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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 201-494-2 | CAS number: 83-67-0
- 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
Eye irritation
Administrative data
- Endpoint:
- eye irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 26 January 2016 - 25 February 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- Test method according to OECD 438. GLP study.
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 016
- Report date:
- 2016
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Theobromine
- EC Number:
- 201-494-2
- EC Name:
- Theobromine
- Cas Number:
- 83-67-0
- Molecular formula:
- C7H8N4O2
- IUPAC Name:
- theobromine
- Test material form:
- solid: particulate/powder
- Remarks:
- migrated information: powder
- Details on test material:
- - Name of test material (as cited in study report): Theobromine.- Physical state: White powder.- Analytical purity: 99.8 %- Purity test date: 12/01/2015- Lot/batch No.: M15015C- Expiration date of the lot/batch: 01/2020- Storage condition of test material: Room temperature
Constituent 1
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Lot/batch No.of test material:M15015C
- Expiration date of the lot/batch: 01/2020
- Purity test date:12/01/2015
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
Test animals / tissue source
- Species:
- other: Eyeballs isolated from chikens killed for human consumption
- Details on test animals or tissues and environmental conditions:
- ANIMALS
The eyeballs used in the experiment were obtained from chickens killed for human consumption (Ex vivo).
- Source: Slaughterhouse, i.e. Zakład Przemysłu Drobiarskiego JAS-DROP in Krzyżowice.
- Age at study initiation: approximately 7 weeks.- Weight at study initiation: 1.5 - 2.5 kg.
- Other: After electric shock and incision of the neck for bleeding, the chicken heads were transported to the laboratory in a plastic container at ambient temperature. During the transport, the heads were humidified with a physiological salt solution by placing moistened paper towels inside the container. The time interval between the collection of the chickens’ heads and the use of their eyeballs in the ICE test was 30 minutes. All eyeballs used in the tests came from the same group of eyes collected on a specific day.Till the moment of their transfer, the eyeballs were kept in special containers at temperature of -18°C.
Test system
- Vehicle:
- unchanged (no vehicle)
- Controls:
- yes
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied: 0.03 g - Duration of treatment / exposure:
- 10 seconds
- Observation period (in vivo):
- 240 minutes
- Number of animals or in vitro replicates:
- The test item and controls (positive and negative) were tested on three eyeballs each.
- Details on study design:
- REMOVAL OF TEST SUBSTANCE
- Washing (if done): With 20 mL physiological saline.
- Time after start of exposure: 10 seconds
SCORING SYSTEM:
Combination of fluoresceing retention, corneal opacity and corneal swelling.
Fluoresceing scores: Score Observations
0 No fluorescein retention
0.5 Very minor single cell staining
1 Single cell staining scattered throughout the treated area of the cornea
2 Focal or confluent dense single cell staining
3 Confluent large areas of the cornea retaining fluorescein
Corneal opacity scores: Score Observations
0 No opacity
0.5 Very faint opacity
1 Scattered or diffuse areas; details of the iris are clearly visible
2 Easily discernible translucent areas; details of the iris are slightly obscured
3 Severe corneal opacity; no specific details of the iris are visible; size of the pupil is barely discernible
4 Complete corneal opacity; iris invisible
Corneal swelling was calculated as % as follows:corneal swelling = [corneal thickness at time t – corneal thickness at time t = 0/ corneal thickness at time t = 0] x100
TOOL USED TO ASSESS SCORE:
- Fluoresceing retention (30 minutes after end of exposure)
- Corneal opacity by assigning appropiate values to opaque areas, the mean corneal opacity value was calculated for all test eyeballs for all observation time points. Based on the highest mean score corneal opacity the final score was given.
- The degree of corneal swelling was determined by measuring corneal thickness using a slit-lamp microscope and an SP-100 pachymeter.The mean percentage of corneal swelling for all test eyeballs was calculated for all observation time points. Based on the highest mean score for corneal swelling, at any time point, the final category score was given.
Results and discussion
In vitro
Resultsopen allclose all
- Irritation parameter:
- fluorescein retention score
- Run / experiment:
- Fist run
- Value:
- 2.3
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Average of 3 eyeballs
- Irritation parameter:
- fluorescein retention score
- Run / experiment:
- Second run
- Value:
- 2
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Average of 3 eyeballs
- Irritation parameter:
- cornea opacity score
- Run / experiment:
- First run
- Value:
- 1
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Average of 3 eyeballs
- Irritation parameter:
- cornea opacity score
- Run / experiment:
- Second run
- Value:
- 1
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Average of 3 eyeballs
- Irritation parameter:
- percent corneal swelling
- Run / experiment:
- First run
- Value:
- -8.1
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Average of 3 eyeballs
- Irritation parameter:
- percent corneal swelling
- Value:
- -8
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Average of 3 eyeballs
- Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system:
In the first run histopathological examinations of the corneas treated with the test item revealed: karyorrhexis (eyeball No. 2), slight vacuolation of the anterior corneal epithelium (eyeballs No. 1, No. 2, No. 3), slight coagulation (eyeball No. 1), exfoliation (eyeball No. 2), necrosis (eyeballs No. 2, No. 3), slight erosions of the superficial layer of the anterior corneal epithelium (eyeballs No. 1, No. 2, No. 3), deposits of the test item on the corneal surface (eyeballs No. 2, No. 3), dissection of the corneal stroma (eyeballs No. 1, No. 2, No. 3). In the second run histopathological examinations of the corneas treated with the test item revealed: slight vacuolation and local coagulation of the anterior corneal epithelium (eyeballs No. 1, No. 2, No. 3), slight exfoliation (eyeballs No. 1, No. 2), necrosis and slight erosions of the superficial layer of the anterior corneal epithelium (eyeballs No. 1, No. 2, No. 3).
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes.
- Acceptance criteria met for positive control: Yes.
- Range of historical values if different from the ones specified in the test guideline: No.
Any other information on results incl. tables
Evaluation of fluorescein retention– the first run.
observation after time t (minutes) |
test item |
positive control imidazole |
negative control physiological saline |
|||
average |
ICE class |
average |
ICE class |
average |
ICE class |
|
30 |
2.3 |
III |
3.0 |
IV |
0.0 |
I |
Evaluation of fluorescein retention – the second run.
observation after time t (minutes) |
test item |
positive control imidazole |
negative control physiological saline |
|||
average |
ICE class |
average |
ICE class |
average |
ICE class |
|
30 |
2.0 |
III |
3.0 |
IV |
0.0 |
I |
Evaluation of corneal opacity– the first run.
observation after time t (minutes) |
test item |
positive control imidazole |
negative control physiological saline |
|||
average |
ICE class |
average |
ICE class |
average |
ICE class |
|
30 |
0.8 |
II |
4.0 |
IV |
0.0 |
I |
75 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
120 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
180 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
240 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
Evaluation of corneal opacity – the second run.
observation after time t (minutes) |
test item |
positive control imidazole |
negative control physiological saline |
|||
average |
ICE class |
average |
ICE class |
average |
ICE class |
|
30 |
0.8 |
II |
4.0 |
IV |
0.0 |
I |
75 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
120 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
180 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
240 |
1.0 |
II |
4.0 |
IV |
0.0 |
I |
Evaluation of corneal swelling (%)– the first run.
observation after time t (minutes) |
test item |
positive control imidazole |
negative control physiological saline |
|||
average |
ICE class |
average |
ICE class |
average |
ICE class |
|
30 |
-0.8 |
I |
53.8 |
IV |
-6.5 |
I |
75 |
-0.5 |
I |
63.2 |
IV |
-5.0 |
I |
120 |
-1.6 |
I |
78.1 |
IV |
-7.9 |
I |
180 |
-4.5 |
I |
94.7 |
IV |
-7.9 |
I |
240 |
-8.1 |
I |
104.8 |
IV |
-5.4 |
I |
“-“ - percentage of corneal thickness decrease, no swelling
Evaluation of corneal swelling (%) – the second run.
observation after time t (minutes) |
test item |
positive control imidazole |
negative control physiological saline |
|||
average |
ICE class |
average |
ICE class |
average |
ICE class |
|
30 |
-0.1 |
I |
52.3 |
IV |
-4.8 |
I |
75 |
-2.4 |
I |
64.1 |
IV |
-4.8 |
I |
120 |
-4.8 |
I |
69.3 |
IV |
-6.6 |
I |
180 |
-8.0 |
I |
79.3 |
IV |
-9.9 |
I |
240 |
-8.0 |
I |
74.2 |
IV |
-9.4 |
I |
“-“ - percentage of corneal thickness decrease, no swelling
Applicant's summary and conclusion
- Interpretation of results:
- irritating
- Remarks:
- Based on EU criteria
- Conclusions:
- The substance can not be classified as severe irritant or corrosive to the eyes and it is not possible to state that the test item is not irritant to the eyes according to the UN GHS classification which is in accordance with the CLP criteria. According to the histopathological findings the test can be regarded as moderately irritating to the eyes.
- Executive summary:
An in vitro (ex vivo) study was conducted in order to determine the potential severe eye damaging effects of the substance theobromine according to the OECD guideline 438 under GLP conditions. Eyeballs were isolated from chickens killed for human consumption and after the appropriate preparation were exposed to 0.03 g of the test item, the same amount of imidazole and 0.03 mL of physiological saline were applied to other eyeballs as positive and negative controls. Three eyeballs were used for the test item and for each control. Fluorescein retention, corneal opacity and corneal swelling were evaluated, then the results of each endpoint were assigned to ICE classes according to OECD guideline 438 recomendations, histopathological evaluation of the corneal layers was conducted. Because the test item was a solid material and the first run lead to a no-classification outcome, a second run of three eyes for the test item was performed as recommended by the guideline, concurrent controls were included. According to the overall in vitro classification (UN GHS), no prediction can be made since the combinations of the 3 endpoints were 2xII and 1xIII in both runs, however taking into account the histopathological findings, the test can be considered as moderately irritating.
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.