Xanthylium, 9-(2-carboxyphenyl)-3,6-bis(diethylamino)-, molybdatesilicate

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

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

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.

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)

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

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

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

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

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

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

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

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

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

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was considered to be non-corrosive to the 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:

25 Mar 1996 - 17 Apr 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: LUMIERE PINK 8135C
- Storage: room temperature
- Description: dark maroon powder

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Age at study initiation: 12 – 16 weeks old
- Weight at study initiation: 2.54 – 2.73 kg
- Housing: suspended metal cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 21
- Humidity (%): 45 - 69
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/ 12

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL (=49 mg)

Duration of post- treatment incubation (in vitro):

1, 24, 48 and 72 hours, additional observatios were made on Days 7, 14 and 21

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): N/A

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 72h

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: no opacity noted

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

not fully reversible within: 21 days

Remarks on result:

positive indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible

Remarks:

14 days

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.3

Max. score:

2

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of weak irritation

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 72h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 21 days

Remarks on result:

probability of mild irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48 h

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 72h

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible within: 21 days

	animal #1				animal #2				animal #3
	24 h	48 h	72 h	mean animal #1	24 h	48 h	72 h	mean animal #2	24 h	48 h	72 h	mean animal #3	mean
Conjunctival Redness	2	1	1	1.3	1	1	0	0.7	2	2	2	2	1.3
Conjunctival Chemosis	1	1	1	1	1	1	0	0.7	2	2	2	2	0.9
Iritis	0	0	0	0	0	0	0	0	1	1	1	1	0.3
Corneal opacity	1	0	0	0.3	0	0	0	0	1	1	1	1	0.6

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The test item was found to irritating to rabbit eye.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Test substance is not classified for irritation to the skin but is classified as a Cat 1 eye irritant.