1-Propanamine, 3-(C9-11-isoalkyloxy) derivs., C10-rich, acetates

Endpoint summary

Description of key information

Etheramine C13i was found to be corrosive in BCOP test, and corrosive to rabbit skin following 3 minutes exposure. Based on read-across similar results can be expected for Etheramine C10i-acetate.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

03 August 2010 to 18 August 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
Available data from Etheramine C13i are considered to be applicable for C10i-etheramine as well. The structure of Etheramine C13i only differs from that of Etheramine C10i, in that the alkyl chain is only 2 carbons longer. This only has limited effects on physical properties, whereas chemical behaviour, reactivity and metabolism are the same. With alkylamines in general, the corrosive properties tend to increase with shortening of the alkyl chain. Therefore the results obtained from Etheramine C13i are therefore also expected to be of similar severity for Etheramine C10i, but certainly not less severe. (See for more information endpoint summary Ch.7.1 Toxicokinetics)

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000, including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

- Source: Harlan, Belton, Leics, England
- Age at study initiation: at least 6 weeks old.
- Weight at study initiation: at least 1.0 kg.
- Housing: Animals were individually housed in labeled cages with perforated floors (Ebeco, Germany, dimensions 67 x 62 x 55 cm) and shelters (Ebeco, Germany, dimensions 40 x 32 x 23 cm).
- Diet (e.g. ad libitum): Pelleted diet for rabbits (Global Diet 2030 from Harlan Teklad®, Mucedola, Milanese, Italy) approximately 100 grams per day. Hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least three times a week.
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: The acclimatization period was at least 5 days before the start of treatment under laboratory conditions.

A health inspection was performed prior to treatment, to ensure that the animals were in a good state of health. Special attention was paid to the skin to be treated, which was intact and free from abnormalities.

Results of analysis for diet (nutrients and contaminants), sawdust, paper and water were assessed and did not reveal any findings that were considered to have affected the study integrity. All certificates and results of analysis are retained in the NOTOX archives.

Animal specifications (sex, age and body weight) are specified in the attached table.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 – 20.3ºC
- Humidity (%): 50 - 82%
Deviations from the maximum level of relative humidity occurred. Laboratory historical data do not indicate an effect of the deviations.
- Air changes (per hr): approximately 15 air changes per hour- Photoperiod (hrs dark / hrs light): 12 hours artificial
fluorescent light and 12 hours darkness per day.

IN-LIFE DATES: From: 03 August 2010 to 18 August 2010

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

other: Adjacent areas of the untreated skin of each animal served as controls.

Amount / concentration applied:

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

Duration of treatment / exposure:

Single application.

Observation period:

14 days.

Number of animals:

1 male.

Details on study design:

STUDY DESIGN
The study was performed in a stepwise manner and started with the treatment of one animal (sentinel) with a stepwise exposure regime. Based on the severity of the skin reactions, no further animals were exposed.

TEST SUBSTANCE PREPARATION
The test substance was applied as delivered by the sponsor.

TEST SITE
Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10x15 cm). To facilitate scoring, treated skin areas were re-clipped at least 3 hours before the observations.

The test substance was applied to the skin of one flank, using a metalline patch# of 2x3 cm. The patch was mounted on Micropore tape#, which was wrapped around the abdomen and secured with Coban elastic bandage#.

REMOVAL OF TEST SUBSTANCE
Four hours after the application, the dressing was removed and the skin cleaned of residual test substance using tap water.

After the final observation, the animal was sacrificed by intra-venous injection of Euthasol® 20% (AST Farma BV, Oudewater, The Netherlands).

OBSERVATIONS
- Mortality/Viability: Twice daily.
- Toxicity: At least once daily.
- Body Weight: Day of treatment (prior to application) and after the final observation.
- Necropsy: The treated animal was subjected to necropsy and a description of all internal macroscopic abnormalities was recorded.
- Irritation:
The skin reactions were assessed at approximately 1, 24, 48 and 72 hours and 7 and 14 days after the removal of the dressings and test substance. The irritation scores and a description of all other (local) effects were recorded.

SCORING SYSTEM:
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

Erythema and eschar formation:
0: No erythema
1: Very slight erythema (barely perceptible)
2: Well-defined erythema
3: Moderate to severe erythema
4: Severe erythema (beet redness) *
*. Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the maximum grade for erythema (= 4) was given.

Oedema formation:
0: No oedema
1; Very slight oedema (barely perceptible)
2: Slight oedema (edges of area well-defined by definite raising)
3: Moderate oedema (raised approximately 1 millimeter)
4: Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure)

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hours, 7 and 14 days

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

(mean)

Time point:

other: 24, 48 and 72 hours, 7 and 14 days

Score:

4

Max. score:

4

Reversibility:

not reversible

Irritant / corrosive response data:

A 3-minute, 1- and 4-hour exposure to 0.5 mL of 3-(Isotridecyloxy)-1-propane amine resulted in erythema and oedema in the treated skin areas. Up to very slight erythema and oedema was observed on Day 1, progressing into severe erythema and oedema from 24 hours after exposure on all treated skin sites. The skin irritation remained present up the end of the observation period (14 days after exposure) on all treated skin sites. Reduced flexibility of the skin was observed between 24 hours and 7 days after exposure on all treated skin sites. A dry wound was visible at 7 days after exposure on the 3-minute skin site, and at 72 hours and 7 days after exposure on the 1- and 4-hour skin site. A bald, noticeable white skin (indicating full thickness destruction of the skin), and superficial red-brown eschar formation was observed at 14-days after exposure on all treated skin sites.

Tables containing individual and mean irritation scores are specified in the attached table.

Other effects:

No staining of the treated skin by the test substance was observed, and no remnants of the test substance were observed on the skin on Day 1.

No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Remarks:

Migrated information

Conclusions:

3-(Isotridecyloxy)-1-propane amine is corrosive to the rabbit skin following 3 minutes exposure.

Executive summary:

Primary skin irritation/corrosion study with 3-(Isotridecyloxy)-1-propane amine in the rabbit (semi-occlusive application) according to OECD 404.

One rabbit was exposed to three samples of 0.5 mL of 3-(Isotridecyloxy)-1-propane amine applied to separate skin-sites on intact, clipped skin using a semi-occlusive dressing. The exposure periods were 3 minutes, 1 hour and 4 hours, respectively. Skin reactions were assessed at least once daily for 4 days after treatment and 7 and 14 days after exposure. Based on the severity of the skin reactions, no further animals were exposed.

Exposure to 3-(Isotridecyloxy)-1-propane amine resulted in erythema and oedema in the treated skin areas. Up to very slight erythema and oedema was observed on Day 1, progressing into severe erythema and oedema from 24 hours after exposure on all treated skin sites. The skin irritation remained present up the end of the observation period (14 days after exposure) on all treated skin sites. Reduced flexibility of the skin was observed between 24 hours and 7 days after exposure on all treated skin sites. A dry wound was visible at 7 days after exposure on the 3-minute skin site, and at 72 hours and 7 days after exposure on the 1- and 4-hour skin site. A bald, noticeable white skin (indicating full thickness destruction of the skin), and superficial redbrown eschar formation was observed at 14-days after exposure on all treated skin sites.

The bald, noticeable white skin observed at 14-days after exposure on all treated skin sites was indicative of full thickness destruction of the skin, and indicated that corrosion of the skin had occurred by dermal application of 3-(Isotridecyloxy)-1-propane amine to the intact rabbit skin.

 

Based on these results and according to the:

-Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations (2007), 3-(Isotridecyloxy)-1-propane amine should be classified as : skin corrosive (Category 1B).

- Regulation (EC) No 1272/2008 on classification, labeling and packaging of substances and mixtures, 3-(Isotridecyloxy)-1-propane amine should be classified as Corrosive (Category 1B) and labeled as H314: Causes severe skin burns and eye damage.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

06-Jul-2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles, but as surfactant should have been probably best been tested as 10% solution.

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH
Available data from Etheramine C13i are considered to be applicable for C10i-etheramine as well. The structure of Etheramine C13i only differs from that of Etheramine C10i, in that the alkyl chain is only 2 carbons longer. This only has limited effects on physical properties, whereas chemical behaviour, reactivity and metabolism are the same. With alkylamines in general, the corrosive properties tend to increase with shortening of the alkyl chain. Therefore the results obtained from Etheramine C13i are therefore also expected to be of similar severity for Etheramine C10i, but certainly not less severe. (See for more information endpoint summary Ch.7.1 Toxicokinetics)

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: OECD guideline 437 “Bovine corneal opacity and permeability (BCOP) test method for identifying ocular corrosives and severe irritants”

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Vehicle:

physiological saline

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µl per cornea

NEGATIVE CONTROL
- Amount(s) applied (volume or weight with unit): 750 µl of physiological saline per cornea

POSITIVE CONTROL
Amount(s) applied (volume or weight with unit): 750 µl per cornea
Concentration (if solution): 10% (w/v) Benzalkonium Chloride

Duration of treatment / exposure:

- Exposure: 10 minutes
- Post incubation period: 120 minutes

Details on study design:

TEST SITE
- Isolated bovine cornea; triplicate per treatment.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes
- Time after start of exposure: 10 minutes

SCORING SYSTEM:
- After the 10 minutes exposure and washing step, the corneas were incubated for 120 +/- 10 minutes at 32 °C. After the completion of the incubation period opacity determination was performed.
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:

In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).


TOOL USED TO ASSESS SCORE:
- opacitymeter and microplate reader

DATA EVALUATION:
A test substance that induces an IVIS ≥ 55.1 is defined as a corrosive or severe irritant

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean

Value:

82

Negative controls validity:

valid

Positive controls validity:

valid

The corneas were turbid after the 10 minutes of treatment with 3-(Isotridecyloxy)-1-propane amine

Interpretation of results:

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

Conclusions:

3-(Isotridecyloxy)-1-propane amine is severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in the report.

Executive summary:

This report describes the ocular irritation properties of 3-(lsotridecyloxy)-1-propane amine on an isolated bovine cornea. The possible ocular irritancy of 3-(lsotridecyloxy)-1-propane amine was tested through topical application for 10±1 minutes. The study procedures described in this report were based on the most recent OECD guideline.

 

3-(lsotridecyloxy)-1-propane amine was a clear colourless liquid with a purity of 95.8%m/m(NMR). The test substance was applied as it is (750 µL) directly on top of the corneas.

The negative control responses of the opacity and permeability values were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (10%(w/v) Benzalkonium Chloride) was 138 and was within the historical positive control data range. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

 

3-(lsotridecyloxy)-1-propane amine induced severe ocular irritation through both endpoints, resulting in a meanin vitroirritancy score of 82 after 10 minutes of treatment.

 

Since 3-(lsotridecyloxy)-1-propane amine induced an IVIS ≥ 55.1, it is concluded that 3-(lsotridecyloxy)-1-propane amine is corrosive or severe irritant in the Bovine Corneal Opacity and Permeability test under the experimental conditions described in this report.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

No data is available on 1-Propanamine, 3-((C9-11-iso-,C10-rich)alkyloxy)-acetate (Etheramine C10i-Acetate). The substance Etheramine C10i-Acetate is produced by neutralising Etheramine C10i with acetic acid.

Etheramine C10i is a strong base with a pKa of about 9.9. When Etheramine C10i-Acetate dissolves in water or physiological media, the solid structure of the salt is broken up by water molecules surrounding the charged particles namely, the positive Etheramine C10i and negative acetate ions.

In physiological circumstances the nitrogen of Etheramine C10i is positively charged, resulting to a cationic surfactant structure, which results to high adsorptive properties to negatively charged surfaces as cellular membranes. The apolar tails easily dissolve in the membranes, whereas the polar head causes disruption and leakage of the membranes leading to cell damage or lysis of the cell content. As a consequence, the whole molecule will not easily pass membrane structures. Cytotoxicity at the local site of contact through disruption of cell membrane will is considered the most prominent mechanism of action for toxic effects. This is a mechanism that is not specifically pH dependent. This is supported with the observation from repeated dose studies of local irritating effects in the stomach at relatively low dose levels where the acidic environment is supposed to have a neutralizing effect. 

 

Etheramines are severe corrosive substances (see below). In in vivo studies effects of corrosion occur following exposure of only 3 minutes. However, contrary in cases where such effects would quickly appear in case they were pH dependant, the corrosive effects developed within the course of a week.

So, although the change of the pH by adding acetate will possibly have a mitigating effect on the severe corrosive properties of etheramines, it can still be expected to be a severe irritant, even likely corrosive.

Further in vitro testing has not been performed as available data shows that in vitro testing using reconstructed skin are not a suitable test system for etheramines, and additional in vivo testing was not considered ethical in view of the expected corrosive properties

Consequently, lacking further information, classification of Etheramine C10i-Acetate as corrosive to the skin is considered the most responsible conclusion.

 

Available study data:

Skin irritation / corrosion:

Data is available on Etheramine C13i that is considered to be applicable for C10i-etheramine as well. The structure of Etheramine C13i only differs from that of Etheramine C10i, in that the alkyl chain is only 2 carbons longer. This only has limited effects on physical properties, whereas chemical behaviour, reactivity and metabolism are the same. With alkylamines in general, the corrosive properties tend to increase with shortening of the alkyl chain. Therefore the results obtained from Etheramine C13i are therefore also expected to be of similar severity for Etheramine C10i, but certainly not less severe.

 

Etheramine C13i was tested in an in vitro skin irritation test using a reconstructed human epidermis model (EU method B.46). However, as the substance caused a non-specific MTT reduction > 30% (39% & 41%), it was conclude that this method is not a suitable test method for C13i-etheramine.

It can be expected that Etheramine C10i will show a similar chemical reactivity towards MTT, and similar as acetate salt.

 

Results from a performed BCOP study indicated that severe effects could be expected following dermal exposure, but as the in vitro dermal irritation study was not technical feasible, an in vivo study was designed specifically to minimize the risk for animal harm. The study was performed in a stepwise manner and started with the treatment of one animal with a stepwise exposure regime.

One rabbit was exposed to three samples of 0.5 mL of Etheramine C13i applied to separate skin-sites on intact, clipped skin using a semi-occlusive dressing. The exposure periods were 3 minutes, 1 hour and 4 hours, respectively. Skin reactions were assessed at least once daily for 4 days after treatment and 7 and 14 days after exposure. Based on the severity of the skin reactions, no further animals were exposed.

All exposures resulted up to very slight erythema and oedema in the treated skin areas on day 1, progressing into severe erythema and oedema from 24 hours after exposure on all treated skin sites. The skin irritation remained present up the end of the 14-day observation period and was accompanied by reduced flexibility of the skin between 24 hours and 7 days. A dry wound as well as a bald, noticeable white skin (indicating full thickness destruction of the skin), and superficial redbrown eschar formation was observed at 14-days after exposure on all treated skin sites.

This in vivo dermal corrosion study in rabbits indicates that Etheramine C13i causes corrosive effects of the skin following 3 minutes exposure, which developed with the course of a week.

 

Eye irritation:

Etheramine C13i induced severe ocular irritation on isolated bovine cornea through, resulting in a mean in vitro irritancy score of 82 after 10 minutes of treatment. As the IVIS is above 55.1, it this means that Etheramine C13i is corrosive or severe irritant in the Bovine Corneal Opacity and Permeability test. One comment could be that this BCOP study was performed using the neat substance, whereas Etheramine C13i being a surfactant, should probably best been tested as 10% solution. However, in view of the corrosive properties to skin this is of minor importance.

 

Etheramine C10i can be expected to have similar effects on skin and eyes as Etheramine C13i, and in vivo testing with corrosive substances should best be avoided.

Justification for classification or non-classification

The in vivo dermal corrosion study in rabbits indicates that Etheramine C13i causes corrosive effects of the skin following 3 minutes exposure, which developed with the course of a week. Consequently, GHS classification 1B is appropriate.

Results for Etheramine C10i can be expected to be similar, and in vivo testing should best be avoided for such corrosive substances. As both the delayed development o fthe effects and the mechanism of action do not suggest this to be pH mediated, the same is expected for Etheramine-C10i-acetate. Classification of Etheramine C10i-acetate is therefore proposed to be the same: GHS classification skin corrosion 1B, with hazard statement H314: Causes severe skin burns and eye damage

 

There is no information is available following exposure via inhalation. However, with a vapour pressure of 0.72 Pa at 20 °C, and for an acetate salt probably much lower, the potential for inhalation of vapours is limited. Inhalation of aerosols may cause respiratory irritation due to the corrosive properties of the substance.