solution. The LNCs were resuspended in RPMI- 1640 culture medium supplemented with different supplements and the total LNC number was determined using an automated cell counter. The LNC suspensions (1 x 106cells) were seeded into 96-well culture plates (five wells per group) and cultured with 0.5 µ Ci [3H] methyl thymidine (3HTdR) for 24 h at 37°C in a humidified atmosphere of 5% CO2, in air. Culture was terminated by a semiautomatic cell harvester, and the 3HTdR incorporation was determined by liquid scintillation counting. The increases in LNC number and3HTdR incorporation relative to controls were derived for each experimental group and expressed as SInand SIp, respectively; SItotalas obtained from SInx SIp, which indicates the total lymph node activation induced by the test chemical. A chemical was regarded as positive if it showed a SItotalvalue of 3 or more. From the results obtained, SItotalvalue for the test chemical was calculated to be 1.55. Hence, it was concluded that the test chemical elicited negative responses under the conditions used and can be considered to be non-sensitizing to mice.
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- data from handbook or collection of data
- Justification for type of information:
- Data is from peer-reviewed scientific journal
- Qualifier:
- according to guideline
- Guideline:
- other: as mentioned below
- Principles of method if other than guideline:
- In the SLNA as well as the LLNA, the lymph node cell (LNC) proliferation of test chemical is measured by incorporation of radioactive (3H) methyl thymidine (3HTdR).
- GLP compliance:
- not specified
- Type of study:
- mouse local lymph node assay (LLNA)
- Species:
- mouse
- Strain:
- Balb/c
- Sex:
- female
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: Japan SLC Inc, Shizuoka,Japan
- Age at study initiation: 6-8 weeks old
- Weight at study initiation: No data available
- Housing: No data available
- Diet (e.g. ad libitum): No data available
- Water (e.g. ad libitum): No data available
- Acclimation period: No data available
ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available
IN-LIFE DATES: From: To: No data available - Vehicle:
- other: Intradermal - Saline; Topical - DMSO
- Concentration:
- Intradermal Injection - 0.25% in saline
Topical Application - 5% in DMSO - No. of animals per dose:
- Groups of mice (n= 3)
- Details on study design:
- PRE-SCREEN TESTS:
- Compound solubility:
- Irritation:
- Systemic toxicity:
- Ear thickness measurements:
- Erythema scores:
MAIN STUDY : Groups of mice (n= 3) were initially injected intradermally with a total of 50 µl of test chemical-FCA emulsion into two sites of the abdominal skin at both sides of the ventral midline. Five days after injection, 25µl of test chemical in vehicle was applied to both sides of each ear for three consecutive days. Control mice were treated by intradermal injection of vehicle-FCA emulsion into the abdomen and then after 5 days they were exposed to vehicle alone on the ears for three consecutive days. The next day following the final topical application, auricular lymph nodes were excised and pooled for each experimental group. A single-cell suspension of LNC was prepared by mechanical disaggregation through a sterile 200-mesh gauge and washed once with Hank's balanced salt solution. The LNCs were resuspended in RPMI- 1640 culture medium supplemented with 25 mM N-2-hydroxyethylpiperazine-N'- 2-ethanesulphonic acid (HEPES), 100 U ml-I penicillin, 100 µg/ml streptomycin and 10% fetal calf serum, and the total LNC number was determined using an automated cell counter. The LNC suspensions (1 x 106 cells) were seeded into 96-well culture plates (five wells per group) and cultured with 0.5 µCi [3H]methyl thymidine ('HTdR) for 24 h at 37°C in a humidifed atmosphere of 5% CO, in air. Culture was terminated by a semiautomatic cell harvester, and the 3HTdR incorporation was determined by liquid scintillation counting. The increases in LNC number and 3HTdR incorporation relative to controls were derived for each experimental group and expressed as SIn and SIp, respectively
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method:
- Criteria used to consider a positive response: SItotal as obtained from SIn x SIp, which indicates the total lymph node activation induced by the test chemical. A chemical was regarded as positive (a sensitizer) if it showed an SItotal value of 3 or more.
TREATMENT PREPARATION AND ADMINISTRATION: - Parameter:
- SI
- Value:
- > 1.15 - <= 1.55
- Test group / Remarks:
- test group
- Cellular proliferation data / Observations:
- CELLULAR PROLIFERATION DATA
:
DETAILS ON STIMULATION INDEX CALCULATION :SIn (stimulation index of LNC number) = 1.35; SIp (stimulation index of LNC proliferation) = 1.15; SI total (stimulation index of total lymph node response) = 1.55
EC3 CALCULATION : The EC3 could not be calculated as the SI values were less than 3.
CLINICAL OBSERVATIONS:
BODY WEIGHTS - Interpretation of results:
- other: not sensitising
- Conclusions:
- The test material was considered as not sensitizing to the skin of BALB/c strain mice in the sensitive mouse lymph node assay (SLNA)
- Executive summary:
A sensitive mouse lymph node assay (SLNA) was performed on Female BALB/c strain mice, 6-8 weeks old to assess the skin sensitization potential of the test chemical.A sensitive mouse lymph node assay (SLNA) developed for the identification of contact allergens and succeeded in enhancement of the sensitivity of the LLNA using Freud’s complete adjuvant (FCA) and two application phases: intradermal injection and repeated topical application. In this method, a group of 3 mice were intra dermally injected with 50 µl of test chemical –FCA emulsion into two sites of the abdominal skin at both sides of the ventral line. After 5 days 25 µl of test chemical in vehicle was applied to both sides of each ear for three consecutive days. Control mice were treated by intradermal injection of vehicle-FCA emulsion into the abdomen and then after 5 days they were exposed to vehicle alone on the ears for three consecutive days. The next day following the final topical application, auricular lymph nodes were excised and a single-cell suspension of LNC was prepared by mechanical disaggregation through a sterile 200-mesh gauge and washed once with Hank's balanced salt solution. The LNCs were resuspended in RPMI- 1640 culture medium supplemented with different supplements and the total LNC number was determined using an automated cell counter.The LNC suspensions (1 x 106cells) were seeded into 96-well culture plates (five wells per group) and cultured with 0.5 µ Ci [3H] methyl thymidine (3HTdR) for 24 h at 37°C in a humidified atmosphere of 5% CO2, in air. Culture was terminated by a semiautomatic cell harvester, and the 3HTdR incorporation was determined by liquid scintillation counting. The increases in LNC number and3HTdR incorporation relative to controls were derived for each experimental group and expressed as SInand SIP, respectively; SItotal as obtained from SInxSIP, which indicates the total lymph node activation induced by the test chemical.A chemical was regarded as positive if it showed anSItotal value of 3 or more. From the results obtainedSItotalvalue for the test chemical was calculated to be 1.55. Hence, it was concluded that the test chemical elicited negative responses under the conditions used and can be considered to be non-sensitizing to mice.
Reference
Results of the SLNA (sensitive mouse lymph node assay) with a range of dyes
Topical application
CONCENTRATION |
Vehicle |
SIn |
SIP |
SItotal |
5 % |
DMSO |
1.35 |
1.15 |
1.55 |
DMSO = dimethylsulphoxide
SIn= stimulation index of LNC number;
SIP= stimulation index of LNC proliferation;
SItotal= stimulation index of total lymph node response.
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
Various studies have been reviewed to determine the dermal sensitization potential of the test chemical. These include in vivo experimental studies performed on mice, guinea pigs as well as humans for the test chemical. The results are mentioned below:
A sensitive mouse lymph node assay (SLNA) was performed on Female BALB/c strain mice, 6-8 weeks old to assess the skin sensitization potential of the test chemical. A sensitive mouse lymph node assay (SLNA) developed for the identification of contact allergens and succeeded in enhancement of the sensitivity of the LLNA using Freund’s complete adjuvant (FCA) and two application phases: intradermal injection and repeated topical application. In this method, a group of 3 mice were intra dermally injected with 50 µl of test chemical –FCA emulsion into two sites of the abdominal skin at both sides of the ventral line. After 5 days 25 µl of test chemical in vehicle was applied to both sides of each ear for three consecutive days. Control mice were treated by intradermal injection of vehicle-FCA emulsion into the abdomen and then after 5 days they were exposed to vehicle alone on the ears for three consecutive days. The next day following the final topical application, auricular lymph nodes were excised and a single-cell suspension of LNC was prepared by mechanical disaggregation through a sterile 200-mesh gauge and washed once with Hank's balanced salt solution. The LNCs were resuspended in RPMI- 1640 culture medium supplemented with different supplements and the total LNC number was determined using an automated cell counter. The LNC suspensions (1 x 106cells) were seeded into 96-well culture plates (five wells per group) and cultured with 0.5 µ Ci [3H] methyl thymidine (3HTdR) for 24 h at 37°C in a humidified atmosphere of 5% CO2, in air. Culture was terminated by a semiautomatic cell harvester, and the 3HTdR incorporation was determined by liquid scintillation counting. The increases in LNC number and3HTdR incorporation relative to controls were derived for each experimental group and expressed as SInand SIp, respectively; SItotalas obtained from SInx SIp, which indicates the total lymph node activation induced by the test chemical. A chemical was regarded as positive if it showed a SItotalvalue of 3 or more. From the results obtained, SItotalvalue for the test chemical was calculated to be 1.55. Hence, it was concluded that the test chemical elicited negative responses under the conditions used and can be considered to be non-sensitizing to mice.
This is result supported by a Skin Prick Test conducted on a patient who had a history of infectious bronchitis. At the age of 8 years, the patient had taken cefaclor suspension (5 ml) (trade name Panacef, Eli Lilly, Florence, Italy, 250 mg/5 ml) for infectious bronchitis. 6 years later, the child was referred to the allergy clinic to determine the cause of this type of adverse reaction. He was now 14 years old, with allergic rhinitis. Panacef suspension contains excipients such as test chemical and strawberry which might cause adverse reactions. A skin prick test (SPT) to test chemical (10 mg/ml) was carried out on the volar side of the forearm with a plastic lancet (1-mm tip) made by Laboratorio Lofarma (Milan, Italy). A positive control with 10 mg/ml histamine was included. SPT results were assessed as positive (wheal response ≥2+) according to the recommendations of the European Academy of Allergology and Clinical Immunology. The Skin Prick Test result for test chemical was negative (0+). Hence the test chemical was considered to be not sensitizing to the skin.
The above result was supported by the Guinea pig maximization test was conducted according to OECD 406 Guidelines with 0.1% solution of test chemical for 3 weeks. Animals received 10 intradermal injections of a 0.1% solution of the colour. The last six injections were given using Freund’s adjuvant. After 14 days, a further intradermal injection was given as challenge. A second epidermal, occlusive challenge was given after additional 10 days. After a further intradermal challenge, positive reactions could be observed in 11 of 19 erythrosine-treated animals. After the epidermal challenge, none of the 19 animals exhibited signs of positive reactions. Therefore, a further intradermal challenge was performed later on after which 15 of 19 animals reacted positive. Epidermal challenge did not cause any effect on skin; hence, the test chemical was not regarded as a skin sensitizer.
Based on the available studies, the test chemical certainly lacks the potential to cause dermal sensitization reactions when tested on humans, guinea pigs as well as mice. Hence it can be concluded to be non-sensitizing to skin and classified under the category “Not Classified” as per CLP Regulation.
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Based on the available studies, the test chemical certainly lacks the potential to cause dermal sensitization reactions when tested on humans, guinea pigs as well as mice. Hence it can be concluded to be non-sensitizing to skin and classified under the category “Not Classified” as per CLP Regulation.
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