Short Chain Fructo Oligosaccharides

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Title:

Unnamed

Year:

2011

Report date:

2011

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:

short chain Fructo-Oligosaccharides

EC Number:

908-300-1

Molecular formula:

C6H11O5(C6H10O5)nOH

IUPAC Name:

short chain Fructo-Oligosaccharides

Test material form:

solid: particulate/powder

Details on test material:

Identification: Actilight 950P
Substance: Short-chain fructo-oligosaccharides
EC No.: 908-300-1
Appearance: White powder
Purity: 96%

COMPOSITION:
scFOS is short chains of fructose molecules linked to a molecule of sucrose (glucose-fructose disaccharide). Thus, scFOS is a multiconstituent substance composed of three oligosaccharides: 1-kestose (GF2), nystose (GF3) and fructosyl-nystose (GF4).
With:
- TOTAL FOS: 96% expressed as dry matter
- Kestose (GF2): 37.1% FOS
- Nystose (GF3): 50.5% FOS
- 1F-Fructofuranosylnystose (GF4:) 12.4% FOS
- Glucose +Fructose + Saccharose : 4.0% expressed as dry matter

Specific details on test material used for the study:

• Sponsor’s identification: ACTILIGHT® 950P
• Form: powder
• Colour: White

In vivo test system

Test animals

Species:

mouse

Strain:

CBA:J

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Elevage Janvier
- Females nulliparous and non-pregnant: yes
- Age at study initiation:8 or 9 weeks old.
- Weight at study initiation: weight range of 20.5 to 24.2 g
- Housing: individually housed in suspended solid-floor polypropylene cages furnished with softwood woodflakes.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least five days under stabling and nutritional conditions identical to those of the test,
- Indication of any skin lesions: No

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19°C to 25°C
- Humidity (%): 30 to 70%,
- Air changes (per hr): approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): the lighting was controlled by a time switch to give twelve hours continuous light (07.00 to 19.00) and twelve hours darkness.

Study design: in vivo (LLNA)

Vehicle:

dimethylformamide

Concentration:

The test item was diluted in dimethylformamide (DMF) at concentrations of 5% (v/v), 10% (w/w) and 25% (w/w).

No. of animals per dose:

Four animals per dose.
A further group of four animals was treated with DMF

Details on study design:

PRE-SCREEN TESTS:
One mouse was treated by daily application of 25 μL of the test item diluted at 50% in dimethylformamide (DMF) to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3). The mouse was observed daily from day 1. Any signs of toxicity or excessive local irritation noted during this period were recorded. Ear thickness was recorded on day 1, day 3 and on day 6. The bodyweight of the mouse was recorded
on Day 1 (prior to dosing) and Day 6.

MAIN STUDY
Groups of four mice were treated with the test item diluted at concentrations of 25% (w/w), 10% (w/w) and 5% (v/v) in the vehicle – dimethylformamide (DMF). The mice were treated by daily application of 25 μL of the appropriate concentration of the test item to the dorsal surface of each ear for three consecutive days (Days 1, 2, 3). The test item formulation was administered using an automatic micropipette and spread over the dorsal surface of the ear using the tip of the pipette.
A further group of four mice received the vehicle alone in the same manner.

ANIMAL ASSIGNMENT AND TREATMENT
the animals were selected at random and given a number unique within the study by indelible ink-marking on the tail and a number written on a cage card.

Positive control substance(s):

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

Results and discussion

Positive control results:

The assay has undergone extensive inter-laboratory validation and has shown to reliably detect test
materials that are moderate to strong sensitisers.
The strain of mouse used in these laboratories had been shown to produce satisfactory responses using known sensitisers and non-sensitisers during the in-house validation.

In vivo (LLNA)

Results

Results 1

Key result

Parameter:

SI

Value:

0.69

Test group / Remarks:

5%

Results 2

Key result

Parameter:

SI

Value:

0.73

Test group / Remarks:

10%

Results 3

Key result

Parameter:

SI

Value:

0.65

Test group / Remarks:

25%

Cellular proliferation data / Observations:

Proliferative response of lymph node cells:
No stimulation index of more than 1.4 was recorded in the study, whatever the tested concentration of the test item.
The Stimulation Index (SI) calculated by pooled approach was 0.69, 0.73 and 0.65 for the treated groups at 5%, 10% and 25%, respectively.
The EC1.4 can not be determined in this study.

Local irritation:
No cutaneous reaction was noted on the treatment site of animals, whatever the administered concentration.
No significant increase in ear thickness and in ear weight was noted in animals treated at 5%, 10% and 25%. Therefore, the test item must be considered as non irritant at the three concentrations.

Any other information on results incl. tables

- Clinical observations and mortality:

No mortality and no signs of systemic toxicity were noted in the test and controls animals during the test.

- Weight evolution:

Bodyweight changes of the test animals between Day 1 and Day 6 were comparable to those observed in the corresponding control group animals over the same period.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under these experimental conditions, the test item ACTILIGHT® 950P must not be classified as a skin sensitiser, in accordance with Regulation EC No. 1272/2008 (CLP).

Executive summary:

The test was performed to assess the skin sensitisation potential of the test item ACTILIGHT® 950P in the CBA/J strain mouse following topical applications to the dorsal surface of the ear.

The experimental protocol was established according the OECD Guideline No.429 dated 22 July 2010 and the test method B42 of the council regulation No. 440/2008. The study was GLP compliant.

Three groups of four animals were treated for three consecutive days (D1, D2, D3) with 50 μL (25 μL per ear) of the test item diluted in dimethylformamide (DMF) at concentrations of 5% (v/v), 10% (w/w) and 25% (w/w). A further group of four animals was treated with DMF.

On D6, the proliferation of lymphocytes in the draining auricular lymph nodes was determined by cell counting.

 

No mortality and no signs of systemic toxicity were noted in the test and control animals during the test.

No cutaneous reaction was noted on the treatment site of animals, whatever the administered concentration.

No significant increase in ear thickness and in ear weight was noted in animals treated at 5%, 10% and 25%. Therefore, the test item must be considered as non irritant at the three concentrations.

The Stimulation Index (SI) calculated by pooled approach was 0.69, 0.73 and 0.65 for the treated groups at 5%, 10% and 25%, respectively. The EC1.4 can not be determined in this study.

 

Under these experimental conditions, the test item ACTILIGHT® 950P must not be classified as a skin sensitiser.