Ethanol, 2,2'-[1,2-ethanediylbis(oxy)]bis-, reaction products with 3-(triethoxysilyl)-1-propanamine

Endpoint summary

Description of key information

Skin Irritation

REACH_not corrosive | Epiderm | OECD 431 | #key study#

REACH_not irritating | Epiderm | OECD 439 | #key study#

Eye Irritation

REACH_no prediction | BCOP | OECD 437 | #key study#

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-01-16 to 2018-01-18

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:

29 July 2016

Qualifier:

according to guideline

Guideline:

other: Council Regulation 440/2008, Method B.40 BIS: “In Vitro Skin Corrosion: Human Skin Model Test”, May 30, 2008

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol for: In Vitro EpiDermTM Skin Corrosion Test (EPI-200-SCT) For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm; Version 07/11/2014

GLP compliance:

yes (incl. QA statement)

Remarks:

(Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany)

Test system:

human skin model

Remarks:

EpiDerm

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

This test uses the EpiDerm™ reconstructed human epidermis model (MatTek) which consists of normal human epidermal keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely mimics the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis.

Vehicle:

unchanged (no vehicle)

Details on test system:

The test was carried out with the reconstituted three-dimensional human skin model EpiDerm (MatTek). This skin model consists of normal (non-cancerous), human-derived epidermal keratinocytes (NHEK) which have been cultured to form a multilayered, highly differentiated model of the human epidermis. The NHEK are cultured on chemically modified, collagen-coated cell culture inserts (Millicell). The EpiDerm skin model exhibits in vivo-like morphological and growth characteristics which are uniform and highly reproducible. It consists of organised basal, spinous, granular and cornified layers analogous to those found in vivo.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 µL

Duration of treatment / exposure:

3 minutes and 60 minutes

Duration of post-treatment incubation (if applicable):

3 h MTT incubation

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 min. treatment

Value:

97.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100.0

Positive controls validity:

valid

Remarks:

1.5

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min. treatment

Value:

106.8

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100.0

Positive controls validity:

valid

Remarks:

5.2

Other effects / acceptance of results:

Value Cut off pass/fail
Mean Absolute OD570 nm NK
(3 min Experiment) 1.609 0.8 ≤ NK ≤ 2.8 pass
Mean Absolute OD570 nm NK
(60 min Experiment) 1.670 0.8 ≤ NK ≤ 2.8 pass
Mean Relative Tissue Viability [%] of
PC (60 min experiment) 1.5 < 15% pass
CV [%]
(in the range of 20 – 100% viability) 7.8% - 20.1% ≤ 30% pass

The test item showed non-specific MTT-reducing potential. Therefore, additional killed tissue controls were treated with the test item to determine the non-specific reduction of MTT (NSMTT) and the results were corrected to the true MTT metabolic conversion (TODTT). The test item showed no water-colouring potential.

The test item showed no corrosive effects. The mean relative tissue viability (% negative control) was ≥ 50% (110.7%, NSMTT-corrected) after 3 min treatment and ≥ 15% (97.6%, NSMTTcorrected) after 60 min treatment.

The controls confirmed the validity of the study. The mean OD570nm of the two negative control tissues was ≥ 0.8 and ≤ 2.8 for each exposure period. The mean relative tissue viability (% negative control) of the positive control was < 15% (1.5%) after 60 min treatment. The coefficient of variation (CV) (in the range of 20 – 100% viability) of replicate tissues of all dose groups was ≤ 30% (7.8% - 20.1%).

Results of 3 minExperiment

Name	Negative Control		Test Item		Positive Control
Tissue	1	2	1	2	1	2
Absolute OD570	1.818	1.381	1.687	1.694	0.130	0.122
	1.854	1.399	1.702	1.704	0.129	0.125
	1.823	1.380	1.712	1.789	0.129	0.125
OD570- Blank Corrected	1.773	1.335	1.642	1.648	0.085	0.077
	1.808	1.354	1.656	1.659	0.083	0.079
	1.777	1.334	1.666	1.744	0.084	0.079
Mean OD570of 3 Aliquots (Blank Corrected)	1.786	1.341	1.655	1.684	0.084	0.079
SD OD570 of 3 Aliquots	0.019	0.011	0.012	0.052	0.001	0.001
Total Mean OD570of 2 Replicate Tissues (Blank Corrected)	1.564*		1.669		0.081
TODTT	-		1.731		-
SD OD570 of 2 Replicate Tissues	0.315		0.021		0.004
Mean Relative Tissue Viability [%]	100.0		106.8		5.2
Mean Relative Tissue Viability [%] - NSMTT Corrected	-		110.7		-
Coefficient Of Variation [%]^***	20.1		1.2		4.8

^*     corrected mean OD₅₇₀ of the negative control corresponds to 100% absolute tissue viability.

^***   coefficient of variation (CV) (in the range of 20 – 100% viability) between two tissues treated identically is ≤ 30%.

Results of the NSMTT control of 3 min Experiment

NSMTT	KU		KT		Negative Control
Tissue	1	2	1	2	1	2
absolute OD570 -values	0.576	0.495	0.449	0.501	1.818	1.381
	0.590	0.490	0.460	0.497	1.854	1.399
	0.597	0.493	0.445	0.497	1.823	1.380
OD570(Blank Corrected)	0.531	0.450	0.404	0.455	1.773	1.335
	0.545	0.445	0.415	0.451	1.808	1.354
	0.552	0.448	0.399	0.452	1.777	1.334
mean OD570 (mean of 2 aliquots)	0.538	0.448	0.409	0.453	1.791	1.344
total mean OD570 (mean of the replicate tissues)	0.493		0.431		1.568*
SD OD570(of the replicate tissues)	0.064		0.031		0.315
NSMTT [%]	-3.92				-
Relative Tissue Viability [%]	-				114.2	85.8
Mean Relative Tissue Viability [%]	-				100.0
SD Tissue Viability [%]	-				20.1
CV [% Viabilities]	-				20.1

Results of 60 min Experiment

Name	Negative Control		Test Item		Positive Control
Tissue	1	2	1	2	1	2
Absolute OD570	1.576	1.710	1.415	1.817	0.070	0.069
	1.579	1.783	1.419	1.816	0.072	0.069
	1.587	1.787	1.494	1.822	0.072	0.069
OD570- Blank Corrected	1.531	1.665	1.370	1.771	0.025	0.024
	1.533	1.738	1.374	1.771	0.027	0.024
	1.542	1.742	1.449	1.776	0.027	0.024
Mean OD570of 3 Aliquots (Blank Corrected)	1.535	1.715	1.397	1.773	0.026	0.024
SD OD570 of 3 Aliquots	0.006	0.043	0.044	0.003	0.001	0.000
Total Mean OD570of 2 Replicate Tissues (Blank Corrected)	1.625*		1.585		0.025
TODTT	-		1.586		-
SD OD570 of 2 Replicate Tissues	0.127		0.266		0.002
Mean Relative Tissue Viability [%]	100.0		97.5		1.5**
Mean Relative Tissue Viability [%] - NSMTT Corrected	-		97.6		-
Coefficient Of Variation [%]^***	7.8		16.8		6.3

^*           corrected mean OD₅₇₀ of the negative control corresponds to 100% absolute tissue viability.

^**             mean relative tissue viability of the 60 min positive control < 15%.

^***            coefficient of variation (CV) (in the range of 20 – 100% viability) between two tissues treated identically is ≤ 30%.

Results of the NSMTT control of 60 min Experiment

NSMTT	KU		KT		Negative Control
Tissue	1	2	1	2	1	2
absolute OD570 -values	0.106	0.095	0.087	0.113	1.576	1.710
	0.107	0.095	0.088	0.112	1.579	1.783
	0.106	0.094	0.087	0.112	1.587	1.787
OD570(Blank Corrected)	0.061	0.049	0.041	0.068	1.531	1.665
	0.062	0.049	0.043	0.067	1.533	1.738
	0.061	0.049	0.042	0.066	1.542	1.742
mean OD570 (mean of the aliquots)	0.061	0.049	0.042	0.067	1.532	1.702
total mean OD570 (mean of the replicate tissues)	0.055		0.055		1.617*
SD OD570(of the replicate tissues)	0.008		0.018		0.120
NSMTT [%]	-0.04				-
Relative Tissue Viability [%]	-				97.7	108.6
Mean Relative Tissue Viability [%]	-				103.1
SD Tissue Viability [%]	-				7.6
CV [% Viabilities]	-				7.4

 

Interpretation of results:

other: not corrosive

Conclusions:

In this study under the given conditions the test item showed no corrosive effects. The test item is classified as “non-corrosive“.

Executive summary:

In the present study the skin corrosivity potential of the test item was analysed. Since corrosive chemicals are cytotoxic after a short time exposure to the stratum corneum of the epidermis the cytotoxic effects of the test item on EpiDerm, a reconstituted three-dimensional human epidermis model, were determined. Hereby, the test item was applied topically. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 3 min and 60 min exposure period and compared to those of the concurrent negative controls.

The test item showed non-specific MTT-reducing potential. Therefore, additional killed tissue controls were treated with the test item to determine the non-specific reduction of MTT (NSMTT) and the results were corrected to the true MTT metabolic conversion (TOD_TT). The test item showed no water-colouring potential.

The test item showed no corrosive effects. The mean relative tissue viability (% negative control) was ≥ 50% (110.7%, NSMTT-corrected) after 3 min treatment and ≥ 15% (97.6%, NSMTT-corrected) after 60 min treatment.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-02-21 to 2018-02-23

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)

Version / remarks:

28 July 2015

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Commission Regulation (EC) No. 640/2012, L 193, Part B.46. “In vitro Skin Irritation: Reconstructed Human Epidermis Test Method” 06-Jul-2012

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: MatTek Corporation Protocol for: In Vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm (EPI-200-SIT)

Version / remarks:

Version 07-Nov-2014

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Test system:

human skin model

Remarks:

EpiDerm

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

This test uses the EpiDerm™ reconstructed human epidermis model (MatTek) which consists of normal human epidermal keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely mimics the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis.

Vehicle:

unchanged (no vehicle)

Details on test system:

This in vitro method is designed to predict and classify the skin irritation potential of a chemical by assessment of its effect on EpiDermTM, a reconstituted three-dimensional human epidermis model. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 60 min exposure period.

The test was carried out with the reconstituted three-dimensional human skin model EpiDerm (MatTek). This skin model consists of normal human epidermal keratinocytes (NHEK) which have
been cultured to form a multilayered, highly differentiated model of the human epidermis. The NHEK are cultured on chemically modified, collagen-coated cell culture inserts (Millicell). The EpiDerm epidermis model exhibits in vivo-like morphological and growth characteristics which are uniform and highly reproducible. It consists of organised basal, spinous and granular layers and a multi-layered stratum corneum analogous to patterns found in vivo.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Dose Groups
1. Negative control 30 µL Dulbecco’s phosphate buffered saline
2. Positive control 30 µL 5% sodium dodecyl sulfate solution
3. Test Item 30 µL (undiluted)

Duration of treatment / exposure:

60 ± 1 min

Duration of post-treatment incubation (if applicable):

24 ± 2 h + 18 ± 2 h (42 ± 2 h)

Number of replicates:

The test was performed on a total of 3 tissues per dose group.

Details on study design:

The test was performed on EpiDerm, an organotypic reconstructed three-dimensional model of the human epidermis. 3 replicate tissues are dosed with the test item, the negative control (30 µL DPBS) and the positive control (30 µL 5% SDS), respectively. After 60 ± 1 min treatment period at 37°C (35 min) and room temperature (25 min) the test item and the controls are rinsed off with DPBS and the tissues are post-incubated for 42 +/- 1 h. Then the tissues are stained via MTT for 3 hours. The extracts are measured photometrically at 570 nm.

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

94.9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100.0

Positive controls validity:

valid

Remarks:

3.0

The mixture of 30 μL test item per 1 mL MTT medium showed no reduction of MTT compared to the solvent. The mixture did not turn blue/purple. Therefore, NSMTT equalled 0%.

The mixture of 30 μL of the test item per 300 μL aqua dest. and per 300 μL isopropanol showed no colouring detectable by unaided eye-assessment. Therefore, NSC equalled 0%.

The test item showed no irritant effects. The mean relative tissue viability (% negative control) was > 50% (94.9%) after 60 min treatment and 42 h post-incubation.

The controls confirmed the validity of the study. The mean absolute OD570 of the three negative control tissues was ≥ 0.8 and ≤ 2.8 (2.002). The mean relative tissue viability (% negative control) of the positive control was ≤ 20% (3.0%). Standard deviation of viability of replicate tissues of all dose groups was ≤ 18% (0.1% - 9.8%).

Result of the Test Item

Name	Negative Control			Positive Control			Test Item
Tissue	1	2	3	1	2	3	1	2	3
Absolute OD570	2.070	1.785	2.187	0.100	0.101	0.100	1.948	1.996	1.686
	2.045	1.789	2.133	0.103	0.107	0.100	1.975	2.028	1.775
OD570(Blank Corrected)	2.027	1.741	2.144	0.057	0.057	0.056	1.905	1.953	1.643
	2.002	1.746	2.090	0.060	0.063	0.057	1.931	1.985	1.731
Mean OD570of the Duplicates (Blank Corrected)	2.014	1.743	2.117	0.058	0.060	0.056	1.918	1.969	1.687
Total Mean OD570of 3 Replicate Tissues (Blank Corrected)	1.958*			0.058			1.858
SD OD570	0.193			0.002			0.150
Relative Tissue Viability [%]	102.9	89.0	108.1	3.0	3.1	2.9	97.9	100.5	86.1
Mean Relative Tissue Viability [%]	100.0			3.0**			94.9
SD Tissue Viability [%]***	9.8			0.1			7.7
CV [% Viabilities]	9.8			3.2			8.1

 

^*          Blank-corrected mean OD_{570 nm} of the negative control corresponds to 100% absolute tissue viability.

^**         Mean relative tissue viability of the three positive control tissues is ≤ 20%.

^***        Standard deviation (SD) obtained from the three concurrently tested tissues is ≤ 18%

Interpretation of results:

GHS criteria not met

Conclusions:

In this study under the given conditions the test item showed no irritant effects. The test item is therefore classified as “non-irritant” in accordance with UN GHS “No Category”.

Executive summary:

In the present study the skin irritant potential of the test item was analysed. The EpiDerm™-Standard Model (EPI-200™), a reconstituted three-dimensional human epidermis model, was used as a replacement for the Draize Skin Irritation Test (OECD TG 404) to distinguish between UN GHS “Category 2” skin irritating test substances and not categorized test substances (“No Category”) which may be considered as non-irritant. Hereby, the test item was applied topically. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 60 min exposure and 42 h post-incubation period and compared to those of the concurrent negative controls.

The mixture of 30 µL test item per 1 mL MTT medium showed no reduction of MTT compared to the solvent. The mixture did not turn blue/purple. Therefore, NSMTT equalled 0%.

The mixture of 30 µL of the test item per 300 µL aqua dest. and/ per 300 µL isopropanol showed no colouring detectable by unaided eye-assessment. Therefore, NSC equalled 0%.

The test item showed no irritant effects. The mean relative tissue viability (% negative control) was > 50% (94.9%) after 60 min treatment and 42 h post-incubation.

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:

2018-02-13 to 2018-05-23

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for the Testing of Chemicals, no. 437 (adopted: 09 October 2017)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Munich, Germany

Species:

other: bovine cornea

Strain:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 μL

Duration of treatment / exposure:

After 10 minutes incubation at 32 ± 1 °C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red).

Duration of post- treatment incubation (in vitro):

2 h before illuminance measurement followed by 90 min before determination of optical density

Number of animals or in vitro replicates:

3 corneas for the test item
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive controls treated with ethanol 100%

Details on study design:

Preparation of the Corneas
The assay uses isolated corneas obtained as a by-product from animals freshly slaughtered at the abattoir A. Moksel AG, Buchloe, Germany.
On the test day, fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories. Immediately after arrival of the eyes, cornea preparation was initiated.
The eyes were carefully examined for defects and any defective eyes were discarded.
The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first. The corneas were incubated for one hour at 32 ± 1 °C.

Calibration of the Opacitometer
The opacitometer (BASF-OP3.0, Duratec) was switched on 15 min before starting the calibration procedure. The filter holder was placed into the opacitometer and the readout was adjusted to 1000 lux ± 10 lux using the “Calibrate”-turning knob. For calibration the glass filter F2 was introduced into the filter holder. The readout should lie in the range between 540-560 lux. To test the linearity of the measurement, two additional calibration filters, glass filter F3 and glass filter F4, were measured. For these glass filters, the opacitometer should display values between 300-310 lux and between 95-105 lux. If this is not the case, the calibration procedure has to be repeated. The calibration procedure is performed before each test and is documented in the raw data.

Treatment of the Corneas
After the equilibration period, the medium was removed from both chambers and replaced with fresh complete RPMI. An initial measurement was performed on each of the corneas using the opacitometer. Three corneas with illuminance readings approximately equivalent to the median illuminance of all corneas were selected as negative-control corneas. The illuminance of each cornea was read and recorded. Only corneas that had an initial illuminance reading I > I0/1.1651 lux were used for the assay. The medium was removed from the anterior chamber and replaced with the test item or control.
750 µL of the test substance or the control substance was introduced into the anterior chamber. After 10 minutes incubation at 32 ± 1 °C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red). Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red). The anterior chamber was refilled with complete RPMI and an illuminance measurement was performed after 2 hours incubation at 32 ± 1 °C. Also, each cornea was observed visually and pertinent observations were recorded.
After the illuminance measurement was performed, the medium was removed from both chambers of the holder. The posterior chamber was refilled with fresh complete RPMI. 1 mL of a 4 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 ± 1 °C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer (Jenway 6405 UV/VIS).



Test Groups
3 corneas for the test item
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive controls treated with ethanol 100%
The BCOP assay is considered to be valid if the in vitro irritation score obtained with the positive control falls within the two standard deviations of the current historical mean.
The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.


Evaluation of Results
The following formula was used to calculate the opacity, whereas the values a and b are equipment-specific variables empirically determined by the manufacturer:

Opacity= ( I0/I-b)/a

with a = 0.025 and b = 0.9894

The value I0 (=I zero) is the illuminance through a holder without cornea, but with windows and liquid. This value is determined by taking the mean for a set of cornea holders and is reevaluated periodically.
The change in opacity for each cornea was calculated by subtracting the initial opacity reading from the final opacity reading. These values were corrected by subtracting from each the average change in opacity observed for the negative-control corneas. The mean opacity value for each treatment was calculated by averaging the corrected opacity values of each cornea for a given treatment.
The mean OD490 for the blank cuvettes was calculated. The mean blank OD490 was subtracted from the OD490 of each cuvette (corrected OD490). Any dilutions that were made to bring the OD490 values into the linear range of the spectrophotometer (OD490 should be less than 1.500), were taken into account by multiplying the OD490 value of the dilution by the dilution factor. The final-corrected OD490 of the test article and the positive control were calculated by subtracting the average-corrected OD490 of the negative-control corneas from the corrected OD490 value of each treated cornea:
Final-corrected OD490 = (OD490 – mean blank OD490) – average-corrected negative control OD490
The mean OD490 value of each treatment group was calculated by averaging the final corrected OD490 values of the treated corneas for that treatment condition.

The following formula was used to determine the in vitro irritation score (IVIS):

IVIS = mean opacity value + (15 x mean permeability OD490 value)

The IVIS cut-off values for identifying test substances as inducing serious eye damage (UN GHS Category 1) and test substances not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given below:
Evaluation of the BCOP Assay
IVIS UN GHS
≤ 3 No Category
> 3; ≤ 55 No prediction can be made
> 55 Category 1

An identification of test substances that should be classified as irritating to eyes (UN GHS Category 2 or Category 2A) or test substances that should be classified as mildly irritating to eyes (UN GHS Category 2B) cannot be made.
For this purpose further testing with another suitable method is required.

Irritation parameter:

in vitro irritation score

Remarks:

mean

Run / experiment:

test item

Value:

46.32

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

0.81

Positive controls validity:

valid

Remarks:

63.38

Other effects / acceptance of results:

No prediction can be made regarding the classification of the test substance according to the evaluation criteria.

Interpretation of results:

study cannot be used for classification

Conclusions:

No prediction can be made regarding the classification of the test substance Ethanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine according to the evaluation criteria.

Executive summary:

Summary Results

The eye irritancy potential of Ethanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine was investigated in the bovine corneal opacity and permeability assay.

Preparation of the test item:  tested as provided by the sponsor

Visual Observation after treatment:  All 3 corneas treated with Ethanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine showed opacity of the tissue.

Mean in vitro irritation score:                    46.32

 

		UN GHS No Category
x		No prediction can be made
		UN GHS Category 1

Classification                                              

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.

The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.

Conclusion

No prediction can be made regarding the classification of the test substanceEthanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine according to the evaluation criteria.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Skin Irritation

The in vitro testing of Ethanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine according to OECD 431 and OECD 439 showed no irritational potential on the skin.

Eye Irritation

The eye irritancy potential of Ethanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine was investigated in the bovine corneal opacity and permeability assay.

All 3 corneas treated with Ethanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine showed opacity of the tissue. Furthermore, the permeability score of the tissues was also significantly increased. An IVIS of 46.32 was determined which is close to the cut-off for classification as severely eye damaging of 55.1.

Due to the severe corneal effects observed, Ethanol, 2,2’[1,2-ethanediylbis(oxy)] bis-, reaction products with 3-(triethoxysilyl)-1-propanamine is classified as severely damaging to eyes.