Registration Dossier

Administrative data

Description of key information

Skin corrosion / irritation: corrosive [UN GHS Cat. 1B] (OECD 431, GLP, K, rel.1), highly irritating (similar to OECD 404, not GLP, SS, rel.2)

Eye irritation/damage: no prediction can be made (OECD 437, GLP, K, rel. 1)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
20 May 2015-10 July 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted according to OECD test Guideline No. 431 and EU Method B.40 BIS. Furthermore, functional model conditions and references to historical control data are included in the report.
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
Adopted 26 September 2014
Deviations:
no
Qualifier:
according to
Guideline:
other: UE Method B.40 BIS (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Specific details on test material used for the study:
4-chloroformylphthalic anhydride
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
Following the REACH bottom-up strategy, the EpiDerm™ Human Skin Model method was used to assess skin corrosion as recommended in the OECD test guideline No. 431.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Epi-200-Kit, MatTek Corporation (Ashland, MA, USA).
- Lot number: 22251 Kit AA
- Production Date: no data
- Shipping date: no data
- Delivery date: no data
- Date received: no data
- Date of initiation of testing: May 26, 2016

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 and 60 minutes at 37 °C in a humidified atmosphere of 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: After the exposure period, the tissues were washed with phosphate buffered saline (Invitrogen Corporation, Breda, The Netherlands) to remove residual test substance. Rinsed tissues were kept in 24 well plates on 300 μl DMEM medium until 6 tissues (= one application time) were dosed and rinsed.
- Observable damage in the tissue due to washing: none reported
- Modifications to validated SOP: none reported

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL MTT solution
- Incubation time: 3 hours at 37 °C, 5% CO2 in air
- Spectrophotometer: TECAN Infinite® M200 Pro Plate Reader
- Wavelength: 570 nm
- Filter: without reference filter
- Linear OD range of spectrophotometer: not reported

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: OD (540-570 nm): 1.852 +/- 0.086 [1.0-3.0]
- Barrier function: ET-50: 7.82 hrs [4.77-8.72 hrs]
- Morphology: normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo.
- Contamination: No contamination

NUMBER OF REPLICATE TISSUES: 4 tissues per test substance together with a negative control and positive control

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
Not needed

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA
A test item is considered corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after 3-minute treatment compared to the negative control tissues is decreased below 50%.
b) In addition, a test item considered non-corrosive (viability >= 50%) after the 3-minute treatment is considered corrosive if the relative tissue viability after 1-hour treatment with the test item is decreased below 15%.
A test item is considered non corrosive in the in vitro skin corrosion test if:
a) The relative mean tissue viability obtained after the 3-minute treatment compared to the negative control tissues is not decreased below 50%.
b) In addition, the relative tissue viability after the 1-hour treatment is not decreased below 15%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 27.1 to 29.9 mg of the solid test substance (skin was moistened with 25 μl Milli-Q water to ensure close contact of the test substance)

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of Milli-Q water was used as supplied

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL of 8.0 N Potassium Hydroxide was used as supplied
Duration of treatment / exposure:
3 and 60 minutes.
Duration of post-treatment incubation (if applicable):
not applicable
Number of replicates:
4 tissues per test substance together with a negative control and positive control
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3-minute exposure
Value:
92
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
100
Positive controls validity:
valid
Remarks:
14
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1-hour exposure
Value:
3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
100
Positive controls validity:
valid
Remarks:
12
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: Not reported
- Direct-MTT reduction: None
- Colour interference with MTT: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: Not included in the report

ACCEPTANCE OF RESULTS:
The in vitro skin corrosion test is considered acceptable if it meets the following criteria:
a) The absolute mean OD570 of the two tissues of the negative control should reasonably be within the laboratory historical control data range.
b) The mean relative tissue viability following 3-minute exposure to the positive control should be ≤ 30%.
c) In the range of 20 – 100% viability, the maximum inter-tissue variability (in viability) is ≤ 30% between two tissues treated identically.
d) In the range of 20 – 100% viability, the maximum difference in percentage between the mean viability of two tissues and one of the two tissues is ≤ 15%.

Mean absorption in the in vitro skin corrosion test with Trimellitic anhydride chloride

     3 -minute application              1 -hour application
 A (OD570)  B (OD570)

 Mean (OD570)

   SD  A (OD570)  B (OD570)  Mean (OD570)    SD
 Negative control 1.740 1.649  1.695   +/- 0.065  1.851  1.794  1.823   +/- 0.041 
 Test item 1.520 1.584  1.552   +/- 0.045  0.054  0.041  0.048  +/- 0.009 
 Positive control 0.285 0.174  0.229   +/- 0.078  0.277  0.155  0.216   +/- 0.086 

OD = Optical Density

SD = Standard Deviation

Duplicate exposures are indicated by A and B.

In this table the values are corrected for background absorption (0.0430). Isopropanol was used to measure the background absorption.

Interpretation of results:
Category 1 (corrosive) based on GHS criteria
Conclusions:
Under the experimental conditions of this study, Trimellitic anhydride chloride is corrosive in the in vitro skin corrosion test under the experimental conditions, therefore, the test substance should be classified category 1 according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations.
Executive summary:

An in vitro skin corrosion study was performed according to the most recent OECD Guideline 431, EU Method B.40 BIS and in compliance with GLP, using the EpiDerm™ Human Skin Model.

 

The test item was applied, after moistening with 25 µL of Milli-Q water (approximately 25 mg for 3-minute and 1 -hour exposures) directly on top of the skin tissue.

The positive control had a mean relative tissue viability of 14% after the 3-minute exposure.

The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The acceptability criteria for the maximum inter-tissue variability in viability between two tissues treated identically and the maximum difference in percentage between the mean viability of two tissues and one of the two tissues were met, indicating that the test system functioned properly.

Skin corrosion is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 3-minute and 1-hour treatments with Trimellitic anhydride chloride compared to the negative control tissues was 92% and 3%, respectively. Because the mean relative tissue viability for Trimellitic anhydride chloride was below 15% after 1 hour treatment it is considered to be corrosive.

Finally, it is concluded that this test is valid and that Trimellitic anhydride chloride is corrosive in the in vitro skin corrosion test under the experimental conditions described in this report

Under the experimental conditions of this study, Trimellitic anhydride chloride is corrosive in the in vitro skin corrosion test under the experimental conditions, therefore, the test substance should be classified category 1 according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations.

This study is considered as acceptable and satisfies the requirement for skin corrosion endpoint.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
From January 31, 1996 to March 5, 1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Remarks:
The study was not performed according to GLP and there are some lack of translation from the original Japanese report. But there is sufficient information and the study was comparable to guideline study with acceptable deviations.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
test on both intact and damaged skin
GLP compliance:
no
Specific details on test material used for the study:
4-chloroformylphthalic anhydride
Species:
rabbit
Strain:
other: Japanese white rabbit
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Kitayama LABES Co., Ltd.
- Weight at the time of administration: 2.84 - 3.04 kg
- Housing: individually stored in a bracket cage made from aluminum
- Diet (e.g. ad libitum): cubed diet (Oriental Yeast, RC-4)
- Water (e.g. ad libitum): tap water from town water
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21- 24
- Humidity (%): 45 - 65
- Air changes (per hr): 15 ventilation frequency/hour
- Photoperiod (hrs dark / hrs light): 12 hours/day of illuminations
Type of coverage:
semiocclusive
Preparation of test site:
other: damaged and intact skin
Vehicle:
other: olive oil
Controls:
no
Amount / concentration applied:
Amount(s) applied (volume or weight with unit): 0.5 g in a little amount of olive oil.
Duration of treatment / exposure:
4 hours
Observation period:
14 days
Number of animals:
3 males
Details on study design:
TEST SITE
- Area of exposure: back of the animals, 4 places (2 for healthy skin and 2 for damaged skin, one place of each skin was used as a control)
- Type of wrap : cotton lint of 2.5cmX2.5cm attached with surgical tape

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: after 4 hours
- Substance removal with a dry cloth

SCORING SYSTEM: Evaluation of skin reaction
- Spiloplaxia and scab formation
0: With No Spiloplaxia
1: Very Slight Spiloplaxia (Grade Accepted at Last)
2: Clear Spiloplaxia
3: Spiloplaxia Strong from Degree of Middle Class
4: Follow Light scab Formation (Trauma Covering Deep Part) on Strong Crimson Spiloplaxia.

- Edema formation
0: With No Edema
1: Very Slight Edema (Grade Accepted at Last)
2: Clear Edema (Distinction is Clearly [ as Circumference ] Possible)
3: Edema of Degree of Middle Class (it is Rising about 1 mm)
4: Strong Edema (it Rises 1 mm or More and, Also around, Spreads)
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Score:
5.67
Reversibility:
not fully reversible within: 14 days
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
animal #1
Score:
4
Reversibility:
not fully reversible within: 14 days
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
animal #2
Score:
6.5
Reversibility:
not fully reversible within: 14 days
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
animal #3
Score:
6.5
Reversibility:
not fully reversible within: 14 days
Irritant / corrosive response data:
Healthy skin:
Spiloplaxia and edema scores from 2 to 3 were observed in all animals since the 30th minute after removal and the spiloplaxia reached the score of 4 after 48 hours of observation. These stimulus reactions were not recovered in the 14th day after pasting elimination.

Damaged skin:
Spiloplaxia and edema scores from 2 to 3 were observed in all animals since the 30th minute after removal and the spiloplaxia reached the score of 4 after 24 hours of observation. These stimulus reactions were not recovered in the 14th day after pasting elimination.

Primary Irritation Index (P.I.I.) Classification:

0: With No Stimulus

0< x<2: A weak stimulus

2<=x<5: Middle degree stimulus

5<=x: A strong stimulus

Interpretation of results:
Category 1 (corrosive) based on GHS criteria
Conclusions:
The substance is considered as highly irritating in the conditions of a Primary Irritation test on rabbits.
Executive summary:

A Skin primary-Irritation Test was performed with trimellitic anhydride mono-chloride on Japanese white rabbit according to a method similar to the OECD 404 Guideline but not in compliance with GLP.

Three male rabbits were exposed to 0.5g of the substance wet with a small amount of olive oil. The paste was applied on the back of the animals on both healthy and damaged skin. On each animal, one place of each healthy and damaged skin was used as control. The tested skin was then covered by cotton lint attached with surgical tape. After 4 hours of exposure, the paste was removed with a dry cloth and the skin was observed during 14 days. Spiloplaxia and edema were scored from 0 (no effects) to 4 (strong effect). The primary irritation index (P.I.I.) was determined for each animal. The mean P.I.I. was found to be 5.67.

Based on the primary irritation index classification used in the study, the substance should be considered as highly irritant.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (corrosive)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 May 2015-12 June 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted according to OECD test Guideline No. 437. Furthermore, functional model conditions and references to historical control data are included in the report.
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Version / remarks:
Adopted July 26, 2013
Deviations:
no
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Version / remarks:
Amended by EC No. 1153/2010 No. L142, 09 December 2010
Deviations:
no
Qualifier:
according to
Guideline:
other: The Ocular Toxicity Working Group (OTWG) of the ICCVAM and the National Interagency Centre for the Evaluation of Alternative Toxicological Methods (NICEATM), Background Review Document (BRD): BCOP Test Method, March 2006
Qualifier:
according to
Guideline:
other: In Vitro Techniques in Toxicology Database (INVITTOX) protocol 127. Bovine Opacity and Permeability (BCOP) Assay, 2006.
Qualifier:
according to
Guideline:
other: Gautheron P., Dukic M., Alix D. and Sina J.F., Bovine corneal opacity and permeability test: An in vitro assay of ocular irritancy. Fundam Appl Toxicol 18:442-449, 1992.
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Specific details on test material used for the study:
4-chloroformylphthalic anhydride
Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco,’s Hertogenbosch, The Netherlands)
- Number of animals: Not specified
- Characteristics of donor animals (e.g. age, sex, weight): Not specified
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Instructions were given to avoid damaging the corneas during excision. Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.
- Time interval prior to initiating testing: as soon as possible after slaughter
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 304.5 to 325.1 mg of test item was directly applied on each cornea (completely covered).
- Concentration (if solution): Not applicable

POSITIVE CONTROL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): 20% (w/v) Imidazole solution

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): 750 µL
- Concentration (if solution): physiological saline
Duration of treatment / exposure:
4 hours ± 10 minutes at 32 ± 1 °C in a horizontal position
Duration of post- treatment incubation (in vitro):
None
Number of animals or in vitro replicates:
Total: 9 corneas - 3 corneas/group for test item, negative and positive controls
Details on study design:
SELECTION AND PREPARATION OF CORNEAS
The eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light. Those exhibiting defects were discarded.
The isolated corneas were stored in a petri dish with cMEM (Eagle’s Minimum Essential Medium (Life Technologies, Bleiswijk, The Netherlands) containing 1% (v/v) L-glutamine (Life Technologies) and 1% (v/v) Foetal Bovine Serum (Life Technologies)). The isolated corneas were mounted in a corneal holder (one cornea per holder) of MC2 (Clermont-Ferrand, France) with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was positioned on top of the cornea and tightened with screws. The compartments of the corneal holder were filled with cMEM of 32 ± 1°C. The corneas were incubated for the minimum of 1 hour at 32 ± 1°C.

QUALITY CHECK OF THE ISOLATED CORNEAS : After the incubation period, the medium was removed from both compartments and replaced with fresh cMEM. Opacity determinations were performed on each of the corneas using an opacitometer (OP-KIT, MC2, Clermont-Ferrand, France). The opacity of each cornea was read against an air filled chamber, and the initial opacity reading thus determined was recorded. Corneas that had an initial opacity reading higher than 7 were not used. Three corneas were selected at random for each treatment group.

NUMBER OF REPLICATES : 3 corneas/group

NEGATIVE CONTROL USED : physiological saline

POSITIVE CONTROL USED : 20% (w/v) Imidazole solution

APPLICATION DOSE AND EXPOSURE TIME : The anterior compartment received the test item (304.5 to 325.1 mg) or negative or positive control at a volume (750 μL) on the surface of the corneae. Since no workable suspension in physiological saline could be obtained, the test substance was used as delivered and added pure on top of the corneas. The corneae were incubated in a horizontal position for 4 hours ± 5 minutes at 32 ± 1 °C.

TREATMENT METHOD: closed chamber

POST-INCUBATION PERIOD: Yes.
Following the final opacity measurement, permeability of the cornea to Na-fluorescein (Merck) was evaluated.
The medium of both compartments (anterior compartment first) was removed. The posterior compartment was refilled with fresh cMEM. The anterior compartment was filled with 1 ml of 5 mg Na-fluorescein/ml cMEM solution (Sigma-Aldrich Chemie GmbH, Germany). The holders were slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the sodium-fluorescein solution over the entire cornea. Corneas were incubated in a horizontal position for 90 ± 5 minutes at 32 ± 1°C.

REMOVAL OF TEST SUBSTANCE:
- Number of washing steps after exposure period: After the incubation the solutions and the test compound were removed and the epithelium was washed at least three times with MEM with phenol red (Eagle’s Minimum Essential Medium Life Technologies).

- POST-EXPOSURE INCUBATION: no

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: The opacitometer determined the difference in the light transmission between each control or treated cornea and an air filled chamber. The numerical opacity value (arbitrary unit) was displayed and recorded. The change in opacity for each individual cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final post-treatment reading. The corrected opacity for each positive control or test substance treated cornea was calculated by subtracting the average change in opacity of the negative control corneas from the change in opacity of each positive control or test substance treated cornea.
The mean opacity value of each treatment group was calculated by averaging the corrected opacity values of the treated corneas for each treatment group.
- Corneal permeability: After the incubation period, the medium in the posterior compartment of each holder was removed and placed into a sampling tube labelled according to holder number. 360 μl of the medium from each sampling tube was transferred to a 96-well plate. The optical density at 490 nm (OD490) of each sampling tube was measured in triplicate using a microplate reader (TECAN Infinite® M200 Pro Plate Reader). Any OD490 that was 1.500 or higher was diluted to bring the OD490 into the acceptable range (linearity up to OD490 of 1.500 was verified before the start of the experiment). OD490 values of less than 1.500 were used in the permeability calculation.
The mean OD490 for each treatment was calculated using cMEM corrected OD490 values. If a dilution was performed, the OD490 of each reading was corrected for the mean negative control OD490 before the dilution factor was applied to the readings.
- Others (e.g, pertinent visual observations, histopathology): Possible pH effects of the test substance on the corneas were recorded. Each cornea was inspected visually for dissimilar opacity patterns.

SCORING SYSTEM: In Vitro Irritancy Score (IVIS) = mean opacity value + (15 x mean OD490 value)

DECISION CRITERIA:
In vitro scrore range: UN GHS:
- =< 3 - No category
- > 3; =< 55 - No prediction can be made
- > 55 - Category 1

ACCEPTABILITY CRITERIA
The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within two standard deviations of the current historical mean.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range.
Irritation parameter:
in vitro irritation score
Run / experiment:
mean/3 corneas incubated 2 hours
Value:
42.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Remarks:
0.0
Positive controls validity:
valid
Remarks:
144.0
Remarks on result:
other: No prediction can be made
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: The corneas were translucent with spots after the 240 minutes of treatment with Trimellitic anhydride chloride. A pH effect of the test substance was observed on the rinsing medium, the corneas were rinsed until no colour change of the medium was observed.

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

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:
The positive control should elicit an In Vitro Irritancy Score that falls within two standard deviations of the historical mean (2 x SD 18.75 to 95.22) for the laboratory.
The negative control mean opacity change value should be ≤2.0 and the permeability mean value ≤0.050 (assays performed from April 2012 to April 2015).

Table 1 - Summary of Opacity, Permeability and In Vitro Scores

 Treatment  Mean Opacity1  Mean Permeability1  Mean In Vitro Score1, 2
 Negative control  0.0  0.000  0.0
 Positive control (20% (w/v) imidazole)  84.3  3.975  144.0
 Test item  42.3  -0.017  42.1

1Calculated using the negative control mean opacity and mean permeability values for the positive control and test item.

2In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

Interpretation of results:
other: no prediction can be made
Conclusions:
Trimellitic anhydride chloride induced ocular irritation through only one endpoint (opacity), resulting in a mean in vitro irritancy score of 42.1 after 240 minutes of treatment.
With an IVIS at 42.1, no prediction can be made as the result is outside the decision criteria (i.e. >3 or =<55)
Executive summary:

The eye hazard potential of the Trimellitic anhydride chloride was evaluated using the Bovine Corneal Opacity and Permeability test (BCOP test), under GLP compliance, according to the OECD Guideline No. 437.

The corneal damage potential of test substance was assessed using fresh bovine cornea. 304.5 to 325.1 mg of test item was applied topically on cornea for 240 minutes at 32 ± 1 °C. After incubation, the solutions and the test compound were removed and the epithelium was washed. Each cornea was inspected visually and then, corneal opacity was measured. Three corneas were used for each treated series (test item; negative control: physiological saline; positive control: 20% (w/v) imidazole). Following the opacity measurement, the permeability of the corneas to sodium fluorescein was evaluated. Two endpoints, corneal opacity and permeability, were measured and combined to give an In Vitro Irritancy Score.

 

The individual in vitro irritancy scores for the negative controls ranged from -0.3 to 0.6. The individual positive control in vitro irritancy scores ranged from 120.9 to 153.6. The corneas treated with the positive control item were turbid after the 240 minutes of treatment. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range (2.0 and 0.050, respectively) indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control was 144.0 and within two standard deviations of the current historical positive control mean (68 - 135). It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

The corneas treated with Trimellitic anhydride chloride were translucent with spots after the 240 minutes of treatment. A pH effect of the test substance was observed on the rinsing medium. Hence, Trimellitic anhydride chloride induced ocular irritation through only one endpoint (opacity), resulting in a mean in vitro irritancy score of 42.1 after 240 minutes of treatment (in vitro irritancy scores ranged from 31.4 to 49.4) and, the mean IVIS was greater than 3 and lower than 55 (i.e. 42.1).

 

Therefore, no prediction on the classification can be made for eye irritation or serious eye damage, according to the GHS Regulation.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion:

One key study and one supporting study were identified for the skin irritation / corrosion.

A key study (WIL Research, 2015, Rel.1) was performed to evaluate the corrosive potential of the test substance for skin. This in vitro skin corrosion study was performed according to the OECD Guideline 431 and in compliance with GLP, using the EpiDerm™ Human Skin Model. The relative mean viability of the test item treated tissues was 92.0% and 3.0% after 3 and 60 minutes exposure, respectively. The quality criteria required for acceptance of results in the test were satisfied. With a tissue viability < 15% after 60 minutes of exposure, the test material was considered to be corrosive to skin. Results of this test using the EpiDerm™ Human Skin Model were >= 50% after 3 min exposure and < 15% after 60 moin exposure, according to the OECD Guideline 431 criteria, the test substance should be classified as Corrosive Cat. 1B or 1C. Therefore, without further information Trimetillic anhydride chloride is classified according to the worse case: UN GHS Cat. 1B.

A supporting study (Environmental Biological Life Science Research Center, 1996, Rel.2) was performed to evaluate the irritant / corrosive potential of the test substance for skin. This skin primary-Irritation Test was performed on Japanese white rabbit according to a method similar to the OECD 404 Guideline but not in compliance with GLP.

Three male rabbits were exposed to 0.5g of the substance wet with a small amount of olive oil. The paste was applied on the back of the animals on both healthy and damaged skin. On each animal, one place of each healthy and damaged skin was used as control. The tested skin was then covered by cotton lint attached with surgical tape. After 4 hours of exposure, the paste was removed with a dry cloth and the skin was observed during 14 days. Spiloplaxia and edema were scored from 0 (no effects) to 4 (strong effect). The primary irritation index (P.I.I.) was determined for each animal. The mean P.I.I. was found to be 5.67.

Based on the primary irritation index classification used in the study, the substance should be considered as highly irritant.

Eye irritation:

A key study was identified for eye irritation. This BCOP assay (WIL Research, 2015, Rel.1) was performed according to the OECD Guideline 437 and in compliance with GLP, using bovine corneas. The quality criteria required for acceptance of results in the test were satisfied. The mean In Vitro Irritancy Score of the test item was 42.1, after the 240-minute exposure period. With an IVIS included between 3 and 55, the test substance cannot be classified for eye irritation or serious eye damage as no prediction can be made, according to the UN GHS Regulation. However, based on the classification for skin irritation (Corrosive, Category 1), the test substance is likely to induce serious eye damage / irreversible effects on the eye (category 1).

Respiratory irritation:

No data was available regarding respiratory irritation, however the substance being classified for skin and eye corrosivity, the substance is likely to be "corrosive to respiratory tract" (labelling EUH 071) according to Regulation (EC) No 1272/2008 (CLP).

Justification for classification or non-classification

Harmonized classification:

Trimetillic anhydride chloride does not have harmonized classification according to the Regulation (EC) No. 1272/2008.

Self-classification:

Based on the available information, the substance should be classified as skin corrosive sub-category 1B (H314 “Causes severe skin burns and eye damage) according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS. Thus, the corrosive substance or mixture is also classified for eye dammage (Serious eye damage Category 1), but the corresponding hazard statement (H318: Causes serious eye damage) is not indicated on the label to avoid redundancy.

No data was available regarding respiratory irritation. In general, a classification for corrosivity is considered to implicitly cover the potential to cause respiratory tract irritation and so the additional Category 3 is considered to be superfluous. However, such substance has to be supplementary labelled with EUH071 if there is a possibility of exposure via inhalation taking into consideration the saturated vapour concentration and the possibility of exposure to dust, particles or droplets of inhalable size. It is also strongly recommended to apply the precautionary statement P260: "Do not breathe dust/fume/gas/mist/vapours/spray." In the case of Trimellitic anhydride chloride (TMAC) this labelling is not applicable as the exposure to TMAC by inhalation is unlikely as it is a hygroscopic solid and presents a low vapour pressure (0.017 Pa at 20°C).