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Acute Toxicity: dermal

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Endpoint:
acute toxicity: dermal
Data waiving:
other justification
Justification for data waiving:
the study does not need to be conducted because the physicochemical and toxicological properties suggest no potential for a significant rate of absorption through the skin
the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)
Justification for type of information:
Dermal Absorption studies by Hallegot et al 2011 and Johnson et al 2013, performed with not treated nanoforms of carbon black indicate lack of potential of these nanoforms to penetrate the skin. Transmission electron microscopy (TEM) analysis of the skin samples treated with eyeliner containing carbon black showed that particulate material (identified as carbon black) was only observed in the outer layers of the Stratum Corneum. There was no evidence of deeper penetration of carbon black material into the Stratum Corneum or into living epidermis Hallegot et al 2011. Johnson et al 2013 applying a test formulation containing not a not treated carbon black (with particle sizes between 20-30 nm) to full-thickness human skin samples in accordance with OECD 428 protocol could demonstrate no permeation of carbon black particles from a cosmetic eyeliner formulation into or beyond the outer layer of the stratum corneum was found.
This data waiving argument is valid for all forms of carbon black (non-nano, not treated and surface treated). Solubility studies in water and dissolution testing in biological fluids indicate a similar lack of insolubility for members of the sets of (nano)forms which also are not prone to release toxic moieties. Given the lack of solubility and particle sizes, treated forms (with particle sizes comparable to that of the not treated forms) and non-nanoforms (with particle sizes greater than that of the treated forms), are not expected to possess a higher skin higher penetration potential than what has been demonstrated for the not-treated forms by Hallegot et al 2011 and Johnson et al 2013.
Cross-referenceopen allclose all
Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
dermal absorption in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
other information
Study period:
before 2011
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Qualifier:
no guideline followed
Principles of method if other than guideline:
Human abdominal full-thickness skin samples from a single donor. About 9.5 mg/cm² of a typical eyeliner formulation containing 6% carbon black was applied to the skin surface and left for 24 hours. The absorption of carbon black particles was estimated by Transmission Electron Microscopy (TEM)
GLP compliance:
not specified
Specific details on test material used for the study:
"Distinctive® Ink Black Lo-AQ" containing 6% pure carbon black
Radiolabelling:
no
Species:
human
Type of coverage:
open
Vehicle:
other: eyeliner formulation
Duration of exposure:
24 hours
Key result
Time point:
24 h
Dose:
9.5mg/cm² of a typical eyeliner formulation containing 6% carbon black
Parameter:
percentage
Absorption:
0 %

Transmission electron microscopy (TEM) analysis of the skin samples treated with eyeliner containing carbon black showed that particulate material (identified as carbon black) was only observed in the outer layers of the Stratum Corneum. There was no evidence of deeper penetration of carbon black material into the Stratum Corneum or into living epidermis

Conclusions:
The results obtained suggest that carbon black as used in a typical eyeliner product has no potential to penetrate into or cross through the skin, thus no potential to produce systemic exposure of consumers.
Executive summary:

The dermal penetratiion of carbon black particles from a typical eyeliner formulatioin was estimated by Transmission Electron Microscopy (TEM) examination of multiple human skin sections exposed for 24 hours (non-occluded). In addition, toluidine blue-stained semi-thin sections of each biopsy from the control and treated skin samples were examined at light microscopy. Carbon black containing particulate matter did not penetrate beyond the outer layers of the stratum corneum.

Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
dermal absorption in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
September - December 2012
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Qualifier:
according to guideline
Guideline:
OECD Guideline 428 (Skin Absorption: In Vitro Method)
Version / remarks:
2004
GLP compliance:
yes
Specific details on test material used for the study:
Test Cosmetic Formulation: Black 2/CI 77266 12%, water 64.14%, gums 3.3%, propylene glycol 15%, trilaureth-4 phosphate 2%, preservatives 1.06%, alcohol denat 2.5%; primary particle size of carbon black: 20-30 nm
Radiolabelling:
no
Species:
human
Type of coverage:
open
Vehicle:
other: see Test Formulation, above
Duration of exposure:
24 hours
Doses:
10 mg/cm2, 2.54 cm2 exposure area, and a receptor volume of approximately 4.5 ml. Since the test material was a thick liquid, the doses were applied to the skin and spread over the surface using pipette tips and the weight of the applied dose recorded after spreading
Dose:
10 mg/cm2 of a test formulation containing 12% carbon black
Parameter:
percentage
Absorption:
0 %
Remarks on result:
other: 24 hours
Conclusions:
Carbon black particles did not penetrate into or beyond the surface layers of human Stratum Corneum following application of a cosmetic formulation.
Executive summary:

A GLP dermal penetration study was performed using the OECD 428 protocol with full-thickness human skin samples and transmission electron microscopy for semi-quantitative evaluation of dermal permeation. No permeation of carbon black particles from a cosmetic eyeliner formulation into or beyond the outer layer of the strateum corneum was found.

Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Justification for type of information:
Endpoint-specific read-across justification" attached under section 13.2
Reason / purpose for cross-reference:
read-across source
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Route of administration:
oral: gavage
Doses:
8000 mg/kg bw (maximum technically achievable concentration)
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 8 000 mg/kg bw
Mortality:
There was no mortality.
Clinical signs:
There were no clinical signs of toxicity.
Gross pathology:
Findings at necropsy were unremarkable
Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral LD50 value in rats is greater than 8000 mg/kg bw (maximum achievable concentration)
Executive summary:

No mortality and no signs of toxicity occured in a OECD 401 guideline study with rats at 8,000 mg/kg bw. Necropsy findings were unremarkable.

Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from June 1978 to July 1978
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: S. Ivanovas GmbH & Co., Med. Versuchstierzuchten K.G., Kißlegg/Allgäu, Germany
- Age at study initiation: 38 days (male), 42 days (female)
- Weight at study initiation: 100 -105 g
- Fasting period before study: 15-16 hours
- Housing: single, in Makrolon cages
- Diet (e.g. ad libitum): ad lib
- Water (e.g. ad libitum): ad lib
- Acclimation period: not reported

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 0.5
- Humidity (%): 55 +/- 5
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): not reported

IN-LIFE DATES: From: June 1978 To: Jui 1978
Route of administration:
oral: gavage
Vehicle:
other: aqueous hydroxypropyl methyl cellulose (0.8%)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: not reported (maximum technically achievable concentration)
- Amount of vehicle (if gavage): 50 mL/kg
- Justification for choice of vehicle:

MAXIMUM DOSE VOLUME APPLIED: not reported
Doses:
8000 mg/kg bw (maximum technically achievable concentration)
No. of animals per sex per dose:
20
Control animals:
no
Details on study design:
TEST ANIMALS: 10 male and 10 female Sprague-Dawley rats, from Ivanovas GmbH & Co., Kisslegg/Allgäu), weight between 100 and 105 g. Age at study begin: 38 days (male animals), 42 days (female animals). Animals were fed standard laboratory diet (Altromin), water was available ad libitum. Animals were held individually in macrolon cages in an air-conditioned room at 22 +/- 0.5 deg centigrade, and a relative humidity of 55% +/- 5%. EXPOSURE TO TEST SUBSTANCE: The test substance was applied via single gavage as suspension in 0.8% aqueous hydroxypropyl methyl cellulose E4M in a constant volumne of 50 mL/kg bw. Food was withdrawn 15-16 hours before administration of the test substance. POST EXPOSURE OBSERVATION PERIOD: 4 weeks. OBSERVATIONS/PARAMETERS: behaviour, food consumption, body weight gain. All animals were necropsied and macroscopically evaluated at the end of the post-observation period.
Statistics:
Not performed
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 8 000 mg/kg bw
Mortality:
There was no mortality.
Clinical signs:
There were no clinical signs of toxicity.
Body weight:
Food consumption was reduced 0, 4, and 6% at days 1, 2, and 14, respectively. Body weight gain was reduced 2, 4, and 4% at days 1, 2, and 14, respectively.
Gross pathology:
Findings at necropsy were unremarkable
Other findings:
Post-dose observation time was 4 weeks. There were no effects with regard to the behaviour of the animals.
Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral LD50 value in rats is greater than 8000 mg/kg bw (maximum achievable concentration)
Executive summary:

No mortality and no signs of toxicity occured in a OECD 401 guideline study with rats at 8,000 mg/kg bw. Necropsy findings were unremarkable.

Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
October 1977
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
other: as suspension in 1% aqueous methyl hydroxy ethyl cellulose
Details on oral exposure:
TEST ANIMALS: 10 male and 10 female Sprague-Dawley rats per group, from Ivanovas GmbH & Co., Kisslegg/Allgäu), weight between 100 and 105 g. Age at study begin: 38 days (male animals), 42 days (female animals). Animals were fed standard laboratory diet (Altromin), water was available ad libitum. Animals were held individually in macrolon cages in an air-conditioned room at 24 +/- 0.5 deg centigrade, and a relative humidity of 60% +/- 3%. EXPOSURE TO TEST SUBSTANCE: The test substance was applied via single gavage as suspension in 1% aqueous methyl hydroxy ethyl cellulose at a constant volume of 50 mL/kg bw. Food was withdrawn 15-16 hours before administration of the test substance. POST EXPOSURE OBSERVATION PERIOD: 4 weeks. OBSERVATIONS/PARAMETERS: behaviour, food consumption, body weight gain. All animals were necropsied and macroscopically evaluated at the end of the post-observation period.
Doses:
6350; 7900; 10000 mg/kg bw
No. of animals per sex per dose:
10
Control animals:
no
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 10 000 mg/kg bw
Mortality:
There was no mortality
Clinical signs:
There were no clincal signs of toxicity
Body weight:
Food consumption was reduced 10, 6, and 6% at days 1, 2, and 7 for the 6350 mg/kg bw group; 6, 8, and 8% at days 1, 2, and 7 for the 7900 mg/kg bw group; and 0, 2, and 6% at days 1, 2, and 7 for the 10000 mg/kg bw group, respectively. Body weight gain was reduced 0, 2, and 6% at days 1, 2, and 7 for the 6350 mg/kg bw group; 0, 0, and 6% at days 1, 2, and 7 for the 7900 mg/kg bw group; and 6, 4, and 6% at days 1, 2, and 7 for the 10000 mg/kg bw group, respectively.
Gross pathology:
Findings at necropsy were unremarkable
Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral LD50 in rats is greater than 10,000 mg/kg bw (maximum technically feasible dose)
Executive summary:

No mortality and no signs of toxicity occured in a OECD 401 guideline study with rats at 10,000 mg/kg bw. Necropsy findings were unremarkable.

Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
December 2011 – March 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
The test item carbon black was suspended in 0.5% aqueous sodium carboxymethylcellulose and given at 10 ml/kg bw.
Analytical verification of doses or concentrations:
not specified
Dose / conc.:
30 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Control animals:
yes
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: neoplastic
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
water consumption and compound intake
Key result
Critical effects observed:
no

Results are summarised by the EU Scientific Committee on Consumer Products (2015) as follows: "Following daily administration of carbon black, there were no deaths, no adverse clinical signs, no ocular findings or changes in body weight gain and food intake when compared to controls. Dark coloured faeces were observed in animals given carbon black at all dose levels. This finding is related to the coloured nature of the test item and hence considered to be non-adverse. There were no relevant changes in laboratory parameters. Some haematology and blood clinical chemistry parameters showed minimal changes when compared to controls. However, as they remained minor and showed no dose-relationship, these changes were considered to be unrelated to the administration of carbon black. There were no changes attributed to carbon black in any of the urinary parameters evaluated. Dosing with carbon black did not produce any changes in organ weights. The only finding at necropsy consisted of black gastro-intestinal tract contents in all animals given carbon black, without any associated gross or microscopic changes in the gastro-intestinal mucosa apart from black discolouration. These findings were also considered to be related to the coloured nature of the test item and hence non adverse. There were no other histopathological findings attributed to carbon black."

Conclusions:
There were no adverse effects found in a modern 90 day GLP and guideline study after repeated oral administration by gavage. The study authors concluded that the repeated daily oral administration to rats of carbon black for 13 weeks at 100, 300 or 1000 mg/kg/day was well tolerated, with some nonadverse findings related to the staining properties of carbon black. Accordingly, under the conditions of this study, the No Observed Adverse Effect Level (NOAEL) was 1000 mg/kg/day.
Executive summary:

The results of a 90 -day GLP and guideline study in rats are summarised by the EU Scientific Committee on Consumer Products (2015) as follows: "Following daily administration of carbon black, there were no deaths, no adverse clinical signs, no ocular findings or changes in body weight gain and food intake when compared to controls. Dark coloured faeces were observed in animals given carbon black at all dose levels. This finding is related to the coloured nature of the test item and hence considered to be non-adverse. There were no relevant changes in laboratory parameters. Some haematology and blood clinical chemistry parameters showed minimal changes when compared to controls. However, as they remained minor and showed no dose-relationship, these changes were considered to be unrelated to the administration of carbon black. There were no changes attributed to carbon black in any of the urinary parameters evaluated. Dosing with carbon black did not produce any changes in organ weights. The only finding at necropsy consisted of black gastro-intestinal tract contents in all animals given carbon black, without any associated gross or microscopic changes in the gastro-intestinal mucosa apart from black discolouration. These findings were also considered to be related to the coloured nature of the test item and hence non adverse. There were no other histopathological findings attributed to carbon black."

Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Endpoint-specific read-across justification" attached under section 13.2
Reason / purpose for cross-reference:
read-across source
Species:
rat
Strain:
Wistar
Sex:
male/female
Route of administration:
oral: gavage
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: neoplastic
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
water consumption and compound intake
Key result
Critical effects observed:
no
Conclusions:
There were no adverse effects found in a modern 90 day GLP and guideline study after repeated oral administration by gavage. The study authors concluded that the repeated daily oral administration to rats of carbon black for 13 weeks at 100, 300 or 1000 mg/kg/day was well tolerated, with some nonadverse findings related to the staining properties of carbon black. Accordingly, under the conditions of this study, the No Observed Adverse Effect Level (NOAEL) was 1000 mg/kg/day.
Executive summary:

The results of a 90 -day GLP and guideline study in rats are summarised by the EU Scientific Committee on Consumer Products (2015) as follows: "Following daily administration of carbon black, there were no deaths, no adverse clinical signs, no ocular findings or changes in body weight gain and food intake when compared to controls. Dark coloured faeces were observed in animals given carbon black at all dose levels. This finding is related to the coloured nature of the test item and hence considered to be non-adverse. There were no relevant changes in laboratory parameters. Some haematology and blood clinical chemistry parameters showed minimal changes when compared to controls. However, as they remained minor and showed no dose-relationship, these changes were considered to be unrelated to the administration of carbon black. There were no changes attributed to carbon black in any of the urinary parameters evaluated. Dosing with carbon black did not produce any changes in organ weights. The only finding at necropsy consisted of black gastro-intestinal tract contents in all animals given carbon black, without any associated gross or microscopic changes in the gastro-intestinal mucosa apart from black discolouration. These findings were also considered to be related to the coloured nature of the test item and hence non adverse. There were no other histopathological findings attributed to carbon black."

Reason / purpose for cross-reference:
data waiving: supporting information
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Endpoint-specific read-across justification" attached under section 13.2
Reason / purpose for cross-reference:
read-across source
Species:
rat
Strain:
Wistar
Sex:
male/female
Route of administration:
oral: gavage
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: neoplastic
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
water consumption and compound intake
Key result
Critical effects observed:
no
Conclusions:
There were no adverse effects found in a modern 90 day GLP and guideline study after repeated oral administration by gavage. The study authors concluded that the repeated daily oral administration to rats of carbon black for 13 weeks at 100, 300 or 1000 mg/kg/day was well tolerated, with some nonadverse findings related to the staining properties of carbon black. Accordingly, under the conditions of this study, the No Observed Adverse Effect Level (NOAEL) was > 1000 mg/kg/day.
Executive summary:

The results of a 90 -day GLP and guideline study in rats are summarised by the EU Scientific Committee on Consumer Products (2015) as follows: "Following daily administration of carbon black, there were no deaths, no adverse clinical signs, no ocular findings or changes in body weight gain and food intake when compared to controls. Dark coloured faeces were observed in animals given carbon black at all dose levels. This finding is related to the coloured nature of the test item and hence considered to be non-adverse. There were no relevant changes in laboratory parameters. Some haematology and blood clinical chemistry parameters showed minimal changes when compared to controls. However, as they remained minor and showed no dose-relationship, these changes were considered to be unrelated to the administration of carbon black. There were no changes attributed to carbon black in any of the urinary parameters evaluated. Dosing with carbon black did not produce any changes in organ weights. The only finding at necropsy consisted of black gastro-intestinal tract contents in all animals given carbon black, without any associated gross or microscopic changes in the gastro-intestinal mucosa apart from black discolouration. These findings were also considered to be related to the coloured nature of the test item and hence non adverse. There were no other histopathological findings attributed to carbon black."

Data source

Materials and methods

Test material

Constituent 1
Chemical structure
Reference substance name:
Carbon black
EC Number:
215-609-9
EC Name:
Carbon black
Cas Number:
1333-86-4
Molecular formula:
C
IUPAC Name:
Carbon black
Test material form:
other: solid: non-nanoform; solid: nanoform, no surface treatment; solid: nanoform, surface-treated

Results and discussion

Applicant's summary and conclusion