Ethyldiisopropylamine

Endpoint summary

Description of key information

EDIPA is moderately irritating to rabbit skin, severely irritating to 
rabbit eyes and irritating to the respiratory tract in rats.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

end 2004-01-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

The temperature and relative humidity recorded in the animal room were somtetimes outside of the target ranges specified in the Study plan. The test item was not protected from light.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CEVAG, Saint Mars d'Egrenne, France
- Age at study initiation: 2 to 4 month old
- Weight at study initiation: 3.1+/-0.1kg
- Housing: indidually in polystyrene cages (48.2x58x36.5cm)
- Diet (e.g. ad libitum): ad libitum 110 pellet diet (SAFE, Villemoisson, Epinay-sur-Orge, France)
- Water (e.g. ad libitum): ad libitum water filtered by a FG Miliipore membrane (0.22µ)
- Acclimation period:at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18+/-3°C
- Humidity (%): 30-70%
- Air changes (per hr): approximately 12 cycles / hour
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: To: 2003-08-12

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: auto-control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5mL

Duration of treatment / exposure:

3 minutes and 4 hours

Observation period:

1 hour, 24, 48, 78 hours after removal of the dressing then daily until D15.

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: no data
- Type of wrap if used: dry gauze pad held in contact by an adhesive hypoallergenic aerated semi occlusive dressing and a restraining bandage.

SCORING SYSTEM: according to the OECD N°404 Guideline scoring scale

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 11 days

Remarks on result:

other: 3min of exposure

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: 3-min exposure

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 11 days

Remarks on result:

other: 4-hour exposure

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: 4-hour exposure

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 13 days

Remarks on result:

other: 4-hour exposure

Irritation parameter:

edema score

Basis:

animal: #1, 2 and 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: 4-hour exposure

Irritant / corrosive response data:

After a 3-minute exposure (one animal), a very slight or slight erythema (grade 1 or 2) was observed from day 1 up to day 14. Dryness of the skin was noted from day 6 up to the end of the observation period (day 15).
After a 4-hour exposure (three animals), a very slight or slight erythema (grade 1 or 2) was recorded in all animals from day 1 up to day 10 or 12. A slight oedema (grade 2) was noted in 1/3 animals on days 5 and 6. Dryness of the skin, sometimes together with crusts in 1/3 animals, was also observed in 2/3 animals between days 5 and 10 or between days 6 and 12.

3 min exposure		1h	24h	48h	72h	D5	D6	D7	D8	D9	D10	D11	D12	D13	D14	D15
Erythema	951	1	1	2	2	2	2	2	2	2	2	1	1	1	1	0
Oedema	951	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
Other	951	*	*	*	*	*	S	S	S	S	S	S	S	S	S	S

* = none

S = dryness of the skin

 

4 h exposure		1h	24h	48h	72h	D5	D6	D7	D8	D9	D10	D11	D12	D13	D14	D15
Erythema	951	1	1	2	2	2	1	1	1	1	1	0	-	-
	926	2	2	2	2	2	2	2	1	1	1	1	1	0
	927	2	2	2	2	2	2	2	2	2	2	1	1	0
Oedema	951	0	0	0	0	0	0	0	0	0	0	0	-	-
	926	0	0	0	0	2	2	0	0	0	0	0	0	0
	927	0	0	0	0	0	0	0	0	0	0	0	0	0
Other	951	*	*	*	*	*	*	*	*	*	*	*	-	-
	926	*	*	*	*	S	S	S	S	S	S	*	*	*
	927	*	*	*	*	*	S	S	S/A	S/A	S/A	S	S	*

 

* = none

S = dryness of the skin

A = crust

- = cutaneous examination performed

Interpretation of results:

moderately irritating

Remarks:

Migrated information not classified according to CLP, R38 according to DSD Criteria used for interpretation of results: EU

Conclusions:

Mean scores over 24, 48 and 72 hours for each animal were 1.7, 2.0 and 2.0 for erythema and 0.0, 0.0 and 0.0 for oedema.

Executive summary:

The dermal irritation of Ethyldiisopropylamine (EDIPA) was evaluated in male New Zealand White rabbits according to OECD N°404 Guideline.

0,5mL of EDIPA was applied during 4 hours in 3 rabbits and during 3 minutes in 1 rabbit on clipped area. The cutaneous reaction was observed one hour, 24, 48, 72 hours after removal of the dressing and then daily to day 15.

After a 3-minute exposure of 0.5mL of EDIPA (one animal), a very slight or slight erythema was observed from day 1 up to day 14. Dryness of the skin was noted from day 6 up to the end of the observation period (day 15). After a 4-hour exposure (three animals), a very slight or slight erythema was recorded in all animals from day 1 up to day 10 or 12. A slight oedema was noted in 1/3 animal on days 5 and 6. Dryness of the skin, sometimes together with crusts in 1/3 animal, was also observed in 2/3 animals between days 5 and 10 or between days 6 and 12.

Mean scores over 24, 48 and 72 hours for each animal were 1.7, 2.0 and 2.0 for erythema and 0.0, 0.0 and 0.0 for oedema.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (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:

12 March 2012 - 11 May 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guidelines; adequate consistence between data, comments and conclusions.

Qualifier:

according to guideline

Guideline:

other: EU method B.47 (In vitro eye irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: OECD guideline 437 (In vitro eye irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Details on test animals or tissues and environmental conditions:

Origin: bovine eyes were obtained from freshly slaughtered cattle at the abattoir (SOCAVIA, Cany Barville - France, SOCAVIA, Beuvillers - France or EVA, Saint-Pierre-sur-Dives - France).
Age: bovine cattle were up to 12 months old.
Reason for choice: bovine corneas are recommended by regulatory authorities for this type of study. They are adapted for the evaluation of potential ocular irritants since they are part of the target organ.
Transport from supplier to CIT: the eyes were transported to CIT at ambient temperature, immerged in buffered Hanks medium containing an antibiotic [Hank’s Balanced Salts Solution (HBSS) plus penicillin/streptomycin (100 units/100 µg/mL final)]. A container with smooth internal surfaces was used for the transport to avoid damage to the corneas.
Origin: bovine eyes were obtained from freshly slaughtered cattle at the abattoir (EVA, Saint-Pierre-sur-Dives - France).
Age: bovine cattle were up to 12 months old.
Reason for choice: bovine corneas are recommended by regulatory authorities for this type of study. They are adapted for the evaluation of potential ocular irritants since they are part of the target organ.
Transport from supplier to CIT: the eyes were transported to CIT at ambient temperature, immerged in buffered Hanks medium containing an antibiotic [Hank’s Balanced Salts Solution (HBSS) plus penicillin/streptomycin (100 units/100 µg/mL final)]. A container with smooth internal surfaces was used for the transport to avoid damage to the corneas.
Selection: a careful macroscopic examination was performed on all eyes to detect the presence of any defects (opacity, scratches, pigmentation, neovascularization, etc). Any eyes with defects were discarded. The examination was performed under a lamp, using HBSS in order to keep the eyes moistened. Particular attention was paid to the corneas and the eyes were swiveled in order to observe the fringe areas and any scratches directly under the light.
Preparation of the selected corneas: the tissues surrounding the eyeball were carefully pulled away and the cornea, surrounded by approximately 2 to 3 mm of sclera, was dissected out. The isolated corneas were stored in HBSS until all corneas had been prepared.
Washing of the corneas: the corneas were washed for 15 minutes, three times, in HBSS plus penicillin/streptomycin (100 units/100 µg/mL final) at room temperature. The corneas were used immediately or within a maximum of 24 hours.

The prepared corneas were stored and used within 24 hours.

Dates of experimental phase: from 07 May 2012 to 11 May 2012.

Vehicle:

unchanged (no vehicle)

Controls:

other: in vitro negative and positive controls

Duration of treatment / exposure:

Exposure period of 10 minutes, followed by rinsing.

Observation period (in vivo):

Opacity measurement:
- before treatment,
- after 2-hour incubation in water.

Permeability measurement:
- after 90-min incubation in water (and other procedures), following the 2nd opacity measurement

Number of animals or in vitro replicates:

Not applicable
Triplicate corneas for each timepoint and tested substance (test item, negative control, positive control)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Rinsing: the anterior part of the eye was emptied and then rinsed 3 times with cMEM.

NEGATIVE CONTROL:
0.9% NaCl.

POSITIVE CONTROL:
10% sodium hydroxide solution (10% NaOH).

SCORING SYSTEM/TOOL
- Opacity:
Using an opacitometer
The average change in opacity during exposure is determined. It is corrected by subtracting the average negative control value from values in positive control and test item.
- Permeability:
Using a spectrophotometer: optical density (OD) at 490 nm wavelength
The optical density is corrected by subtracting the average negative control value from values in positive control and test item.
- Scoring:
In vitro irritancy score (IVIS) = Corrected Opacity + (15 x Corrected OD)

Interpretation: see below

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

66.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

In vitro, the test item was corrosive or severe irritant to the eye.

Executive summary:

The objective of this study was to evaluate the potential corrosive or severe irritant properties of ethyldiisopropylamine on bovine eyes. The study was performed according to the guideline OECD 437, in compliance with CIT standard operating procedures and with the principles of Good Laboratory Practices. Corneas obtained from freshly slaughtered calves were mounted in corneal holders. Both chambers of the corneal holder were filled with complemented MEM culture media (cMEM) and pre-incubated for 1 hour at 32°C. Three corneas were used for each treated series (test item, positive control and vehicle control). Before the treatment, a first opacity measurement was performed using an opacitometer. The test item was tested at the concentrations of 100%, in a single experiment using a treatment time of 10 minutes. At the completion of the treatment period, the test item was removed from the front opening of the anterior chamber and the epithelium was rinsed. The corneas were then incubated for 2 hours at 32°C before a second opacity measurement was performed. After the second opacity measurement, the medium of the anterior chamber was removed and filled with a fluoresceine solution. The holders were then incubated vertically for 90 minutes at 32°C. At the end of the incubation, the optical density of the solution from the posterior chamber of each holder was measured in order to determine the permeability of the cornea. Then the cornea was observed for opaque spots and other irregularities.

No notable opaque spots or irregularities were observed on negative control corneas following the treatment. Opacity and fluoresceine fixation were observed on the corneas treated with the test item following treatment. The In Vitro Irritancy Score (IVIS) was: 66.6. As the test item induced an IVIS = 55.1, it was considered as ocular corrosive/severe irritant.

Ethyldiisopropylamine is considered to be corrosive or severely irritant to the eye.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

Skin irritation

 

The dermal irritation of Ethyldiisopropylamine (EDIPA) was evaluated in male New Zealand White rabbits according to OECD N°404 Guideline (Klein 2004). After a 3-minute exposure of 0.5mL of EDIPA (one animal), a very slight or slight erythema was observed from day 1 up to day 14. Dryness of the skin was noted from day 6 up to the end of the observation period (day 15). After a 4-hour exposure (three animals), a very slight or slight erythema was recorded in all animals from day 1 up to day 10 or 12. A slight oedema was noted in 1/3 animal on days 5 and 6. Dryness of the skin, sometimes together with crusts in 1/3 animal, was also observed in 2/3 animals between days 5 and 10 or between days 6 and 12. Mean scores over 24, 48 and 72 hours for each animal were 1.7, 2.0 and 2.0 for erythema and 0.0, 0.0 and 0.0 for oedema. Therefore, the test item EDIPA is classified as mild irritant when applied topically.

 

Eye irritation

 

The objective of this study was to evaluate the potential corrosive or severe irritant properties of ethyldiisopropylamine on bovine eyes. The study was performed according to the guideline OECD 437, in compliance with CIT standard operating procedures and with the principles of Good Laboratory Practices. Corneas obtained from freshly slaughtered calves were mounted in corneal holders. Both chambers of the corneal holder were filled with complemented MEM culture media (cMEM) and pre-incubated for 1 hour at 32°C.Three corneas were used for each treated series (test item, positive control and vehicle control).Before the treatment, a first opacity measurement was performed using an opacitometer. The test item was tested at the concentrations of 100%, in a single experiment using a treatment time of 10 minutes. At the completion of the treatment period, the test item was removed from the front opening of the anterior chamber and the epithelium was rinsed. The corneas were then incubated for 2 hours at 32°C before a second opacity measurement was performed. After the second opacity measurement, the medium of the anterior chamber was removed and filled with a fluoresceine solution. The holders were then incubated vertically for 90 minutes at 32°C.At the end of the incubation, the optical density of the solution from the posterior chamber of each holder was measured in order to determine the permeability of the cornea. Then the cornea was observed for opaque spots and other irregularities. No notable opaque spots or irregularities were observed on negative control corneas following the treatment. Opacity and fluoresceine fixation were observed on the corneas treated with the test item following treatment. The In Vitro Irritancy Score (IVIS) was: 66.6.As the test item induced an IVIS = 55.1, it was considered as ocular corrosive/severe irritant.

 

Respiratory tract irritation

Clinical signs of respiratory tract irritation were observed in rat exposed for 4 hours to concentrations of 0.48 mg ethyldiisopropylamine/L and upwards (Nagy, 2013).

Justification for selection of skin irritation / corrosion endpoint:
Key study

Justification for selection of eye irritation endpoint:
Key study

Effects on skin irritation/corrosion: moderately irritating

Effects on eye irritation: highly irritating

Effects on respiratory irritation: irritating

Justification for classification or non-classification

Skin irritation:

In the skin irritation assay in rabbits, mean scores over 24, 48 and 72 hours for each animal were 1.7, 2.0 and 2.0 for erythema and 0.0, 0.0 and 0.0 for oedema. On this basis, EDIPA does not merit classification as a skin irritant in accordance with Regulation (EC) No 1272/2008, but warrant a classification H316 according to GHS. 

Eye irritation:

In a BCOP assay, EDIPA is considered to be corrosive or severely irritant to the eye. On this basis, EDIPA is classified as eye irritant category 1 (H318 Causes serious eye Damage) in accordance with Regulation (EC) No 1272/2008.

 

Respiratory tract irritation:

EDIPA induced clinical signs of respiratory tract irritation in rat in an acute inhalation toxicity study. On this basis, EDIPA is classified as STOT-SE cat. 3 (H335 May cause respiratory irritation) in accordance with Regulation (EC) No 1272/2008.