Registration Dossier

Administrative data

Description of key information

Skin irritation:    
Not irritating (OECD TG 404, GLP), neat test substance, read-across MDEA-Esterquat C16-18 and C18 unsatd.
Very slight erythema was observed 1 h after exposure in one animal and in 2/3 animals at the 24 hour reading and was reversible within 48 hours after exposure. No oedema was observed.
Eye irritation:
Not irritating (OECD 405; GLP), 100 mg neat test substance, read-across MDEA-Esterquat C16-18 and C18 unsatd. (IV = 50)
Mild conjunctival redness (score 1) was observed in 3/3 rabbits after 1 hour and was present in 2/3 animals at the 24 hour reading. Reversibility was shown within 48 hours. Chemosis (score 1) was observed in 2/3 rabbits and was reversible within 24 hours.
Not irritating (OECD 405 LVET; GLP), 10 mg neat test substance, read-across MDEA-Esterquat C16-18 and C18 unsatd.
In the main group no eye irritation was observed, all readings were zero. Mild conjunctival redness (score 1) was observed in 2/3 rabbits of subgroup (rinsed group) after 24 hours and was reversible within 48 hours.

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:
1992-06-30 to 1992-07-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
The study was conducted in compliance with the following GLP Regulations: GLP in Switzerland, Procedures and Principles, March 1986; OECD Principles of GLP, Paris, France, adopted May 12, 1981 and EPA, GLP Standards, Final Rule, U.S.A., Federal Register,
Species:
rabbit
Strain:
other: Chbb: NZW (SPF)
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Dr. Karl Thomae GmbH
- Age at study initiation: 19 weeks
- Weight at study initiation: Male: 3.2 kg and Females: 3.1 - 3.3 kg
- Housing: Individually housed in stainless steel cages (size: 86 X 54 X 33 cm) equipped with feed hoppers and drinking water bottles
- Diet (e.g. ad libitum): Pelleted standard Kliba 341, Batch 70/92 rabbit maintenance diet, ad libitum
- Water (e.g. ad libitum): Community tap water from Fullinsdorf, ad libitum
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/-3
- Humidity (%): 40-70 %
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light (approx. 100 lux)/ 12 hours dark, music during the light period


IN-LIFE DATES: From: 1992-06-30 To: 1992-07-05
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 gm
- Concentration (if solution): Not applicable


VEHICLE
- Amount(s) applied (volume or weight with unit): Not applicable
- Concentration (if solution): Not applicable
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable
Duration of treatment / exposure:
Three Chbb: NZW (SPF) rabbits were dosed with 0.5 gm of Diethyl ester dimethyl ammonium chloride (DEEDMAC, E-4429.01). The test article was applied moistened with tap water to approximately 6 cm2 of the intact skin of the clipped area. It was covered with a 3.0 cm X 3.0 cm patch of surgical gauze. The gauze was covered with a semi-occlusive dressing. The duration of treatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm-tap water. The skin reaction was assessed at 1, 24, 48 and 72 hours after removal of the dressing, gauze patch and test article.
Observation period:
1, 24, 48 and 72 hours after removal of the dressing, gauze patch and test article.
Number of animals:
3 animals (one male and two females)
Details on study design:
TEST SITE
- Area of exposure: Approximately 6 cm2
- % coverage: Covered with a 3.0 cm X 3.0 cm patch of surgical gauze
- Type of wrap if used: The gauze was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with an elastic bandage


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Skin was flushed with lukewarm-tap water
- Time after start of exposure: 4 hours


SCORING SYSTEM: Erythema and escher scoring from0-4 with 0 = no erythema, 1= very slight erythema, 2 = well defined erythema, 3 = moderate to severe erythema, 4 = severe erythema to slight eschar formation.
Oedema scoring from0-4 with 0 = no oedema, 1= very slight oedema, 2 = slight oedema, 3 = moderate oedema, 4 = severe oedema.
Irritation parameter:
erythema score
Basis:
animal: #1, #3
Time point:
24/48/72 h
Score:
0.33
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Remarks on result:
no indication of irritation
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritation parameter:
edema score
Basis:
animal: #1, #2, #3,
Time point:
24/48/72 h
Score:
0
Max. score:
4
Remarks on result:
no indication of irritation
Irritant / corrosive response data:
PDII 0.33 at 1 hour for intact skin site
PDII 0.67 at 24 hours for intact skin site
PDII 0 at 48 and 72 hours for intact skin site
Other effects:
No corrosive effect occurred on the treated skin of any animal at each measuring interval

GHS Classification Evaluation

 Animal # Degree of erythema after...[observation time]              Degree of oedema after...[observation time]        GHS ClassificationØ 24/48/72 h>/= 2.3?  
   1h  24h  48h  72h  1h  24h  48h  72h  Erythema  Oedema
 40 (M)  1  1  0  0  0  0  0  0  No  No
         Ø 24/48/72 h =  0.33           Ø 24/48/72 h =  0      
 41 (F)  0  0  0  0  0  0  0  0  No  No
         Ø 24/48/72 h =  0           Ø 24/48/72 h =  0    
 42 (F)  0  1  0  0  0  0  0  0  No  No
          Ø 24/48/72 h =  0.33            Ø 24/48/72 h =  0    
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: GHS Regulation EC No 1272/2008
Conclusions:
MDEA-Esterquat C16-18 and C18 unsatd. is slightly irritating to the skin of rabbits. The mean grades of skin reactions at 24, 48 and 72 h reading correspond to values classified as not irritant by theDirective 67/548/EEC as well as GHS Regulation EC No 1272/2008.
Executive summary:

In a primary dermal irritation study comparable to OECD guideline 404, 12 May 1981, three White New Zealand rabbits were dermaly exposed to 0,5 g  of MDEA-Esterquat C16-18 and C18 unsatd. undiluted for 4 hours to an intact area of approximately 6 cm² covered by a semi-occlusive dressing. Animals then were observed for 3 days.  Irritation was scored by the method of Draize 1;  24; 48 and 72 hours after exposure.

Very slight erythema was observed 1 h after exposure in one animal and in 2/3 animals at the 24 hour reading and was reversible within 48 hours after exposure. No oedema was observed.

The mean grades of skin reactions at 24, 48 and 72 h reading correspond to values classified as not irritant by the Directive 67/548/EEC as well as GHS Regulation EC No 1272/2008.

When applied to the skin, the test article MDEA-Esterquat C16-18 and C18 unsatd. is classified as "not irritant" in this study.

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

Eye irritation

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Human or animal in vivo or in vitro data data on local irritation/corrosion are not availablefor the target substance MDEA Esterquat C18 satd.. Therefore for the assessment of irritating effects of MDEA Esterquat C18 results from the following studies are taken into consideration:

- an OECD 404 primary dermal irritation study in vivo of the source substance MDEA-Esterquat C16-18 and C18 unsatd.

- an additional OECD 404 primary dermal irritation in vivo study of thesource substanceMDEA-Esterquat C16-18 and C18 unsatd. 32% in corn oil

- an OECD 402 acute dermal toxicity studyof thesource substanceMDEA-Esterquat C16-18 and C18 unsatd

- an OECD 439 in vitro skin irritation test with thethesource substanceMDEA-Esterquat C16-18 and C18 unsatd

- an OECD 405 primary eye irritation study in vivo of the source substance MDEA-Esterquat C16-18 and C18 unsatd. (IV = 50)

- an OECD 405 low volume eye irritation test in vivo of the source substance MDEA-Esterquat C16-18 and C18 unsatd.

 

 

Skin irritation

In a primary dermal irritation study comparable to OECD Guideline 404 (24 April 2002), three White New Zealand rabbits were dermally exposed to 0.5 g of undiluted MDEA-Esterquat C16-18 and C18 unsatd. An intact skin area of approximately 6 cm² was covered by a semi-occlusive dressing for a duration of 4 hours. Animals were observed for 3 days. Irritation was scored using the method of Draize at the following timepoints: 1; 24; 48 and 72 hours after exposure.

Very slight erythema was observed in one animal 1 hour after exposure and in 2/3 animals after 24 hours and was fully reversible within 48 hours after exposure. No edema was observed.

 

In an additional study comparable to OECD Guideline 404 (24 April 2002), three White New Zealand rabbits were dermally exposed to 0.4 mL of MDEA-Esterquat C16-18 and C18 unsatd. 32 % of a.i. diluted in corn oil was administered to an intact and abraded skin area of approximately 2.5 cm² each under an occlusive dressing for a duration of 4 hours. Animals were observed for 3 days. Irritation was scored by the method of Draize at the following time points: 4.5; 24 and 72 hours after exposure.

Abraded and intact skin areas showed exactly the same scores. Very slight erythema was observed 4.5 h after exposure in all three animals. It was still apparent in 2/3 animals at the 24 hour reading and was fully reversible within 72 hours after exposure. No edema was observed.

Due to the very detailed description of irritation scores in the acute dermal toxicity study, additional data on the irritation endpoint were available. In an acute dermal toxicity study (comparable to OECD Guideline 402), groups of 3 male and 3 female New Zealand White rabbits were exposed dermally to MDEA-Esterquat C16-18 and C18 unsatd. (40 % a.i.) in oleum arachidis. Treatment lasted for 24 hours under an occlusive dressing. The applied dose of 2000 mg/kg bw resulted in an average concentration of 36 mg/cm² (assumed density 1; area of 10 x 15 cm).

Slight (1) to moderate (2) erythema and mainly moderate (2) edema with partly decreased intensity were observed at the treatment sites from 24 hours up to the 72 hours after treatment and were fully reversible within 7 days. The mean score were 1.3 for erythema and 1.75 for edema (after 24 and 72 hours). The applied scores of 1 and 2 for erythema and edema were comparable to the scores of the Draize method.

Furthermore an in vitro skin irritation study according to OECD Guideline 439 (In Vitro Skin Irritation) (22 July 2010) and EU method B.46 (In vitro skin irritation: reconstructed human epidermis model test) (20 July 2012) is available. 10.0 to 11.7 mg MDEA-Esterquat C16-18 and C18 unsatd. (100% a.i.) were applied in triplicates for 15 min to a three-dimensional human epidermis model (EPISKIN Small Model (EPISKIN-SM, 0.38 cm², Batch no.: 12-EKIN-047)). After 15 minutes exposure at room temperature, the tissues were washed with phosphate buffered saline to remove residual test substance. Subsequently the skin tissues were incubated for 43 h at 37°C. Cytotoxicity (irritancy) was expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

Positive (5% SDS) and negative (PBS) control gave the appropriate responses.

The relative mean tissue viability obtained after 15 minutes treatment with MDEA-Esterquat C16-18 and C18 unsatd. compared to the negative control tissues was 108%. Since the mean relative tissue viability for the test substance was above 50%, MDEA-Esterquat C16-18 and C18 unsatd. is considered to be not irritating.

 

Eye irritation

In a primary eye irritation LVET (Low Volume Eye Test) study comparable to OECD Guideline 405 (24 April 2002), 10 mg of MDEA-Esterquat C16-18 and C18 unsatd. was directly applied to the cornea of one eye of six young adult New Zealand White rabbits. Reactions were observed for 7 days afterwards. Irritation was scored by the method of Draize.

No eye irritation was observed, all readings were zero. No information on quantity of active ingredient contained in the test substance was given in this study report. However, according to information given by the producer, the test substance contained 80 % of a.i. (in isopropanol).

Additional data exist for a substance with the same structure but a different distribution of the fatty acid moiety which contains a higher amount of unsaturated C18, in particular, a primary eye irritation study is available according to OECD Guideline 405 (12 May 1981). 100 mg of MDEA-Esterquat C16-18 and C18 unsatd. (Iodine value of 50 instead of 25, the highest value specified for the source substance MDEA-Esterquat C16-18 and C18 unsatd ) was instilled into the conjunctival sac of the left eye of 3 young adult New Zealand White rabbits, 1 male and 2 females. Animals were then observed for 72 hours. Irritation was scored by the method of Draize.

Mild conjunctival redness (score 1) was observed in 3/3 rabbits after 1 hour and was still present in 2/3 animals at the 24 hour reading. Full reversibility was shown within 48 hours. Chemosis (score 1) was observed in 2/3 rabbits and was fully reversible within 24 hours.

Following the Integrated Testing Strategy (ITS) according to Guidance on information requirements and chemical safety assessment, Endpoint specific guidance, Chapter R. 7.2.6 there is sufficient information available to proceed with classification and labelling using a weight of evidence approach. According to this guidance, for existing substances, the use of methods other than those specified in Annex V of Directive 67/548/EEC, or corresponding OECD methods, such as LVET (Griffith et al., 1980) may be accepted on a case-by-case basis.

The low volume eye test (LVET) is an alternative to the standard Draize eye irritation method. In contrast to the OECD Guideline 405, 10 µl/mg of test substance is directly applied to the cornea. Except for these two modifications, the methods are identical including the scoring method and data interpretation.

The LVET is an approved ASTM method (Standard test method for evaluation of eye irritation in Albino Rabbits E 1055-99) and has been widely used for detergent and cleaning preparations. The method is judged to be suitable to demonstrate toxicological effects on man of substances potentially hazardous to the eye (Freeberg et al. 1984).

Especially for detergent and cleaning preparations, the LVET is considered to be the best available predictor of effects on the human eye (ECETOC monograph 32, 2002). The acceptance of the LVET by national and international regulatory bodies and a further update of OECD Guideline 405 are under investigation. A retrospective validation study performed by ECVAM will shortly be peer-reviewed by the ESAC (Grindon et al. 2008).

MDEA-Esterquat C16-18 and C18 unsatd. and MDEA Esterquat C18 satd. are cationic surfactants which are used as a fabric conditioner and therefore the substance fits exactly into the domain of applicability of detergent and cleaning products. In this context the results of the Low Volume Eye Test are of prime importance for the weight of evidence approach. Additional results are available from an eye irritation study according to OECD Guideline 405 with a structurally identical substance differing only in the fatty acid moiety. As compared to the one submitted for registration, this substance contains a higher amount of the unsaturated C18 fatty acids, therefore the hazard prediction derived from this study is more likely to be an over- than an underestimation.

The results of the LVET give no evidence of an eye irritating potential of the structurally similar source substance MDEA-Esterquat C16-18 and C18 unsatd. This is supported by observations made with a similar substance with higher iodine value. Mild conjunctival redness and chemosis (scores 1) were observed for the period of up to 24 hours after treatment and were fully reversible after the 48 hours.

 

Respiratory irritation

No data on the respiratory irritation of MDEA Esterquat C18 satd. is available.

 

There are no data gaps for the endpoint irritation/corrosion. No human information is available for this endpoint. However, there is no reason to believe that these results would not be applicable to humans.

 

Justification for read-across

 For details on substance identity, toxicokinetics and detailed toxicological profiles, please refer also to the general justification for read-across given in chapter 5 of the CSR and attached as pdf document to section 7 of the IUCLID file.

 

Structural similarity

a. Structural similarity and functional groups

The target substance, MDEA Esterquat C18 satd., consists of an amine backbone (MDEA = Methyldiethanol amine) esterified with the long chain fatty acid stearic acid (C18 satd.; IV (iodine value) < 1). The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The source substance, MDEA EsterquatC16-18 and C18 unsatd., consists of the same amine backbone (MDEA = Methyldiethanol amine) but esterified with a mixture of the long chain fatty acids C16, C18 and C18 unsaturated (in general IV < 25; IV = 50 for OECD 405 eye irriation study). The main reaction product is the dialkylester compound, next to that small amounts of the monoalkylester may be formed. The amine function is quaternised with two methyl groups. The counter ion is Chloride.

The source and the target substance share structural similarities with common functional groups (quaternary amines, esters, and fatty acid chains with comparable length and degree of saturation).

b. Common breakdown products

The metabolism expected to occur is hydrolysis of the ester-bond by esterases. However, the rate of hydrolysis is assumed to be low. The fraction of metabolised molecules would result in free fatty acids and Dimethyl-DEA (DEA = Diethanolamine). The carboxylic acids are further degraded by the mitochondrial beta-oxidation process (for details see common text books on biochemistry). The fatty acids enter normal metabolic pathways and are therefore indistinguishable from fatty acids from other sources including diet. The quaternary ammonium ions are not expected to be further metabolised, but excreted unchanged via the urine. 

c. Differences

The differences in fatty acid chain length (higher percentage of C16 in the source substance MDEA Esterquat C16 -18 and C18 unsatd.) and degree of saturation (higher degree of unsaturation of the C18 fatty acid chains in the source substance substance MDEA Esterquat C16-18 and C18 unsatd.) could be relevant for the endpoint skin and eye irritation. Tests in animals and human show that the skin and eye irritation potential of fatty acids decreases with increasing chain length (HERA Targeted Risk Assessment Report of Fatty Acid Salts, June 2002), while irritation potential of the unsaturated oleic acid (C18:1) was shown to be higher as of the saturated stearic acid (C18)(CIR 2005).

Therefore the hazard prediction for MDEA Esterquat C18 satd. derived from studies with test substances with higher degree of unsaturation and shorter alkyl chains are likely to be an over- rather than an underestimation. 

Comparison of molecular structures and local toxicity data of MDEA Esterquat C18 satd. and MDEA-Esterquat C16-18 and C18 unsatd.

 

 

Source substances

Target substance

 

Endpoints

 

MDEA-Esterquat C16-18 and C18 unsatd.

 

MDEA Esterquat C18 satd.

Skin Irritation

OECD 404, RL 1, GLP

 

grading at 24, 48 and 72 hours after removal of the test material

 

mean scores erythema: 0.33/0.00/0.33

mean scores edema: 0.00/0.00/0.00.

effects were fully reversible within 48 hours

 

 Not irritating

read-across from MDEA-Esterquat C16-18 and C18 unsatd.

 

OECD 404, RL 2, GLP

 

40% in corn oil, occlusive dressing

 

grading at 24 and 72 hours after application of the test material

 

mean scores erythema: 1.00/< 1.00/0.00

mean scores edema: 0.00/0.00/0.00.

effects were fully reversible within 72 hours

 

 Not irritating

OECD 405, RL 1, GLP

 

40% in corn oil, 24 hours under occlusive dressing, 6 rabbits

 

grading at 24 and 72 hours after application of the test material

 

mean scores erythema: 1.3

mean scores edema:1.75

effects were fully reversible within 7 days

 

 Not irritating

OECD 439, RL 1, GLP

 

relative mean tissue viability obtained after 15 minutes treatment: 108%

 

 Not irritating

Eye Irritation

OECD 405, RL 1, GLP

 

Grading after 24, 48 and 72 hours

 

mean cornea score: 0.00/0.00/0.00

mean iris score: 0.00/0.00/0.00

mean conjunctivae score:0.00/0.33/0.33.

mean chemosis score: 0.00/0.00/0.00

effects were fully reversible within 48 hours

 

 

not irritating

read-across from MDEA-Esterquat C16-18 and C18 unsatd.

 

OECD 405 (LVET), RL 2, GLP

 

Grading after 24, 48 and 72 hours

 

mean cornea score: 0.00

mean iris score: 0.00

mean conjunctivae score:0.00

mean chemosis score: 0.00

 

 

not irritating

Fatty acids

<C16      <7%                                            

C16, 16‘  26-35%

C18         42-52%

C18‘        15-20%

C18‘‘,18‘‘‘ ≤ 1.5%

>C18       ≤ 2%

C16 8%

C18 92%

IV

< 25

< 1

Headgroup

MDEA

MDEA

Anion

Chloride

Chloride

 

References

Cosmetic Ingredient Review; Final Report of the Cosmetic Ingredient Review Expert Panel; Final Report on the Safety Assessment of Oleic Acid, Lauric Acid, Palmitic Acid, Myristic Acid, and Stearic Acid; June 2005.

HERA Targeted Risk Assessment Report of Fatty Acid Salts, June 2002


Justification for selection of eye irritation endpoint:
No single key study has been selected, since both studies (OECD guideline studies, no deviations, GLP) are used in a weight of evidence approach

Justification for classification or non-classification

Based on the available data MDEA Esterquat C18 satd. do not require to be classified as irritating to the skin or irritating to the eye according to Directive 67/548/EEC as well as GHS Regulation EC No 1272/2008 and labelling is not necessary.