Registration Dossier

Administrative data

Description of key information

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-12-03 to 2009-12-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted to current accepted guidelines.
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Hillcrest, Belton, Loughborough, UK
- Age at study initiation: 12 to 20 weeks old
- Weight at study initiation: 2.28 to 2.64 kg
- Housing: Individually housed in suspended cages.
- Diet: 2030 Teklad Global Rabbit diet (Harlan Teklad, Blackthorn, Bicester, Oxon, UK) available ad libitum.
- Water : Mains drinking water available ad libitum.
- Acclimation period: Minimum of 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): A minimum of 15 changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hour cycle
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied : 0.5 g moistened sufficiently with 0.5 mL of distilled water to achieve a paste
Duration of treatment / exposure:
Animal were exposed for 4 hours.
Observation period:
Animals were observed for 72 hours for skin reactions
Number of animals:
3 animals in total (1 animal used in the initial test, with a further 2 animals in the main test.)
Details on study design:
TEST SITE
- Area of exposure: The test material was introduced a 2.5 cm x 2.5 cm cotton gauze patch
- Type of wrap if used: The trunk of each rabbit was wrapped in an elasticated corset.


REMOVAL OF TEST SUBSTANCE
- Washing : Any residual test material was removed by gentle swabbing with cotton wool soaked in distilled water.
- Time after start of exposure: 4 hours post administration of test material.


SCORING SYSTEM: Draize JH (1959) “Dermal Toxicity” In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. Associations of Food and Drug Officials of the United States, Austin, Texas p.46-59
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Remarks:
of three animals
Time point:
other: 24 and 72 hours post dosing
Score:
0
Max. score:
8
Reversibility:
other: Not applicable, no effects noted during the study.
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
68702
Time point:
24/48/72 h
Score:
0
Max. score:
0
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
68703
Time point:
24/48/72 h
Score:
0
Max. score:
0
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
68704
Time point:
24/48/72 h
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
68702
Time point:
24/48/72 h
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
68703
Time point:
24/48/72 h
Score:
0
Max. score:
0
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
68704
Time point:
24/48/72 h
Score:
0
Max. score:
0
Irritant / corrosive response data:
The scores for erythema and oedema at the 24 and 72-hour readings were totalled for the three test rabbits (12 values) and this total was divided by six to give the primary irritation index of the test material.

The test material produced a primary irritation index of 0.0.
Other effects:
Not reported

Measurement of pH

The pH of the test material was determined prior to commencement of the study and found to be as follows:

 

Preparation

pH Measurement

immediately

after 10 minutes

after 20 minutes

 10 % w/w aqueous preparation of the test material

 6.2

6.9

6.9 

Table 1: Individual Skin Reactions

 

Skin Reaction

Observation Time (Following Patch Removal)

Individual Scores – Rabbit Number and Sex

Total

68702 Male

68703 Male

68704 Male

Erythema/Eschar Formation

Immediately

0

0

0

(0)

1 Hour

0

0

0

(0)

24 Hours

0

0

0

0

48 Hours

0

0

0

(0)

72 Hours

0

0*

0*

0

Oedema Formation

Immediately

0

0

0

(0)

1 Hour

0

0

0

(0)

24 Hours

0

0

0

0

48 Hours

0

0

0

(0)

72 Hours

0

0*

0*

0

Sum of 24 and 72-hour Readings (S): 0

Primary Irritation Index (S/6): 0/6 = 0.0

Classification: Non-Irritant

() = total values not included in calculating the primary irritation index

 

Table 2: Individual Bodyweights and Bodyweight Changes

 

Rabbit Number and Sex

Individual Bodyweight (kg)

Bodyweight Change (kg)

Day 0

Day 3

68702 Male

2.28

2.33

0.05

68703 Male

2.64

2.65

0.01

68704 Male

2.28

2.31

0.03

Interpretation of results:
not irritating
Remarks:
Criteria used for interpretation of results: OECD GHS
Conclusions:
MnSO4 is not irritating to skin.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 December 2009 to 14 December 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP and to a current guideline
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: One New Zealand White rabbit was supplied by Harlan Laboratories UK Ltd, Bicester, Oxon, UK.
- Age at study initiation: Twelve to twenty weeks old.
- Weight at study initiation: 2.14 kg
- Housing: The animal was housed in a suspended cage.
- Diet (e.g. ad libitum): Free access to food (Certified Rabbit Diet).
- Water (e.g. ad libitum): Free access to mains drinking water.
- Acclimation period: At least five days.

ENVIRONMENTAL 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): Lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Volume of 0.1 ml of the test material, which was found to weigh approximately 80 mg.

Duration of treatment / exposure:
Single application of the test material, up to 72 hours exposure.
Observation period (in vivo):
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment.
Number of animals or in vitro replicates:
1
Details on study design:
APPLICATION OF TEST SUBSTANCE
Immediately before the start of the test, both eyes of the provisionally selected test rabbit were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. An animal free of ocular damage was used.

A volume of 0.1 ml of the test material, which was found to weigh approximately 80 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test material, an assessment of the initial pain reaction was made.

After consideration of the ocular responses produced in this animal, no additional animals were treated.


SCORING SYSTEM:
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).

Any other ocular effects were also noted. Any clinical signs, if present, were also recorded.

Interpretation of results:
The numerical values corresponding to tissue and observation time were recorded. The data relating to the conjunctiva 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 conjunctiva = (A + B + C) x 2
Score for iris = D x 5
Score for cornea = (E x F) x 5


Using the numerical data obtained a modified version of the system described by Kay J H and Calandra J C (1962), J. Soc. Cosmet. Chem. 13, 281-289 was used to classify the ocular irritancy potential of the test material. This was achieved by adding together the scores for the cornea, iris and conjunctiva for each time point for the rabbit. The total scores for each observation were calculated. The maximum total score together with the persistence of the reactions enabled classification of the eye irritancy potential of the test material.

If evidence of irreversible ocular damage is noted, the test material will be classified as corrosive to the eye.

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.


Irritation parameter:
maximum mean total score (MMTS)
Basis:
animal #1
Time point:
other: 24 and 48 hours
Score:
36
Max. score:
110
Reversibility:
not reversible
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
1
Reversibility:
not fully reversible within: 72 hours
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.7
Max. score:
1
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.7
Max. score:
3
Reversibility:
not fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.7
Max. score:
3
Reversibility:
not fully reversible within: 72 hours
Irritant / corrosive response data:
Individual scores for ocular irritation are given in Table 1.

Scattered or diffuse corneal opacity was noted in the treated eye at the 34, 48 and 72 hour observations.

Iridial inflammation was noted in the treated eye at the 24 and 48 hour observations.

Moderate conjunctival irritation was noted in the treated eye one hour after treatment with severe conjunctival irritation noted at the 48 and 72 hour observations and moderate conjunctival irritation at the 72 hour observation.

Haemorrhage scattered over the nictitating membrane and upper and lower conjunctival membranes were noted in the treated eye at the 24, 48 and 72 hour observations.

Signs of pain, distress and ill health including bodyweight loss, lethargy, distressed respiration and uncoordinated movement were noted at the 7 day observation. Due to the observed signs the animal was humanely killed 7 days after treatment, in accordance with company policy and UK Home Office guidelines.
Other effects:
None reported.

Table 1: Individual Scores for Ocular Irritation

Rabbit Number and Sex

68710 Male

IPR= 3

Time After Treatment

1 Hour

24 Hours

48 Hours

72 Hours

CORNEA

 

 

 

 

E = Degree of Opacity

0

1

1

1

F = Area of Cornea Involved

0

3

3

2

Score (E x F) x 5

0

15

15

10

IRIS

 

 

 

 

D

0

1

1

0

Score (D x 5)

0

5

5

0

CONJUNCTIVA

 

 

 

 

A = Redness

2

3H

3H

2H

B = Chemosis

2

3

3

2

C = Discharge

2

2

2

2

Score (A + B + C) x 2

12

16

16

12

Total Score

12

36

36

22


IPR=   Initial pain reaction

H =      Haemorrhage scattered over the nictitating membrane and upper and lower conjunctival membranes

Interpretation of results:
irritating
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
MnSO4 should be classified as an irritant. Based upon the results MnSO4 should be classified under CLP as Cat. 1 for eye damage.
Executive summary:

Introduction. 

The study was performed to assess the irritancy potential of the test material to the eye of the New Zealand White rabbit. The method was designed to meet the requirements of the following:

- OECD Guidelines for the Testing of Chemicals No. 405 “Acute Eye Irritation/Corrosion” (adopted 24 April 2002)

- Method B5 Acute Toxicity (Eye Irritation) of Commission Regulation (EC) No. 440/2008

Result.

A single application of the test material to the non-irrigated eye of one rabbit produced scattered or diffuse corneal opacity, iridial inflammation, severe conjunctival irritation and haemorrhage over the nictitating and conjunctival membranes. Signs of pain, distress and ill health, including weight loss, lethargy, respiratory distress and uncoordinated movement were noted at the 7 day observation. Due to the observed signs the animal was humanely killed 7 days after treatment, in accordance with company policy and current UK Home Office guidelines.

Conclusion.

The test material produced a maximum total score of 36 and was considered to be at least a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system (based on one rabbit only).

The test material was considered to be irritant and due to irreversible ocular damage (based on one rabbit only), is assigned a category 1 classification with the hazard phrase H318.

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

MnSO4 is not irritating to skin, based on an in vivo study in rabbit (IUCLID section 7.3.1, Pooles A). MnSO4 is irritating to the eyes and should be classified as Eye Damage Cat. 1 on the basis of the severity of the effects seen on a single animal (IUCLID section 7.3.2, Pooles A)