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Diss Factsheets

Administrative data

Description of key information

eye irritation:

- study conducted according to Luepke, N.P. (1985) Hen's egg chorio allantoic membrane test for irritation potential Fd. Chem. Toxic. 23, 287-291, HET-CAM, Lohmann Selected Leghorn chicken eggs were exposed to 126 to 175 mg of Partially unsaturated IQAC, DMS quarternised for 300 sec, haemorrhage, coagulation and lysis of the blood vessels were recorded, no irritating effects were observed during 5 min incubation with the test substance (no solvent). The calculated mean irritancy index is 0.00, not irritating, read-across, key-study.

- study conducted according to OECD guideline 405, GLP, 0.1 mL of Partially unsaturated IQAC, DMS quarternised were instilled into the conjunctival sac of the right eye of 3 New Zealand White rabbits for 24h, the eyes were washed with slaine solution thereafter and the animals further observed for 7 days, not irritating , read-across, key-study

- study conducted according to Luepke, N.P. (1985) Hen's egg chorio allantoic membrane test for irritation potential Fd. Chem. Toxic. 23, 287-291, HET-CAM, chicken eggs of the breed SPAFAS were exposed to 200 mg of Partially unsaturated IQAC, DMS quarternised as 87.5 % (w/w) preparation for 5 min, haemorrhage, coagulation and lysis of the blood vessels were recorded, no effect was detected after treatment with the test substance preparation (0.0 scores of total 21 possible scores), not irritating, read-across, supporting study

- study conducted according to Luepke, N.P. (1985) Hen's egg chorio allantoic membrane test for irritation potential Fd. Chem. Toxic. 23, 287-291, HET-CAM, chicken eggs of the breed SPAFAS were exposed to Partially unsaturated IQAC, DMS quaternised (75 % in PEG) for 5 min, haemorrhage, coagulation and lysis of the blood vessels were recorded, treatment with the test substance resulted in the appearance of haemorrhage on the CAM of 3 eggs 5 minutes post application (1.5 scores of total 21 possible scores), slightly irritating, read-across, supporting study

- study conducted according to Draize test - Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics, FDA (1959), 5% a.i. (i.e. 0.1 mL) of Partially unsaturated IQAC, DMS quaternised were instilled into the conjunctival sac of

6 New Zealand Albino rabbits, the animals were observed for 7 days after application at least daily, there were no indication of irritation, not irritating, read-across, supporting study

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:
1984-02-02 to 1984-03-19
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
other: HLD Versuchsplan P 2/152, Revision 5, regulations of US Food and Drug Administration (Appraisal of the Safety of Chemicals in Food, Drugs and Cosmetics, 1959), den FDA Good Laboratory Practice Regulations
Version / remarks:
1959
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Richtlinien fiir Hautverträglichkeitsprüfungen am Kaninchen (Januar 1981) der US lnteragency Regulatory Liaison Group (IRLG)
Version / remarks:
1981
Deviations:
no
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Züchter H. Fortkamp, 4540 Lengerich
- Age at study initiation: 11-17 weeks
- Weight at study initiation: 1.90 - 2.80 kg
- Housing: individually in Steel cages with perforated metal floors over trays with bedding
- Diet (e.g. ad libitum): ad libitum, SSniff K, Alleindiät für Kanninchen, Ssniff Versuchstier-Diäten GmbH, 4770 Soest
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-23
- Humidity (%): 40-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light):12/12

Type of coverage:
occlusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

Duration of treatment / exposure:
24 h
Observation period:
72 h, with reading intervals of 24 h and 72 h
Number of animals:
6
Details on study design:
TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: Gauze covered with a self-adhesive, impermeable dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none
- Time after start of exposure: After 24 h the substance was removed with disposable wipes.

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) 24 and 72 h

SCORING SYSTEM:
- Method of calculation: Draize, J. H., (1959), Association of Food and Drug Officials of the US, Austin, Texas, "The Appraisal of the Safety of Chemicals in Food, Drugs and Cosmetics"
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timpoints measured
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #4
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #4
Time point:
72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #5
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #5
Time point:
72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #6
Time point:
24 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #6
Time point:
72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Remarks on result:
positive indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other:
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other:
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other:
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #4
Time point:
24 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other:
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #5
Time point:
24 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other:
Remarks:
only two time points measured
Irritation parameter:
edema score
Basis:
animal #6
Time point:
24 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 h
Remarks on result:
other:
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #1
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no indication for irritation
Remarks on result:
no indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #2
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no indication for irritation
Remarks on result:
no indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #3
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no indication for irritation
Remarks on result:
no indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #4
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no indication for irritation
Remarks on result:
no indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #5
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no indication for irritation
Remarks on result:
no indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
edema score
Basis:
animal #6
Time point:
72 h
Score:
0
Max. score:
4
Reversibility:
other: no indication for irritation
Remarks on result:
no indication of irritation
Remarks:
only two timepoints measured
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
erythema score
Basis:
animal #4
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
erythema score
Basis:
animal #5
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
erythema score
Basis:
animal #6
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
edema score
Basis:
animal #1
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
edema score
Basis:
animal #2
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
edema score
Basis:
animal #3
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
edema score
Basis:
animal #4
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
edema score
Basis:
animal #5
Time point:
48 h
Remarks on result:
not measured/tested
Irritation parameter:
edema score
Basis:
animal #6
Time point:
48 h
Remarks on result:
not measured/tested

Table 1: Irritant/corrosive response data for each animal at each observation time up to removal of each animal from the test

Score at time point / Reversibility

Erythema

Edema

Max. score: 4

Max. score: 4

24 h

 2/2/2/2/2/2

1/1/1/1/1/1 

72 h

2/2/2/2/2/2 

0/0/0/0/0/0 

Average 24h, 72h

 2/2/2/2/2/2

0.5/0.5/0.5/0.5/0.5/0.5 

Reversibility*)

 n.c.

 c.

Average time (h) for reversion

 n.d.

72 h 

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible, n.d. = not determined

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
In the present test 6 New Zealand rabbits were dermally exposed to 0.5 g of the test item for 24h under occclusive conditions. All animals showed moderate erythema up to 72h after patch removal. Edema formation was observed in all six animals but was fully reversible within 72h. Based on these results the substance needs to be classified as Category 2 - irritant according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).
Executive summary:

In a primary dermal irritation study according to HLD Versuchsplan P 2/152, Revision 5, regulations of US Food and Drug Administration (Appraisal of the Safety of Chemicals in Food, Drugs and Cosmetics, 1959), den FDA Good Laboratory Practice Regulations) and Richtlinien für Hautverträglichkeitsprüfungen am Kaninchen (Januar 1981) der US lnteragency Regulatory Liaison Group (IRLG),6 young adult New Zealand rabbitswere dermally exposed to 0.5 gofhydrogenated tallow/nortallow based IQACfor24hours under occlusive conditions.  Animals then were observed for72 hours.  Irritation was scored by the method of(Draize, JH, (1959)).

 All animals showed moderate erythema up to 72h after patch removal. Edema formation was observed in all six animals but was fully reversible within 72h.In this study,hydrogenated tallow/nortallow based IQAC isa dermal irritant. Based on these results the substance needs to be classified as Category 2 - irritant according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010-03-23 to 2010-03-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Age at study initiation: no data
- Weight at study initiation: 2818.2 9 and 2709.8 9 (female), 2912.6 9 (male). In deviation to the study plan the animals were slightly overweight (weight > 2500 g). However, this deviation is considered not affecting the outcome of the study.
- Housing: in steel cages with a plastic bottom, a non-barrier system with air conditioning was used.
- Diet: ad libitum, food of type "2023 Maintenance diet for rabbits " frorn Altramin, Lage, Germany as pelleted diet (Batch Nos.: 0926 and 1354; expiry dates: 2010-07-07 and 2010-12-02)
- Water: ad libitum, tap water from municipal source (SWK AQUA GmbH, Krefeld, Germany)
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C ± 3°C
- Humidity (%): 30 % -70 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data
Vehicle:
unchanged (no vehicle)
Controls:
other: the left eye remained untreated and served as a control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml of the test substance
- Concentration (if solution): unchanged
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
3 animals: 2-females, 1 male
Details on study design:
REMOVAL OF TEST SUBSTANCE
After recording the observations at 24 hours, the eyes of all rabbits were further examined with the aid of fluorescein. One drop of a 0.5 % (w/v) fluorescein solution in saline was poured into both eyes and washed off with saline solution 30 seconds later.

SCORING SYSTEM: the ocular reactions were assessed using the following numerical scoring system:
I.) Cornea opacity
No ulceration or opacity 0
Scattered or diffuse areas of opacity, details of iris clearly visible 1
Easily discernible translucent areas, details of iris slightly obscured 2
Nacreous areas, no details of iris visible, size of pupil barely discernible 3
Opaque cornea, iris not discernible thraugh the opacity 4

I.) Cornea area
No lesion 0
One quarter (or less) but not zero 1
Greater than one quarter, but less than one half 2
Greater than one half, but less than three quarters 3
Greater than three quarters, up to whole area 4

lI.) Iris
Normal 0
Markedly deepened rugae, congestion, swelling, moderate
circumcorneal hyperaemia or injection, iris still reactive to light 1
No reaction to light, haemorrhage, or grass destruction 2
III.) Conjunctivae redness
Vessels normal 0
Some blood vessels hyperaemic ( injected) above normal 1
More diffuse, crimson colour, individual vessels not easily discernible 2
Diffuse beefy red 3

III.) Conjunctivae chemosis
Normal 0
Some swelling above normal 1
Obvious swelling with partial eversion of lids 2
Swelling with lids about half closed 3
Swelling with lids more than half closed 4

III.) Conjunctivae discharge
No discharge 0
Any amount different from normal ( does not include
small amounts observed in inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to the lids 2
Discharge with moistening of the lids and hairs and affecting
a considerable area around the eye 3

TOOL USED TO ASSESS SCORE: fluorescein (Fluorescein sodium C.I. 45350, Merck KGaA, Darmstadt, Germany)
Irritation parameter:
cornea opacity score
Basis:
animal: #1,#2,#3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
animal: #1,#2,#3
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
animal: #1,#2,#3
Time point:
24/48/72 h
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.67
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
All three animals showed moderate ocular reaction 1 h after application of the test substance (two scores for conjunctivae redness and chemosis). 24 h after application moderate ocular reactions were still observed in all three animals. Cornea opacity or iris irritation was not observed in any of the three animals. Slight irritation of the conjunctivae persists until 72 h after treatment in all animals but 7 days after application all three animals were without any sign of ocular irritation.
Other effects:
Symptoms of systemic toxicity caused by the test substance were not observed during the whole study. Body weight gain was positive and within normal range.

Examination of the treated eyes was carried out 1 hour and 24, 48, 72 hours and 7 days after application of the test substance.

 

1 h

24 h

48 h

72 h

7 d

Mean

(24h-72h)

I. Cornea opacity

   Cornea area

A

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

B

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

II. Iris

A

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

0/0/0

III. Conjunctivae redness

     Conjunctivae chemosis

     Conjunctivae discharge

A

2/2/2

2/2/2

1/1/1

1/1/1

0/0/0

1.33/1.33/1.33

B

2/2/2

2/2/1

2/2/1

1/2/1

0/0/0

1.67/2/1

C

1/1/1

1/1/1

0/0/0

0/0/0

0/0/0

0.33/0.33/0.33

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)
Conclusions:
Moderate ocular reactions were observed within 24 hour following application of 0.1 mL of the test substance to the eyes. Effects on the cornea and the iris did not occur. The severity of the observed effects declined with time and the lesions were completely reversible within 7 days.
Executive summary:

In a primary eye irritation study according to OECD Guideline 405 (2002), 0.1 mL of partially unsaturated IQAC, DMS quaternised

(75 % in propylene-glycol) was instilled into the conjunctival sac of three New Zealand White rabbits. The eyes were not rinsed after substance application. Animals then were observed for 7 days. Irritation was scored by the method of Draize.

At 24 hours post application the eyes of all animals were further examined with fluorescein for cornea damage. All three animals showed moderate ocular reaction 1 h after application of the test substance (two scores for conjunctivae redness and chemosis). 24 h after application moderate ocular reactions were still observed in all three animals. Cornea opacity or iris irritation was not observed in any of the three animals. Slight irritation of the conjunctivae persisted until 72 h after treatment in all animals but 7 days after application all three animals were without any sign of ocular irritation.

 

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2007-10-10 to 2007-10-12
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to guideline
Guideline:
other: Luepke, N.P. (1985) Hen's egg chorio allantoic membrane test for irritation potential Fd. Chem. Toxic. 23, 287-291
GLP compliance:
yes (incl. QA statement)
Species:
other: chicken eggs
Strain:
other: Lohmann Selected Leghorn
Details on test animals or tissues and environmental conditions:
Test System
The HET-CAM was carried out with Lohmann Selected Leghorn chicken eggs.

The eggs were obtained from the LSL Rhein-Main Geflügelvermehrungsbetriebe (0-64807 Oieburg, Germany) on day one. Afterwards they were
incubated at 37,5 ± 0.5 °C for eight days. While incubation the eggs were rotated to prevent an attachment of the embryo to one side of the egg.
On day eight the eggs were candled and non-viable eggs were discarded.
The rest of the eggs were placed upward in the incubator and were incubated for another day without rotation.
Vehicle:
water
Controls:
yes
Amount / concentration applied:
Test item: pure, as delivered by the sponsor (purity 100%)
Negative control: physiological sodium chloride solution (0.9% (v/v))
Positive control / SDS (sodium dodecyl sulfate (purity> 99%), Merck (64293 Darmstadt, Germany): 1% solution in deion. water
Positive control / NaOH (sodium hydroxide solution c(NaOH) = 0.1 mol/l (0.1 N), Merck (64293 Darmstadt, Germany): 0.1 N
Duration of treatment / exposure:
300 sec.
Observation period (in vivo):
300 sec.
Number of animals or in vitro replicates:
negative control: 3 eggs
positive control 0.1 N NaOH: 3 eggs
positive control 1 % SDS: 3 eggs
Test substance: 6 eggs
Irritation parameter:
other: HET-CAM: mean irritation score
Remarks:
with test item
Run / experiment:
mean of all eggs
Value:
0
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
Score was 0 of maximum 21
Irritation parameter:
other: HET-CAM: mean irritation score
Remarks:
negative control (0.9% (v/v) NaCl in H2O)
Run / experiment:
mean of all eggs
Value:
0
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
Score was 0 of maximum 21
Irritation parameter:
other: HET-CAM: mean irritation score
Remarks:
positive control (0.1 N NaOH)
Run / experiment:
mean of all eggs
Value:
19.21
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
reversibility not applicable/ score 19.21 of maximum 21
Irritation parameter:
other: HET-CAM: mena irritation score
Remarks:
positive control (1 % SDS)
Run / experiment:
mean of all eggs
Value:
8.82
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
reversibility not applicable/score was 8.82 of maximum 21
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: not specified
- Test item (pure): No irritating effects were observed during 5 min incubation with the test substance (no solvent). The calculated mean irritancy index is 0.00.
- Negative control (physiological sodium chloride solution (0.9% (v/v)): The negative control showed no irritating effect on the blood vessels under the membrane. The calculated mean irritancy index is 0.00.
- Positive control / SDS (sodium dodecyl sulfate, 1% solution in deion. water): The positive control SDS induced severe irritation on the blood vessels. The calculated mean irritancy index is 8.82.
- Positive control / NaOH (sodium hydroxide solution, 0.1 N): The positive control 0.1 N NaOH induced severe irritation on the blood vessels. The calculated mean irritancy index is 19.21.

The supplement of OECD Guideline 405 (2002) claims a sequential testing strategy for eye irritation and corrosion under inclusion of reliable in vitro tests to minimise the need for in vivo testing. Therefore investigators have been searching for alternatives to the Draize rabbit eye test for assessment of the irritating potential. One alternative model is the chorioallantoic membrane (CAM), a highly vascularized membrane that forms to surround a developing chick embryo and which is insensitive to pain. For evaluation of the acute irritation potential of a test substance in analogy to the Draize rabbit eye irritation test a definitive dose is applied onto the CAM and the possible reactions are recorded. According to French regulations the hen's egg chorioallantoic membrane test (HET-CAM) is an accepted alternative method to the Draize test for the control of innocuity of cosmetics and toiletries (Journal Officiel de la Republique Francaise, 1996, pp. 1913719138).

In addition, German regulatory authorities agreed to accept the classification of severely eye irritating chemicals as R41 according EU regulations by using the HET-CAM test (Spielmann et ai, 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. 741858).

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
No irritating effects were observed during 5 min incubation with partially unsaturated IQAC, DMS quaternised (no solvent). The calculated irritancy
mean index is 0.00. Thus, the test substance was graded as not irritating according to the evaluation scheme.
Executive summary:

In this hen's egg chorioallantoic membrane test (HET-CAM), an validated alternative method to the Draize test, the potential to cause irritation upon first contact with eyes or mucous membranes was assessed by application of the test substance onto the chorioallantoic membrane (CMA). Six eggs were treated with the partially unsaturated IQAC, DMS quaternised (no solvent, 100 % pure). Amounts approximately between 126 and 175 mg of the test item were applied to the membranes so that at least 25 % of the membranes were covered by the test item. During the observation for 300 seconds lesions in close proximity to the covered membranes were noted. The membranes of the eggs were observed for 300 seconds. Lesions of the underlying blood vessels were noted. In general there were three endpoints which were observed and the time when that particular endpoint was observed was noted. The three endpoints were haemorrhage, coagulation and lysis of the blood vessel. From these data a mean irritancy index is calculated (maximal possible score = 21).

No irritating effects were observed during 5 min incubation with the partially unsaturated IQAC, DMS quaternised (no solvent, 100 % pure). The calculated mean irritancy index is 0.00.

Parallel testing of negative and positive control substances (3 eggs per substance) affirmed the sensitivity and reliability of the experimental technique used. Negative control (physiological sodium chloride solution (0.9 % (v/v)) showed no irritating effect on the blood vessels under the membrane. The calculated mean irritancy index is 0.00. Positive controls sodium dodecyl sulfate (1 % solution in deion. water) and sodium hydroxide solution (0.1 N) induced mean irritancy indices of 8.82 and 19.21, respectively.

In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item partially unsaturated IQAC, DMS quaternised (no solvent, 100 % pure) does not possess any irritating potential.

 

 

 

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2009-11-06 to 2009-11-06
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to guideline
Guideline:
other: Luepke, N.P. (1985) Hen's egg chorio allantoic membrane test for irritation potential Fd. Chem. Toxic. 23, 287-291
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
other: chicken eggs
Strain:
other: strain SPAFAS
Details on test animals or tissues and environmental conditions:
Test orqanism
Fresh, intact chicken eggs of the breed SPAFAS supplied by Charles River, 97633 Sulzfeld, Germany were used. The eggs were delivered less than two days after laying. The weight of the chicken eggs was 55 to 59 g.

lncubation of eggs
After delivery eggs were stored at least for 24h with blunt ends upwards at temperatures between 15°C and 22°C to ensure that the air space is adjusted under the blunt end.
The incubation of the eggs took place in an incubator with an automatic rotating device under continuous ventilation at a temperature of 37.5 °C (± 0.5 °C) and a relative humidity of 62.5 % (±7.5 %). Temperature and humidity were recorded every day. The eggs were turned automatically every hour. No deviations from pre-set conditions were recorded during incubation period.
The whole incubation period amounted to 9 days. Up to this time point it can be expected that the embryo is insensitive to pain. From the 6th incubation day on the eggs were illuminated every second day with a candeling lamp. Eggs, which were not fertilized or not developed, were rejected.

Preparation and dosage of test substance
Before application the test substance was warmed up to 37.5 °C (± 0.5 °C) in an incubator. 0.2 mL of the test substance were applied onto each CAM of six eggs.
Vehicle:
unchanged (no vehicle)
Controls:
other: Yes, tap water
Amount / concentration applied:
Test item: 0.2 mL
Negative control: tap water
Positive control: Texapon ASV 70 Spezial (sodium magnesium lauryl-myristyl-6-ethoxy-sulfate, Henkel AG & Co. KGaA, Düsseldorf, Lot. No.: CS 90250040) (5 % (w/w) active substance content in tap water)
Duration of treatment / exposure:
5 minutes
Observation period (in vivo):
Observed reactions at 0.5, 2 and 5 minutes p.a. were scored.
Number of animals or in vitro replicates:
For testing according to standard operating procedure 6 eggs were used. For test validation according to standard operating pracedure 6 eggs each were inserted for the positive contral Texapon ASV 70 Spezial (as 5 % (w/w) active substance preparation) and for the negative / vehicle control tap water.
Details on study design:
The test was carried out 9 days after insertion of the eggs into the incubator. For testing the egg shell around the air space was peeled away and the now visible white egg membrane was wetted with physiological saline (not colder than room temperature). After approximately 30 seconds the saline was poured off and the membrane was carefully removed with forceps. Eggs which showed small lesions or hemorrhages after preparation were not used for the test.
Prepared eggs were placed under a cold light lamp (KL 1500 LCD, Schott Glas, Wiesbaden, Germany) and immediately the test substance was applied onto the CAM. After application of the test substance, the positive and negative control, respectively, the blood vessels of the CAM were continuously observed over a period of 5 minutes. Observed reactions at 0.5, 2 and 5 minutes p.a. were scored. For evaluation 2 minutes and 5 minutes p.a. parts of the dosed test substance were moved aside with a spatula. Untreated areas served as control. After the end of the observation period the next freshly prepared eggs were exposed to the test substance in the same manner.
Irritation parameter:
other: HET-CAM, mean irritation score
Remarks:
test item
Run / experiment:
mean of all eggs
Value:
1.5
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: slight irritation
Remarks:
score 1.5 of maximum 21
Irritation parameter:
other: HET-CAM, mean irritation score
Remarks:
negative control
Run / experiment:
mean of all eggs
Value:
0
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
score 10 of maximum 21
Irritation parameter:
other: HET-CAM, mean irritation score
Remarks:
positive control/(sodium magnesium lauryl-myristyl-6-ethoxy-sulfate)
Run / experiment:
mean of all eggs
Value:
10
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
score 10 of maximum 21
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: No other effect was detected after treatment with the test substance.

- Test item (pure): treatment with the test substance resulted in the appearance of haemorrhage on the CAM of 3 eggs 5 minutes post application (1.5 scores of total 21 possible scores).
- Negative control:Application of the negative control tap water onto the CAM showed no effects; the irritation index therefore was 0.0 of 21 total possible scores.
- Positive control:Treatment with a 5 % active substance preparation of the reference substance Texapon ASV 70 Spezial resulted in an irritation index of 10.0 of 21 total possible scores.

Summary of the observed effects after application of the test substance preparation onto the CAM.

 

 

0.5 min

2 min

5 min

Sum

Vascular Injection

0/0/0/0/0/0

0/0/0/0/0/0

0/0/0/0/0/0

 

 

0/3/3/0/3/0

Haemorrhage

0/0/0/0/0/0

0/0/0/0/0/0

0/3/3/0/3/0

Coagulation

0/0/0/0/0/0

0/0/0/0/0/0

0/0/0/0/0/0

 

 

Summary of the observed effects after application of the reference substance Texapon ASV 70 Spezial (5%(w/w)active substance preparation in tap water) onto the CAM

 

 

0.5 min

2 min

5 min

Sum

Vascular Injection

5/5/5/5/5/5

  -

 -

 

 

10.0

Haemorrhage

0/0/0/0/0/0

5/5/5/5/5/5

 -

Coagulation

0/0/0/0/0/0

0/0/0/0/0/0

0/0/0/0/0/0

 

 

 

Interpretation of results:
slightly irritating
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
Treatment with the test substance resulted in the appearance of haemorrhage on the CAM of 3 eggs 5 minutes post application. No other effect was detected after treatment (1.5 scores of total 21 possible scores).
Thus, the partially unsaturated IQAC, DMS quaternised was graded as slightly irritating according to the evaluation scheme presented in this report..
Executive summary:

 Partially unsaturated IQAC, DMS quaternised (75 % in PEG) was tested in the hen's egg chorioallantoic membrane test (HET-CAM), an validated alternative method to the Draize test, to evaluate the acute irritation potential. The potential to cause irritation upon first contact with eyes or mucous membranes was assessed by application of the test substance onto the chorioallantoic membrane (CAM) of 6 fertilized and incubated eggs (strain SPAFAS).

The occurrence of vascular injection, haemorrhage and coagulation was recorded 30 seconds, 2 minutes and 5 minutes after application of 200 µL of the test substance preparation. For evaluation 2 minutes and 5 minutes p.a. parts of the dosed test substance were moved aside with a spatula.

Treatment with the test substance resulted in the appearance of haemorrhage on the CAM of 3 eggs 5 minutes post application. No other effect was detected after treatment (1.5 scores of total 21 possible scores). Application of the negative control tap water onto the CAM showed no effects; the irritation index was 0.0 of 21 total possible scores. Parallel to the experiment the reference substance Texapon ASV 70 Spezial was tested as 5% (w/w) active substance preparation in water. Treatment with this Texapon ASV 70 Spezial preparation resulted in an irritation index of 10.0 of 21 total possible scores.

Under the conditions of this study the test substance partially unsaturated IQAC, DMS quaternised can be graded as slightly irritating (1.5 scores of total 21 possible scores).

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2009-11-27 to 2009-11-27
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to guideline
Guideline:
other: Luepke, N.P. (1985) Hen's egg chorio allantoic membrane test for irritation potential Fd. Chem. Toxic. 23, 287-291
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Species:
other: chicken eggs
Strain:
other: strain SPAFAS
Details on test animals or tissues and environmental conditions:
Test orqanism
Fresh, intact chicken eggs of the breed SPAFAS supplied by Charles River, 97633 Sulzfeld, Germany were used. The eggs were delivered less than two days after laying. The weight of the chicken eggs was 55 to 59 g.

lncubation of eggs
After delivery eggs were stored at least for 24h with blunt ends upwards at temperatures between 15°C and 22°C to ensure that the air space is adjusted under the blunt end.
The incubation of the eggs took place in an incubator with an automatic rotating device under continuous ventilation at a temperature of 37.5 °C (± 0.5 °C) and a relative humidity of 62.5 % (±7.5 %). Temperature and humidity were recorded every day. The eggs were turned automatically every hour. No deviations from pre-set conditions were recorded during incubation period.
The whole incubation period amounted to 9 days. Up to this time point it can be expected that the embryo is insensitive to pain. From the 6th incubation day on the eggs were illuminated every second day with a candeling lamp. Eggs, which were not fertilized or not developed, were rejected.

Preparation and dosage of test substance
Before application the test substance was warmed up to 40 °C in a water bath and mixed with preheated 40°C tap water. Therefore 0.63 g tap water were added to 4.39 g test substance and mixed on a magnetic stirrer. The test substance preparation was stored at 37.5 °C (± 0.5 °C) in an incubator to avoid solidification. 200 mg of the test substance were applied onto each CAM of six eggs to assess the irritation potential of the test substance as 87.5 % (w/w) preparation.
Due to the creamy consistence of the test substance 200 mg were applied onto each CAM instead of 0.2 mL.
Vehicle:
water
Remarks:
tap water
Controls:
yes, concurrent vehicle
Amount / concentration applied:
Test item: 200 mg
Negative control: tap water
Positive control: Texapon ASV 70 Spezial (sodium magnesium lauryl-myristyl-6-ethoxy-sulfate, Henkel AG & Co. KGaA, Düsseldorf, Lot. No.: CS 90250040) (5 % (w/w) active substance content in tap water)
Duration of treatment / exposure:
5 minutes
Observation period (in vivo):
Observed reactions at 0.5, 2 and 5 minutes p.a. were scored.
Number of animals or in vitro replicates:
For testing according to standard operating procedure 6 eggs were used. For test validation according to standard operating procedure 6 eggs each were inserted for the positive contral Texapon ASV 70 Spezial (as 5 % (w/w) active substance preparation) and for the negative / vehicle contral tap water.
Details on study design:
The test was carried out 9 days after insertion of the eggs into the incubator. For testing the egg shell around the air space was peeled away and the now visible white egg membrane was wetted with physiological saline (not colder than room temperature).
After approximately 30 seconds the saline was poured off and the membrane was carefully removed with forceps. Eggs which showed small lesions or hemorrhages after preparation were not used for the test.
Prepared eggs were placed under a cold light lamp (KL 1500 LCD, Schott Glas, Wiesbaden, Germany) and immediately the test substance was applied onto the CAM.
After application of the test substance, the positive and negative/vehicle control, respectively, the blood vessels of the CAM were continuously observed over a period of 5 minutes. Observed reactions at 0.5, 2 and 5 minutes p.a. were scored. For evaluation the dosed test substance was Iifted shortly from the CAM. Untreated areas served as control. After the end of the observation period the next freshly prepared eggs were exposed to the test substance in the same manner.
Irritation parameter:
other: HET-CAM, mean irritation score
Remarks:
test item
Run / experiment:
mean of all eggs
Value:
0
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
score of 0 of maximum 21
Irritation parameter:
other: HET-CAM, mean irritation score
Remarks:
negative control/ tap water
Run / experiment:
mean of all eggs
Value:
0
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
score of 0 of maximum 21
Irritation parameter:
other: HET-CAM, mean irritation score
Remarks:
positive control/ 5% Texapon
Run / experiment:
mean of all eggs
Value:
10.3
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
score 10.3 of maximum 21
Other effects / acceptance of results:
- Test item: No effect was detected after treatment with the test substance preparation (0.0 scores of total 21 possible scores).
- Negative control: Application of the negative control tap water onto the CAM showed no effects; the irritation index therefore was 0.0 of 21 total possible scores.
- Positive control:Treatment with a 5 % active substance preparation of the reference substance Texapon ASV 70 Spezial resulted in an irritation index of 10.3 of 21 total possible scores.
No other effect was detected after treatment with the test substance

 Summary of the observed effects after application of the reference substance Texapon ASV 70 Spezial (5%(w/w) active substance preparation in tap water) onto the CAM

 

 

0.5 min

2 min

5 min

Sum

Vascular Injection

5/5/5/5/5/5

  -

 -

 

 

10/10/10/10/10/12

Haemorrhage

0/0/0/0/0/7

5/5/5/5/5/ -

 -

Coagulation

0/0/0/0/0/0

0/0/0/0/0/0

0/0/0/0/0/0

 

 

 

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
No effect was detected after treatment with the test substance preparation of partially unsaturated IQAC, DMS quaternised (lPA/Wasser) (0.0 scores of total 21 possible scores).
Thus, the test substance partially unsaturated IQAC, DMS quaternised (IPA/Wasser) was graded as not irritating according to the evaluation scheme.
Executive summary:

Partially unsaturated IQAC, DMS quaternised (IPA/Wasser) was tested in the hen's egg chorioallantoic membrane test (HET-CAM), an validated alternative method to the Draize test, to evaluate the acute irritation potential. The potential to cause irritation upon first contact with eyes or mucous membranes was assessed by application of a preheated 87.5 % (w/w) preparation of the test substance in tap water onto the chorioallantoic membrane (CAM) of 6 fertilized and incubated eggs (strain SPAFAS).

Before application the test substance was warmed up to 40°C in a water bath and mixed with preheated 40°C tap water. The test substance preparation was stored at 37.5 °C (± 0.5 °C) in an incubator to avoid solidification. The occurrence of vascular injection, haemorrhage and coagulation was recorded 30 seconds, 2 minutes and 5 minutes after application of 200 mg of the test substance preparation. For evaluation the dosed test substance was lifted shortly from the CAM.

After treatment with the preparation of the test substance partially unsaturated IQAC, DMS quaternised (IPA/Wasser) no effect was detected (0.0 scores of total 21 possible scores). Application of the negative/vehicle control tap water onto the CAM showed no effects; the irritation index was 0.0 of 21 total possible scores. Parallel to the experiment the reference substance Texapon ASV 70 Spezial was tested as 5 % (w/w) active substance preparation in tap water. Treatment with this Texapon ASV 70 Spezial preparation resulted in an irritation index of 10.3 of 21 total possible scores.

Under the conditions of this study the preparation of the test substance partially unsaturated IQAC, DMS quaternised (IPA/Wasser) can be graded as not irritating (0.0 scores of total 21 possible scores).

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: - Acceptable, well-documented study report which meets basic scientific principles.
Qualifier:
according to guideline
Guideline:
other: Draize test - Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics, FDA (1959)
GLP compliance:
no
Species:
rabbit
Strain:
other: New Zealand Albino
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 2.4 - 2.6 kg
- Housing: single caging
- Diet: ad libitum, standard diet, (Muemml Z, Plange)
- Water: ad libitum
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/+ 1
- Humidity (%): 45 - 55
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12

Vehicle:
water
Controls:
other: untreated eye served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml
- Concentration (if solution): 5 % in Aqua dest.
Observation period (in vivo):
7 days
(observation after 1, 2, 8 h, 1, 2, 3, 4, 5, 6, and 7 days of treatment)
Number of animals or in vitro replicates:
6
Details on study design:
SCORING SYSTEM: according the Draize scoring system


TOOL USED TO ASSESS SCORE: no data
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
other: mean 24, 48, 72 hours
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Remarks:
5 % test substance
Irritation parameter:
iris score
Basis:
mean
Time point:
other: mean 24, 48, 72 hours
Score:
0
Max. score:
2
Remarks on result:
no indication of irritation
Remarks:
5 % test substance
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
other: mean 24, 48, 72 hours
Score:
0
Max. score:
3
Remarks on result:
no indication of irritation
Remarks:
5 % test substance
Irritation parameter:
chemosis score
Basis:
mean
Time point:
other: mean 24, 48, 72 hours
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Remarks:
5 % test substance
Irritant / corrosive response data:
CLINICAL EYE OBSERVATION
- At 1, 2, and 8 h conjunctivea of all animals showed slight diffuse crimson colour, and some blood vessels were hyperaemic (conjunctival score 1). The conjunctival effects of test substance were fully reversivle after 24 h.
- Cornea and iris were at all scorings without any effects.
Interpretation of results:
not irritating
Remarks:
Migrated information 5 % dilution of test substance Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)
Conclusions:
Judgment is based on effects on the cornea, iris and conjunctiva calculated as mean scores following grading at 24, 48 and 72 hours after instillation of the test material, in consideration of reversibility. Mean scores for all animals were zero.
Executive summary:

In a primary eye irritation study partially unsaturated IQAC, DMS quaternised (5 % a.i. in Aqua dest.,according to producer information substance has a a.i. of 74 - 77 %) was instilled into the conjunctival sac of the left eye of 6 New Zealand Albino rabbits. The eyes were not rinsed after substance application. Animals were then observed for 7 days. Irritation was scored by the method of Draize.

At 1, 2, and 8 hours p.a. all animals showed slight diffuse crimson colour, and some blood vessels were hyperaemic (conjunctival score 1). Corneal or iris were not involved. Conjunctival effects of partially unsaturated IQAC, DMS quaternised were fully reversible after 24 h.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

skin irritation:

In a skin irritation test according to OECD guideline 404, 6 New Zealand rabbits were dermally exposed to 0.5 g of hydrogentaed tallow/nortallow based IQAC for 24h under occclusive conditions. Occurrence of erythema and edema was recorded at 24h and 72 h after application. All animals showed moderate erythema up to 72h after patch removal which was not reversible within the experimental time. Edema formation was observed in all six animlas but was fully reversible within 72h. Thus, the substance is considered to be a mild irritant to the skin.

In a supporting primary dermal irritation study performed according to OECD Guideline 404 (Apr. 24, 2002), three young adult New Zealand White rabbits were dermally exposed to 0.5 mL of partially unsaturated IQAC, DMS quaternised (100 % a.i.) moistened with 0.5 mL purified water for 4 hours under semi-occlusive dressing. Animals then were observed for 14 days. The skin reaction was scored according to OECD Guideline 404.

One hour after removal of the dressing a well-defined erythema was observed in all animals which progressed into moderate to severe and persisted as very slight up to 10 or 14 days after treatment. A very slight to moderate oedema was recorded in all animals from the 1-hour up to the 72-hour reading or up to 10 days after treatment. Scaling was present in all animals 7 to 14 days after removal of the application patch. No alterations and no corrosive effects were observed on the treated skin.

In this study, partially unsaturated IQAC, DMS quaternised is a dermal irritant. The test substance is irritating to the skin based on results of the topical semi-occlusive application in rabbits.

eye irritation:

There are no data for hydrogenated tallow/nortallow based IQACs regarding eye irritation potential, however, there are information for the structurally similar substance partially unsaturated IQAC, DMS quarternised. A detailed justification for read-across is attached to IUCLID section 13.

In a hen's egg chorioallantoic membrane test (HET-CAM), an validated alternative method to the Draize test, the potential to cause irritation upon first contact with eyes or mucous membranes was assessed by application of partially unsaturated IQAC, DMS quarternised onto the chorioallantoic membrane (CMA). Six eggs were treated with the partially unsaturated IQAC, DMS quaternised (no solvent, 100 % pure). Amounts approximately between 126 and 175 mg of the test item were applied to the membranes so that at least 25 % of the membranes were covered by the test item. During the observation for 300 seconds lesions in close proximity to the covered membranes were noted. The membranes of the eggs were observed for 300 seconds. Lesions of the underlying blood vessels were noted. In general there were three endpoints which were observed and the time when that particular endpoint was observed was noted. The three endpoints were haemorrhage, coagulation and lysis of the blood vessel. From these data a mean irritancy index is calculated (maximal possible score = 21).

No irritating effects were observed during 5 min incubation with the partially unsaturated IQAC, DMS quaternised (no solvent, 100 % pure). The calculated mean irritancy index is 0.00.

Parallel testing of negative and positive control substances (3 eggs per substance) affirmed the sensitivity and reliability of the experimental technique used. Negative control (physiological sodium chloride solution (0.9 % (v/v)) showed no irritating effect on the blood vessels under the membrane. The calculated mean irritancy index is 0.00. Positive controls sodium dodecyl sulfate (1 % solution in deion. water) and sodium hydroxide solution (0.1 N) induced mean irritancy indices of 8.82 and 19.21, respectively. In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item partially unsaturated IQAC, DMS quaternised (no solvent, 100 % pure) does not possess any irritating potential.

In a second study, a primary eye irritation study according to OECD Guideline 405 (2002), 0.1 mL of partially unsaturated IQAC, DMS quaternised (75 % in propylene-glycol) was instilled into the conjunctival sac of three New Zealand White rabbits. The eyes were not rinsed after substance application. Animals then were observed for 7 days. Irritation was scored by the method of Draize.

At 24 hours post application the eyes of all animals were further examined with fluorescein for cornea damage. All three animals showed moderate ocular reaction 1 h after application of the test substance (two scores for conjunctivae redness and chemosis). 24 h after application moderate ocular reactions were still observed in all three animals. Cornea opacity or iris irritation was not observed in any of the three animals. Slight irritation of the conjunctivae persisted until 72 h after treatment in all animals but 7 days after application all three animals were without any sign of ocular irritation.

Further supporting information are available for the source substance partially unsaturated IQAC, DMS quarternised. Partially unsaturated IQAC, DMS quaternised (IPA/Wasser) was tested in the hen's egg chorioallantoic membrane test (HET-CAM), an validated alternative method to the Draize test, to evaluate the acute irritation potential. The potential to cause irritation upon first contact with eyes or mucous membranes was assessed by application of a preheated 87.5 % (w/w) preparation of the test substance in tap water onto the chorioallantoic membrane (CAM) of 6 fertilized and incubated eggs (strain SPAFAS). Before application the test substance was warmed up to 40°C in a water bath and mixed with preheated 40°C tap water. The test substance preparation was stored at 37.5 °C (± 0.5 °C) in an incubator to avoid solidification. The occurrence of vascular injection, haemorrhage and coagulation was recorded 30 seconds, 2 minutes and 5 minutes after application of 200 mg of the test substance preparation. For evaluation the dosed test substance was lifted shortly from the CAM.

After treatment with the preparation of the test substance partially unsaturated IQAC, DMS quaternised (IPA/Wasser) no effect was detected (0.0 scores of total 21 possible scores). Application of the negative/vehicle control tap water onto the CAM showed no effects; the irritation index was 0.0 of 21 total possible scores. Parallel to the experiment the reference substance Texapon ASV 70 Spezial was tested as 5 % (w/w) active substance preparation in tap water. Treatment with this Texapon ASV 70 Spezial preparation resulted in an irritation index of 10.3 of 21 total possible scores. Under the conditions of this study the preparation of the test substance partially unsaturated IQAC, DMS quaternised (IPA/Wasser) can be graded as not irritating (0.0 scores of total 21 possible scores). 

Also, partially unsaturated IQAC, DMS quaternised (75 % in PEG) was tested in the hen's egg chorioallantoic membrane test (HET-CAM), an validated alternative method to the Draize test, to evaluate the acute irritation potential. The potential to cause irritation upon first contact with eyes or mucous membranes was assessed by application of the test substance onto the chorioallantoic membrane (CAM) of 6 fertilized and incubated eggs (strain SPAFAS). The occurrence of vascular injection, haemorrhage and coagulation was recorded 30 seconds, 2 minutes and 5 minutes after application of 200 µL of the test substance preparation. For evaluation 2 minutes and 5 minutes p.a. parts of the dosed test substance were moved aside with a spatula. Treatment with the test substance resulted in the appearance of haemorrhage on the CAM of 3 eggs 5 minutes post application. No other effect was detected after treatment (1.5 scores of total 21 possible scores). Application of the negative control tap water onto the CAM showed no effects; the irritation index was 0.0 of 21 total possible scores. Parallel to the experiment the reference substance Texapon ASV 70 Spezial was tested as 5% (w/w) active substance preparation in water. Treatment with this Texapon ASV 70 Spezial preparation resulted in an irritation index of 10.0 of 21 total possible scores. Under the conditions of this study the test substancepartially unsaturated IQAC, DMS quaternised can be graded as slightly irritating (1.5 scores of total 21 possible scores).

In a third supporting report partially unsaturated IQAC, DMA quarternised was applied in a primary eye irritation study, (5 % a.i. in Aqua dest.,according to producer information substance has a a.i. of 74 - 77 %) was instilled into the conjunctival sac of the left eye of 6 New Zealand Albino rabbits. The eyes were not rinsed after substance application. Animals were then observed for 7 days. Irritation was scored by the method of Draize.

At 1, 2, and 8 hours p.a. all animals showed slight diffuse crimson colour, and some blood vessels were hyperaemic (conjunctival score 1). Corneal or iris were not involved. Conjunctival effects of partially unsaturated IQAC, DMS quaternised were fully reversible after 24 h.

Justification for classification or non-classification

Based on all available data hydrogenated tallow/nortallow based IQACs needs to be classified as Castegory 2 - irritant according the Globally Harmonized System for Classification and Labelling of Chemicals (GHS). According to Regulation (EC) No 1272/2008 (CLP) the test item does not need to be classified with respect to skin irritation.

The eye irritation studies presented here were conducted with the structurally similar substance partially unsaturated IQAC; DMS quarternised. The test substance is regarded more reactive than the target substance based on the occurrence of unsaturated alkyl-moieties. Furthermore, the presented tests were performed with different concentration of the test substance in diverging solvents, thus, it cannot be excluded that the transient effects that were observed in some studies result from the type of solvent and the test item concentrations.

Based on the available data regarding eye irritation hydrogenated tallow/nortallow based IQACs does not need to be classified according to Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System for Classification and Labelling of Chemicals (GHS). Further information is also available in the detailed justification for read-across attached to IUCLID section 13.