Registration Dossier

Administrative data

Description of key information

Skin irritation

In a K1 in vivo skin irritation study in New Zealand White Rabbits equivalent to OECD Guideline 404, imazalil sulfate was found to be a mild irritant with a primary irritation index of 0.7. Based on CLP Regulation (EC) No 1272/2008, the test item is considered not to be classified (score < 2.3) (Moore, 1998, K1).

Eye irritation

From a K1 in vivo eye irritation study in New Zealand White Rabbits equivalent to OECD Guideline 405 it could be concluded that a 10% w:v solution of the test item in distilled water is classified as severely irritating to the eye based on CLP Regulation (EC) No 1272/2008 (Moore, 1998, K1).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
April 13 to April 17, 1998
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Remarks:
Study performed similar to OECD Guideline 404.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes
Remarks:
The study meets the requirements of 40 CFR Part 160 with the following exception: stability, characterization, identity and verification of the test item concentration as received and tested are the responsbility of the sponsor.
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: ZR027180PUE201/ PSL Code Number E71118-2H
- Expiration date of the lot/batch: October 15, 2003
- Purity test date: no data


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: no data
- Stability under test conditions: Expected to be stable for the duration of testing.
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in water (>500 g/L)

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Prior to application, the test item was moistened to achieve a dry paste by preparing a 90% w/w mixture in distilled water. Five tenths of a gram of the test item (dry weight basis) was placed on a 1 inch x 1inch, 4-ply gauze pad and applied to one 6 cm2 intact dose site.
- Final preparation of a solid: The sample was applied as a 90% w/w mixture in distilled water.
Species:
rabbit
Strain:
other: New Zealand albino
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Davidson's Mill Farm, South Brunswick, NJ
- Age at study initiation: Adult
- Weight at study initiation: no data
- Housing: The animals were singly housed in suspended stainless steel caging with mesh floors which conform to the size recommendations in the Guide for the Care and Use of Laboratory Animals DHEW (NIH) No. 86.23. Litter paper was placed beneath the cage and was changed at least three times per week.
- Diet (e.g. ad libitum): Pelleted Purina Rabbit Chow #5326.
- Water (e.g. ad libitum): Filtered tap water was supplied ad libitum by automatic water dispensing system.
- Acclimation period: 18 or 46 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.8-22.8° C
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 hour light/12 dark cycle

IN-LIFE DATES: no data
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
water
Remarks:
distilled
Controls:
other: Untreated skin areas were used as control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 gram of test item (dry weight basis)


Duration of treatment / exposure:
4 hours
Observation period:
up to 72 hours
Number of animals:
2 males and 4 females
Details on study design:
TEST SITE
- Area of exposure: Dorsal area and trunk. On the day prior to application, a group of animals was prepared by clipping (Oster model #A2-small) the dorsal area and the trunk.
- % coverage: one 6 cm2 area
- Type of wrap if used: Micropore tape
The test material was applied to approximately to 1 inch x 1 inch, 4-ply gauze pad and applied to one 6 cm2 intact dose site on each animal. The pad and entire trunk of each animal were then wrapped with a semi-occlusive 3 inch Micropore tape to avoid dislocation of the pad. Elizabethan collars were placed on each rabbit and they were returned to their designated cages.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): The test sites were gently wiped with water and a clean towel to remove residual test item.
- Time after start of exposure: 4 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days)
- Skin reactions were recorded at 1, 24, 48 and 72 hours after administration
- Cage side observations: once daily during the test period.

SCORING SYSTEM:
The dermal irritation was assessed according to the Draize Standard Skin Irritation Grading Scale
- Primary Dermal Irritation Index (PDII) was classifed as follows:
PDII Classification
Less than 2.0 Slightlly irritating
2.0-5.0 Moderately irritating
Greater than 5.0 Severly irritating






Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
24/48/72 h
Score:
0.7
Max. score:
8
Reversibility:
fully reversible within: 72 hours
Remarks on result:
other: PDII reported in the study and included in this IUCLID entry was calculated across the 24, 48, and 72 h timepoints
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
0.37
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
0.13
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritant / corrosive response data:
One rabbit was free from dermal irritation throughout the study. Within twenty-four hours of patch removal, very slight to well-defined erythema was noted at five of six treated sites. Two rabbits also exhibited ver slight edema. The overall incidence and severity of irritation decreased with time. All affected animals were free from erythema and edema within 72 hours.

All animals appeared active and healthy.  Apart from the dermal irritation noted, there were no other signs of gross toxicity, adverse pharamologic effects or abnormal  behavior.

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the classification system used, the Imazalil sulfate, is classified as slightly irritating to the skin when moistened with distilled water prior to application.

Under the conditions of this study, the Primary Irritation Index (PII) was 0.7, indicating that the test substance is a mild irritant. Based on the mean scores for erythema and edema at 24, 48 and 72 hours and the criteria of the CLP Regulation, the substance is considered as non irritant to the skin (score < 2.3).
Endpoint:
skin irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro skin irritation study does not need to be conducted because adequate data from an in vivo skin irritation study are available
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1998-07-8 to 1998-9-9
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
OECD Guideline 405
Principles of method if other than guideline:
Based on Draize test and similar to OECD Guideline 405 (Acute Eye Irritation/Corrosion)
GLP compliance:
yes (incl. QA statement)
Remarks:
The study meets the requirements of 40 CFR Part 160 with the following exception: stability, cha racterization, identity and verification of the test item concentration as received and tested are the responsbility of the sponsor.
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: #ZR027180PUE201
- Expiration date of the lot/batch: October 15 2003
- Purity: 75%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Not specified
- Stability under test conditions: Expected to be stable for the duration of testing
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in water (>500 g/l).

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: A 10% w/v solution of the active ingredient in the test substance was prepared by dissolving 13.3 g of the test substance (10g of imazalil) in distilled water.


Species:
rabbit
Strain:
New Zealand White
Remarks:
Albino
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Davidson's Mill Farm, South Brunswick, NJ
- Age at study initiation: Adult
- Weight at study initiation: Not specified
- Housing: The animals were singly housed in suspended stainless steel caging with mesh floors which conform to the size recommendations in the Guide for the Care and Use of Laboratory Animals DHEW (NIH) no 86.23. Litter paper was palced beneath the cage and was changed at least 3 times/ week.
- Diet (e.g. ad libitum): Pelleted Purina Rabbit Chow #5326
- Water (e.g. ad libitum): Filtered Tap water was supplied ad libitum by automatic water dispensing system.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16.7 - 21.1 °C
- Humidity (%): Not specified
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): Not specified

In Life dates : 8-17 July 1998
Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): one tenth of a millimeter of the test solution
- Concentration (if solution):10 %w/v solution of active ingredient in the test substance in distilled water.

Duration of treatment / exposure:
One application
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
6 (3 males and 3 females)
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No.

SCORING SYSTEM:
The eye irritation scores were evaluated according to the scoring system by Draize (1944)
- Cornea
a. Opacity-degree of density (Area most dense taken for reading)
No opacity 0
Scattered or diffuse areas details of iris clearly visible 1
Easily discernible translucent area, details of iris slightly obscured 2
Opalescent areas, no details of iris visible, size of pupil barely discernible 3
Opaque iris invisible 4

b. Area of cornea involved
One quarter (or less), but not zero 1
Greater than one quarter, but less than half 2
Greater than half, but less than three quarters 3
Greater than three quarters, up to whole area 4

- Iris
a. Values
Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these ot combination of any thereof) iris still reacting to light (sluggish reaction is positive) 1
No reaction to light, Hemorrhage, gross destruction 2

- Conjunctivae
a. Redness (Palpebral and bulbar)
Vessels normal 0
Vessels definately injected above normal 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2
Diffuse beefy red 3

b. Chemosis
No swelling 0
Any swelling above normal (includes nictitating membrane) 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half-closed 3
Swelling with lids about half-closed to completely closed 4

c. Discharge
No discharge 0
Any amount different from normal (does not include small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs, and considerable area around the eye 3

Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
other: Cornea Area score
Basis:
animal #1
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not reversible
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
not fully reversible within:
Remarks:
the observed period
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
animal #1
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
not fully reversible within:
Remarks:
the observed period
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
other: Cornea Area score
Basis:
animal #2
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
not fully reversible within:
Remarks:
The observed period
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #2
Time point:
24/48/72 h
Score:
2.33
Max. score:
3
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
discharge
Basis:
animal #2
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
not fully reversible within:
Remarks:
The observed period
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.67
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
other: Cornea area score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.67
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: The observed period
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Discharge
Basis:
animal #3
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
not reversible
Irritation parameter:
cornea opacity score
Basis:
animal #4
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
other: Cornea area score
Basis:
animal #4
Time point:
24/48/72 h
Score:
3.67
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #4
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #4
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
not fully reversible within:
Remarks:
The observed period
Irritation parameter:
chemosis score
Basis:
animal #4
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Discharge
Basis:
animal #4
Time point:
24/48/72 h
Score:
2.67
Max. score:
3
Reversibility:
not fully reversible within:
Remarks:
The observed period
Irritation parameter:
cornea opacity score
Basis:
animal #5
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
other: Cornea area score
Basis:
animal #5
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #5
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #5
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within:
Remarks:
The observed period
Irritation parameter:
chemosis score
Basis:
animal #5
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Discharge
Basis:
animal #5
Time point:
24/48/72 h
Score:
2.67
Max. score:
3
Reversibility:
fully reversible
Irritation parameter:
cornea opacity score
Basis:
animal #6
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
other: Cornea area score
Basis:
animal #6
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
iris score
Basis:
animal #6
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible
Irritation parameter:
conjunctivae score
Remarks:
Redness
Basis:
animal #6
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible
Irritation parameter:
chemosis score
Basis:
animal #6
Time point:
24/48/72 h
Score:
2.33
Max. score:
4
Reversibility:
not fully reversible within:
Remarks:
The observed period
Irritation parameter:
conjunctivae score
Remarks:
Discharge
Basis:
animal #6
Time point:
24/48/72 h
Score:
3
Max. score:
3
Reversibility:
not fully reversible within:
Remarks:
The observed period
Irritant / corrosive response data:
One hour after test solution instillation, all six treated eyes exhibited corneal opacity, iritis and conjonctivitis. The overall incidence and severity of irritation persisted in all animals through Day 7, with pannus evident in two rabbits. Due to the severity and irreversible nature of irritation, the sponsor authorized termination of the study and the test animals were euthanized on Day 9.
Other effects:
Apart from eye irritation, there were no other signs of gross toxicity, adverse pharmacologic effects or abnormal behavior.
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Based on the classification system used, a 10% w:v solution of the active ingredient (Imazalil) in Fungaflor 75 SP, lot #ZR027180PUE201 in distilled water is classified as severely irritating to the eye. The classification was raised from moderately to severely irritating due to scores greater than 10 for all rabbits at the 24 hour evaluation interval.
Endpoint:
eye irritation: in vitro / ex vivo
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
an in vitro eye irritation study does not need to be conducted because adequate data from an in vivo eye irritation study are available
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation

In a K1 in vivo skin irritation study in New Zealand White Rabbits equivalent to OECD Guideline 404, Imazalil Sulfate was found to be a mild irritant with a primary irritation index of 0.7. Based on CLP Regulation (EC) No 1272/2008, the test item is considered not to be classified (score < 2.3).

An in vitro skin corrosion study was waived based on the justification that adequate data from an in vivo skin irritation study is available.

In a K1 in vivo skin irritation study equivalent to OECD guideline 404, imazalil base -R 23979 was tested on New Zealand White Rabbits. The results concluded that the dermal application of imazalil base produced no dermal irritation.

This study is mentioned as supporting information but is not entered in IUCLID.

Eye irritation

In a primary eye irritation study (Moore G., 1998, K1), a 10% w:v solution of the test item in distilled water was instilled into the conjunctival sac of 6 (3 males and 3 females). Ocular irritation was evaluated using a high-intensity white light (Mag lite) in accordance with Draize et al. at 1, 24, 48, and 72 hours and 4 and 7 days post-installation.

One hour after test solution instillation, all six treated eyes exhibited corneal opacity, iritis and conjunctivitis. The overall incidence and severity of irritation persisted in all animals through Day 7, with pannus evident in two rabbits. Due to the severity and irreversible nature of irritation, the sponsor authorized termination of the study and the test animals were euthanized on Day 9.

 

An in vitro eye irritation study was waived based on the justification that adequate data from an in vivo eye irritation study are available.

Although the eye irritation endpoint is covered with available data on Imazalil sulfate, an additional study conducted with the source substance Imazalil base is added to the endpoint, in order to provide additional information to include in the read-across justification document used to cover other endpoints.

Imazalil base was administered once in the left eye conjunctival sac. All animals were observed daily and were scored according to the Draize scoring system at 1h and 1, 2, 3, 4, 7, 10, 14, 21 days after application (Teuns et al., 1990b, K1). The exposure to the test item resulted in opaque or translucent lesions of the cornea, redness of the conjunctiva, chemosis and changes in the iris. Corneal lesions persisted over at least 21 days and were described as opacities covering more than one quarter but less than one half of the area During the 21 days observation period, this irritation showed good reversibility, which however was not yet complete after 21 days for 2/3 animals. Therefore based on the criteria laid out in the CLP regulation, Imazalil is considered to be corrosive to the eye based on the irreversibility of the corneal opacity observed in these rabbits.

Justification for classification or non-classification

Skin irritation

According to the in vivo skin irritation study, mild irritation was noted. However, the test item did not meet the criteria for classification as irritant or corrosive according to the criteria of the CLP regulation (EC) No 1272/2008.

Eye irritation

Based on the results of a primary eye irritation study, a 10% w:v solution of the test item in distilled water is classified as severely irritating to the eye according to the criteria of the CLP regulation (EC) No 1272/2008 (Eye Dam. Cat .1).