Reaction product of "Fatty acids, C16-18 (even numbered) and C18-unsaturated" and "reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol

Endpoint summary

Description of key information

The LLNA skin sensitisation study was performed according to OECD Guideline 429 and GLP principles. In addition, company data and read across data is available.

For a NICNAS notification in Australia further data was required, as the available information (see below) was not considered sufficient to reverse the classification that was based on the LLNA study. The GPMT skin sensitisation study was performed according to OECD Guideline 406 and GLP principles.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 August, 2012 - 09 November 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

(1992)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

(2008)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2600 (Skin Sensitisation)

Version / remarks:

(2003)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Previously, a LLNA study was performed. However, for a NICNAS notification in Australia further data was required, as the available information was not considered sufficient to reverse the classification that was based on the LLNA study. Therefore a GPMT study was conducted.

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany.
- Age at study initiation: Young adult animals (approx. 4 weeks old)
- Weight at study initiation: 239 - 306 g
- Housing: Group housing of maximally 5 animals per labeled Noryl cage.
- Diet: Complete maintenance diet for guinea pigs (MS-H ered, SSNIFF® Spezialdiäten, GmbH, Soest, Germany). In addition, hay (TecniLab-BMI BV, Someren, The Netherlands) was provided at least twice a week.
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 24
- Humidity (%): 40 - 70
- Air changes (per hr): Approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

Route:

intradermal and epicutaneous

Vehicle:

corn oil

Concentration / amount:

20% for the intradermal induction and 100% for the epidermal induction.
50% for the challenge phase.

Route:

epicutaneous, occlusive

Vehicle:

corn oil

Concentration / amount:

20% for the intradermal induction and 100% for the epidermal induction.
50% for the challenge phase.

No. of animals per dose:

Test animals: 20
Control animals: 10

Details on study design:

RANGE FINDING TESTS: (4 animals, age: between 4 and 9 weeks old)

Intradermal injections:
A series of four test substance concentrations was used (10, 20, 50 and 100%). Each of two animals received two different concentrations in duplicate (0.1 mL/site) in the clipped scapular region. The injection sites were assessed for irritation 24 and 48 hours after treatment.
- Result: A 20% test substance concentration was selected for the main study: necrosis was observed at 100 and 50%, while only erythema was observed at 20% (grade 2 at 24h, grade 1 at 48h).

Epidermal application:
A series of four test substance concentrations was used (10, 20, 50 and 100%) Two different concentrations were applied (0.5 mL each) per animal to the clipped flank, using Metalline patches (2x3 cm) mounted on Medical tape which were held in place with Micropore tape and subsequently Coban elastic bandage. The animals receiving intradermal injections were treated with the lowest concentrations and two further animals with the highest concentrations. After 24 hours, the dressing was removed and the skin cleaned of residual test substance using water. The treated skin areas were assessed for irritation 24 and 48 hours after exposure.
- Result: A 100% concentration was selected for the epidermal induction exposure as no signs of necrosis were present but only slight irritation was observed.

MAIN STUDY
The Main study was performed in a step wise fashion. Initially, ten experimental and five control animals were treated. Based on the results, the study was extended with another set of ten experimental and five control animals.
A. INDUCTION EXPOSURE
- No. of exposures: 1

1) Intradermal injections on day 1:
- Site: scapular region. One of each pair was on each side of the midline and from cranial to caudal:
Three pairs of intradermal injections:
1) 0.1 mL: FCA (50% in water for injection)
2) 0.1 mL: test substance at a 20% concentration (control animals: 0.1 mL corn oil)
3) 0.1 mL: 1:1 mixture of the test substance at a 40% concentration + FCA (undiluted)
- Readings: on day 3 (48 hrs after the injections)

2) Topical application on day 8:
- Amount: 0.5 mL (control animals: 0.5 mL corn oil) 100% test substance
- Area: approximately 6 cm^2
- Exposure period: 48 hours (occlusive)
- Readings: scores were rated directly after patch removal

B. CHALLENGE EXPOSURE (all animals, with the 50% test substance and the vehicle)
- Day of challenge: day 21 or 22
- Exposure period: 24 hours (occlusive)
- Site: flank
- Amount: 0.1 mL
- Readings: scores were rated 24 and 48 hours after patch removal

RECHALLENGE: Day 30, like first challenge, in first set of animals.

OBSERVATIONS
Mortality/Viability: Twice daily
Toxicity: At least once daily.
Body weights: Prior to start and after (each) challenge.
Necropsy: No necropsy was performed.
Irritation: Skin reactions were graded according to OECD 406. The intradermal reactions were assessed for erythema only or, if necrosis is present, the diameter of necrosis. A description of all other local effects was recorded.
To facilitate scoring, the epidermally treated skin-areas were clipped at least 3 hours before the next reading.

Challenge controls:

Not applicable.

Positive control substance(s):

yes

Remarks:

the results of the latest reliability check, performed in May/June 2012 with Alpha- Hexylcinnamaldehyde, are reported.

Positive control results:

The latest reliability check shows a sensitisation rate of 80%

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

50%

No. with + reactions:

1

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50%. No with. + reactions: 1.0. Total no. in groups: 20.0.

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

vehicle

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: vehicle. No with. + reactions: 0.0. Total no. in groups: 10.0.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

50%

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 20.0.

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

vehicle

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: vehicle. No with. + reactions: 0.0. Total no. in groups: 10.0.

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

test chemical

Dose level:

50%

No. with + reactions:

1

Total no. in group:

10

Clinical observations:

(first set of animals were rechallenged)

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 50%. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: (first set of animals were rechallenged).

Key result

Reading:

rechallenge

Hours after challenge:

24

Group:

negative control

Dose level:

vehicle

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

(first set of animals were rechallenged)

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 24.0. Group: negative control. Dose level: vehicle. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: (first set of animals were rechallenged).

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

50%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

(first set of animals were rechallenged)

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: (first set of animals were rechallenged).

Key result

Reading:

rechallenge

Hours after challenge:

48

Group:

negative control

Dose level:

vehicle

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

(first set of animals were rechallenged)

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: vehicle. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: (first set of animals were rechallenged).

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

20%

No. with + reactions:

8

Total no. in group:

10

Remarks on result:

positive indication of skin sensitisation

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

20%

No. with + reactions:

7

Total no. in group:

10

Remarks on result:

positive indication of skin sensitisation

Interpretation of results:

other: Not classified.

Remarks:

According to Regulation (EC) 1272/2008

Conclusions:

In a guinea pig maximisation test method the potential of Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol for skin sensitisation was tested according to OECD 406 guideline and GLP principles, showing a sensitization rate of 5 per cent.
Based on these results Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol does not have to be classified and has no obligatory labelling requirement for sensitization by skin contact according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Executive summary:

In a guinea pig maximisation test method the potential of Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol for skin sensitisation was tested according to OECD 406 guideline and GLP principles.

Slight to moderate erythema was observed during the intradermal induction (conc. 20%) at all injection sites in control and test animals.

Slight erythema was observed following the epidermal induction (conc. 100%) in 12 test animals and 3 control animals.

No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study.

In the first set of ten experimental and five control animals, the first challenge resulted in skin reactions of discrete or patchy erythema in one experimental animal in response to the 50% test substance concentration. No skin reactions were evident in the control animals. To confirm the results of the first challenge, the first challenge was repeated in a second challenge one week later. The second challenge results were the same as for the first challenge.

Based on the test guidelines, a second set of ten experimental and five control animals were treated in the same way as the first set. No skin reactions were evident after the challenge exposure in this second set of animals. These results indicate a sensitization rate of 5 per cent.

Based on these results Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol does not have to be classified and has no obligatory labelling requirement for sensitization by skin contact according to Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

LLNA study:

In a mouse lymph node assay, performed according to OECD, EC and EPA test guidelines, mouse were treated with 0%, 25%, 50%, or 100% of Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol on three consecutive days by open application on the ears. Three days after the last exposure all animals were injected with 3H-methyl thymidine. Radioactivity measurements of the lymph nodes were performed. Slight erythema was noted among the animals treated at 50% and in all animals treated at 100%. No oedema was observed in any of the animals examined. The irritation of the ears as shown by the animals was considered not to have a toxicologically significant effect

on the activity of the nodes. The majority of nodes were considered normal in size, except for the enlarged nodes of three animals treated at 50% and three animals treated at 100%. No macroscopic abnormalities of the surrounding area were noted. The SI values calculated for the substance concentrations 25, 50 and 100% were 1.4, 4.7 and 6.9 respectively. These results indicate that the test substance could elicit an SI >= 3. The data showed a dose-response and an EC3 value of 37.1% was calculated. Based on these results the substance is a skin sensitiser.

Company health records:

For Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol a medical record on skin sensitisation is provided by the manufacturing company. From this it is clear that operators and lab technicians might be exposed during sampling and analysis of these samples, although operators have RMMs such as gloves and overall in place. Sevenhundred and five sampling times over a period of 7 years are recorded, showing that mainly the same operators and technicians have dealt with this work, so if exposure might have taken place, this did not result in skin sensitisation.

GPMT study:

In a guinea pig maximisation test method the potential of Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol for skin sensitisation was tested according to OECD 406 guideline and GLP principles.

Slight to moderate erythema was observed during the intradermal induction (conc. 20%) at all injection sites in control and test animals. Slight erythema was observed following the epidermal induction (conc. 100%) in 12 test animals and 3 control animals. No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study.

In the first set of ten experimental and five control animals, the first challenge resulted in skin reactions of discrete or patchy erythema in one experimental animal in response to the 50% test substance concentration. No skin reactions were evident in the control animals. To confirm the results of the first challenge, the first challenge was repeated in a second challenge one week later. The second challenge results were the same as for the first challenge. Based on the test guidelines, a second set of ten experimental and five control animals were treated in the same way as the first set. No skin reactions were evident after the challenge exposure in this second set of animals.These results indicate a sensitization rate of 5 per cent. Based on these results Radia 7853 does not have to be classified.

Discussion:

In a human patch test with Radiagreen RA, no or only a minimal risk of skin sensitization is present when this substance is coming into contact with human skin. Radiagreen RA is a reaction product of fatty acids (C8-C10) and triglycerol, while Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol is a reaction product of fatty acids C16-18 and C18 unsatd and rape-oil fatty acids (CAS nrs 67701-08-0 and 93165-31-2), and Polyol PX (reaction mass of1,3-Propanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- (CAS nr 68658-38-8) and 1,3-dioxane-5,5-dimethanol (CAS nr 6228-25-7), overall EC nr 911-819-6).All the starting products of Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol are currently not classified for sensitization according to GHS.

In addition, a literature search has been performed on all constituents. This shows that palmitic (C16), stearic (C18) and oleic (C18 unsatd) acid do not show sensitising properties in human patch tests and in other tests with humans (see Toxnet, mainly HSDB). Also pentaerythritol is considered not sensitizing (Toxnet, HSDB). No public data on Polyol PX are available or its complete constituents. A study report on Polyol PX (2010, acc to OECD 429 and GLP) has been submitted for REACH registration, showing a LLNA study with negative results (access is permitted).

Several publications have shown that for some type of substances, LLNA studies gave positive results while GPMT studies gave negative results. The publications of e.g. Kreiling et al. (Food Chem Tox, 46, p. 1896 -1904, 2008) and Garcia et al. (Reg Tox Pharm, 58, p. 301 -7, 2010) describe the results of LLNA and GPMT testing for several (unsaturated) fatty acids having a chain length from C4 to (>) C18. For most substances the LLNA

gave positive results (12 out of 19), while the GPMT did give only one positive result (1 out of 19). As these test substances were either endogenous physiological constituents, common and widely used natural constituents of food and/or cosmetics, or nonionic sugar lipid surfactants, the high number of positives in the LLNA test was unexpected. Radia 7853 does contain (unsaturated) fatty acids C16-C18 as starting products.

For a NICNAS notification in Australia further data was required, as the available information (see above) was not considered sufficient to reverse the classification that was based on the LLNA study. The GPMT skin sensitisation study was performed according to OECD Guideline 406 and GLP principles and was shown to be not sensitising.

 

Taking all this information into account, it is therefore concluded that Reaction products of Fatty acids, C16-18 and C18-unsatd. and reaction mass of 1,3-alkanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]- and 1,3-heteromonocycle-5,5-dimethanol is not expected to have sensitising properties and thus does not have to be classified accordingly.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on all information available, the substance is not classified as skin sensitizer according to CLP Regulation (EC) No. 1272/2008.