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Administrative data

Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study in compliance with guidelines

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)

Test material

Constituent 1
Reference substance name:
Wheat, ext., hydrolyzed
EC Number:
309-358-5
EC Name:
Wheat, ext., hydrolyzed
Cas Number:
100209-50-5
Molecular formula:
UVCB substance, not applicable.
IUPAC Name:
Wheat, ext., hydrolyzed
Details on test material:
Identification: Wheat Solubles, Hydrolyzed
Description: Light brown-yellow powder with lumps
Batch: Cargill WSH Batch A
Purity: treated as 100% pure
Stability under storage conditions: Stable

In vivo test system

Test animals

Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
Species:Mouse, CBA/J strain, inbred, SPF-Quality.

Source: Harlan, Horst, The Netherlands

Age and body weight:Young adult animals (approx. 11 weeks old) were selected. Body weight variation was within +/- 20% of the sex mean.

Identification: Tail mark with marker pen.

Health inspection: A health inspection was performed prior to treatment, to ensure that the animals were in a good state of health. Special attention was paid to the ears, which were intact and free from any abnormality.

Conditions: Animals were housed in a controlled environment, in which optimal conditions were considered to be approximately 15 air changes per hour, a temperature of 21.0 ± 3.0ºC (actual range: 21.0 - 21.9ºC), a relative humidity of 40-70% (actual range: 40 - 80%) and 12 hours artificial fluorescent light and 12 hours darkness per day.

Accommodation: Individual housing in labeled Macrolon cages (MI type; height 12.5 cm) containing sterilized sawdust as bedding material (Litalabo, S.P.P.S., Argenteuil, France). Paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom) was supplied as cage-enrichment.
The paper was removed on Day 1 prior to dosing and was supplied again after scoring of the ears on Day 3.

Acclimatization period: The acclimatization period was at least 5 days before the start of treatment under laboratory conditions. Accommodation was as described above except that the animals were group housed in Macrolon cages (MIII type; height 18 cm).

Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).

Water: Free access to tap water.

Study design: in vivo (LLNA)

Concentration:
In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 5, 10 or 25% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Propylene glycol).
No. of animals per dose:
20 females (nulliparous and non-pregnant), five females per group.
Details on study design:
Test substance concentrations selected for the main study were based on the results of a preliminary study.

In the main study, three experimental groups of five female CBA/J mice were treated with test substance concentrations of 5, 10 or 25% w/w on three consecutive days, by open application on the ears. Five vehicle control animals were similarly treated, but with vehicle alone (Propylene glycol).

Three days after the last exposure, all animals were injected with 3H-methyl thymidine and after five hours the draining (auricular) lymph nodes were excised and pooled for each animal.
After precipitating the DNA of the lymph node cells, radioactivity measurements were performed. The activity was expressed as the number of Disintegrations Per Minute (DPM) and a stimulation index (SI) was subsequently calculated for each group.
Positive control substance(s):
hexyl cinnamic aldehyde (CAS No 101-86-0)
Statistics:
no data

Results and discussion

Positive control results:
The six-month reliability check with Alpha-hexylcinnamicaldehyde indicates that the Local Lymph Node Assay is an appropriate model for testing for contact hypersensitivity.

In vivo (LLNA)

Resultsopen allclose all
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
The SI values calculated for the substance concentrations 5, 10 and 25% were 1.0, 1.3 and 1.1 respectively. Since there was no indication that the test substance elicits an SI ≥ 3 when tested up to 25%, Wheat Solubles, Hydrolyzed was considered not to be a skin sensitizer. It was established that the EC3 value (the estimated test substance concentration that will give a SI =3) (if any) exceeds 25%.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: Mean DPM/animal values for the experimental groups treated with test substance concentrations 5, 10 and 25% were 925, 1231 and 1023 DPM respectively. The mean DPM/animal value for the vehicle control group was 922.

Any other information on results incl. tables

No irritation of the ears was observed in any of the animals examined.

 

All auricular lymph nodes of the animals of the experimental and control groups were considered normal in size. No macroscopic abnormalities of the surrounding area were noted in any of the animals.

 

Mean DPM/animal values for the experimental groups treated with test substance concentrations 5, 10 and 25% were 925, 1231 and 1023 DPM respectively.

The mean DPM/animal value for the vehicle control group was 922.

 

 

The SI values calculated for the substance concentrations 5, 10 and 25% were 1.0, 1.3 and 1.1 respectively.

 

Since there was no indication that the test substance elicits an SI3 when tested up to 25%, Wheat Solubles, Hydrolyzed was considered not to be a skin sensitizer. It was established that the EC3 value(the estimated test substance concentration that will give a SI =3)(if any) exceeds 25%.

 

The six-month reliability check withAlpha-hexylcinnamicaldehyde indicates that the Local Lymph Node Assay as performed at NOTOX is an appropriate model for testing for contact hypersensitivity.

 

Based on these results Wheat Solubles, Hydrolyzed would not be regarded as a skin sensitizer according to the recommendations made in the test guidelines. It does not have to be classified and has no obligatory labelling requirement for sensitization by skin contact according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2007) and theRegulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

 

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information

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