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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 223-095-2 | CAS number: 3734-33-6
- 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 Sensitization:
Denatonium benzoate[BITREX] produced a 0% (0/20) sensitization rate and was considered to be non- sensitizing to guinea pig skin.
Key value for chemical safety assessment
Skin sensitisation
Link to relevant study records
- Endpoint:
- skin sensitisation: in vivo (non-LLNA)
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with generally accepted scientific standards and described in sufficient detail
- Justification for type of information:
- data is from experimental reports following standard procedures
- Qualifier:
- according to guideline
- Guideline:
- EPA OPP 81-6 (Skin Sensitisation)
- Principles of method if other than guideline:
- To assess the skin contact sensitization potential of the test material in the albino guinea pig
- GLP compliance:
- yes (incl. QA statement)
- Type of study:
- Buehler test
- Justification for non-LLNA method:
- not specified
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): N-benzyl-2-[(2,6-dimethylphenyl)amino]-N,N-diethyl-2-oxoethanaminium benzoate hydrate
-Common name: Denatonium benzoate
- Molecular formula: C21H29N2O.C7H5O2
- Molecular weight: 446.58 g/mol
- Substance type: organic
- Physical state: Solid
- Form: white granules
- Batch Number: 22362
- Storage conditions: room temperature, room temperature in the dark from 22 February, 1995 - Species:
- guinea pig
- Strain:
- Dunkin-Hartley
- Sex:
- male
- Details on test animals and environmental conditions:
- TEST ANIMALS
- Source: David Hall Limited, Staffordshire
- Females (if applicable) nulliparous and non-pregnant: [yes/no/not specified]: no
- Microbiological status of animals, when known:no data available
- Age at study initiation:8-12 weeks old
- Weight at study initiation:336-449g
- Housing: the guinea pigs were housed in groups of upto two in solid floor polypropylene cages furnished with woodflakes
- Diet (e.g. ad libitum): Guinea pig FDI diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: minimum acclimatisation period of 5 days for each animal
- Indication of any skin lesions: no data available
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22 deg C
- Humidity (%): 44-65%
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours darkness - Route:
- epicutaneous, occlusive
- Vehicle:
- water
- Concentration / amount:
- 0.5 ml of 75%w/w in distilled water
- Day(s)/duration:
- 3 , 6 hour exposures
- Adequacy of induction:
- highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
- No.:
- #1
- Route:
- epicutaneous, occlusive
- Vehicle:
- water
- Concentration / amount:
- 0.5ml of the test material in 75% w/w in distilled water
- Day(s)/duration:
- 6 hours
- Adequacy of challenge:
- highest non-irritant concentration
- No. of animals per dose:
- twenty test animals and 10 control animals
- Details on study design:
- RANGE FINDING TESTS:
The concentrations for the topical induction and topical challenge exposure were determined by the 'sighting tests' in which groups of guinea pigs were treated with various concentrations of the test chemical. The procedure followed is as follows:
a] Selection of concentration for topical induction exposure
2 untreated guinea pigs were treated with 0.5 ml of the four concentrations of test chemical in distiled water (75%, 50%, 25 and 10% w/w). The highest concentration causing mild to moderate irritation after 6 hour occlusive exposure was selected as the concentration for the topical induction.
b] Selection of concentration for the topical challenge exposure:
2 guinea pigs were treated with 0.5 ml of two concentrations of test chemical in distilled water (50,75% w/w).These animals had been treated identically to the control animals of the main study on days 0,7 and 14. The highest concentration of the test material which produced no evidence of irritation 24 or 48 hours after a 6 hour occlusive exposure were selected for the topical challenge exposure.
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 - 6 hours exposure
- Exposure period: 6 hours
- Test groups: 20 guinea pigs
- Control group: 10 guinea pigs
- Site: left flank
- Frequency of applications: on day 0,7 and 14 - 6 hours exposure
- Duration: 6 hours exposure
- Concentrations: 0.5 ml of 75%w/w in distilled water
B. CHALLENGE EXPOSURE
- No. of exposures: single exposure
- Day(s) of challenge: day 28
- Exposure period: 24 hours
- Test groups: 20 guinea pigs
- Control group: 10 guinea pigs
- Site: right flank
- Concentrations: 0.5 ml of 75%, 50% w/w in distilled water
- Evaluation (hr after challenge): approximately 24 and 48 hours after removal, the degree of erythema and edema was quantified
OTHER: Classification of the Sensitization response
A comparison of the intensities and durations of the reactions at the test material challenge sites in both test and control animals permits the identification of sensitization reactions.
If the test material at the maximum non-irritant concentration produces reactions in test animals at 24 or 48 hour reading, these reactions are attributable to contact sensitization.If irritation is observed in the control animals, only reactions in the test animals that exceed the most severe reaction seen in the control animals are attributable to contact sensitization. The results are expressed in terms of incidence and severity of responses:
Severity score: sum of the skin responses at each observation divided by the number of animals in the group
Incidence score: total number of animals showing sensitization responses divided by the number of animals in the group - Challenge controls:
- no data available
- Positive control substance(s):
- yes
- Remarks:
- 2-mercaptobenzothiazole was used a control
- Positive control results:
- Contact sensitization reactions were elicited in 3 of the 20 previously induced test group animals in topical challenge.The known contact sensitizer,
2 -Mercaptobenzothiazole, therefore produced a 15% sensitization rate. This was considered to be a satisfactory sensitization response under the conditions of the test. - Key result
- Reading:
- 1st reading
- Hours after challenge:
- 48
- Group:
- test chemical
- Dose level:
- 75% w/w
- No. with + reactions:
- 0
- Total no. in group:
- 20
- Clinical observations:
- no dermal reactions apart from fur loss was reported at 24 and 48 hour observation period
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- 1st reading
- Hours after challenge:
- 48
- Group:
- other: vehicle control
- Dose level:
- 0.5ml distilled water
- No. with + reactions:
- 0
- Total no. in group:
- 10
- Clinical observations:
- No dermal reactions were observed in the control animals
- Remarks on result:
- no indication of skin sensitisation
- Reading:
- 1st reading
- Hours after challenge:
- 48
- Group:
- positive control
- Dose level:
- 50% w/w in acetone
- No. with + reactions:
- 3
- Total no. in group:
- 20
- Clinical observations:
- contact sensitization was elicited in 3 of 20 previously induced test animals in challenge exposure
- Remarks on result:
- positive indication of skin sensitisation
- Interpretation of results:
- other: not sensitizing
- Conclusions:
- Denatonium benzoate[BITREX] produced a 0% (0/20) sensitization rate and was considered to be non- sensitizing to guinea pig skin.
- Executive summary:
A study was performed to assess the dermal sensitization potential of Denatonium benzoate[BITREX] in guinea pigs. Buehler delayed contact hypersensitivity method following EPA Guidelines 81 -6 was conducted in a GLP certified laboratory (Sustainability Support Services (Europe) AB has letter of access). Twenty test and 10 control Dunkin Hartley male guinea pigs were used for the main study, 4 guinea pigs were used for the sight finding study.
The concentrations for the topical induction and topical challenge exposure were determined by the 'sighting tests' in which groups of guinea pigs were treated with various concentrations of the test chemical. The procedure followed is as follows:
a] Selection of concentration for topical induction exposure
2 untreated guinea pigs were treated with 0.5 ml of the four concentrations of test chemical in distiled water (75%, 50%, 25 and 10% w/w). The highest concentration causing mild to moderate irritation after 6 hour occlusive exposure was selected as the concentration for the topical induction.
b] Selection of concentration for the topical challenge exposure:
2 guinea pigs were treated with 0.5 ml of two concentrations of test chemical in distilled water (50,75% w/w).These animals had been treated identically to the control animals of the main study on days 0,7 and 14. The highest concentration of the test material which produced no evidence of irritation 24 or 48 hours after a 6 hour occlusive exposure were selected for the topical challenge exposure.
The concentrations obtained from these studies for the topical induction and challenge are as follows:
Topical Induction: 75% w/w in distilled water
Topical Challenge: 50, 75% w/w in distilled water
During Induction phase, 0.5 ml of the test material at concentration of 75% w/w in distilled water was applied to the left flank of the test animals under an occlusive dressing for 6 hours. The induction exposure procedure was repeated on the same site on days 7, 14 for a total of 3 6 -hour exposures. Approximately 24 hours after each induction exposure (days 1, 8, 15), the degree of erythema and edema were quantified. The same procedure was followed for the control animals except that the vehicle alone was applied. On day 28, the right flank of the test animals was clipped free of hair and 0.5 ml of the test material at concentrations of 75%,50 w/w in distilled water was applied to right flank under occlusive conditions for 6 hours. After 6 hours, the patches were cut and the treatment sites were rinsed with distilled water and the test sites were marked with an indelible marker. Approximately 24, 48 hours after patch removal, the degree of erythema and edema were quantified. 2 -Mercaptobenzothiazole was used as a positive control. Contact sensitization reactions were elicited in 3 of the 20 previously induced test group animals in topical challenge. The known contact sensitizer, 2 -Mercaptobenzothiazole, therefore produced a 15% sensitization rate. This was considered to be a satisfactory sensitization response under the conditions of the test.
Denatonium benzoate[BITREX] produced a 0% (0/20) sensitization rate and was considered to be non- sensitizing to guinea pig skin.
Reference
Table 1: Buehler delayed contact hypersensitivity study in the guinea pigs
Individual reactions in test animals during induction
Induction Concentration: 75%w/w, Vehicle: Distilled water
Animal number |
Evaluation of induction sites on day |
||||||||
1 |
8 |
15 |
|||||||
Erythema |
Edema |
Other |
Erythema |
Edema |
Other |
Erythema |
Edema |
Other |
|
1 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
2 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
3 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
4 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
5 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
6 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
7 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
8 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
9 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
10 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
11 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
12 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
13 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
14 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
15 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
16 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
17 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
18 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
19 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
20 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
Table 2: Buehler delayed contact hypersensitivity study in the guinea pigs
Individual reactions in test animals during challenge
Induction Concentration: 75%, 50 w/w, Vehicle: Distilled water
Animal number |
Skin reactions (Hours after removal of dressing) |
|||||||||||
75% w/w |
50% w/w |
|||||||||||
24 hours |
48 hours |
24 hours |
48 hours |
|||||||||
Erythema |
Edema |
Other reactions |
Erythema |
Edema |
Other reactions |
Erythema |
Edema |
Other reactions |
Erythema |
Edema |
Other reactions |
|
1 |
0 |
0 |
FL |
0 |
0 |
FL |
0 |
0 |
- |
0 |
0 |
- |
2 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
3 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
4 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
5 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
6 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
7 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
8 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
9 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
10 |
0 |
0 |
FL |
0 |
0 |
FL |
0 |
0 |
- |
0 |
0 |
- |
11 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
12 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
13 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
14 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
15 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
16 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
17 |
0 |
0 |
FL |
0 |
0 |
FL |
0 |
0 |
- |
0 |
0 |
- |
18 |
0 |
0 |
FL |
0 |
0 |
FL |
0 |
0 |
- |
0 |
0 |
- |
19 |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
0 |
0 |
- |
20 |
0 |
0 |
FL |
0 |
0 |
FL |
0 |
0 |
- |
0 |
0 |
- |
Sum of the ski n responses (A) |
0+0 |
0+0 |
0+0 |
0+0 |
||||||||
Severity (A/20) |
0 |
0 |
0 |
0 |
||||||||
Number showing sensitization response (N) |
0 |
0 |
0 |
0 |
||||||||
Incidence (N/20) |
0 |
0 |
FL - fur loss
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (not sensitising)
- Additional information:
Skin Sensitization:
Denatonium benzoatehas been investigated in various studies for potential for dermal sensitization to a greater or lesser extent. The studies are based on in vivo experiments in humans and guinea pigs.
A study was performed to assess the dermal sensitization potential of Denatonium benzoate[BITREX] in guinea pigs. Buehler delayed contact hypersensitivity method following EPA Guidelines 81 -6 was conducted in a GLP certified laboratory (Sustainability Support Services (Europe) AB has letter of access). Twenty test and 10 control Dunkin Hartley male guinea pigs were used for the main study, 4 guinea pigs were used for the sight finding study.
The concentrations for the topical induction and topical challenge exposure were determined by the 'sighting tests' in which groups of guinea pigs were treated with various concentrations of the test chemical. The procedure followed is as follows:
a] Selection of concentration for topical induction exposure
2 untreated guinea pigs were treated with 0.5 ml of the four concentrations of test chemical in distiled water (75%, 50%, 25 and 10% w/w). The highest concentration causing mild to moderate irritation after 6 hour occlusive exposure was selected as the concentration for the topical induction.
b] Selection of concentration for the topical challenge exposure:
2 guinea pigs were treated with 0.5 ml of two concentrations of test chemical in distilled water (50,75% w/w).These animals had been treated identically to the control animals of the main study on days 0,7 and 14. The highest concentration of the test material which produced no evidence of irritation 24 or 48 hours after a 6 hour occlusive exposure were selected for the topical challenge exposure.
The concentrations obtained from these studies for the topical induction and challenge are as follows:
Topical Induction: 75% w/w in distilled water
Topical Challenge: 50, 75% w/w in distilled water
During Induction phase, 0.5 ml of the test material at concentration of 75% w/w in distilled water was applied to the left flank of the test animals under an occlusive dressing for 6 hours. The induction exposure procedure was repeated on the same site on days 7, 14 for a total of 3 6 -hour exposures. Approximately 24 hours after each induction exposure (days 1, 8, 15), the degree of erythema and edema were quantified. The same procedure was followed for the control animals except that the vehicle alone was applied. On day 28, the right flank of the test animals was clipped free of hair and 0.5 ml of the test material at concentrations of 75%,50 w/w in distilled water was applied to right flank under occlusive conditions for 6 hours. After 6 hours, the patches were cut and the treatment sites were rinsed with distilled water and the test sites were marked with an indelible marker. Approximately 24, 48 hours after patch removal, the degree of erythema and edema were quantified. 2 -Mercaptobenzothiazole was used as a positive control. Contact sensitization reactions were elicited in 3 of the 20 previously induced test group animals in topical challenge. The known contact sensitizer, 2 -Mercaptobenzothiazole, therefore produced a 15% sensitization rate. This was considered to be a satisfactory sensitization response under the conditions of the test.
Denatonium benzoate[BITREX] produced a 0% (0/20) sensitization rate and was considered to be non- sensitizing to guinea pig skin.
This is supported by the experimental study performed by Bert Bjorkner (Contact Dermatitis, 1980: 6: 466-471) to assess the sensitization potential of Denatonium benzoate in human volunteers. A 30-year-old male developed asthma and pruritus after using an insecticidal spray (Pyrex®). The same symptoms appeared with denatonium benzoate (Bitrex®).
For all the above reasons, Open patch test was performed on the volunteer to assess the sensitizing potential of insecticidal spray (Pyrex®) and denatonium benzoate (Bitrex®).
Denatonium benzoate was tested at a concentration from 2 X 10-2 mg/l in water and diluted till 2x10-7mg/l. Water was used as a control. 0.1 ml of Denatonium benzoate was spread with a glass rod on normal skin on the volar aspect of the forearms. The test reactions were read after 20 and 30 min. Ten control subjects were tested with the same chemicals and in the same way.
Denatonium benzoate was sensitizing to the skin at a concentration of 2 X 10-2 mg/L, 2 X 10-3 mg/L, 2 X 10-4 mg/L, 2 X 10-5 mg/L, 2 X 10-6 mg/L. No sensitizing reaction was observed when the test solution was diluted to 2x10-7 mg/l.
Hence, Denatonium benzoate can be considered to be not sensitizing to human skin when exposed to concentrations below 2x10-7mg/l.
Based on the available data, Denatonium benzoate was considered to be not sensitizing to skin.Comparing the above annotations with the criteria of CLP regulation, test chemical can be classified under the category “Not Classified”.
Respiratory sensitisation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Justification for classification or non-classification
Available studies indicate a strong possibility of Denatonium benzoate being not sensitizing to skin. Hence, it can be classified under the Category "Not Classified" as per CLP regulation.
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.