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Description of key information

In conclusion, under the present test conditions 2,3-Epoxypropyl neodecanoate, oligomeric reaction products with toluene-4-sulfonic acid revealed sensitising properties in the local lymph node assay (LPT, 2012).


Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2011-10-13 to 2011-11-03
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
2010
Deviations:
yes
Remarks:
Measurement of the cell proliferation by cell counting instead of radioactive labeling. In addition, measurements of ear swelling and ear weights were done to discriminate the irritating potential from the sensitizing potential of the test substance.
Qualifier:
according to
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Version / remarks:
2008
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
1998
Qualifier:
according to
Guideline:
other: Note for Guidance on Non-Clinical Tolerance Testing of Medicinal Products, CPMP/SWP/2145/00, adopted March 1, 2001
Principles of method if other than guideline:
Modified LLNA (IMDS; Integrated Model for the Differentiation of Skin Reactions). Modifications are authorized in the OECD TG 429 and the Note of
Guidance SWP/ 2145/00 of the CPMP (2001). Information on validation of IMDS and scientific justification is given in: Ehling G et al., Toxicology 212, 60-68
and 69-79 (2005).
GLP compliance:
yes (incl. certificate)
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
NMRI
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS: 
- Source: Charles River Laboratories Germany GmbH, 97633 Sulzfeld, Germany
- Sex: female
- Age: 62 days
- Weight at study initiation: 29 - 34 g
- Housing: single
- Diet: ad libitum, ssniff R/M-H V 1534 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany)
- Water: ad libitum, tap water
- Acclimatisation period: at least 5 days
- Controls: 6 female mice, treated with vehicle
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C +/- 3 °C
- Humidity (%): 55 % +/- 15 %
- Photoperiod (hrs dark / hrs light): 12 hour light/12 hour dark cycle
- Air exchange: 12 to 18 times per hour
Vehicle:
acetone/olive oil (4:1 v/v)
Concentration:
25 %, 50% w/w, 100%
No. of animals per dose:
6 female mice
Details on study design:
RANGE FINDING TESTS:
- 3 animals/test item to examine irritating potential and handling/application to select the appropriate concentrations. Two concentrations of 25%
and 50% of DYNAPOL KATALYSATOR 1203 in acetone / olive oil (3 + 1, v/v), w/w, and the undiluted test item were examined.
TREATMENT PREPARATION AND ADMINISTRATION:
- Three concentrations of 25% and 50 % of DYNAPOL KATALYSATOR 1203 in acetone / olive oil (3 + 1, v/v), w/w and the undiluted test item were
examined.
- Vehicle: olive oil (3 + 1 v/v), w/w.
- Weight of each animal, clinical observations and ear swelling measurements at the helical edge of both ears using an Oditest micrometer are
recorded
Open application of 25 µl
- vehicle alone
- test item in 3 concentrations
- positive control onto the dorsal part of both ears of the animals.
This treatment was repeated on three consecutive days (d1, d2 and d3).
24 h after last application (day 4), ear swelling measurements are carried out at the helical edge of both ears using an Oditest micrometer
Immediatly after measurement animals were sacrificed under ether anaesthesia.
The appropriate organs were then removed.
Lymphatic organs (the auricular lymph nodes) were then stored on ice in PBS /0.5% BSA.
Investigations:
- ear swelling
- ear weight (Punch biopsies of 8 mm of the apical area)
- weight of lymph nodes
- cell counts in lymph nodes
- stimulation index is calculated by dividing the absolute number of weight or cell counts of the substance treated lymph nodes by the vehicle treated ones.
Animals were observed once daily for clinical signs of local systemic irritation at the application site or of systemic toxity. Body weight were recorded on day 1 and day 4.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
The so-called stimulation (or LLN-) index to establish the sensitising potential is calculated by dividing the average absolute lymph node weight or
lymph node cell counts per group of the test item treated lymph nodes by the vehicle treated ones. Thus, in case of no stimulating effect the index
for the lymph node cell count is always below 1.4 (cut-off value). An index above 1.4 is considered positive (sensitising properties). For lymph node
weight a significance at p ≤ 0.01 is considered positive, however, an increase in lymph node weight is an indication for possible irritating properties
not sensitising properties.
In addition, the average ear weights per group and the average ear thickness per group will be compared to the vehicle control group as an indication for possible irritating properties.
Parameter:
SI
Remarks on result:
other: see below
Key result
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: modified LLNA; measurement of cell counts instead of radioactive labeling

Main study results: stimulation indices (SI):

Parameter

Group 1, negative control

Group 2,

25%

Group 3, 50%

Group 4, 100%

Group 5,

positive control

Lymph node cell count

1.000

1.531

1.620

1.655

1.659

Lymph node weight

1.000

1.441

1.729

1.769

1.576

Ear weight

1.000

1.028

1.023

0.972

1.102

Difference of

ear thickness

1.000

0.980

0.984

1.020

1.082

- significantly different from control at p ≤ 0.01

Conclusions:
Under the present test conditions, test item at concentrations of 25% or 50% in acetone / olive oil (3 + 1, v/v), w/w, and the undiluted test item revealed sensitising properties in the local lymph node assay.
Executive summary:

The purpose of this study was to determine the sensitising potential of test item in the local lymph node assay in mice.

Two concentrations of 25% and 50% of 2,3-Epoxypropyl neodecanoate, oligomeric reaction products with toluene-4-sulfonic acid in acetone / olive oil (3 + 1, v/v), w/w, and the undiluted test item were tested in six female NMRI mice per group and compared to a vehicle control group.

In addition, a positive control group(25% solution v/v ofa-hexyl cinnamic aldehyde inacetone/olive oil (3+1 v/v)) was employed.

In a preliminary experiment, concentrations of 25%, 50% and 100% of test item, employing 1 animal per concentration, were examined. No pronounced irritating properties were observed in this preliminary experiment at concentrations of 25%, 50% or 100%, no differences in ear weight and ear thickness were noted.

Stimulation indices (SI):

Parameter

Group 1, negative control

Group 2,

25%

Group 3, 50%

Group 4, 100%

Group 5,

positive control

Lymph node cell count

1.000

1.531

1.620

1.655

1.659

Lymph node weight

1.000

1.441

1.729

1.769

1.576

Ear weight

1.000

1.028

1.023

0.972

1.102

Difference of

ear thickness

1.000

0.980

0.984

1.020

1.082

____ significantly different from control at p ≤ 0.01

Treatment with test item at concentrations of 25%, 50% or 100% revealed statistical significantly increased values for lymph node cell count and lymph node weight (statistically significant at p ≤ 0.01). The stimulation indices of the lymph node cell count exceeded the threshold level of 1.4.

The threshold level for the ear weight of 1.1 was not exceeded, i.e. no irritating properties were noted.

Hence, the test item showed sensitising properties at concentrations of 25%, 50% or 100%.

The positive control group caused the expected increases in lymph node cell count and lymph node weight (statistically significant at p ≤ 0.01). Therefore, the study could be regarded as valid.

No signs of local or systemic intolerance were recorded. The animal body weight was not affected by the treatment.

In conclusion, under the present test conditions, 2,3-Epoxypropyl neodecanoate, oligomeric reaction products with toluene-4-sulfonic acid at concentrations of 25% or 50% in acetone / olive oil (3 + 1, v/v), w/w, and the undiluted test item revealed sensitising properties in the local lymph node assay.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

In conclusion, under the present test conditions,2,3-Epoxypropyl neodecanoate, oligomeric reaction products with toluene-4-sulfonic acid at concentrations of 25% or 50% in acetone / olive oil (3 + 1, v/v), w/w, and the undiluted test item revealed sensitising properties in the local lymph node assay.

Justification for selection of skin sensitisation endpoint:
Only one study available

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

Justification for selection of respiratory sensitisation endpoint:
According to REACH TGD chapter R7 A, Figure 7.3-2 no testing proposal for respiratory sensitisation is needed to perform Annex VII or VIII studies.

Justification for classification or non-classification

The results of the skin sensitizing study in the Local Nymph Node Assay (LPT, 2012) for 2,3-Epoxypropyl neodecanoate, oligomeric reaction products with toluene-4-sulfonic acid indicate skin sensitisation at concentrations of 25% or 50% in acetone / olive oil (3 + 1, v/v), w/w, and the undiluted test item under the conditions employed and may cause an allergic skin reaction. It must therefore be classified according to the criteria of EC Directive 67/548/EECand EC Regulation 1272/2008.