Reaction mass of copper digluconate, sodium sulphate and copper sulphate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10.Apr. 2007 - 21. Aug. 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

OECD Guidelines for Testing of Chemicals, Section 4, number 405 "Acute Eye lrritation 1 Corrosion", adopted April 24, 2002.

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Directive 2004/73/EC

Version / remarks:

Commission Directive 2004/73/EC, B.5. "Acute Toxicity: Eye lrritation/Corrosion", April 29, 2004.

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Directive 2001/59/EC

Version / remarks:

Commission Directive 2001/59/EC of 6 August 2001, Annex VI

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Copper di-D-gluconate

EC Number:

208-408-2

EC Name:

Copper di-D-gluconate

Cas Number:

527-09-3

Molecular formula:

C12H22CuO14

IUPAC Name:

copper(2+) bis(2,3,4,5,6-pentahydroxyhexanoate)

Constituent 2

Reference substance name:

Sodium sulphate

EC Number:

231-820-9

EC Name:

Sodium sulphate

Cas Number:

7757-82-6

Molecular formula:

H2O4S.2Na

IUPAC Name:

disodium sulfate

Constituent 3

Reference substance name:

Copper sulphate

EC Number:

231-847-6

EC Name:

Copper sulphate

Cas Number:

7758-98-7

Molecular formula:

CuSO4

IUPAC Name:

copper(2+) ion sulfate

Test material form:

liquid

Specific details on test material used for the study:

TEST ITEM PREPARATION
0.1 mL (per animal) of the test item was measured with a syringe and applied undiluted as it was delivered by the Sponsor.
The pH of the test ítem was measured befare administration and was found to be 2.67.
According to Commission Directive 2004/73/EC, B.5. and OECD Guidelines 405, a test item needs not to be tested if the pH-value is less than 2 or greater than 11.5, owing to its predictable corrosive properties.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST SYSTEM
- Test system: Young Adult New Zealand White Rabbit
- Rationale: Recognized by the international guidelines as the recommended test system
- Source: Granja San Bernardo, Ctra. de Tarazana, s/n, 31522-Tulebras, Navarra (Spain)
- Total number of animals: 2
- Sex: Males
- Body weight at start of treatment: 3.0-3.2 kg
- ldentification: By numbered tags in the ears
- Acclimatization: From 27 to 34 days
- Veterinary inspection: During acclimatization the animals were examined by a veterinary surgeon to assure their satisfactory health status for use in the study.

HUSBANDRY
- Room number: Room 107
- Conditions: Standard Laboratory Conditions
Air-conditioned with ranges for room temperatura 22-25 °C, relative humidity 40-70% and 10-20 air changes per hour. Room temperatura and relative humidity were monitored continuously. There was a 12-hour fluorescent light/12-hour dark cycle.
- Accommodation: lndividually in stainless steel cages (52 x 58 x 43 cm). The cages were identified by a card stating the animal's number, sex and date of arrival, Study Number, Study Director's name, test ítem identification, dese level, administration route, and date of treatment.
- Diet: Pelleted standard TEKLAD 2030 Global Rabbit diet ad libitum, (supplied by Harlan lnterfauna Ibérica, S.L., Ctra. Sant Miquel del Fai, km. 3, Apartado 38, 08182-Sant Feliu de Codines, Barcelona (Spain), Batch No. N495 and N516, Expiry Date: 4 and 26 June 2007), was offered.
- Water: Tap water ad libitum by means of an automatic drinking system. Results of bacteriological, chemical and contaminant analyses were archived at RCC CIDA S.A.

Test system

Vehicle:

not specified

Controls:

not specified

Amount / concentration applied:

The test item was applied at 0.1 m/L animal

Observation period (in vivo):

OBSERVATIONS
Viability/Mortality: Daily from acclimatization of the animals to the termination of test.
Clinical signs: Daily from acclimatization of the animals to the termination of test.
Body weights:At start of acclimatization, on the day of application and in each observation point.
Eye reaction: See sections 4. 7 and 4.8.

EYE OBSERVATION TIME POINTS ANO TERMINATION
lmmediately after the administration, a plastic collar was placed on the rabbits for 1 hour to avoid their rubbing their eyes.
The eyes of each animal were examined approximately 1, 24, 48, 72 hours and 7 days, as well as 10, 14, 17 and 21 days after administration in one of the animals.
The observation of the cornea was always completad, except for the reading at 1 hour after administration, with an examination following instillation of an aqueous solution of 2% sodium fluorescein and subsequent washing with 0.9% physiological saline. lf in two consecutíve readings no corneal lesion was recorded, the use of sodium fluorescein was not required in subsequent observations. Once the excess fluorescein had been removed, the corneal alterations were observad with the aid of a transilluminator with a cobalt blue filter.
The treated eyes were not rinsed after instillation of the test item.

Details on study design:

TREATMENT
The eyes of the animals were examinad before test item administration. The eyes of the animals used in the study were in good condition.
On the day of treatment, 0.1 ml of the test item was placed in the conjunctival sac of the right eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test item. The left eye remained untreated and served as the reference control.
As it was suspected that the test ítem might produce irritancy, a single animal (one male) was treated first.
As an irritant effect was observad in the first animal, a second animal was administered, instead of exposing two additional animals simultaneously. As the second animal showed irritant effects, the test was suspended and no more additional animals were necessary.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

Nº 2815

Time point:

24/48/72 h

Score:

0.67

Reversibility:

not specified

Results 2

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

Nº 2815

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not irritation

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #1

Remarks:

Nº 2815

Time point:

24/48/72 h

Score:

3

Reversibility:

not specified

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

Nº 2815

Time point:

24/48/72 h

Score:

2.33

Reversibility:

not specified

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

Nº 2818

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not irritation

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

Nº 2818

Time point:

24/48/72 h

Score:

0

Reversibility:

not specified

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #2

Remarks:

Nº 2818

Time point:

24/48/72 h

Score:

2

Reversibility:

not specified

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

Nº 2818

Time point:

24/48/72 h

Score:

1.33

Reversibility:

not specified

Irritant / corrosive response data:

IRRITATION
The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, reddening and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity were 0.67 and 0.00 respectively. The individual mean scores for iris were 0.00 for both animals. The individual mean scores for the conjunctivae were 3.00 and 2.00 for reddening and 2.33 and 1.33 for chemosis, respectively.
The mean values at 24, 48 and 72 hours following administration for each type of lesion for both animals together were:
Corneal opacity: 0.33
Lesions in the iris: 0.00
Reddening: 2.50
Chemosis: 1.83
Very slight opacity (grade 1) was observed in the cornea of one animal (no. 2815) 24 and 48 hours after administration. No corneal opacity was recorded in this animal 72 hours after administration.
No abnormal findings were observed in the iris of any animal at any of the measurement intervals.
One animal showed (no. 2815) markedly reddened; beefy red eye (grade 3) 24, 48 and 72 hours after treatment. Reddening diminished progressively to moderate reddening (grade 2) on days 7 and 10, and slight reddening (grade 1) on days 14 and 17 after administration.
The other animal (no. 2818) showed moderate reddening (grade 2) at 24, 48 and 72 hours post-administration.
Marked swelling (chemosis) (grade 4) of the conjunctivae was observad in both animals 60 minutes after administration. At 24, 48 and 72 hours marked (grade 3) or obvious (grade 2) swelling was observad in one animal (no. 2815), and obvious or slight (grade 1) swelling was observed in the other animal (no. 2818).
Moderate or slight ocular discharge was observed in animal 2815 during all the observation period, as well as marked or moderate discharge in animal 2818 during observations made 60 minutes, 24, 48 and 72 hours after administration.
No abnormal findings except slight ocular discharge were observad in the treated eye of animal no. 2815 21 days after treatment, the end of the observation period for this animal. No abnormal findings were observad in the treated eye of the remaining animal (no. 2818) 7 days after treatment, the end of the observation period for this animal.

Other effects:

VIABILITY/MORT ALITY ANO CLINICAL SIGNS
No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
COLORATION
No staining of the treated eyes produced by the test item was observad.
CORROSION
No corrosion of the cornea was observed at any of the reading times.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Conclusion of the study report attach
Based upon the referred classification (Commission Directiva 2001/59/EC of August 06, 2001 ), SERGOMILL60 is considerad to be "IRRITATING" to the rabbit eye, thus it is assigned the risk phrase R36.
Conclusion according actual legislation
The mean values at 24, 48 and 72 hours following administration for each type of lesion for both animals together were:
Corneal opacity: 0.33
Lesions in the iris: 0.00
Reddening: 2.50
Chemosis: 1.83
The product clasifies as Category 2* (irritating to eyes) according to REGULATION (EC) No 1272/2008 OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 16 December 2008
*Classification extracted from table 3.3.2., page 126 of CLP regulation according to which a conjuctival rednes value above 2 triggers substance classification as Eye Irrit. 2

Executive summary:

The primary eye irritation potential of the test item was investigated according to OECD test guideline no. 405. The test ítem was applied by instillation of 0.1 ml into the right eye of each of two young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48, 72 hours and 7 days, as well as 10, 14, 17 and 21 days after test ítem instillation in one of the animals.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity were 0.67 and 0.00 respectively. The individual mean scores for iris were 0.00 for both animals. The individual mean scores for the conjunctivae were 3.00 and 2.00 for reddening and 2.33 and 1.33 for chemosis, respectively.

The mean values at 24, 48 and 72 hours following administration for each type of lesion for both animals together were:

Corneal opacity: 0.33

Lesions in the iris: 0.00

Reddening: 2.50

Chemosis: 1.83

The instillation of the test item into the eye resulted in moderate, early-onset ocular changes, such as reddening of the conjunctivae, discharge and chemosis. Very slight and transient opacity was also observed in one animal (no. 2815). Marked swelling (grade 4) was observed in both animals 60 minutes after administration, which remitted progressively to obvious swelling (grade 2) in one animal and to slight swelling (grade 1) in the other. After 7 days, the edema has remitted completely in both animals. Reddening observed in both animals was moderate (grade 2) or marked (grade 3), and !asted up to 72 hours in one animal (no. 2818) and up to 17 days in the other (no. 2815), although reddening observed on days 14 and 17 in this last animal was slight (grade 1 ).

No abnormal findings were observed in the treated eye 21 days after the administration (in animal no. 2815) and 7 days afterthe administration (in animal no. 2818).

No corrosion was observed at any of the measuring intervals. No staining of the treated eyes by the test ítem was observed and no clinical signs were observed.

Based upon the referred classification criteria (Commission Directiva 2001/59/EC of August 06, 2001 ), the test item is considerad to be "IRRITATING" to the rabbit eye, thus it is assigned the risk phrase R36.