Chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminium

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Remarks:

in the Bovine Corneal Opacity and Permeability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

adopted
09. Oct. 2017: “Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage”

Deviations:

not specified

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

adopted 14. Feb. 2017

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminium

EC Number:

237-998-4

EC Name:

Chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminium

Cas Number:

14154-42-8

Molecular formula:

C32H16AlClN8

IUPAC Name:

31h-phthalocyaninato(2-)-n29,n39,n31,n32]-chloro[29(sp-5-12)-aluminu; chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-,(SP-5-12)-Aluminum;CHLOROALUMINUM PHTHALOCYANINE;CHLORO(29H,31H-PHTHALOCYANINATO)ALUMINUM; CHLORO(PHTHALOCYANINATO)ALUMINIUM;ALUMINUM PHTHALOCYANINE CHLORIDE;PHTHALOCYANINE CHLOROALUMINUM; chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminium

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

other: Bovine

Strain:

other: Bos primigenius Taurus (fresh bovine corneas)

Details on test animals or tissues and environmental conditions:

Fresh bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and 60 months old. The eyes were transported to the test facility in Hanks’ Balanced Salt Solution with 1% Penicillin-Streptomycin solution (Penicillin 100 U/mL, Streptomycin 100 µg/mL) in a suitable cooled container within 1 hour.

Test system

Vehicle:

other: • Schott-bottles, glass vials, and culture flasks for solutions and media

Controls:

yes

Amount / concentration applied:

Negative control HBSS
Positive control imidazole, 20 % solution in HBSS

Duration of treatment / exposure:

4h at 32 ± 1 °C

Number of animals or in vitro replicates:

3

Details on study design:

-SELECTION AND PREPARATION OF CORNEAS : The fresh bovine eyes were obtained from cattle which were between 12 and 60 months old. The eyes were transported to the test facility in Hanks’ Balanced Salt Solution(HBSS) with 1% Penicillin-Streptomycin solution (Penicillin 100 U/mL, Streptomycin 100 µg/mL) in a suitable cooled container within 1 hour.
After having carefully cleaned and sterilised the cornea holders, they were kept in the incubation chamber at 32 ± 1 °C.
On the day of the assay, the MEM without phenol red was supplemented with sodium bicarbonate, L-glutamine and 1% fetal calf serum (= complete MEM) and stored in a water bath at 32 ± 1 °C.
The same was performed with the MEM with phenol red, but without addition of sodium bicarbonate.

-QUALITY CHECK OF THE ISOLATED CORNEAS : The corneas were examined and only corneas which were free from damages were used

-NUMBER OF REPLICATES : 3

-NEGATIVE CONTROL USED : Yes (HBSS)

-POSITIVE CONTROL USED : Yes (imidazole, 20 % solution in HBSS)

-APPLICATION DOSE AND EXPOSURE TIME : 750 µL, 4h

-TREATMENT METHOD: Open chamber

-POST-INCUBATION PERIOD: no

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: Rinsing the anterior chamber with cMEM with phenol red and final rinsing with cMEM without phenol red.
- POST-EXPOSURE INCUBATION: 90 minutes at 32 ± 1 °C.

-METHODS FOR MEASURED ENDPOINTS:
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry / microtiter plate reader] (OD490)

-SCORING SYSTEM: In Vitro Irritancy Score (IVIS) :
The IVIS of each replicate of the negative control: IVIS = opacity difference + (15 x corrected OD492 value).
The IVIS of each replicate of the positive control and of the test item: IVIS = (opacity difference – mean opacity difference of the negative control) + [15 x (OD492 – mean OD492 of the negative control)].

-DECISION CRITERIA: According to OECD Guideline no. 437 (Oct. 2017).

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Remarks:

Bovine Corneal

Value:

ca. 0.74

Vehicle controls validity:

not specified

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the test item Chloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminium showed no effects on the cornea of the bovine eye. The calculated IVIS was 0.74.
According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS ≤ 3 re-quires no classification for eye irritation or serious eye damage.

Executive summary:

One valid experiment was performed.

Bovine corneas were used. They were collected from slaughtered cattle that were between 12 and 60 months old.

The test itemChloro[29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]aluminiumwas tested as 20% suspension. The test item was applied to the cornea of a bovine eye which had been previously incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been measured.

The test item was incubated on the cornea for 4 hours at 32 ± 1 °C. After removal of the test item, opacity and permeability values were measured.

 

Hank’s Balanced Salt Solution (HBSS) was used as negative control. The negative control showed no irritating effect on the cornea and the calculated IVIS (In VitroIrritancy Score) was 2.57.

20% imidazole solution was used as positive control. The positive control induced serious eye damage on the cornea and was within two standard deviations of the current historical mean. The calculated IVIS was 102.27.