TRACER YELLOW D-100

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation / corrosion

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: According to OECD guideline 431 and EC method B40

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

Harlan cytotest cell research GmbH

Details on test animals or test system and environmental conditions:

Human skin model test.

Amount / concentration applied:

25 mg test material per tissue.

Duration of treatment / exposure:

3 minutes and 1 hour.

Irritation parameter:

other: Relative absorbance

Basis:

mean

Time point:

other: 3

Score:

> 50

Max. score:

80

Reversibility:

no data

Remarks on result:

other: In vitro study

Irritant / corrosive response data:

Not corrosive to skin according UN GHS / EU CLP Cat.1 / DSD

Dose group	Exposure interval	Absorbance 570 nm Tissue 1*	Absorbance 570 nm Tissue 2*	Mean absorbance of 2 tissues	Rel. Absorbance [% of negative control]**
Negative control	3 min	1.345	1.377	1.361	100
Positive control	3 min	0.073	0.090	0.081	6.0
D-100	3 min	1.074	0.904	0.989	72.7
Negative control	1 hour	1.152	1.229	1.190	100.0
Positive control	1 hour	0.021	0.011	0.016	1.3
D-100	1 hour	1.005	0.916	0.960	80.7

* Mean of three replicate wells after blank correction

**relative absorbance: = 100 x (absorbance of test item)/ (absorbance of negative control)

The optical evaluation of the MTT-reducing capacity of the test item after one hour incubation with MTT-reagent did not show evidence of a blue colour and thereby was not considered to be a MTT reducer.

The test item D-100 is considered to be non-corrosive to skin:

- since the viability after 3 minutes exposure is greater than 50 % and

- the viability after 1 hour exposure is greater than 15%.

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information

Conclusions:

Not corrosive to skin according UN GHS / EU CLP Cat.1 / DSD

Executive summary:

This in vitro study was performed to assess the corrosive potential of D-100 by means of the Human Skin Model Test. Independent duplicate tissues of the human skin model EST-1000™ were exposed to the test item, the negative control or the positive control for 3 minutes and 1 hour, respectively. About 25 mg of the solid test material were applied to the tissues and wetted with 50 μL deionised water. A volume of 50 μL of either the negative control (deionised water) or the positive control (8.0 N KOH) was applied to each tissue. After exposure to the negative control the absorbance values exceeded the required acceptability criterion of mean OD570 ≥ 0.8 for both treatment intervals thereby confirming the acceptable quality of the tissues. Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control, both for the 3 minutes exposure period and for the 1 hour exposure period thus confirming the validity of the test system. After exposure to the test item D-100 the relative absorbance values decreased to 72.7% after 3 minutes. After the 1 hour exposure relative absorbance values were reduced to

80.7%. Nevertheless, both values are well above the threshold of 50% for the 3 minutes exposure and above 15% for the 1 hour exposure. Therefore, the test item was considered to be not corrosive. In conclusion, it can be stated that in this study and under the reported experimental conditions, the test item D-100 was non corrosive to skin according to UN GHS / EU CLP Cat.1 / DSD (67/548/EEC).

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:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: According to OECD guideline 437 and EC test method B47

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

GLP compliance:

yes (incl. QA statement)

Remarks:

Harlan Cytotest cell research GmbH

Details on test animals or tissues and environmental conditions:

isolated bovine cornea according to OECD guideline 437

Amount / concentration applied:

0.75 mL

Duration of treatment / exposure:

240 minutes

Irritation parameter:

other: In vitro score

Basis:

mean

Time point:

other: 240

Score:

< 55.1

Max. score:

0.19

Reversibility:

not specified

Irritant / corrosive response data:

Not corrosive / not severe irritant to the eye

Table of results.

Results after 240 minutes incubation time.

*corrected values

**negative values are only calculated values that do not reflect reality

Test group	opacity value = difference (t240 -t0 ) of opacity	Mean	permeability at 490 nm (OD490)	Mean	in vitro score	mean in vitro irritation score	proposed in vitro irritation scale
Negative control	231	2.00	0.0630.0560.055	0.058	2.953.841.83	2.87	Non corrosive / non severe irritant
Positive control	192.00*172.00*185.00*		0.046*0.036*0.017*		192.69172.54185.26	183.50	Corrosive / severe irritant
D-100	-1.00*-2.00*-2.00*		0.079*0.002*-0.002*		0.19-1.97-2.03	0.00 (-1.27**)	Non corrosive / non severe irritant

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

Not corrosive / not severe irritant to the eye

Executive summary:

This in vitro study was performed to assess the corneal irritation and damage potential of D- 100 by means of the BCOP assay using fresh bovine corneae. After a first opacity measurement of the fresh bovine corneae (t0), the 20% (w/v) suspension in saline (0.9% (w/v) NaCl in deionised water) of the test item D-100, the positive, and the negative controls were applied to corneae and incubated for 240 minutes at 32 ± 1 °C. The posterior chamber contained MEM medium supplemented with sodium bicarbonate and Lglutamine and 1% fetal calf serum (FCS) (complete medium = cMEM). After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae and opacity was measured again (t240). After the opacity measurements permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C. With the negative control (saline) neither an increase of opacity nor permeability of the corneae could be observed. The positive control (10% (w/v) Benzalkonium chloride in saline) showed clear opacity effects corresponding to a classification as corrosive / severe irritant to the eye (CLP/EPA/GHS (Cat 1)). Relative to the negative control, the test item D-100 did not cause any increase of the corneal opacity or permeability. The calculated mean in vitro irritation score was 0.00. According to OECD 437 the test item is classified as not corrosive / not severe irritant to the eye. In conclusion, according to the current study and under the experimental conditions reported, the test item D-100 is not corrosive / not severe irritant to the eye (CLP/EPA/GHS (Cat 1)).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Justification for selection of skin irritation / corrosion endpoint:
Key study reveiled the product is not irritant and not corrosive to skin.

Justification for selection of eye irritation endpoint:
Key study reveiled the product is not irritant and not corrosive to the eye.

Justification for classification or non-classification

Key test report reveiles that the test substance is not irritant and not corrosive to the skin and not irritant / corrosive to the eye.