Xanthylium, 9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethyl-, molybdatesilicate

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 Jul - 06 Jul 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

The test item was found to have the potential to cause color interference with the MTT endpoint, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.

Deviations:

yes

Remarks:

This deviation was considered to have not affected the integrity or validity of the study.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Version / remarks:

The test item was found to have the potential to cause color interference with the MTT endpoint, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.

Deviations:

yes

Remarks:

This deviation was considered to have not affected the integrity or validity of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Xanthylium, 9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethyl-, molybdatesilicate

EC Number:

263-793-4

EC Name:

Xanthylium, 9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethyl-, molybdatesilicate

Cas Number:

63022-06-0

Molecular formula:

Not applicable as the substance is UVCB

IUPAC Name:

reaction products of ethyl 2-[3-(ethylamino)-6-ethylimino-2,7-dimethylxanthen-9-yl]benzoate with silicomolybdic acid

Test material form:

solid: nanoform

Details on test material:

Shape
Shape Category: spheroidal
Shape: spherical
Pure Shape: Yes
Typical Composition: ≤100%
range: >0; ≤100%

Particle size distribution & range
Shape Category: spheroidal
Percentile D10, typical value: 35nm
Percentile D10, range: ≥10; ≤50nm
Percentile D50, typical value: 50nm
Percentile D50, range: ≥35; ≤100nm
Percentile D90, typical value: 85nm
Percentile D90, range: ≥50; ≤150nm
Fraction in size range 1-100nm: ≥50; ≤100%

Crystallinity
structure: Amorphous
Pure structure: Yes

Specific Surface Area
Typical specific surface area: ca. 45m2/g
Range: ≥10; ≤200m2/g
Skeletal density: ca. 1,8 g/cm3

Surface Functionalisation/treatment
surface treatment applied: no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
Identification: Lumière Pink S.M. 8135N
Batch: 021340
Purity: >90%
Physical state/Appearance: Red powder
Expiry Date: 01 July 2022
Storage Conditions: Room temperature in the dark

Negative Control
Identification: sterile distilled water
Batch 3012436
Purity: Not supplied
Expiry Date: 01 October 2018
Storage Conditions: Room temperature

Positive Control
Identification: 8.0N Potassium Hydroxide
Batch: SLBM9898V
Purity: 7.92N
Expiry Date: 07 April 2020
Storage Conditions: Room temperature

EpiDerm™ Reconstructed Human Epidermis Model Kit
Supplier: MatTek
Date received: 04 July 2017
EpiDermTM Tissues (0.63cm2) lot number: 25828
Assay Medium lot number: 062917TMA

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Source strain:

other: epithelial, derived from human skin, and formed into a stratified, cornified epithelium

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
Supplier: MatTek
Date received: 04 July 2017
EpiDermTM Tissues (0.63cm2) lot number: 25828
Assay Medium lot number: 062917TMA

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
- Temperature of post-treatment incubation: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Rinsing was achieved by filling and emptying each tissue under a constant soft stream of DPBS to gently remove any residual test item.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 60
- Optical density – 570nm

Control samples:

yes, concurrent negative control

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):25 mg

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL


POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 μL
- Concentration (if solution): 8.0 N

25 μL of sterile water was added for wetting of the test item to increase tissue surface contact.

Duration of treatment / exposure:

3 and 60 minutes

Duration of post-treatment incubation (if applicable):

3 h at 37 °C, 5% CO2

Number of replicates:

2

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Nean Negative Control 3 min

Value:

100

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean Negative Control 30 min

Value:

100

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean Positive Control 3 min

Value:

5.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 4

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean Positive Control 60 min

Value:

4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 5

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean Test Item 3 min

Value:

120.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 6

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean Test Item 60 min

Value:

100.1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Direct-MTT reduction:
- Colour interference with MTT:
The solution containing the test item was a purple color, therefore additional color correction tissues were incorporated into the testing procedure. However, the results obtained showed that no color interference occurred.


ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements:yes

Any other information on results incl. tables

The relative mean viabilities for each treatment group :

Exposure Period	Percentage Viability
	Negative Control	Positive Control	Test Item
3 minute	100	5.6	120.4
60 minute	100	4	100.1

 

Mean OD570Values and Viabilities for the Negative Control Item, Positive Control Item and Test Item

Tissue	Exposure Period	Mean OD570 of individual tissues	Mean OD570 of duplicate tissues	Standard Deviation	Coefficient of Variation (%)	Relative Mean Viability (%)
Negative Control	3 Minutes	1.524	1.568	0.062	4	100
		1.612
	60 Minutes	1.759	1.795	0.051	2.8
		1.831
Positive Control	3 Minutes	0.097	0.088	0.013	N/A	5.6
		0.079
	60 Minutes	0.071	0.072	0.001	N/A	4
		0.073
Test Item	3 Minutes	1.796	1.888	0.13	6.9	120.4
		1.98
	60 Minutes	1.777	1.796	0.026	1.5	100.1

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was considered to be non-corrosive to the skin.