Octasodium 2,2'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino(1-hydroxy-3,6-disulphonatonaphthalene-2,8-diyl)azo]]bisnaphthalene-1,5-disulphonate

Endpoint summary

Description of key information

Skin irritation:

A study was performed for the assessment of the skin irritancy of the test item Bayscript Magenta BB with reconstructed human epidermis (RhE). The experiment was carried out in vitro using the commercially available test method epiCS®. The study was conducted in accordance with OECD TG 439 and EU Test Method B.46.The results of the assay show no skin irritant properties of the test item Bayscript Magenta BB and thus, the test item requires no classification according to UN GHS.

Eye irritation:

Based on the experimental conditions reported in an OECD TG 437 study the test item is classified as not seriously damaging the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

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

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)

Test system:

artificial membrane barrier model

Source species:

human

Cell type:

other: reconstructed human epidermis

Cell source:

other: reconstructed human epidermis

Source strain:

other: reconstructed human epidermis

Justification for test system used:

Human full thickness skin models and reconstituted epidermal equivalents are in vitro engineered tissue cultures that provide a three dimensional architecture which is biochemically, morphologically and functionally comparable to human epidermal tissue/skin in vivo. In contrast, organotypic skin explant systems are based on ex vivo skin removed from human or mouse and cultured in toto, afterwards. According to the literature all the models are useful in screening for topically applied irritant, corrosive or photocytotoxic substances.

Vehicle:

unchanged (no vehicle)

Details on test system:

The model used for this study has a functional stratum corneum with an underlying layer of living cells as recommended by the test guidelines. The barrier function of the stratum corneum is adequate, as has been shown by the supplier.

The viability of the living cells in the model must be sufficiently high to discriminate well between the positive and negative control substances. Cell viability is measured by the amount of MTT reduction, i.e. an OD value, following exposure to the negative control substance or the test item.

Control samples:

yes, concurrent vehicle

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

Three replicates of the epiCS inserts were exposed to the test item (30 mg), negative or positive control (each 30 µl). For the solid test item 30 µl of 0.9% NaCl were used to moisten and ensure good contact with the epidermis surface. A piece of nylon mesh was used as a spreading aid.

Duration of treatment / exposure:

The incubation of the treated epiCS inserts was carried out at RT for 20 min.

Duration of post-treatment incubation (if applicable):

After a post treatment incubation period of 42 h of the rinsed tissue in the incubator a MTT assay was performed

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean of 3 replicates

Value:

117

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Interpretation of results:

GHS criteria not met

Executive summary:

A study was performed for the assessment of the skin irritancy of the test item Bayscript Magenta BB with reconstructed human epidermis (RhE). The experiment was carried out in vitro using the commercially available test method epiCS®.

The study was conducted in accordance with OECD TG 439 and EU Test Method B.46.

The test item was applied undiluted topically to the RhE tissue construct in triplicates and incubated for 20 minutes, followed by a 42 hours post-treatment incubation period.

Cell viability was measured in a photometer by the amount of MTT (methylthiazole tetrazolium) reduction. The optical density value obtained for the test item was used to calculate the percentage of viability relative to the negative control, which is set at 100%.

The results of the concurrent negative control (NC, 0.9 % NaCl) and positive control (PC, 5 % SDS) demonstrated the viability (NC) and sensitivity (PC) of the test model.

The following value of cell viability was recorded for the test item: 118 % (rounded).

In conclusion the results of the assay used show no skin irritant properties of the test item Bayscript Magenta BB and thus, the test item requires no classification according to UN GHS.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

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

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Test system: isolated cornea from eyes of slaughtered cattle

Vehicle:

physiological saline

Controls:

yes, concurrent vehicle

yes, concurrent positive control

Amount / concentration applied:

750 µl of the test material formulations were applied to the corneas.

Duration of treatment / exposure:

4 hours

Duration of post- treatment incubation (in vitro):

90 min

Number of animals or in vitro replicates:

3

Details on study design:

In this test method, eye damaging properties of a test item is assessed by quantitative measurements of changes in corneal opacity and permeability with an opacitometer and a visible light spectrophotometer, respectively. Both measurements are used to calculate an IVIS, which is used to assign an in vitro irritancy hazard classification
category for prediction of the in vivo ocular irritation potential of a test item.

The BCOP test method uses isolated corneas from the eyes of freshly slaughtered cattle. Corneal opacity is measured quantitatively as the amount of light transmission through the cornea. Permeability is measured quantitatively as the amount of sodium fluorescein dye that passes across the full thickness of the cornea, as detected in the medium in the posterior chamber of the corneal holder. Test items are applied to the epithelial surface ofthe cornea by addition to the anterior chamber of the corneal holder.

Irritation parameter:

cornea opacity score

Run / experiment:

Mean

Value:

9

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein leakage

Run / experiment:

Mean

Value:

0.012

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

other: IVIS (in vitro irritation score)

Run / experiment:

Mean

Value:

9.2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Irritant / corrosive response data:

Based on OECD TG 437 and the experimental conditions reported the test item is classified as not seriously damaging the eye

Tabular In vitro irritancy score

	Cornea No.	Opacity per cornea	Permeability per cornea	IVIS per cornea	IVIS per group; mean (SD)
Negative control isotonic saline solution	1 2 3	0.7 -0.2 0.4	0.009 0.007 0.008	0.8 -0.1 0.5	0.4 (0.5)
Positive Control 20% Imidazole	4 5 6	76.4  77.4 93.3	0.980 0.806 0.891	91.1  89.5 106.6	95.7 (9.5)
Test item 20 % Bayscript Magenta BB	10 11 12	10.1  4.1 12.8	0.020 0.020 -0.003	10.4 4.4 12.8	9.2 (4.4)

IVIS < 55: classification as Category 1 is not warranted

Interpretation of results:

GHS criteria not met

Executive summary:

Based on the experimental conditions reported in an OECD TG 437 study the test item is classified as not seriously damaging the eye (no Category 1).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

No skin irritation was reported in an OECD TG 439 study (EU Test Method B.46) and no severe eye irritation was reported in an OECD TG 437 study.