Registration Dossier

Administrative data

Description of key information

Skin Sensitisation in vivo (WoE, OECD 406/EU Method B.6, Buehler and GPMT): not sensitising

RA from source substances Glycine, N-methyl-, N-coco acyl derivatives, sodium salts (CAS 61791-59-1), N-methyl-N-[C18-(unsaturated)alkanoyl]glycine (EC 701-177-3) and Sodium N-lauroylsarcosinate (CAS 137-16-6)

Skin Sensitisation in vitro (WoE, OECD 442C (DPRA), OECD 442D (LuSens), OECD 442E (h-CLAT)): not sensitising

RA from source substance Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3)

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation, other
Remarks:
in vitro / in chemico and in vivo (non-LLNA)
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Remarks:
Summary of available data used for the endpoint assessment of the target substance
Adequacy of study:
weight of evidence
Justification for type of information:
Please refer to analogue justification report provided in IUCLID section 13
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
induction: 0.05 and 5%; challenge: 5%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Source: CAS 137-16-6, Inversek, 1987, GPMT
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
challenge: 5%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Source: CAS 137-16-6, Inversek, 1987, GPMT
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
induction: 0.05 and 5%; challenge: 5%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Source: CAS 137-16-6, Inversek, 1987, GPMT
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
challenge: 5%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Source: CAS 137-16-6, Inversek, 1987, GPMT

Additional studies taken into account in the Weight-of-Evidence approach:

in vitro / in chemico:

CAS 30364-51-3, BASF, 2016: negative for dendritic cell activation (h-CLAT)

CAS 30364-51-3, BASF, 2016: positive for keratinocyte activation (LuSens)

CAS 30364-51-3, BASF, 2016: negative for peptide reactivity (DPRA)

in vivo:

CAS 61791-59-1, Frey-Tox, 2005: not sensitising (Buehler)

EC 701-177-3, Ciba Geigy, 1981b: not sensitising (GPMT)

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.
Executive summary:

.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

No data on the skin sensitisation potential are available for the registered substance Reaction mass of N-(1-oxooctadecyl)sarcosine and N-hexadecanoyl-N-methylglycine (EC 947-850-7). Therefore, reliable study data from analogue substances were considered. In fact, with respect to in vivo data, a weight of evidence approach was applied, referring to following substances: Glycine, N-methyl-, N-coco acyl derivatives, sodium salts (CAS 61791-59-1), N-methyl-N-[C18-(unsaturated)alkanoyl]glycine (EC 701-177-3) and Sodium N-lauroylsarcosinate (CAS 137-16-6). Additonally, in vitro data as available from the analogue substance Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) also were taken into account.

 

in vivo studies

The skin sensitisation properties of Glycine, N-methyl-, N-coco acyl derivatives, sodium salts (CAS 61791-59-1) were tested in a study performed according to the OECD TG 406 under GLP conditions, using the test for delayed contact hypersensitivity in guinea pigs (Buehler test, Frey-Tox, 2005). The Buehler test was performed on 30 female Albino guinea pigs (Crl:HA). For the dermal inductions the initially test item concentration was 100 and 75% (v/v), respectively (1st induction 100% (v/v), 2nd and 3rd induction 75% (v/v)), whereas a 50% (v/v) preparation of the test item was selected for the challenge application. Topical application of the appropriate test substance concentration (20 test animals) or vehicle (10 control animals) was performed once a week for 6 hours at the flanks of each animal for three consecutive weeks. Seven days following the last topical application of the test substance or vehicle all animals were challenged with the test substance at a concentration of 50% (v/v) for 6 hours. Skin reactions were evaluated 24 and 48 h after challenge application. Only 1/20 animals (5%) responded with a slight erythema formation after challenge application. As a limitation of this study, no data on periodic reliabilty checks with an appropriate positive control were mentioned in the study report. In conclusion, the test material was not skin sensitising under the conditions of this Buehler test.

 

The skin sensitising properties of N-methyl-N-[C18-(unsaturated)alkanoyl]glycine (EC 701-177-3) were tested in a Guinea Pig Maximisation Test (GPMT) similar to OECD Guideline 406 (Ciba Geigy, 1981b). In the study, Pirbright white guinea pigs (20/group) were induced with a single intradermal injection of the test substance at 5% (in 20% ethanol, 80% saline) using the adjuvant complete Freund and an epicutaneous application of the test substance at 30% (Vaseline PhH VI) on the injection site. The negative control group was included in the study but no information about treatment was given. Epicutaneous challenge exposure was conducted 20 days after the first induction for 24 h under occlusive conditions. 3% of the test substance was applied on the flank. Evaluation of skin reactions was carried out 24 h after challenge. No positive control substance was included in the study. No skin reactions were observed in any animal in the negative control group. In the treatment group, three animals with very slight erythema and two animals with well-defined erythema were observed (corresponding to 25% positive results). Thus, the available data on skin sensitisation do not provide evidence for sensitising properties of N-methyl-N-[C18-(unsaturated)alkanoyl]glycine (EC 701-177-3) under the conditions of this GPMT.

 

The skin sensitising properties of Sodium N-lauroylsarcosinate (CAS 137-16-6) were tested in a Guinea Pig Maximisation Test (GPMT) according to EU method B.6 under GLP conditions (Inversek, 1987). A preliminary range finding test was conducted to determine suitable concentrations for the main study for the intradermal injection and the patch testing. In the main study, female Dunkin-Hartley guinea pigs (20/group) were induced on Day 0 with 3 pairs of intradermal injections of FCA, 0.05% (v/v) of the test substance alone and 0.05% test substance in FCA/water. Epicutaneous induction was done by application of the test substance at 5% in water on Day 8. The negative control group was treated with FCA, water or water/FCA alone. Six days after the injection phase the injection site of each of the test and control group animals was shaved again and then wetted with 10% aqueous SLS to provoke a mild inflammatory response to enhance the possibility of sensitisation. Epicutaneous challenge exposure was conducted 20 days after the first induction for 24 h under occlusive conditions. Test substance was applied at a concentration of 5% on the right flank, and the vehicle was applied on the left flank, respectively. Evaluation of skin reactions was carried out 48 and 72 h after challenge exposure. No positive control substance was included in the study. Moderate erythema formation was noted in test group animals, and slight erythema was noted in control group animals after induction. After challenge exposure, all test and vehicle control animals showed no skin reactions after 48 and 72 h. No changes in body weight were observed in any animal during the study period. No further clinical signs were noted at any time point during the study. Thus, the available data on skin sensitisation do not provide evidence for sensitising properties of Sodium N-lauroylsarcosinate under the conditions of this GPMT.

 

in vitro Studies

The reactivity of the analogue substance Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) towards synthetic cysteine (C)- or lysine (K)-containing peptides was evaluated in the Direct Peptide Reactivity Assay (DPRA) according to OECD TG 442C (BASF, 2016). For this purpose the test substance was incubated with synthetic peptides for ca. 24 hours at ca. 25°C and the remaining non-depleted peptide concentrations were determined by high performance liquid chromatography (HPLC) with gradient elution and UV-detection at 220 nm. The test substance was dissolved at a 100 mM concentration in de-ionized water. Three samples of the test substance were incubated with each peptide in ratios of 1:10 (for C-peptide) or 1:50 (for K-peptide). Visual observation after the 24 -hour incubation time did not reveal precipitates in any samples of the test substance with the peptides. Additionally triplicates of the concurrent vehicle control were incubated with the peptides. Further, a co-elution control was performed in order to detect possible interference of the test substance with the peptides. The samples consisted of the test substance, vehicle and the respective peptide buffer but without peptide. Moreover the samples were analyzed by measuring UV absorbance at 258 nm and the area ratio 220 nm / 258 nm was calculated as a measure of peak purity. No co-elution of test substance and peptides was noticed. The mean C-peptide depletion, caused by the test substance was determined to be -8.52%. The mean K-peptide depletion, caused by the test substance was determined to be 0.23%. Negative depletions were considered to be “zero” for calculation of the mean peptide depletion, which was thus calculated to be 0.11%. Based on the observed results it was concluded that Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) shows a minimal chemical reactivity in the DPRA under the test conditions chosen.

 

The keratinocyte activating potential of the source substance Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) was evaluated in the LuSens assay according to OECD TG 442D (BASF, 2016). For this purpose the test substance was incubated with a Luciferase reporter cell line (LuSens cells) for ca. 48 hours at 37°C and antioxidant response element (ARE) dependent luciferase activity was measured in a luminometer. In order to determine the concentrations suitable for the main experiment a pre-test was performed. Cells were exposed to 9 concentrations of the test substance and cytotoxicity was determined by MTT assay. The CV75 value (= estimated concentration that affords 75% cell viability) was determined by linear regression from the concentration response curve. In the main test luciferase activity was measured after 48 hour exposure. In parallel a MTT assay was performed to assess cytotoxicity of the test substance. A total of 3 valid experiments were performed. At concentrations used in the main experiment the test substance was soluble in 1% DMSO in culture medium 3 (based on good homogeneity of the preparation). No precipitates were noticed in any concentration after 48 hours. The EC1.50 (the concentration resulting in a 1.50-fold luciferase induction) was calculated to be 19 μg/mL (experiment 3). In summary, after 48 hours of exposure to test substance Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) luciferase activity in LuSens cells was induced in at least two consecutive concentrations with statistical significance affording at least 70% viability in at least two independent experiments. Therefore, it was concluded that the substance has a keratinocyte activating potential.

 

The potential of the test substance Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) to induce the cell membrane markers CD86 and CD54 expression was evaluated in the Human Cell Line Activation Test (h-CLAT) according to OECD TG 442E (BASF, 2016). For this purpose the test substance was incubated with human pro-monocytic cell line THP-1 for ca. 24 hours at 37°C and membrane markers expression was measured by flow cytometry. In order to determine the concentrations suitable for the main experiment a pre-test was performed. Cells were exposed to 10 concentrations of the test substance and cytotoxicity was determined by propidium iodide (PI) intercalation into the DNA. The CV75 value (=estimated concentration that affords 75% cell viability) was determined by linear regression from the concentration response curve. In the main test after 24 hour exposure THP-1 cells were stained with FITC labeled anti-human-CD86/ anti-human-CD54 antibody and propidium iodide and the fluorescence intensity was analyzed using flow cytometry. A total of 2 valid experiments was performed. At concentrations used in the main experiment the test substance was soluble in culture medium. No precipitates were noticed in any concentration after 24 hours. Calculation of an EC150 (the concentration resulting in a RFI of 150) for CD86 and an EC200 (the concentration resulting in a RFI of 200) for CD54 was not applicable. In summary, after 24 hours of exposure to Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) CD86 and CD54 expression was not induced in THP-1 cells affording at least 50% viability in at least two independent experiments. Therefore, it was concluded that the test substance did not induce dendritic cell activation.

 

Based on the results of the 3 in vitro studies, the analogue substance Sodium N-methyl-N-(1-oxotetradecyl)aminoacetate (CAS 30364-51-3) is not peptide reactive, activates keratinocytes and does not activate dendritic cells. Therefore, the test substance is not considered to be a skin sensitiser.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The available skin sensitisation data from analogue substances are conclusive but not sufficient for classification, and thus, the criteria for classification according to Regulation (EC) No. 1272/2008 (CLP) are not met. Therefore, Reaction mass of N-(1-oxooctadecyl)sarcosine and N-hexadecanoyl-N-methylglycine (EC 947-850-7) is also considered not to meet the criteria for classification for skin sensitisation according to Regulation (EC) No 1272/2008 (CLP).