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EC number: 812-222-2 | CAS number: 4443-23-6
Skin sensitisation: not sensitising based on weight of evidence from 3 in chemico/in vitro studies.
- positive in DPRA (OECD 442C, GLP)
- negative in KeratinoSens (OECD 442D, GLP)
- negative in U-SENS (OECD 442E, GLP)
Respiratory sensitisation: no study available
Upon preparation of the SPCC test item samples a precipitate was observed while upon preparation of the SPCL test item samples no precipitate or phase separation was observed. After incubation of the SPCC and SPCL test item samples, no precipitate or phase separation was observed in any of the samples.An overview of the individual results of the cysteine and lysine reactivity assays as well as the mean of the SPCC and SPCL depletion are presented in the table below. In the cysteine reactivity assay the test item showed 1.2% SPCC depletion while in the lysine reactivity assay the test item showed 30.4% SPCL depletion. The mean of the SPCC and SPCL depletion was 15.8% and as a result the test item was positive in the DPRA and was classified in the “lowreactivityclass” when using the Cysteine 1:10 / Lysine 1:50 prediction model. In conclusion, this DPRA test is valid. TA-3 was positive in the DPRA and was classified in the “low reactivity class” when using theCysteine 1:10 / Lysine 1:50 prediction model.
Table 2: Overview Luminescence Induction and Cell Viability Positive Control EDMG in Experiment 1 and 2
Exp 1 luminescence
Exp 1 viability (%)
Exp 2 luminescence
Exp 2 viability (%)
Table 3: Overview Luminescence Induction and Cell Viability of TA-3 in Experiment 1 and 2
Overview Stimulation index of CD86 and Cell Viability in Experiment 1 and 2 of the Positive, Negative and Vehicle Control
% Viability (Mean)*
IgG1 value (%)
CD86 basal expression (%)
RPMI Mean Viability
* Red values are either below 70% viability, above 150 S.I..
In chemico/in vitro skin sensitisation
There are three studies available, in which the substance was tested in chemico and in vitro for potential induction of skin sensitisation. All three studies were conducted under GLP conditons and in accordance with OECD Guidelines 442C (DPRA), 442D (KeratinoSens) and 442E (U-SENS), respectively.
- Direct Peptide Reactivity Assay (DPRA)
The objective of this study was to determine the reactivity of the test item towards model synthetic peptides containing either cysteine (SPCC) or lysine (SPCL). After incubation of the test item with either SPCC or SPCL, the relative peptide concentration was determined by High-Performance Liquid Chromatography (HPLC) with gradient elution and photodiode array (PDA) detection at 220 nm and 258 nm. SPCC and SPCL Percent Depletion Values were calculated and used in a prediction model which allows assigning the test item to one of four reactivity classes used to support the discrimination between sensitizers and non-sensitizers.
The study procedures described in this report were based on the most recent OECD guideline.
In the cysteine reactivity assay the test item showed 1.2% SPCC depletion while in the lysine reactivity assay the test item showed 30.4% SPCL depletion. The mean of the SPCC and SPCL depletion was 15.8% and as a result the test item was considered to be positive in the DPRA and classified in the “low reactivity class” when using the Cysteine 1:10 / Lysine 1:50 prediction model.
- KeratinoSens™ Assay
The objective of this study was to evaluate the ability of the test item to activate the antioxidant/electrophile responsive element (ARE)-dependent pathway in the KeratinoSens™assay.
Cells were incubated with the test item in a concentration range of 0.49 – 1000 µM (2-fold dilution series) for 48 hours ± 1 h.
The activation of the ARE-dependent pathway was assessed by measuring the luminescence induction compared to the vehicle control. In addition, the viability was assessed with an MTT assay.
The test item showed no toxicity (no IC 30 and IC 50 value) and no biologically relevant induction of the luciferase activity (no EC 1.5 value) was measured at any of the test concentrations in both experiments. The maximum luciferase activity induction (I max ) was 1.17-fold and 1.15-fold in experiment 1 and 2, respectively. The test item is classified as negative in the KeratinoSens™ assay since negative results (<1.5-fold induction) were observed at test concentrations up to 1000 µM.
In conclusion, the test item is classified as negative (no activation of the antioxidant/electrophile responsive element (ARE)-dependent pathway in keratinocytes) under the experimental conditions described.
- U-SENS ™assay
The objective of this study is to evaluate the ability of the test item to increase the expression levels of CD86 cell surface marker in the U937 cell line in the U937 cell line activation Test (U-Sens™) assay. The cell viability before incubation with the test item was > 90% (99% in experiment 1 and 2). Cells were incubated with the test item in a concentration range of 1.0 – 200 μg/mL and 20 – 200 μg/mL in experiment 1 and 2, respectively. The increase of CD86 cell surface marker expression was assessed by measuring the amount fluorescent cell staining of the CD86 cell surface marker compared to the vehicle control. In addition, the viability was assessed with propidium iodide.The test item showed no toxicity (No CV70value) and no biologically relevant induction of the CD86 activity (No EC150 value) was measured at any of the test concentrations in both experiments. The test item is classified as negative in the U-Sens™ assay since negative results (< 150% increase) were observed at all test concentrations with a cell viability of >70% compared to the vehicle control. In conclusion, the test item is classified as negative (no increase in the expression levels of CD86 cell surface marker in the U937 cell line) under the experimental conditions of the study. Based on the weight of evidence from the three available in chemico/in vitro studies, the substance is not considered to be skin sensitising.
The available data indicate that the substance does not meet the classification criteria for skin sensitisation in accordance with Regulation (EC) No 1272/2008 (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).
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