Monalazone disodium

Endpoint summary

Description of key information

Skin irritation

OECD439 (Epiderm): Irritant Cat 2

ISO (rabbit): Mild Irritant Cat 3

Eye irritation

OECD 492 (in vitro): Irritant (Cat 2) or Corrosive (Cat 1)

OECD 437 (in vitro): Corrosive (Cat1)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

GLP compliance:

yes

Specific details on test material used for the study:

20% Monalazone Disodium
5% Monalazone Disodium in water.
Stored at room temperature in the absence of light. The dilutions were prepared immediately prior to exposures.

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

The 3-Dimensional human epithelial skin model (EpiDerm™, MatTek, Ashland, MA) is made up of normal human keratinocytes cultured on a permeable synthetic membrane at the air-liquid interface in a chemically defined medium. The cells form a multilayered, highly differentiated model of the human epidermis that consists of organized basal, spinous, granular, and cornified layers and closely resembling native epidermis. Each lot of tissues is Quality Assured by MatTek, Inc. according to specific QC standards including: histology (cell layers), tissue viability (MTT mean optical density) and reproducibility (SD).

Upon receipt, the MatTek EpiDerm™ tissue cultures were placed in 0.9 mL of fresh Maintenance medium (6-well plate) for one hour. The tissues were then transferred to 6-well plates containing 0.9 mL of fresh Maintenance medium and they were incubated overnight. 30 µL of the negative control, DPBS, and positive control, 5% SDS, were added to the apical surface of tissues. 30 µL of each test material was added to the apical surface of tissues. All tissues were placed into the 37°C incubator with 5% CO2. The exposure time was 1 hour, with 35 minutes exposure in the incubator and 25 minutes at room temperature. After the 1 hour exposure, the tissues were rinsed 20 to 25 times with 1 mL of DPBS. The apical surface was gently blotted with a cotton swab. The tissues were placed in 0.9 mL of fresh Maintenance medium (6-well plate) for 24 hours. After 24 hours, the basal media was replaced with 0.9 mL of fresh Maintenance medium (6-well plate) and incubated for another 18 hours prior to performing the MTT assay.

Tissues were then evaluated for cell viability using an MTT Assay where yellow MTT is reduced to purple formazan primarily by enzymes (reductases) located in the mitochondria of living cells. According to established Cyprotex procedure, a stock solution of MTT (Sigma, M-5655) was prepared in Maintenance medium (provided with tissues) just prior to use and warmed to 37°C in a water bath. Tissues inserts were transferred to 24-well plates containing 300 µL MTT medium (1 mg/mL). After 3 hour MTT incubation, the formazan salt was extracted with 2 mL isopropanol per tissue and the optical density of the extracted formazan is determined using a spectrophotometer at 570 nm. Viable cells had the greatest amount of MTT reduction and hence the highest absorbance values. Relative cell viability was calculated for each tissue as % of the mean of the negative control tissues.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

30 ul

Duration of treatment / exposure:

1 hour

Duration of post-treatment incubation (if applicable):

42 h

Number of replicates:

Three

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

20% Monalazone disodium

Value:

11.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

5% Monalazone Disodium

Value:

42

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

	Mean	SD	SEM	%CV	p-value	Significant?
DPBS	100.0	3.6	2.1	3.6
5% SDS	5.0	0.7	0.4	13.1	<0.001	YES
20% monalazone disodium	11.9	5.2	3.0	43.7	<0.001	YES
Dilute (5% monalazone disodium) in Water	42.0	8.3	4.8	19.8	0.001	YES

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

The test articles (20% monalazone disodium and 5% monalazone Disodium) were determined to be irritants to human skin as the viability of the exposed cells were <50% of control.

Executive summary:

The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article under three conditions by determining the viability of the tissues following exposure via MTT. The objective of this study was to assess the dermal irritation potential of the Sponsor’s submitted test article under three conditions. Pre-testing showed the test articles/conditions were not colored though one test article, 20% Monalazone Disodium did auto-reduce MTT. A small data correction was necessary as the auto-reduction was observed during the study. Tissues were exposed to test articles and controls for one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay.

The assay passed all the quality controls. The negative control tissue OD was between 1.651 and 1.942, the positive control was within the confidence limits and was deemed an irritant, and the test article SDs were all ≤18. The MTT data show the positive control, 5% sodium dodecyl sulfate (SDS), reduced tissue viability to 5.0% of control. The mean viability of tissues after exposure to the test articles, 20% monalazone disodium and Dilute 5% monalazone disodium in water were <50% of control. Therefore, the test articles were determined to be irritants to human skin according to the OECD test guideline followed for this study.

 

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

June 26,2016-July 16, 2016

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

other: ISO 10993-10. Biological evaluation of medical devices- Part 10

GLP compliance:

not specified

Specific details on test material used for the study:

Identification: Lot #52716BC13
Physical Description of the Test Article: Liquid with 20% active component (Monalazone disodium)
Storage Conditions: Room Temperature

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Source: Robinson Services, Inc.
Sex: Male
Body weight range 2.4 kg to 2.5 kg at selection
Age: Young adult
Acclimation period: Minimum 5 days
Number of animals: Three
Identidication method: Ear tag

Animal Management:

Husbandry, Housing and Environment
Conditions conformed to NAMSA Standard Operating Procedures that are based on the "Guide for the Care and Use of Laboratory Animals." Animals were individually housed in stainless steel or plastic suspended cages identified by a card indicating the lab number, animal number,
test code, sex, and date dosed.
The animal housing room temperature and relative humidity were monitored daily. The temperature for the room was set to 61-72°F and the relative humidity was set to 30-70%. There were no significant temperature or relative humidity excursions that adversely affected
the health of the animals.
The light cycle was controlled using an automatic timer (12 hours light, 12 hours dark).

Food, Water and Contaminants
A commercially available rabbit feed, Laboratory Rabbit Diet - 5326, was provided daily. Potable water was provided ad libitum through species appropriate water containers or delivered through an automatic watering system. No contaminants present in the feed and water impacted the results of this study.

Accreditation
NAMSA is an AAALAC International accredited facility and is registered with the United States Department of Agriculture. Additionally, NAMSA maintains an approved Animal Welfare Assurance on file with the National Institutes of Health, Office for Laboratory Animal Welfare.

Personnel
Associates involved were appropriately qualified and trained.

Veterinary Care
Standard veterinary medical care was provided in this study.

IACUC
This procedure has been approved by the NAMSA Institutional Animal Care and Use Committee (IACUC), and is reviewed at least annually by the same committee.

Selection
Only healthy, previously unused, animals free from irritation or other dermatological lesions that could interfere with the test were selected.

Type of coverage:

occlusive

Preparation of test site:

abraded

Controls:

yes, concurrent no treatment

Amount / concentration applied:

20% monalazone

Duration of treatment / exposure:

24 h

Observation period:

1, 24, 48 and 72 hours

Number of animals:

Three

Details on study design:

Test Procedure
The animals were weighed and the fur on the back of each animal was clipped with an electric clipper 4 to 24 hours prior to treatment. On the day of treatment, four sites, two on each side of the back and positioned cranially and caudally, were designated on each animal. The sites were free of blemishes that could interfere with the interpretation of results. Just prior to application, the test and control application sites on the right side of the back were abraded. Each site to be abraded received four parallel epidermal abrasions with a sterile needle within an approximate 25 mm x 25 mm area. The application sites on the left side of the back remained intact. The test article was applied to one cranial site non-abraded on the left side and one caudal site abraded on the right side (two sites per animal) by introduction under a 4 ply gauze layer to an area of skin approximately 25 mm x 25 mm square. The patches were covered with a nonreactive tape. The control was similarly applied to the opposite cranial and caudal sites. The trunk of each animal was wrapped with an elastic binder to maintain the test patches in position. Animals were returned to their cages after treatment. After a minimum of 23 hours and a maximum of 24 hours exposure, the binders, tape, and patches were removed. The perimeter of each site was marked with nontoxic ink. The sites were gently wiped with a gauze sponge dampened with deionized water in an attempt to remove any remaining residue.

Laboratory Observations
1. Animals were observed daily for general health.
2. Body weights were recorded for each animal at pretreatment.
3. Dermal observations for erythema and edema were recorded at 1, 24, 48 and
72 hours after patch removal in accordance with the criteria in Appendix 1.
All times and temperatures reported herein are approximate and are within ranges established by the external standards described in the References section of this report and/or NAMSA standard operating procedures.

Evaluation and Statistical Analysis
The Primary Irritation Index of the test was calculated following test completion for each animal. The erythema and edema scores obtained at the 24, 48 and 72 hour intervals were added together and divided by the total number of observations. The 1 hour score was not included in the calculation. This calculation was conducted separately for the test and control article for each animal. The score for the control was subtracted from the score for the test article to obtain the Primary Irritation Score. The Primary Irritation Score for each animal was added together to determine the Combined Primary Irritation Score. The Combined Primary Irritation Score was divided by the number of animals to obtain the Primary Irritation Index. The Primary Irritation Index was characterized based on the definitions outlined in Appendix 1.

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.5

Max. score:

8

Reversibility:

not fully reversible within: 72 h

Remarks on result:

probability of mild irritation

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

8

Reversibility:

not fully reversible within: 72 h

Remarks on result:

probability of mild irritation

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.5

Max. score:

8

Reversibility:

not fully reversible within: 72 h

Remarks on result:

probability of mild irritation

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24/48/72 h

Score:

1.3

Max. score:

8

Remarks on result:

probability of mild irritation

Interpretation of results:

Category 3 (mild irritant) based on GHS criteria

Conclusions:

The primary Irritation index for 20% Monalazone Disodium was calculated to be 1.3 in the ISO 10993-10 method under the experimental conditions described in this report. The response of the test article was categorized as slight.

Executive summary:

The test article, (20% monalazone disodium), was evaluated for primary skin irritation in rabbits. This study was conducted in accordance with the guidelines of ISO 10993-10, Biological evaluation of medical devices - Part 10: Tests for irritation and skin sensitization. Two 0.5 mL portions of the test article and control (4 ply, 25 mm x 25 mm gauze patch) were topically applied to the skin of each of three rabbits and left in place for a minimum of 23 hours and a maximum of 24 hours. The sites were graded for erythema and edema at 1, 24, 48 and 72 hours after removal of the single sample application. There was no to well-defined erythema and no to very slight edema observed on the skin of the animals

treated with the test article. The Primary Irritation Index for the test article was calculated to be 1.3. The response of the test article was categorized as slight.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

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:

Jan 18-Jan 25, 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The EpiOcular tissue construct is a nonkeratinized epithelium prepared from normal human keratinocytes (MatTek). It models the cornea epithelium with progressively stratified, but not cornified cells. The ability to expose the tissue topically is essential to model the same kind of progressive injury expected in vivo. It also allows both solid and liquid test materials to be applied directly to the tissue.

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

human

Details on test animals or tissues and environmental conditions:

The EpiOcular™ human cell construct for eye irritation testing (OCL-200-EIT) (Lot No.27020) was obtained from MatTek In Vitro Life Science Laboratories, SR.

Vehicle:

unchanged (no vehicle)

Controls:

yes

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

50 mg

Duration of treatment / exposure:

6 hours

Duration of post- treatment incubation (in vitro):

18 hours

Number of animals or in vitro replicates:

Two

Details on study design:

Treatment
After pre-incubation tissues were pre-wetted with 20 L of Dulbecco's Phosphate Buffered Saline (DPBS), then approximately 50 mg of the test item and 50 μL control items were applied topically onto the tissue surface for 6 hours. Two tissues were used per treatment, negative and positive controls. After exposure period, each tissue was rinsed gently with DPBS to remove any residual test item. After post-soak immersion at room temperature for
25 minutes, tissues than were transferred to fresh medium and incubated for 18 hours. Afterwards, the MTT assay was performed by transferring the tissues to 24-wells plate containing MTT medium (1 mg/mL) and incubated at 37±1°C in a humidified atmosphere of 5±1% CO2 in air for 3 hours.
After incubation, the blue formazan salt formed by cellular mitochondria was extracted with 2 mL/tissue of isopropanol for 2 hours at room temperature.
Each extraction solutions in a volume of 200 μL were transferred to a 96-well plate and the optical density of extracted formazan was determined using a spectrophotometer at 540 nm. Before testing, to identify the possible interference with MTT endpoint, test item was checked for its ability to reduce MTT directly.

Assessment of Direct Test Item Reduction by MTT
Test items may have the ability to directly reduce MTT and to form a blue/purple reaction product which could have an impact on the quantitative MTT measurement. Therefore, it is necessary to assess this ability for each test item prior to conducting any assays with viable tissues. For this purpose, a 1.0 mg/mL MTT solution (in DMEM) was prepared. Approximately 50 mg of the test item was added to 1 mL of the MTT solution in a 6-well plate and the
mixture was incubated in the dark at 37  1°C in a humidified atmosphere of 5  1% CO2 in air (standard culture conditions) for three hours. A negative control (50 μL of aqua pro injectione) was run concurrently. At the end of the exposure time the colour of treated MTT remained unchanged and it was concluded that the test item did not reduce MTT directly.

Assessment of Colored or Staining Materials
Colored test items or test items which become colored after application to the tissues may interfere with the quantitative photometric MTT measurement if the colorant binds to the tissue and is extracted together with MTT. Therefore, each test item has to be checked for its colorant properties.
For this purpose, test item in the amount approximately 50 mg was added to 2 mL of isopropanol, the same amount as used for MTT extraction and incubated in 6-well plates for 3 hours at room temperature. Two 200 μL aliquots of isopropanol solutions and of pure isopropanol were transferred to a 96-well plate and the absorbance was measured with a plate reader at the MTT measurement wavelength. After subtraction of the OD for isopropanol, the OD of the test item solution was < 0.08 and the test item has not to be considered as possibly interacting with the MTT measurement. The test item in the amount approximately 50 mg was added into 1 mL of purified water. The mixture was incubated in 6-well plate in the incubator at 37±1°C in a humidified atmosphere of 5±1% CO2 in air for 60 min. At the end of the exposure time the presence of the staining was evaluated. The colour of mixture was unchanged and the test item had not the potential to stain tissue.

Receipt and Preparation of Cultures
EpiOcular™ was delivered one day before the pre-incubation of tissues and was stored in original package at 2-8°C. At day 1, each culture was removed from the agarose gel using a sterile forceps, inspected and transferred to a pre-labeled 6-well plates containing 1 mL of assay medium per well. The EpiOcularTM cultures were pre-incubated at 37±1°C in a humidified atmosphere of 5±1% CO2 for 60 min. At the end of the first pre-incubation
period, the medium was replaced by 1 mL of fresh assay medium. Further, the tissues were pre-incubated in incubator overnight prior to dosing for release of transport stress related compounds and debris.

Assay Procedure
After the overnight incubation, the tissues were pre-wetted with 20 uL of DPBS. The tissues were incubated at standard culture conditions for 30 minutes. After pre-wetting, the negative and positive controls were tested by applying 50 uL topically on the tissues. The test item was applied topically onto the tissue surface at amount approximately 50 mg. Two tissues were used per treatment, negative and positive controls. The cultures were returned to the incubator for 6 hours. After treatment time, tissues were rinsed with DPBS (in three glass beakers) to remove any residual test material. After rinsing, the tissue was immediately transferred to and immersed in 5 mL of previously-warmed assay medium in 12-well plate for a 25-minute immersion incubation (Post-Soak) at room temperature. At the end of the Post-Soak immersion, each insert was removed from the medium and the insert was blotted on absorbent material and transferred to the appropriate well of the pre-labelled 6-well plate containing 1 mL of warm medium. The tissues were post-incubated for an additional 18 hours. Then, the cultures were transferred to 24-well plate containing 0.3 mL/well of MTT reagent (1 mg/mL) and incubated at 37±1°C in a humidified atmosphere of 5±1% CO2 in air for 3 hours. After incubation, the cultures were blotted on absorbent paper and transferred to a pre-labeled 6-well plate and extracted in 2 mL of isopropanol at room temperature for 2 hours. Volume of 2x 200 μL of each extraction solution were transferred to a 96-well plate and the absorbances (ODs) were recorded.

Handling of Results
Data include cytotoxicity and viability determination. The optical densities (ODs) were read in a 96-well plate spectrophotometer using a wavelength 540 nm without a reference filter.Data files of optical densities (ODs) generated by spectrophotometer (without blank substraction) were manually transported into the first spreadsheet of the EXCEL workbook (Import). The control of data transmission was performed by the responsible person. The
blank corrections, calculation of results and statistical parameters were done automatically in the second part of the workbook (Results). For each individual tissue treated with test item, PC and NC, the individual relative tissue viability was calculated. For each test item, PC and NC, the mean relative viability of the two individual tissues were calculated and used for classification according to the Prediction Model. The spreadsheet was shown a graph of the results (% of relative viability ± SD).

Irritation parameter:

other: % viability

Value:

2.9

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The assay is considered valid if the following criteria were met:
Negative Control (NC):
The negative control OD > 0.8 and < 2.5.

Positive Control (PC):
The mean relative viability of the positive control is at 6 hr exposure below 50% of control viability.

Standard Deviation (SD):
The difference of viability between the two relating tissues of a single chemical is < 20% in the same run (for positive and negative control tissues and tissues of single chemicals). This applies also to the killed controls (single chemicals and negative killed control) and the colorant controls which are calculated as percent values related to the viability of the relating negative control.

Table 1. Eye irritation potential of monalazone disodium after 6hr exposure in human model.

Substance	OD mean	SD of OD	Viability mean (%)	SD of viabilities	in vivo prediction
Negative control (H2O)	1,560	0,102	100,0	6,54	NI
Postive control	0,552	0,019	35,4	1,19	I
Monalazone disodium	0,045	0,007	2,9	0,42	I

Interpretation of results:

study cannot be used for classification

Conclusions:

In conclusion, Monalazone disodium was irritating to the eye in the EpiOcular™ test under the experimental conditions described in this report. The test item is identified as requiring classification and labelling according to UN GHS (Category 2 or Category 1).
This Test Guideline does not allow discrimination between eye irritation/reversible effects on the eye (Category 2) and serious eye damage/irreversible effects on the eye (Category 1), nor between eye irritants (optional Category 2A) and mild eye irritants (optional Category 2B), as defined by UN GHS. For these purposes, further testing with other in vitro test methods is required.

Executive summary:

Monalazone disodium was examined for eye irritation potential in EpiOcularTM Eye Irritation Test in compliance with OECD Guideline 492. MatTek EpiOcular™ tissue samples were treated in duplicate with the test article, negative control and postive control (exposure time of 6 hours). The magnitude of viability was quantified by MTT test. The absolute mean OD of negative control (OD570 = 1.560) was within the laboratory historical control data range. The positive control had a mean cell viability of 35.4% after 6 hours exposure. The positive control met the acceptance criterion: mean tissue viability less than 50%. The viability of culture treated by Monalazone disodium was 2.9%. Based on the results of the study (mean tissue viability ≤60%)

, the test item Monalazone disodium is considered to be irritant (I).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

EpiDerm

The MatTek EpiDerm™ model was used to assess the potential dermal irritation of the test article under three conditions (similair to OECD 439) by determining the viability of the tissues following exposure via MTT. The objective of this study was to assess the dermal irritation potential of the Sponsor’s submitted test article under three conditions. Pre-testing showed the test articles/conditions were not colored though one test article, (20% monalazone Disodium) did auto-reduce MTT. A small data correction was necessary as the auto-reduction was observed during the study. Tissues were exposed to test articles and controls for one hour, followed by a 42 hour post-exposure recovery period. The viability of each tissue was determined by MTT assay.

The assay passed all the quality controls. The negative control tissue OD was between 1.651 and 1.942, the positive control was within the confidence limits and was deemed an irritant, and the test article SDs were all ≤18. The MTT data show the positive control, 5% sodium dodecyl sulfate (SDS), reduced tissue viability to 5.0% of control. The mean viability of tissues after exposure to the test articles, (20% active) and Dilute (5% active) in water were <50% of control. Therefore, the test articles were determined to be irritants to human skin according to the OECD test guideline followed for this study.

ISO method

The test article, (20% Monalazone disodium), was evaluated for primary skin irritation in rabbits. This study was conducted in accordance with the guidelines of ISO 10993-10, Biological evaluation of medical devices - Part 10: Tests for irritation and skin sensitization. Two 0.5 mL portions of the test article and control (4 ply, 25 mm x 25 mm gauze patch) were topically applied to the skin of each of three rabbits and left in place for a minimum of 23 hours and a maximum of 24 hours. The sites were graded for erythema and edema at 1, 24, 48 and 72 hours after removal of the single sample application. There was no to well-defined erythema and no to very slight edema observed on the skin of the animals treated with the test article. The Primary Irritation Index for the test article was calculated to be 1.3. The response of the test article was categorized as slight.

Eye Irritation

EIT

Monalazone disodium was examined for eye irritation potential in EpiOcularTM Eye Irritation Test by Lazova, 2018 in compliance with OECD Guideline 492. MatTek EpiOcular™ tissue samples were treated in duplicate with the test article, negative control and postive control (exposure time of 6 hours). The magnitude of viability was quantified by MTT test. The absolute mean OD570 of negative control (1.560) was within the laboratory historical control data range. The positive control had a mean cell viability of 35.4% after 6 hours exposure. The positive control met the acceptance criterion as the mean tissue viability was less than 50%. The viability of culture treated by Monalazone disodium was 2.9%. Based on the results of the study (mean tissue viability ≤60%), the test item is identified as requiring classification and labelling according to UN GHS (Category 2 or Category 1).

This Test Guideline does not allow discrimination between eye irritation/reversible effects on the eye (Category 2) and serious eye damage/irreversible effects on the eye (Category 1), nor between eye irritants (optional Category 2A) and mild eye irritants (optional Category 2B), as defined by UN GHS. For these purposes, further testing with other in vitro test methods was required.

 

BCOP

The evaluation of the ocular irritancy of Monalazone Disodium was carried out by Palaskar, 2018 using the Bovine Corneal Opacity and Permeability assay. A 0.75 mL (20%w/v) of test item, positive control (Imidazole) and negative control (distilled water) was applied by means of closed chamber method. The control and treated corneaswere then subjected to the opacity and permeability measurements. 

In this study, the negative control (distilled water) was classified as UN GHS No Category (IVIS score 1.503) and the positive control (Imidazole) and the test item was Monalazone Disodium classified under Category 1 (IVIS score 127.108 & 71.783 respectively).

It can be concluded that the test item Monalazone Disodium is predicted as agent causing eye irritation/serious eye damage (Cat 1) by the BCOP test method.

Justification for classification or non-classification

The studies by Willoughby, J.A. (2017) (OECD 439) and Severhof, L.A. (2016) (ISO method) shows that Monalazone disodium needs to be classified as Skin irrit. 2 according to CLP Regulation 1272/2008.

The results of the key studies (OECD 492; Lazova, J. 2018) and (OECD 437; Palaskar, G.S. 2018) indicates that Monalazone disodium needs to be classified as Eye Dam Cat 1 according to CLP Regulation 1272/2008.