Reaction mass of 2,2',2''-nitrilotriethanol and disodium succinate and sodium acetate and sodium bromide

Endpoint summary

Description of key information

The test substance was found to be not irritating to skin, and not irritating to the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation / corrosion

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-10-21 to 2012-11-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study according to OECD TG 431 (2004) and under GLP

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: EpiDerm™ tissues model

Strain:

other: EpiDerm™

Details on test animals or test system and environmental conditions:

EXPERIMENTAL DETAILS- Source: Epi-200 kits and MTT-100 assays were purchased from MatTek Corporation (Ashland, MA 01721, USA).- Age at study initiation: EpiDerm™ tissues were shipped at 4 °C on medium-supplemented agarose gels in a 24- well plate on October 23, 2012. After receipt of the EpiDermTM tissues before starting the assay, the tissues were transferred to 6-well plates with assay medium, which was immediately replaced before the test is started. Test start was October 24, 2012. At least 1 hour before dosing, EpiDerm™ tissues were removed from the refrigerator. Under sterile conditions using sterile forceps, the inserts were transferred into 6-well plates containing the pre-warmed assay medium. A 2 -well plate was prepared as holding plate containing 300 μL assay medium. The holding plate was pre warmed in an incubator (37 ± 1.5 °C, 5 ± 0.5 % CO2) until use.- Acclimation period: 1 dayENVIRONMENTAL CONDITIONS- Temperature (°C): 37 ± 1.5 °C- Humidity (%): not applicable for this in vitro test- Air changes (per hr): not applicable for this in vitro test- Photoperiod (hrs dark / hrs light): not applicable for this in vitro test- CO2 concentration (%): 5 ± 0.5 % CO2IN-LIFE DATES: 2012-10-24 to 2012-10-25

Type of coverage:

other: EpiDerm™ tissues model

Preparation of test site:

other: EpiDerm™ tissues model

Vehicle:

unchanged (no vehicle)

Controls:

other: EpiDerm™ tissues treated with water

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied (volume or weight with unit): Each approximately 25 – 28 mg of the test item was applied to each tissue, wetted with 50 μL of deionised water, and spread evenly to the surface- Concentration: 25-28 mg/0.050 mL = 500-560 g/LVEHICLE- Amount(s) applied (volume or weight with unit): no vehicle

Duration of treatment / exposure:

3 minutes and 1 hour

Observation period:

after exposure period 3 hour inmcubation period in MTT-solution (MTT=(3-4,5-dimethyl thiazol 2-yl) 2,5-diphenyl-tetrazolium bromide) rinsing with DBPS (Dulbecco's Phosphate Buffered Saline) immersing within isopropanole transfering to blue formazan solution

Number of animals:

Duplicates of EpiDerm™ tissues were exposed to the test item, positive or negative control for each of two different exposure periods: 3 minutes and 1 hour.

Details on study design:

TEST SITE- Area of exposure: EpiDerm™ tissues- % coverage: 100 %REMOVAL OF TEST SUBSTANCE- Washing (if done): yes- Time after start of exposure: 3 minutes or 1 hourSCORING SYSTEM: determination of optical density

Irritation parameter:

other: corrosion of EpiDerm™ tissues models

Basis:

other: relative absorbance value

Time point:

other: exposure of 3 minutes

Score:

> 50

Reversibility:

other: viability

Irritation parameter:

other: corrosion of EpiDerm™ tissues models

Basis:

other: relative absorbance value

Time point:

other: exposure of 1 hour

Score:

> 15

Reversibility:

other: viability

Interpretation of results:

not classified

Remarks:

Migrated informationnon corrosiveCriteria used for interpretation of results: other: EU CLP and UN GHS

Conclusions:

In this valid, reliable and conclusive study according to OECD TG 431 and under the reported experimental conditions, the test item Reaction mass of CXN1-55 was non corrosive to skin according to EU CLP and UN GHS.

Executive summary:

This valid, reliable and conclusive in vitro study was performed to assess the corrosive potential of Reaction mass of CXN1-55 by means of the Human Skin Model Test with EpiDerm™ tissues models according to OECD TG 431.

Independent duplicate tissues of EpiDermTM were exposed to either the test item Reaction mass of CXN1-55 (each approximately 25 – 28 mg, each wetted with 50 μL of deionised water, and spread evenly to the surface), the negative control (deionised water) or the positive control (8.0 N KOH) for 3 minutes and 1 hour, respectively.

After exposure to the negative control the absorbance values met the required acceptability criterion of mean OD570 ≥ 0.8 for both treatment intervals thereby confirming the acceptable quality of the tissues.

Exposure to the positive control induced a decrease in the relative absorbance compared to the negative control, both for the 3 minutes exposure period (22.9 %) and for the 1 hour exposure period (9.4 %) thus confirming the validity of the test system and the specific batch of tissue models.

After exposure to the test item Reaction mass of CXN1-55 the relative absorbance value did not decrease at all after 3 minutes exposure (104.9 %). After 1 hour exposure the relative absorbance value was reduced to 87.3 %. Both values did not exceed the threshold for corrosivity which is defined to be 50 % after the 3 minutes exposure and 15 % after the 1 hour exposure. Therefore, the test item was not considered to be corrosive.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-11-07 to 2012-11-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study according to OECD TG 439 (2010) and under GLP

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: EpiDerm™ tissues model

Strain:

other: EpiDerm™

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Epi-200 kits and MTT-100 assays diluent were purchased from MatTek Corporation (82105 Bratislava, Slovakia)- Age at study initiation: EpiDerm™ tissues were shipped at 4 °C on medium-supplemented agarose gels in a 24- well plate and arrived on November 06, 2012. On day of receipt EpiDerm™ tissues were kept in the refrigerator at 4 °C until starting the pre-incubation phase.Prewarming: One day prior to the performance, the plastic bag containing the 24-well plate with epidermal tissues was opened under sterile conditions. The inserts with the EpiDerm™ tissues were placed in the upper wells, and were pre-incubated for 60 minutes in the incubator (37 ± 1.5 °C, 5 ± 1% CO2, 95 ± 5% RH). Following, the inserts were transferred from upper wells into the lower wells of the 6-well plates, and, the pre-incubation was continued for further about 24 hours (37 ± 1.5 °C, 5 ± 1% CO2, 95 ± 5% RH). After pre-incubation of EpiDerm™ tissues was completed, the negative and positive control and the test item were added into the insert atop the corresponding EpiDerm™ triplicate tissues.- Acclimation period: 1 dayENVIRONMENTAL CONDITIONS- Temperature (°C): 37 ± 1.5 °C- Humidity (%): not applicable for this in vitro test- Air changes (per hr): not applicable for this in vitro test- Photoperiod (hrs dark / hrs light): not applicable for this in vitro test- CO2 concentration (%): 5 ± 0.5 % CO2IN-LIFE DATES: From: To: 2012-11-07 to 2012-11-12

Type of coverage:

other: EpiDerm™ tissues model, the test item was spread to match size of tissue

Preparation of test site:

other: EpiDerm™ tissues model

Vehicle:

unchanged (no vehicle)

Controls:

other: EpiDerm™ tissues treated with DPBS (Dulbecco's Phoasphate Buffered Saline)

Amount / concentration applied:

Each approximately 25 mg (using a weighing spoon) of the test item were applied to the tissues, wetted with 25 μL DPBS, and spread to match size of tissue. For the positive and negative controls 30 μL were dosed per tissue.

Duration of treatment / exposure:

60 minutes

Observation period:

After the end of the treatment interval the inserts were removed immediately from the 6-well plate and tissues were gently rinsed with DPBS at least 15 times in order to remove any residual test material. After the rinsing the inserts were submerged in DPBS at least three times. Afterwards the inserts were once again rinsed with DPBS from the inside and the outside. Excess DPBS was removed by gently shaking the inserts and blotting the bottom with sterile blotting paper.The tissues were then transferred into new 6-well plates with 0.9 mL of fresh assay medium in the upper row. The surface of the tissues was carefully dried using sterile cotton tipped swap. Tissues were incubated for nearly 18 hours at 37 ± 1.5 °C, 5 ± 0.5 % CO2.New 6-well plates (or lower row of the same plates) were filled with 0.9 mL of fresh assay medium, and the inserts were transferred to the new plates. The wells were incubated for another 3 hours post-incubation at 37 ± 1.5 °C, 5 ± 0.5 % CO2.After the treatment procedure was completed for all tissues of each time point cell culture inserts were transferred from the holding plates to the MTT-plates. After a 3 hour incubation period (37 ± 1 °C, 5 ± 0.5 % CO2) MTT solution was aspirated from the wells and wells were rinsed three times with DPBS. Inserts were transferred into new 24 well plates. The inserts were immersed into extractant solution by gently pipetting 2 mL extractant solution (isopropanol) into each insert. The level rose above the upper edge of the insert, thus the tissue was completely covered from both sides. The 24 well plate was sealed to inhibit isopropanol evaporation. The formazan salt was extracted for about 69 hours at room temperature.

Number of animals:

Triplicates of EpiDerm™ tissues were exposed to the test item, positive or negative control for the one exposure period: 60 minutes.

Details on study design:

TEST SITE- Area of exposure: EpiDerm™ tissues- % coverage: 100 %REMOVAL OF TEST SUBSTANCE- Washing (if done): yes- Time after start of exposure: 60 minutesSCORING SYSTEM: determination of optical densitynegative control: DPBS (Dulbecco's Phosphate Buffered Saline)positive control: 5 % Sodium lauryl sulfate

Irritation / corrosion parameter:

other: other: irritation of EpiDerm™ tissues models

Value:

> 50

Remarks on result:

other:

Remarks:

Basis: other: relative absorbance value. Time point: 60 minutes. Max. score: 84.0. Reversibility: other: viability. (migrated information)

Table 1 Results after treatment with Reaction mass of CXN1-55 and the controls

Dose group	Exposure Interval	Absorbance 570 nm Tissue 1*	Absorbance 570 nm Tissue 2*	Absorbance 570 nm Tissue 3*	Mean Absorbance of 3 Tissues	Relative Absorbance [%] Tissue 1, 2 + 3**	Coefficient of Variation (=Standard deviation/Mean * 100)	Rel. Absorbance [% of Negative Control]**
Negative Control	60 min.	2.117	2.423	2.175	2.238	94.6	7.3	100.0
						108.3
						97.2
Positive Control	60 min.	0.091	0.088	0.077	0.085	4.1	8.6	3.8
						3.9
						3.4
Test item	60 min.	2.019	1.752	1.857	1.876	90.2	7.2	83.8
						78.3
						83.0

* Mean of three replicate wells after blank correction

** Relative absorbance [rounded values] = 100 x absorbance of test item / absorbance of negative control

Interpretation of results:

not irritating

Remarks:

Migrated information

Conclusions:

In this valid, relaible and conclusive study according to OECD TG 439 and under the reported experimental conditions, the test item Reaction mass of CXN1-55 was not irritant to skin.

Executive summary:

This valid, reliable and conclusive in vitro study was performed to assess the irritation potential of Reaction mass of CXN1-55 by means of the Human Skin Model Test with EpiDerm™ tissues models according to OECD TG 439 and under GLP.

Three tissues of the human skin model EpiDerm™ were treated with the test item, the negative or the positive control for 60 minutes. About 25 mg of the test item were applied to each tissue, wetted with 25 μL DPBS, and spread to match the tissue size. 30 μL of either the negative control (DPBS) or the positive control (5 % Sodium lauryl sulfate) were applied to each tissue. After treatment with the negative control the absorbance values were well above the required acceptability criterion of mean OD ≥ 1.0 and ≤ 2.5 for the 60 minutes treatment interval thus showing the quality of the tissues. Treatment with the positive control induced a decrease in the relative absorbance to 3.8 % as compared to the negative control for the 60 minutes treatment interval thus ensuring the validity of the test system. After treatment with the test item Reaction mass of CXN1-55 the mean relative absorbance value decreased to 83.8 % compared to the relative absorbance value of the negative control. This value is above the threshold for irritancy of ≤ 50 %. Therefore, the test item is not considered to possess an irritant potential. In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item Reaction mass of CXN1-55 is not irritant to skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-02-07 to 2013-03-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study according to OECD TG 437 (2009) and EU Method B.47 and under GLP

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: bovine eyes from at least 9 month old donor cattle

Strain:

other: strain of cattle not necessary to be specified in this study

Details on test animals or tissues and environmental conditions:

TEST ANIMALS- Source: Odenwaldschlachthof Brensbach, 64395 Brensbach, Germany- Age at study initiation: Freshly isolated bovine cornea (at least 9 month old donor cattle)- Acclimation period: Freshly isolated bovine eyes of at least 9 month old donor cattle were collected from the abattoir. Excess tissue was removed from the excised eyes. The isolated eyes were transported to the laboratory in Hank’s BSS supplemented with streptomycin / penicillin at ambient temperature. The corneae were isolated on the same day after delivery of the eyes. The corneae were inserted in pre-cooled preservation medium (Medium 199 supplemented with L-glutamine, Nabicarbonate and Taurine). Shortly before use, Dextran was added to the medium and stored in the refrigerator at 2 - 8 °C until use on the following day.Each isolated cornea was mounted in a specially designed cornea holder according to the description given in OEDC guideline 437, annex III, that consists of anterior and posterior compartments, which interface with the epithelial and endothelial sides of the cornea, respectively. For equilibration, the corneae in the holder were incubated in a vertical position for about one hour at 32 ± 1 °C in a water-bath.ENVIRONMENTAL CONDITIONS- Temperature (°C): 32 ± 1 °C- Humidity (%): not applicable for this study since the corneae are submersed in a solution within the cornea holder.- Air changes (per hr): not applicable for this study since the corneae are submersed in a solution within the cornea holder.- Photoperiod (hrs dark / hrs light): not applicable for this study.IN-LIFE DATES: 2013-02-07 to 2013-02-07

Vehicle:

physiological saline

Remarks:

0.9 % (w/v) NaCl (saline)

Controls:

yes

Amount / concentration applied:

731.17 mg of the test item were solved in 3.7 mL of saline to reach a weight/volume ratio of 20 %.Positive Control: 10 % (w/v) Benzalkonium chloride (purity not indicated by the producer) in 0.9 % (w/v) NaCl (saline) served as positive control.Negative Control: Saline served as negative control.The anterior compartment received the test item solution or negative or positive control at a volume of 0.75 mL each on the surface of the corneae.

Duration of treatment / exposure:

240 minutes

Observation period (in vivo):

At the end of the incubation period, the basal opacity was determined (t0).Each corneae with a value of the basal opacity > 7 was discarded. Sets of three corneae were used for treatment with the test item and the negative and positive controls.After the exposure time, the test item or control items, respectively, were rinsed off from the application side with saline. Afterwards, fresh incubation medium (MEM, supplemented with sodium bicarbonate and L-glutamine. Immediately before starting the test, MEM was supplemented with 1 % fetal calf serum (FCS)) was added into the anterior compartment and opacity was measured (t240). The opacitometer determines changes in the light transmission passing through the corneae, and displays a numerical opacity value. This value was recorded in a table. The opacitometer (OP_KiT opacitometer (Electro Design, 63-Riom, France)) was calibrated and the opacity of each of the corneae was determined by reading each holder placed in the photoreceptor compartment for treated cornea.Following to the opacity readings, the permeability was measured as an indication of the integrity of the epithelial cell sheets. After the final opacity measurement was performed, the incubation medium was removed from the anterior compartment and replaced by 1 mL of a 0.5 % (w/v) sodium fluorescein solution in HBSS (Hank’s Buffered Salt Solution). Corneae were incubated again in a horizontal position for 90 ± 10 minutes in a water-bath at 32 ± 1 °C. Incubation medium from the posterior compartment was removed, well mixed and transferred into a 96 well plate and the optical density at 490 nm (OD490) was determined with a spectrophotometer (Versamax® Molecular Devices). The absorbance values will be determined using the software SoftMax Pro Enterprise (version 4.7.1).

Number of animals or in vitro replicates:

3 corneae each for the negative control, the positive control and the test item treatment

Details on study design:

REMOVAL OF TEST SUBSTANCE- Washing (if done): after the exposure period of 240 minutes- Time after start of exposure: 240 minutesSCORING SYSTEM:Opacity: The change of opacity value of each treated cornea or positive and negative control corneae is calculated by subtracting the initial basal opacity from the post treatment opacity reading (t240 – t0), for each individual cornea.The average change in opacity of the negative control corneae is calculated and this value is subtracted from the change in opacity of each treated cornea or positive control to obtain a corrected opacity.Permeability: The corrected Optical density OD490 value of each cornea treated with positive control and test item is calculated by subtracting the average negative control cornea value from the original permeability value for each cornea.IVIS (In Vitro Irritation Score): The following formula is used to determine the IVIS of the negative control: IVIS = opacity value + (15 x OD490 value).The following formula is used to determine the IVIS of the positive control and the test item: IVIS = (opacity value – opacity value mean negative control) + (15 x corrected OD490 value).The mean IVIS value of each treated group was calculated from the IVIS values.TOOL USED TO ASSESS SCORE:Opacity: measurement of the light transmission passing through the corneae (OP_KiT opacitometer (Electro Design, 63-Riom, France)).Permeability: measurement of sodium fluorescein in the medium from the posterior part of the eye holder after incubation of the anterior part with 0.5 % (w/v) sodium fluorescein solution in HBSS for 90 ± 10 minutes in a water-bath at 32 ± 1 °C (spectrophotometer (Versamax® Molecular Devices), determination of absorbance values using the software SoftMax Pro Enterprise (version 4.7.1)).

Irritation parameter:

cornea opacity score

Basis:

other: opacity value IVIS

Time point:

other: 240 minutes

Score:

4.08

Reversibility:

not specified

With the negative control (saline) neither an increase of opacity nor permeability of the corneae could be observed (mean IVIS = 2.71).

The positive control (10% (w/v) Benzalkonium chloride in saline) induced distinct opacity on the corneae (mean IVIS =229.67)corresponding to a classification as corrosive /severe irritant to the eye (CLP/EPA/GHS (Cat 1)).

Relative to the negative control, the test item Reaction mass of CXN1-55 caused only a slight increase of the corneal opacity. Permeability effects did not occur. The calculated mean IVIS was 4.08 (threshold for corrosivity / severe irritancy: IVIS ≥ 55.1) . According to OECD 437 the test item is classified as not corrosive / not severe irritant to the eye.

Interpretation of results:

other: not corrosive

Remarks:

Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, according to the current study and under the experimental conditions reported, the test item Reaction mass of CXN1-55 is not corrosive / not severely irritating to the eye (CLP/EPA/GHS (Cat 1)).

Executive summary:

A valid, reliable and conclusive in vitro study was performed to assess the corneal irritation and damage potential of Reaction mass of CXN1-55 by means of the BCOP assay using fresh bovine corneae. Testing was performed compliant to OECD testing guideline no. 437 and EU Method B.47 and under GLP.

After a first opacity measurement of the fresh bovine corneae (t0), the 20 % (w/v) solution in saline (0.9 % (w/v) NaCl in deionised water) of the test item Reaction mass of CXN1-55, the positive, and the negative controls were applied to corneae and incubated for 240 minutes at 32 ± 1 °C. The posterior chamber contained incubation medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae and opacity was measured again (t240).

After the opacity measurements permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C.

With the negative control (physiological saline) neither an increase of opacity nor permeability of the corneae could be observed.

The positive control (10 % (w/v) Benzalkonium chloride in saline) caused distinct opacity of the corneae corresponding to a classification as corrosive / severe irritant to the eye (CLP/EPA/GHS (Cat 1)).

Relative to the negative control, the test item Reaction mass of CXN1-55 caused a very slight increase of the corneal opacity. Permeability could not be observed. The calculated mean IVIS was 4.08. According to OECD 437 the test item is classified as not corrosive / not severe irritant to the eye.

In conclusion, according to the current study and under the experimental conditions reported, the test item Reaction mass of CXN1-55 is not corrosive / not severe irritant to the eye (CLP/EPA/GHS (Cat 1)).

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-03-28 to 2013-05-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study under GPL according to OECD TG 405 (2012)

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS- Source: Harlan Laboratories UK Ltd., Leicestershire, UK- Age at study initiation: twelve to twenty weeks old- Weight at study initiation: 2.96 to 3.31 kg- Housing: individually housed in suspended cages. Free access to mains drinking water and food.- Diet: ad libitum, food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK)- Water: ad libitum- Acclimation period: at least five daysENVIRONMENTAL CONDITIONS- Temperature (°C): 17 to 23 °C - Humidity (%): 30 to 70 % - Air changes (per hr): at least fifteen changes per hour- Photoperiod (hrs dark / hrs light): twelve hours continuous light (06:00 to 18:00) and twelve hours darknessIN-LIFE DATES: From: To: 2013-03-18 to 2013-04-29

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL- Amount(s) applied: 100 mgVEHICLE: no

Duration of treatment / exposure:

14 days

Observation period (in vivo):

14 days: 1, 24, 48, 72 hours, 7 days, 14 days after application

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE- Washing (if done): noSCORING SYSTEM:The numerical values corresponding to each animal, tissue and observation time were recorded. The data relating to the conjunctivae were designated by the letters A (redness), B (chemosis) and C (discharge), those relating to the iris designated by the letter D and those relating to the cornea by the letters E (degree of opacity) and F (area of cornea involved). For each tissue the score was calculated as follows:Score for conjunctivae=(A + B + C) x 2Score for iris=D x 5Score for cornea=(E x F) x 5Using the numerical data obtained a modified version of the system described by Kay J.H. and Calandra J.C. (1962) was used to classify the ocular irritancy potential of the test item. This was achieved by adding together the scores for the cornea, iris and conjunctivae for each time point for each rabbit. The group means of the total scores for each observation were calculated. The highest of these group means (the maximum group mean score) together with the persistence of the reactions enabled classification of the eye irritancy potential of the test item. If evidence of irreversible ocular damage is noted, the test item will be classified as corrosive to the eye.TOOL USED TO ASSESS SCORE: not explicitly cited

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 1 hour

Score:

19

Max. score:

19

Reversibility:

fully reversible within: 14 days

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 24 hours

Score:

10

Max. score:

12

Reversibility:

fully reversible within: 14 days

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 48 hours

Score:

8

Max. score:

8

Reversibility:

fully reversible within: 14 days

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 72 hours

Score:

5.3

Max. score:

6

Reversibility:

fully reversible within: 14 days

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 7 days

Score:

2.7

Max. score:

4

Reversibility:

fully reversible within: 14 days

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 14 hours

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 14 days

Irritant / corrosive response data:

Ocular Reactions:No corneal effects were noted during the study.Iridial inflammation was noted in all treated eyes one hour after treatment.Moderate conjunctival irritation was noted in all treated eyes one hour after treatment and at the 24 and 48 Hour observations. Minimal conjunctival irritation was noted in all treated eyes at the 72 Hour observation and in two treated eyes at the 7 Day observation.One treated eye appeared normal at the 7 Day observation and two treated eyes appeared normal at the 14 Day observation.

Other effects:

All animals showed expected gain in body weight during the study.

Interpretation of results:

irritating

Remarks:

Migrated informationCriteria used for interpretation of results: OECD GHS

Conclusions:

In a valid, reliable and conclusive study according to OECD TG 405 (2012) and EU Method A.5 (2008), the test item Reaction mass of CXN1-55 was classified:moderate irritant to the rabbit eye according to a modified Kay & Calandra classification system,Category 2A (irritating to eyes) according to the Globally Harmonized System of Classification and Labeling of Chemicals,Irritant (Category 2) according to the Regulation (EC) No 1272/2008, relating to the Classification, Labeling and Packaging of Substances and Mixtures.

Executive summary:

An Eye Irritation test was performed to assess the irritancy potential of the test item “Reaction Mass of CXN1-55” following topical application to the eye of the New Zealand White rabbit in compliance with OECD No. 405 and EU Method A.5 (2008) and under GLP.

A single application of the test item to the non-irrigated eye of three rabbits produced iridial inflammation and moderate conjunctival irritation. One treated eye appeared normal at the 7 Day observation and two treated eyes appeared normal at the 14 Day observation.

The test item produced a maximum group mean score of 19.0 and was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.

The test item was classified as Category 2A (irritating to eyes) according to the Globally Harmonized System of Classification and Labeling of Chemicals.

The test item was also classified as Irritant (Category 2) according to the Regulation (EC) No 1272/2008, relating to the Classification, Labeling and Packaging of Substances and Mixtures.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

This valid, reliable and conclusive in vitro study was performed to assess the irritation potential of Reaction mass of CXN1-55 by means of the Human Skin Model Test with EpiDerm™ tissues models according to OECD TG 439 and under GLP. The test item Reaction mass of CXN1-55 is not irritating to skin.

Skin corrosion:

This valid, reliable and conclusive in vitro study was performed to assess the corrosive potential of Reaction mass of CXN1-55 by means of the Human Skin Model Test with EpiDerm™ tissues models according to OECD TG 431 and under GLP. The test item was not considered to be corrosive.

Eye irritation/corrosion:

An Eye irritation test was performed to assess the irritancy potential of the test item “Reaction Mass of CXN1-55” following topical application to the eye of the New Zealand White rabbit. Testing was performed in compliance with OECD No. 405 and under GLP. The test item is considered to be irritating to the eye according to Regulation 1272/2008 (EU CLP)

Justification for classification or non-classification

Based on the current data, CNX1-55 is not classified for local skin effects according to Regulation (EC) No. 1272/2008 on Classification, Labelling and Packaging of Substances and Mixtures.

CNX1 -55 is classified category 2A (irritating to eyes) according to the Globally Harmonized System of Classification and Labeling of Chemicals, Irritant (Category 2) according to the Regulation (EC) No 1272/2008, relating to the Classification, Labeling and Packaging of Substances and Mixtures.