Registration Dossier

Administrative data

Description of key information

Skin irritation: Three reliable studies were available for this endpoint (Two Klimisch 1, one Klimisch 2 study). The recent Huntsman study (Vasquez, 2012) was considered as the key study for this endpoint and indicated that the substance should be classified as a mild irritant (CLP Cat 3). The substance was observed to be slightly irritating in the DOT Corrosivity test in rabbits (Mallory VT, 1982) and non-corrosive in the Epiderm Test from BASF (2003).

Eye irritation: One reliable study was available for the eye irritation endpoint (Klimisch 1 study from BASF, 2002). The test substance was found to be irritating in the HET-CAM in vitro corrosion test (equivalent to OECD 405).

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-02-28 to 2012-03-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 1L708

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temp (21-26°C)

OTHER SPECIFICS:
- Analytical purity: Certificate of Analysis was provided by the Sponsor. No additional analysis was performed.
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Millbrook Breeding Labs
- Age at study initiation: 19 weeks
- Weight at study initiation: 3.3 - 3.5 kg
- Housing: individually
- Diet (e.g. ad libitum): access to PMI certified Hi Fiber Rabbit diet
- Water (e.g. ad libitum): ad libitum via an automatic watering device
- Acclimation period: minimum of 5 days prior dosing

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 22 °C
- Humidity (%): 36 - 63 %
- Photoperiod (hrs dark / hrs light): 12h light / 12h dark

IN-LIFE DATES: From: 28 Feb 2012 To: 9 Mar 2012
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
Duration of treatment / exposure:
3 minutes, 1 hour, 4 hours
Observation period:
10 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: 5 cm x 5 cm
- Type of wrap if used: gauze patch

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test article was removed using water and gauze

SCORING SYSTEM: Draize
Irritation parameter:
primary dermal irritation index (PDII)
Basis:
mean
Time point:
other: 30-60 min, 24h, 48h, 72h after exposure
Score:
1.7
Max. score:
8
Reversibility:
fully reversible within: 10 days
Remarks on result:
other: after 4h exposure
Irritation parameter:
erythema score
Basis:
animal: 646M
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 10 days
Remarks on result:
other: after 4 h treatment
Irritation parameter:
erythema score
Basis:
animal: 647M
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 9 days
Remarks on result:
other: after 4h treatment
Irritation parameter:
erythema score
Basis:
animal: 648M
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 9 days
Remarks on result:
other: after 4h treatment
Irritation parameter:
edema score
Basis:
animal: 646M
Time point:
24/48/72 h
Score:
0.67
Max. score:
4
Reversibility:
fully reversible within: 10 days
Remarks on result:
other: after 4h treatment
Irritation parameter:
edema score
Basis:
animal: 647M
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: after 4h treatment
Irritation parameter:
edema score
Basis:
animal: 648M
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
other: after 4h treatment
Irritant / corrosive response data:
After 3 min exposure, the following is observed: very slight erythema without edema observed starting on day 3. Flaking of the skin observed on day 7. After 60 min exposure, the following is observed: well-defined erythema without edema observed starting on day 3. Flaking of the skin observed starting on day 8. After 60 min exposure, the following is observed: well-defined erythema and/or very slight edema observed starting at 24h post unwrap. Flaking of the skin observed starting on day 6.
Other effects:
No mortality is observed.
No clinical signs were recorded.
Interpretation of results:
Category 3 (mild irritant) based on GHS criteria
Conclusions:
The substance is found to be a mild irritant (cat 3). The primary irritation index was 1.7.
Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2002-01-23 to 2002-06-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
OECD 431 (Draft) "In vitro Skin Corrosion: Human Skin Model Test" 2002
Principles of method if other than guideline:
The potential of the test substance to cause dermal corrosion was assessed by a single topical application of 50 µl of the test substance to a reconstructed three dimensional human epidermis model (EpiDerm).
GLP compliance:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Abl. Nr. 85-4113
- Date of production: November 16, 2000

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: was guaranteed by the sponsor

OTHER SPECIFICS:
- Name of test material (as cited in study report): Polyetheramin T 403; Substance No.: 01/0618-1
- Analytical purity: 93.9 %
- Homogeneity: Homogeneous by visual inspection
Species:
other: EpiDerm TM Skin Corrosivity Test
Strain:
other: not applicable
Type of coverage:
other: not applicable
Preparation of test site:
other: not applicable
Vehicle:
unchanged (no vehicle)
Controls:
other: in vitro
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µl of the test substance
Duration of treatment / exposure:
3 min and 1 hour(s)
Observation period:
not applicable
Number of animals:
not applicable
Details on study design:
TEST SYSTEM
- The EpiDerm System consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues (surface 0.6 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs®, 10 mm QS) and shipped world-wide as kits (EpiDermTM 200), containing 24 tissues on shipping agarose.
- Tissue model: Epi-200
- Origin: MatTek Corporation, Ashland MA, USA


TEST PROCEDURE
Two tissues per exposure time (3 minutes at room temperature or 1 hour in the incubator, as a rule) and test group (test material, negative control and positive control (12 tissues per test) are used. If applicable, 1 killed tissue per exposure time is treated to control for direct MTT reduction.
The tissues are washed with PBS to remove residual test material 3 minutes or 1 hour after start of treatment. Rinsed tissues are kept in 24-well plates (holding plates) at room temperature on assay medium until all tissues per application time are dosed and rinsed. The assay medium is then replaced by MTT solution and tissues are incubated for 3 hours.
After incubation, tissues are washed with PBS and the formazan produced by the tissues is extracted with Isopropanol over night at room temperature. The optical density at a wavelength of 570 nm (OD570) of the extracts is determined spectrophotometrically. Blank values are established of 3 microtiter wells filled with Isopropanol for each microtiter plate.



CONTROLS
- Negative control (NC): Doubly distilled water
- Positive control (PC): 8 N potassium hydroxide (Sigma-Aldrich, Munich, Germany)


ASSAY ACCEPTANCE CRITERIA:
- Negative control (NC ): mean OD570 of the NC per exposure time > = 0.8
- Positive control (PC ): mean relative tissue viability of the 3 min positive control is <= 30 %
- Tissue variability: The inter tissue variability is considered to be acceptable if the difference of the OD570 values of
the two tissues is <= 0.3
- Killed controls (KC ): OD570 of the killed control tissues treated as negative control should be <= 0.35

Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minutes
Value:
ca. 94
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1h
Value:
ca. 62
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid

The test substance is able to directly reduce MTT. However, this ability of direct MTT reduction did not impair the study result as demonstrated by the concurrently performed exposure of control tissues inactivated by freezing.
Viability of the test substance treated tissues determined after an exposure period of 3 minutes was 94%. Viability of the test substance treated tissues determined after an exposure period of 1 hour was 62% .These results do not indicate a corrosive potential of the test substance.

 

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the observed results and applying the evaluation criteria cited in chapter 3.8 it was concluded, that the substance does not have a corrosive potential in the EpiDermTM skin corrosivity test under the test conditions chosen.
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
No data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
deviation in scoring time; occlusive coverage
GLP compliance:
not specified
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: # J-153

OTHER SPECIFICS:
- Name of test material (as cited in study report): 5601-8-20
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Perfection Breeders Douglassville, Pensylvania
- Age at study initiation: not relevant
- Weight at study initiation: 2-3 kg
- Housing: Rabbits were housed individually in cages sized in accordance with the ''Guide for the Care and Use of Labolatory Animals'' of the Institute of Labolatory Resources, National Research Council.
- Diet (e.g. ad libitum): ad libitum, checked daily and added or replaced as needed. Feeders were designed to reduce soiling,bridging, and scattering.
- Water (e.g. ad libitum):fresh tap water, fit for human consumption, ad libitum, using 16 ounce glass bottels with rubber stopper and stainless steel sipper tube or an automatic watering system supplied by Edstrom Industries Inc., Waterford, Wisconsin.
- Acclimation period:5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C±3°C
- Humidity (%): 30 to 70%
- Photoperiod (hrs dark / hrs light): 12 hours light, 12hours dark


IN-LIFE DATES: From: To:
Type of coverage:
occlusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
not specified
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5ml/site

Duration of treatment / exposure:
The test substance was kept in contact with the skin site for 4 hours.
Observation period:
44 hours
Number of animals:
Six (6)
Details on study design:
TEST SITE
- Area of exposure: trunk (clipped free of fur )
- % coverage:
- Type of wrap if used: rubber dam and an Ace bandage to retard evaporation.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): only wiped to remove remaining material
- Time after start of exposure: 4h


Irritation parameter:
erythema score
Basis:
other: all animals considered individually
Time point:
other: 4 and 44 hrs
Remarks on result:
other: No scoring system used; slight erythema visible at 4hrs after treatment
Irritation parameter:
edema score
Basis:
other: all animals considered individually
Time point:
other: 4 and 44 hrs
Remarks on result:
other: No scoring system used; no signs of edema visible during observation period
Irritant / corrosive response data:
observation after 4hrs: 3 out of 6 rabbits: slight erythema; 3/6 : no erythema
observation after 44hrs: 6/6 : no erythema
No signs of edema or necrosis were visible at either observation period.
Interpretation of results:
GHS criteria not met
Conclusions:
Based upon the results of the DOT Corrosivity Study in Rabbits, 5601-8-20, was not considered to be corrosive.
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:
No data
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
Due to animal welfare reasons, the potential of severe irritation is determined in the HET-CAM in vitro system before deciding on a possible study in the rabbit as suggested in OECD Guideline 405.
Deviations:
no
Remarks:
The HET-CAM Test is an alternative in vitro method for testing of severe eye/mucous membrane damage using the chorionallantoic membrane of fertilized, incubated hen eggs.
Principles of method if other than guideline:
The study was performed according to the methods described in the following publications:
- Lüpke N.P. (1985): Hen's Egg Chorioallantoic Membrane Test for Irritation Potential. Fd. Chem. Toxic. 23, pp. 287 -291.
- Spielmann, H. (1995): HET-CAM Test. In: Methods in Molecular Biology, 43 (eds.:O'Hare, S. and Atterwill, 0. K.) pp. 199 -204
- Spielmann, H. et al. (1996): Results of a Validation Study in Germany on Two In Vitro Alternatives to the Draize Eye Irritation Test the HET-CAM Test and the 3T3 NRU Cytotoxicity Test. ATLA 24, pp. 741 -858.
In addition the study follows the testing strategy tor determination of eye irritation/corrosion as given in the following guideline:
Organization for Economic Co-operation and Development (OECD), OECD Guidelinestor testing of chemicals, Guideline No. 405:
Acute Eye Irritation/Corrosion", adopted April 24, 2002.
GLP compliance:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: ABl. Nr. 85-4113
- Date of production: November 16, 2000

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: was guaranteed by the sponsor

OTHER SPECIFICS:
- Name of test material (as cited in study report): Polyetheramin T 403; Substance No.: 01/0618-1
- Analytical purity: 93.9 %
- Homogeneity: Homogeneous by visual inspection
Species:
other: HET-CAM in vitro corrosion test
Strain:
other: hen eggs
Vehicle:
water
Controls:
no
Amount / concentration applied:
10, 100 %
Duration of treatment / exposure:
up to a maximum time period of 3.5 minutes
Observation period (in vivo):
210 seconds
Number of animals or in vitro replicates:
3 eggs for 10 and 100 %
Details on study design:
Type of eggs: Fresh, fertilized hen eggs produced under controlled SPF conditions.
Strain/quality: White Leghorn, SPAFAS Inc., USA, SPF Premium.
Origin: Charles River Deutschland GmbH, Extertal.
Identification: At start of incubation period continuous numbering of the eggs with a felt pen.
Reasons for the selection: Hen eggs are recommended as preferred type of eggs in the quoted references. Analogous to test animals, controlled SPF housing guarantees a defined microbiological status of the eggs.
Climate: Breeding in an incubator at constant temperature of 37.5°C (± 0.5°C) and a relative humidity of 62.5% (± 7.5%).
Automatic rotating device: Until including incubation day 8 and/or day 9, the eggs were rotated automatically 5 times a day. On the day before application the eggs were placed with the blunt end upward and were not rotated until preparation. The incubation conditions were checked daily. Deviations were recorded.
Candling of the eggs: The eggs were candled before the start of incubation and on the 9th and/or 10th day. Any defective or unfertilized eggs were discarded.
Irritation parameter:
in vitro irritation score
Run / experiment:
egg no. 1
Value:
19.2
Irritation parameter:
in vitro irritation score
Run / experiment:
egg no. 2
Value:
19.1
Irritation parameter:
in vitro irritation score
Run / experiment:
egg no. 3
Value:
18.9
Irritation parameter:
in vitro irritation score
Run / experiment:
egg no. 1
Value:
19.9
Irritation parameter:
in vitro irritation score
Run / experiment:
egg no. 2
Value:
19.6
Irritation parameter:
in vitro irritation score
Run / experiment:
egg no. 3
Value:
19.4

Concentration

Egg No.

Time (seconds) until appearance of:

Index for Irritation

 

 

Haemorrhagia:

Lysis:

Coagulation:

 

Undiluted test substance

1

5.0

38.0

30.0

19.2

2

6.0

39.0

33.0

19.1

3

9.0

41.0

37.0

18.9

Mean

n= 3

6.7

39.3

33.3

19.0

Concentration

Egg No.

Time (seconds) until appearance of:

Index for Irritation

 

 

Haemorrhagia:

Lysis:

Coagulation:

 

Test substance 10% in doubly distilled water

1

8.0

22.0

17.0

19.9

2

13.0

25.0

21.0

19.6

3

19.0

29.0

23.0

19.4

Mean

n= 3

13.3

25.3

20.3

19.6

Evaluation: May cause severe eye damage.

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
Classification: category 1 (serious eye damage)
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The data used in the first draft of the IUCLID dossier for the test substance is of uneven quality and is often conflicting with regard to toxicity testing results for the same mammalian tox endpoints. To clarify the hazard profile for the test substance, follow-up toxicity studies were conducted in 2012 using contemporary OECD study protocols. These follow-up studies were designed with exposure parameters (dosage and/or duration) that are complementary to the UN GHS classification criteria. These studies also used a test sample of the substance which was well-characterized as to chemical identity and the presence of impurities. Due to the high quality of data obtained from these studies, they have been incorporated in the IUCLID dossier as our "key studies" for the CSR and CLP endpoints.

Skin irritation:

The test substance was found to be a mild irritant after exposure for resp. 3 minutes, 1 hour and 4 hours (Huntsman, 2012). The observed effects range from very slight erythema without edema on day 3 with flaking of the skin observed on day 7 (after 3 minutes of exposure) to well-defined erythema and/or very slight edema 24h after exposure with flaking of the skin observed on day 6 (after 4 hours of exposure).

The test substance was found to be non corrosive after 3 minutes and 1 hour exposure in the in vitro EpiDerm skin corrosivity test (BASF AG, 2003). Similar observations were made by Mallory et al (1982) after 4 hours of exposure: 3 out of 6 rabbits showed slight erythema, while the other 3 showed no erythema after 4h observation. 48h after exposure, all animals reverted to normal (no erythema).

In a Klimisch 4 study of Moreno (1978) the test substance was found to be corrosive. As only a qualitative assessment of individual animal data was performed instead of Draize scoring, the study could not be taken into account for the classification of this substance.

Finally, the primary skin irritation test in albino rabbits, performed according to OECD guideline 404, resulted in a Mean Primary Irritation Score of 7.2/8.0 (after 24h exposure to intact and abraded skin). The DOT corrosivity test in albino rabbits, described in the same study, resulted in a Mean Primary Irritation Score of 5.8/8.0 (after 4h exposure) and the test substance was considered to be non corrosive (Industrial Bio-Test Laboratories Inc., 1979). However, as the study was performed on a derivate of the substance and not on the substance itself, the study should be disregarded.

In addition, moderate to severe skin irritation is also observed in the 90 days repeated dose study in the mid (50 mg/kg) and high (160 mg/kg) dose groups. The skin irritation was reversible after discontinuation of treatment during the recovery period.

Based on the available information, the substance is considered to be a mild irritant to the skin according to UN GHS criteria, but not classified under EU CLP.

Eye irritation:

BASF (2002), performed an in vitro HET-CAM study according to OECD guideline 405, in order to investigate the eye damage potential in albino rabbits and observed that the substance may cause serious eye damage under the conditions of the test. According to the CLP criteria, the substance should be classified into category 1 for eye irritation (category 1: irreversible irritation to eye). Industrial Bio-Test Laboratories, Inc (1979), performed according to OECD guideline 405, investigated the eye irritation potential in albino rats and observed a maximum Mean Irritation Score of 89.6 /110. However, as the study was performed on a derivate of the substance and not on the substance itself, the study was disregarded.


Effects on skin irritation/corrosion: slightly irritating

Effects on eye irritation: irritating

Justification for classification or non-classification

- Based on the available data and according to the criteria laid down in UN GHS the test substance should be considered as a category 3 mild skin irritant substance. Based on the available data and according to the criteria laid down in the CLP Regulation (EC) 1272/2008, the substance doesn't need to be classified.

- Based on the available data and according to the criteria laid down in CLP Regulation (EC) 1272/2008 the test substance should be considered as a category 1 eye irritant (irreversible irritation to eye).