[No public or meaningful name is available]

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26th September 2016 - 25th October 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

D-fructose and D-glucose and reaction products of D-fructose and D-glucose and hexamethylenediamine

IUPAC Name:

D-fructose and D-glucose and reaction products of D-fructose and D-glucose and hexamethylenediamine

Test material form:

liquid

In vivo test system

Test animals

Species:

mouse

Strain:

CBA/Ca

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Envigo RMS B.V.
- Females (if applicable) nulliparous and non-pregnant: yes]
- Age at study initiation: 8 to 12 weeks old
- Weight at study initiation: 15 to 23 g
- Housing: Randomly allocated to cages
- Diet (e.g. ad libitum): Free access to food (2014C Teklad Global Rodent diet)
- Water (e.g. ad libitum): Free access to water
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25
- Humidity (%): 30 to 70%
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours continuous light and 12 hours darkness

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

50%, 25% or 10% v/v

No. of animals per dose:

Groups of 4 mice

Details on study design:

PRE-SCREEN TESTS:
- Compound solubility: 25µL of the undiluted test item at a concentration of 50% v/v in DMF
- Irritation: Suggested to not produce excessive local skin irritation at the highest suitable concentration
- Systemic toxicity: Suggested to not produce systemic toxicity
- Ear thickness measurements: pre-dose on day 1, post dose on Day 3 and Day 6.

MAIN STUDY

ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response: The test item will be regarded as a sensitizer if at least one concentration of the test item results in a threefold or greater increase in 3HTdR incorporation compared to control values.

TREATMENT PREPARATION AND ADMINISTRATION: the mice were treated by daily application of 25µL of the appropriate concentration of the test item to the dorsal surface of each ear for three consecutive days. This was completed using an automatic micropipette. A further group of 4 mice received the vehicle alone in the same manner. Five days after the first application all mice were injected via the tail vein with 250uL of phosphate buffered saline containing 3F-methyl thymidine giving a total of 20uCi to each mouse

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Results and discussion

Positive control results:

a-Hexylcinnamaldehyde, tech., 85% was considered to be a sensitizer under the conditions of the test

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks:

10% v/v concentration in DMF

Value:

1.17

Results 2

Parameter:

SI

Remarks:

25% v/v concentration in DMF

Value:

1.04

Results 3

Key result

Parameter:

SI

Remarks:

50% v/v concentration in DMF

Value:

1.29

Cellular proliferation data / Observations:

DETAILS ON STIMULATION INDEX CALCULATION
Expressed as the mean radioactive incorporation for each treatment group divided by the mean radioactive incorporation of the vehicle group

CLINICAL OBSERVATIONS:
There were no deaths. No signs of systemic toxicity were noted in the test or control animals during the test.

BODY WEIGHTS
Body weight change of the test animals between day 1 and day 6 was comparable to that observed in the corresponding control group animals over the same period

Any other information on results incl. tables

See attached document for tabulated information

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was considered to be a non-sensitizer under the conditions of the test

Executive summary:

Pre-reacted hexamethylenediamine and glucose/fructose in high solids binder formulated in DMF (dimethylformamide) was assessed for its possible skin sensitising potential in accordance with the OECD Testing Guideline 429. The study was GLP-compliant.

For this purpose a local lymph node assay (LLNA) was performed using groups of four female mice exposed to concentrations of 0, 10, 25, and 50% of test substance in DMF. These concentrations were determined based on the results of a pre-study.

The animals did not show any signs of systemic toxicity or local skin irritation during the course of the study and no cases of mortality were observed.

In this study Stimulation Indices (S.I.) of 1.17, 1.04, and 1.29 were determined with the test item at concentrations of 10, 25, and 50% in DMF, respectively. The EC3 value could not be calculated, since all S.I.´s are below the threshold value of 3.

The test item Pre-reacted hexamethylenediamine and glucose/fructose in high solids binder was not a skin sensitiser under the test conditions of this study.