Potassium; 2-[bis[2-[bis(carboxylatomethyl)amino]ethyl]amino]acetate; iron(3+)

Endpoint summary

Description of key information

An in vitro and an in vivo OECD skin and eye irritation tests were available.  

Test results showed that both DTPA-FeHNa and EDTA-FeNa are not irritating to skin and eyes. Based on read across with comparable metal-chelates like EDTA-FeNa no further testing was performed (see also section 13 for read across) and those results are also valid for DTPA-FeK.

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:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

August-September 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well performed and reported GLP study

Justification for type of information:

A substantial body of evidence exists that the toxicity profiles of chelates depends mainly on metal ion, its affinity to this metal, and their ability to supply or to sequester it from the body/environment. Both EDTA and DTPA are chelates with similar structure and physico-chemical properties tham their toxicity are also similar. Please see RA statement in section 13.

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: in vitro

Details on test animals or test system and environmental conditions:

Test system: EPISKIN Small ModelTM (EPISKIN-SMTM, 0.38 cm2, Batch no.: 12-EKIN-031).
This model is a three-dimensional human epidermis model, which consists of adult human-derived epidermal keratinocytes which have been seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. The keratinocytes were cultured for 13 days, which results in a highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.

Rationale: In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimise the need of in vivo testing. One of the validated in vitro skin irritation tests is the EPISKIN test, which is recommended in international guidelines (e.g. OECD and EC).

Source: SkinEthic Laboratories, Lyon, France.

Type of coverage:

other: not applicable

Preparation of test site:

other: not applicable

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

The solid test substance (11.0 to 11.7 mg) was applied directly on top of the skin tissue. DTPA-FeHNa was spread to match the size of the tissue.

Duration of treatment / exposure:

See below

Observation period:

Not applicable

Number of animals:

Not applicable

Details on study design:

See below

Irritation / corrosion parameter:

% tissue viability

Value:

95

Negative controls validity:

valid

Positive controls validity:

valid

DTPA-FeHNa was checked for possible direct MTT reduction by adding the test substance to MTT medium. Because no colour change was observed it was concluded that DTPA-FeHNa did not interact with MTT.

The mean absorption at 570 nm measured after treatment with DTPA-FeHNa and controls are presented in Table 1.

Table 2 shows the mean tissue viability obtained after 15 minutes treatment with DTPA-FeHNa compared to the negative control tissues. Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with DTPA-FeHNa compared to the negative control tissues was 95%. Since the mean relative tissue viability for DTPA-FeHNa was above 50% DTPA-FeHNa is considered to be non-irritant.

The positive control had a mean cell viability after 15 minutes exposure of 4%. The absolute mean OD570 of the negative control tissues was within the laboratory historical control data range (See APPENDIX 3). The standard deviation value of the percentage viability of three tissues treated identically was less than 8%, indicating that the test system functioned properly.

Table1            Mean absorption in thein vitroskin irritation test with DTPA-FeHNa

	A (OD570)	B (OD570)	C (OD570)	Mean (OD570)		SD
Negative control	0.883	1.018	0.969	0.957	±	0.069
DTPA-FeHNa	0.982	0.849	0.907	0.913	±	0.067
Positive control	0.029	0.035	0.044	0.036	±	0.007

OD = optical density

SD = Standard deviation

Triplicate exposures are indicated by A, B and C.

In this table the values are corrected for background absorption (0.042). Isopropanol was used to measure the background absorption.

 

Table2            Mean tissue viability in thein vitroskin irritation test with DTPA-FeHNa

	Mean tissue viability (percentage of control)
Negative control	100
DTPA-FeHNa	95
Positive control	4

 

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test substance is not irritating in an in vitro test (OECD 439).

Executive summary:

This report describes the ability of DTPA-FeHNa to induce skin irritation on a human three dimensional epidermal model (EPISKIN Small Model (EPISKIN-SM^TM)). The possible skin irritation potential of DTPA-FeHNa was tested through topical application for 15 minutes.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch CFC-10333 (429H0352) of DTPA-FeHNa was a yellow-green powder with a purity of ~97%. Skin tissue was moistened with 5 µl of Milli-Q water and 11.0 to 11.7 mg of DTPA-FeHNa was applied directly on top of the skin tissue for 15 minutes. After a 42 hour post-incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

Skin irritation is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 15 minutes treatment with DTPA-FeHNa compared to the negative control tissues was 95%. Since the mean relative tissue viability for DTPA-FeHNa was above 50% after 15 minutes treatment DTPA-FeHNa is considered to be non-irritant.

The positive control had a mean cell viability of 4% after 15 minutes exposure. The absolute mean OD₅₇₀(optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 8%, indicating that the test system functioned properly.

Finally, it is concluded that this test is valid and that DTPA-FeHNa is non-irritant in thein vitroskin irritation test under the experimental conditions described in this report.

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

July 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well conducted study according to GLP

Justification for type of information:

A substantial body of evidence exists that the toxicity profiles of chelates depends mainly on metal
ion, its affinity to this metal, and their ability to supply or to sequester it from the body/environment.
Both EDTA and DTPA are chelates with similar structure and physico-chemical properties tham their t
oxicity are also similar. Please see RA statement in section 13.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

Himalayan

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: LPT, 24601 Löhndorf, germany
- Age at study initiation: 3-5 months
- Weight at study initiation: 1.8 - 2.3 kg
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 20 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20+/- 3°C
- Humidity (%): 30-70%
- Air changes (per hr): 60 per hour
- Photoperiod (hrs dark / hrs light):12/12


IN-LIFE DATES: From: 2. To: 6.July 2007

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 mg/animal

Duration of treatment / exposure:

4 h

Observation period:

72 h

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 6 cm²
- % coverage:
- Type of wrap if used: gauze patch


REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1 hour

Score:

1

Reversibility:

fully reversible

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Irritant / corrosive response data:

Very slight erythema (grade 1) was observed in one animal at 1 h after removal of the patch. There were no further skin reactions and no systemic adverse effects.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

The test iterm is not irritating to the skin.

Executive summary:

In a skin irritation study according to OECD guideline 404 a dose of 500 mg FeNaEDTA was applied to the shaved skin of three male rabbits for 4 hours. Very slight erythema was noted for one animal at 1 hour after patch removal. No other skin effects or systemic effects were noted during the observation period of 72h.

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:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

August 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well performed and reported GLP study

Justification for type of information:

The underlying hypothesis for the read-across is that chelates have the same mode of action based on their ability to chelate, remove or add metal cations to body causing perturbation of body’s micronutrients balance.
The source substance is a chelating agent in a target substance. The only difference between the target and the main source substance is presence of iron (Fe) and potassium (K) cations instead H+ cations. As iron and potassium are an essential macro- and microelements required by all forms of life, is considered not to influence the toxicological activity.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: in vitro

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

Since no workable suspension of DTPA-FeHNa in physiological saline could be obtained, the test substance was used as delivered by the sponsor and added pure on top of the corneas.

Duration of treatment / exposure:

See below

Observation period (in vivo):

See below

Number of animals or in vitro replicates:

Not applicable

Details on study design:

Negative control: a negative control, physiological saline (Merck, Darmstadt, Germany) was included to detect non-specific changes in the test system and to provide a baseline for the assay endpoints.

Positive control: 20% (w/v) Imidazole (Merck Schuchardt DHG, Germany) [CAS Number 288-32-4] solution prepared in physiological saline.

Test System: bovine eyes were used as soon as possible after slaughter on the same day.
Rationale: in the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimise the need of in vivo testing (1-6). As a consequence a validated and accepted in vitro test for eye irritation should be performed before in vivo tests are conducted. One of the proposed validated in vitro eye irritation tests is the Bovine Corneal Opacity and Permeability (BCOP) test.
Source: bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands), where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter.
Transport: eyes were collected and transported in physiological saline in a suitable container.

All eyes were carefully examined for defects by holding the eyes submersed in physiological saline. Those exhibiting unacceptable defects, such as opacity, scratches, pigmentation and neovascularization were discarded.

The isolated corneas were stored at 32 +/- 1 degrees C in a petri dish with cMEM (Eagle’s Minimum Essential Medium (Invitrogen Corporation, Breda, The Netherlands) containing 1% (v/v) L-glutamine (Invitrogen Corporation) and 1% (v/v) Foetal Bovine Serum (Invitrogen Corporation)). The isolated corneas were mounted in a corneal holder (one cornea per holder) of MC2 (Clermont, 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 degrees C. The corneas were incubated for the minimum of 1 hour at 32 +/- 1 degrees C.

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, 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.

The medium from the anterior compartment was removed and 750 uL of the negative control, 20% (w/v) Imidazole solution (positive control) or 20% (w/w) test substance solution were introduced onto the epithelium of the cornea. The holder was slightly rotated, with the corneas maintained in a horizontal position, to ensure uniform distribution of the solutions over the entire cornea. Corneas were incubated in a horizontal position for 240 +/- 10 minutes at 32 +/- 1 degrees C. After the incubation the solutions were removed and the epithelium was washed at least three times with MEM with phenol red (Eagle’s Minimum Essential Medium, Invitrogen Corporation). Possible pH effects of the test substance on the corneas were recorded. The anterior and the posterior compartment were refilled with fresh cMEM and an opacity determination was performed without any further incubation. After the completion of the incubation period each cornea were inspected visually for dissimilar opacity patterns and the opacity determination was performed.

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.

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. 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 degrees C.

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 uL 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.

Irritation parameter:

in vitro irritation score

Value:

>= 23 - <= 24

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Value:

>= 21 - <= 23

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

No

DTPA-FeHNa was tested as it is. Table 1 summarizes the opacity, permeability and in vitro irritancy scores of  DTPA-FeHNa and the controls. The individual in vitro irritancy score for the negative controls was -1.2. The individual positive control in vitro irritancy scores ranged from 105 to 118. The corneas treated with the positive control were turbid after the 240 minutes of treatment.

The corneas treated with DTPA-FeHNa showed opacity values ranging from 21 to 23 and permeability values ranging from 0.080 to 0.187. The corneas were clear after the 240 minutes of treatment with DTPA-FeHNa. No pH effect of the test substance was observed on the rinsing medium Hence, the in vitro irritancy scores ranged from 23 to 24 after 240 minutes of treatment with DTPA-FeHNa.

Table1            Summary of opacity, permeability andin vitroscores

Treatment	Mean Opacity^1	Mean Permeability^1	Mean In vitro Irritation Score^1, 2
Negative control	-1	0.000	-1.0
Positive control (Benzalkonium Chloride)	88	1.562	111
DTPA-FeHNa	22	0.118	24

 

1     Calculated using the negative control mean opacity and mean permeability values.

2     In vitroirritancy score (IVIS) = mean opacity value + (15 x mean OD₄₉₀value).

Table 2            Historical control data for the BCOP studies

 

	Negative control			Positive control
	Opacity	Permeability	In vitro Irritancy Score	In vitro Irritancy Score
Range	-5 – 1	-0.03 – 0.059	-4.7 – 1.1	70 – 180
Mean	-0.1	0	0	111
SD	0.73	0.01	0.71	20
n	90	90	90	90

SD = Standard deviation

n = Number of observations

The above mentioned historical control data range of the controls were obtained by collecting all data over the period of January 2009 to January 2012.

 

 

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test substances was not irritating in OECD test 437.

Executive summary:

This report describes the ocular irritation properties of DTPA-FeHNa on an isolated bovine cornea. The possible ocular irritancy of DTPA-FeHNa was tested through topical application for approximately 240 minutes.

The study procedures described in this report were based on the most recent OECD and EC guideline.

Batch CFC-10333 (429H0352) of DTPA-FeHNa was a yellow-green powderwith a purity of ~97%. 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 (301 to 306 mg).

The mean negative control responses for opacity and permeability were less than the upper limits of the laboratory historical rangeindicating that the negative control did not induce irritancy on the corneas, except the opacity of one of the corneas. However, since the opacity values of the other two corneas were within the historical control data range, the validity of the test was considered not affected.The meanin vitroirritancy score of the positive control (20% (w/v) Imidazole) was 111 and within the historical positive controldata range.It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

The meanin vitroirritancy score was 24 after 240 minutes of treatment with DTPA-FeHNa.

Since the meanin vitroirritancy score for DTPA-FeHNa was below 55.1 after 240 minutes treatment DTPA-FeHNa is considered to be not severe irritant or corrosive.

Finally, it is concluded that this test is valid and that DTPA-FeHNa is not severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

 

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

July 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well conducted study according to GLP

Justification for type of information:

A substantial body of evidence exists that the toxicity profiles of chelates depends mainly on metal
ion, its affinity to this metal, and their ability to supply or to sequester it from the body/environment.
Both EDTA and DTPA are chelates with similar structure and physico-chemical properties tham their t
oxicity are also similar. Please see RA statement in section 13.

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:

Himalayan

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: LPT germany
- Age at study initiation: 4-5 months
- Weight at study initiation: 2.2-2.4
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 20 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/-3°C
- Humidity (%): 30-70%
- Air changes (per hr): 60
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 4. To: 12.July 2007

Vehicle:

unchanged (no vehicle)

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

iris score

Basis:

animal: 1,2,3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48 h

Score:

1

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Reversibility:

fully reversible within: 4 days

Irritation parameter:

chemosis score

Basis:

animal: 1,2,3

Time point:

24/48/72 h

Score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Under the present test conditions a single administration of 100 mg FeNaEDTA per animal into the conjunctival sac of the right eye of three rabbits caused the following effects:
Corneal opacity (grad 1) was observed in animal no. three 24 hours to 6 days after instillation.
The fluorescein test performed 24 hours after instillation revealed corneal staining in animal no. 3 (up to 1/4 of the surface).
Conjunctival redness (grad 1) was observed in animal no. one 60 minutes, in animal no. two 60 minutes to 48 hours and in animal no. three 24 hours to 4 days after instillation.
In addition, secretion was observed in all 3 animals 1 hour after instillation.
The irises were not affected by instillation of the test item.
There were no systemic intolerance reactions.

Time after administration	C O R N E A	I R I S	C O N J U N C T I V A E	C H E M O S I S
	Opacity		Redness
	A n i m a l  n o. : 1 / 2 / 3
before dosing	0/0/0	0/0/0	0/0/0	0/0/0
60 minutes	0/0/0	0/0/0	1/1/0	0/0/0
24 hours	0/0/1	0/0/0	0/1/1	0/0/0
48 hours	0/0/1	0/0/0	0/1/1	0/0/0
72 hours	0/0/1	0/0/0	0/0/1	0/0/0
4 days	-/-/1	-/-/0	-/-/1	-/-/0
5 days	-/-/1	-/-/0	-/-/0	-/-/0
6 days	-/-/1	-/-/0	-/-/0	-/-/0
7 days	-/-/0	-/-/0	-/-/0	-/-/0

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

Although the test item induced slight irritation, no classification is needed according to OECD-GHS.

Executive summary:

An eye irritation study with three male Himalyan rabbits according to OECD guideline 405 and GLP was performed.

A single administration of 100 mg FeNaEDTA per animal into the conjunctival sac of the right eye of three rabbits caused the following effects:

Corneal opacity (grade 1) was observed in animal no. three 24 hours to 6 days after instillation.

The fluorescein test performed 24 hours after instillation revealed corneal staining in animal no. 3 (up to 1/4 of the surface).

Conjunctival redness(grade 1) was observed in animal no. one 60 minutes, in animal no. two 60 minutes to 48 hours and in animal no. three 24 hours to 4 days after instillation. In addition, secretion was observed in all 3 animals 1 hour after instillation.

Irises were not affected by instillation of the test item.

There were no systemic intolerance reactions.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for selection of eye irritation endpoint:

Well performed and reported GLP study

Justification for classification or non-classification

Based on the test results, no classification for skin and eye irritation is needed.