Fatty acids, tall-oil, reaction products with pentaethylenehexamine

Endpoint summary

Description of key information

- DPRA test: negative

- ARE reporter assay: negative

- MUSST assay: positive

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Reference 1

Endpoint:

skin sensitisation: in chemico

Remarks:

DPRA test

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2012/13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 442C (In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA))

Principles of method if other than guideline:

Gerberick GF, Vassallo JD, 8ailey RE, Chaney JG, Morrall SW, Lepoittevin JP.
Development of a Peptide Reactivity Assay for Screening Contact Allergens.
Toxicological Seiences 81 ,332-343, 2004.

Gerberick GF, Vassallo JD, Foertsch LM, Price 88, Chaney JG, Lepoittenvin JP.
Quantificationn of Chemical Peptide Reactivity for Screening Contact Allergens: A
Classification Tree Model Approach. Toxicological Seiences 97(2), 417-427, 2007.

Bauch C, Kolle SN, Fabian E, Pachel C, Ramirez T, Wiench B, Wruck CJ , van
Ravenzwaay B, Landsiedei R. lntralaboratory validation of four in vitro assays for
the prediction of the skin sensitizing potential of chemicals. Toxicology in Vitro 25,
1162 - 1168, 2011.

Maxwell G, Aeby P, Ashikaga T, Bessou-Touya S, Diembeck W, Gerberick F,
Kern P, Marrec-Fairley M, Ovigne JM, Sakaguchi H, Schroeder K, Tailhardat M,
Teissier S, Winkler P. Skin sensitisation: the Colipa strategy for developing and
evaluating non-animal test methods for risk assessment. AL TEX 28(1 ): 50-5,
2011.

GLP compliance:

yes (incl. QA statement)

Type of study:

other: direct peptide reactivity assay

Parameter:

cysteine depletion

Remarks:

mean depletion (%)

Value:

0 %

Result: minimal reactivity

mean peptide depletion = 0%

Interpretation of results:

study cannot be used for classification

Reference 2

Endpoint:

skin sensitisation: in vitro

Remarks:

ARE reporter gene assay

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 442D (In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method)

Principles of method if other than guideline:

At the time of study conduct, there were no official national or international guidelines for the LuSens Assay; however, the study was performed according to the methods described in the following publication:
Bauch C, Kolle SN, Ramirez T, Eltze T, Fabian E, Mehling A, Teubner W, van Ravenzwaay B, Landsiedel R, (2012), Putting the parts together: Combining in vitro methods to test for skin sensitizing potentials, Regul Toxicol Pharmacol, 63(3):489-504.

GLP compliance:

yes (incl. QA statement)

Type of study:

activation of keratinocytes

Details on the study design:

- Cell line: LuSens, transgenic keratinocyte cell line derived from HaCaT cells, prepared in collaboration with Christoph J. Wruck, RWTH Aachen, Germany
- LuSens cells from the working cell bank were thawed and cultured in culture medium 1 under standard culture conditions (37°C, ca. 5% CO2, ≥ 90% humidity) for at least 2 weeks at passage >5 but not longer than 15 passages prior to testing.
- Before substance incubation, cells were seeded in 96-well microtiter plates (120 μL of 0.83 x 10^5 cells/mL cell suspensions) in culture medium 2.
- Positive control: Ethylene glycol dimethacrylate (EGDMA 18 μg/mL) PSN 08/0496

Positive control results:

The fold induction (luciferase activity) of the positive control was 5.62 in the first experiment and 6.69 in the second experiment at concentrations that did not reduce viability below 70%.

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of skin sensitisation

After 48 hours of exposure to test substance luciferase activity in LuSens cells was not induced at concentration affording at least 70% viability. From this it has to be concluded that test substance does not have a keratinocyte activating potential.

Interpretation of results:

study cannot be used for classification

Conclusions:

Based on the observed results it was concluded that the test item does not induce luciferase activity in LuSens cells under the test conditions chosen.

Reference 3

Endpoint:

skin sensitisation: in vitro

Remarks:

MUSST assay

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2012/2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

no guideline available

Principles of method if other than guideline:

In Vitro Sensitization: Dendritic Cell Line Activation Assay Myeloid U937 Skin Sensitization Test (MUSST)
There are no official national or international guidelines for the MUSST Assay; however, the study was performed according to the methods described in the following publications:
 Python F, Goebel C, Aeby P. (2007) Assessment of the U937 cell line for the detection of contact allergens. Toxicol Appl. Pharmacol. 220(2), 113-24.
 Bauch C, Kolle SN, Fabian E, Pachel C, Ramirez T, Wiench B, Wruck CJ, van Ravenzwaay B, Landsiedel R. (2011) Intralaboratory validation of four in vitro assays for the prediction of the skin sensitizing potential of chemicals. Toxicology in Vitro 25, 1162 – 1168.

GLP compliance:

yes (incl. QA statement)

Type of study:

activation of dendritic cells

Details on the study design:

- Preparation of the cells:
U937 cells from the working cell bank were thawed and cultured in suspension using complete RPMI 1640 medium with 25 mM HEPES buffer and 2 mM L-glutamine supplemented with 10% fetal bovine serum and 100 U/mL penicillin and 100 μg/mL streptomycin under standard culture conditions (37°C, ca. 5% CO2, ≥ 90% humidity) until for 5 passages but not longer than passage 13 prior to testing.
For substance incubation, cells were seeded in 96-well microtiter plates (100 μL of 0.5x10^6 cells/mL cell suspensions).

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of skin sensitisation

	1st experiment		2nd experiment		3rd experiment
c [ug/ml]	CD 86 induction	rel. viability	CD 86 induction	rel. viability	CD 86 induction	rel. viability
VC	1.00	100	1.00	100	1.00	100
0.3	0.99	99.7	1.36	100.4	1.15	99.9
0.53	1.06	100	1.72	100.5	1.26	100.1
1.06	1.17	99.8	1.94	100.4	1.29	100
2.11	1.14	99.6	2.18	100	1.28	99.9
4.22	1.14	99.3	3.54	98.3	1.63	99.5
VC	1.00	100	1.00	100	1.00	100
LA 200 ug/ml	0.83	100.1	1.00	100.7	1.02	99.9
EDA 70 ug/ml	2.26	93.6	2.86	94	2.33	92.8

Interpretation of results:

study cannot be used for classification

Conclusions:

Based on the observed results it was concluded that the test item does induce dendritic cell activation in the MUSST assay under the test conditions chosen.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

A combination of several in vitro methods addressing key steps of the adverse outcome pathway (AOP) for skin sensitization as defined by OECD, has been conducted to assess the skin sensitizing potential of the test material:

• protein reactivity (DPRA),

• activation of keratinocytes (luSens), and

• activation of dendritic cells (MUSST).

The combination of test methods and the evaluation of their results has been evaluated and published by Bauch et al., 2012. Based on the performance standards of the OECD test guideline no. 429 (Local Lymph Node Assay, LLNA, OECD 2010) the evaluation based on the DPRA, LuSens and MUSST methods yields an overall accuracy of 95% compared to results in humans (for comparison: for the same data set the LLNA yielded an overall accuracy of 86%). A skin sensitizing (quantitative) potency assessment using the reported results was not validated at the time of writing of this report. Based on the results and applying the evaluation criteria described the test item is predicted not to be a skin sensitizer.

According to the evaluation criteria: "ln the test battery evaluation a weight of evidence (WoE) approach is used: Any two of the three tests determine the overall results, i.e. any two positive test results drive the prediction of a sensitizer, while any two negative test results drive the prediction of a test substance to be a non-sensitizer (Bauch et al. 2012)." the test item is predicted not to be a skin sensitizer.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for skin sensitization under Regulation (EC) No. 1272/2008, as amended for the 14th time in Regulation (EU) 2020/217.