Saccharomyces cerevisiae cell wall, extracted

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 09 May 2011 to 19 December 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24 April 2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

30 May 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Saccharomyces cerevisiae cell wall, extracted

EC Number:

949-711-6

Molecular formula:

Not applicable (UVCB substance)

IUPAC Name:

Saccharomyces cerevisiae cell wall, extracted

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage de Gerome (Quartier Labaste­ F40260 Linxe)
- Age at study initiation: 13 weeks old
- Weight at study initiation: 2.66 - 3.15 kg
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet (e.g. ad libitum): foodstuff (SDS - C15) ad libitum
- Water (e.g. ad libitum): tap-water from public distribution system ad libitum
Microbiological and chemical analyses of the water were carried out once evety six months by IPL, Sante, Environnement Durables - Atlantique
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23
- Humidity (%): 30 to 70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12 (continuous light (07.00 to 19.00))

IN-LIFE DATES: not specified.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the other eye remained untreated serving as control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g
- Concentration (if solution): not applicable

Duration of treatment / exposure:

0.1 g of the test item was instilled into the conjunctival sac of one eye after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second in order to prevent loss of the test item. The other eye remained untreated serving as control.

Observation period (in vivo):

Ocular examinations were performed on both right and left eyes 1h, 24, 48 and 72 hours following treatment. If no reaction is observed 72 hours after instillation, the study is terminated. In case of persistent reactions, additional observations can be carried out from day 4 to day 21 in order to determine the reversible character of the lesions observed.

Number of animals or in vitro replicates:

3 female rabbits (initially, a single animal was treated. After consideration of the responses produced on day 1, two additional animals were treated).

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no
- Time after start of exposure: not applicable.

SCORING SYSTEM:
Eye examinations were carried out using the scale of lesion scores in the following order:

CHEMOSIS (A)
No swelling ....................................................................................................................................0
Slight swelling, including the nictitating membrane...................................................................... 1
Swelling with eversion of the eyelid..............................................................................................2
Swelling with eyelid half-closed....................................................................................................3
Swelling with eyelid more than half-closed...................................................................................4


DISCHARGE (B)
No discharge.................................................................................................................................0
Slight discharge (normal slight secretions in the inner corner not to be taken into account)........1
Discharge with moistening of the eyelids and neighbouring hairs...............................................2
Discharge with moistening of the eyelids and large areas around the eye..................................3


REDNESS (C)
Blood vessels normal....................................................................................................................0
Vessels significantly more prominent than normal......................................................................1
Vessels individually distinguishable with difficulty:
Generalised red coloration..........................................................................................................2
Generalised deep red coloration.................................................................................................3


IRIS (D)
Normal ...........................................................................................................................................0
Iris significantly more wrinkled than normal, congestion,
swelling of the iris which continues to react to light, even slowly................................................1
No reaction to light, haemorrhage, significant damage (any or all of these characteristics)........2


CORNEA: DEGREE OF OPACITY (E)
No modification visible either directly or after instillation of fluorescein (no loss of glint or polish)...0
Translucent areas (diffuse or disseminated), iris details clearly visible ...........................................1
Easily identifiable translucent area, iris details slightly obscured......................................................2
Opalescent area, no iris details visible, pupil outline scarcely distinguishable...................................3
Total corneal opacity, completely obscuring the iris and pupil............................................................4


CORNEA: EXTENT OF OPACITY (F)
Opaque area present but covering one quarter or less.................................................................... 1
Between one quarter and half......................................................................................................... 2
Between half and three quarters........................................................................................................3
Between three quarters and the entire surface................................................................................ 4


TOOL USED TO ASSESS SCORE: Examination of reactions can be facilitated by use of a binocular loupe, hand slit-lamp, biomicroscope, or other suitable device. After recording the observations at 24 h, the eyes may be further examined with the aid of fluorescein.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

mean

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 2

Irritation parameter:

cornea opacity score

Remarks:

mean

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 4

Irritation parameter:

iris score

Remarks:

mean

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 5

Irritation parameter:

iris score

Remarks:

mean

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 6

Irritation parameter:

iris score

Remarks:

mean

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 7

Irritation parameter:

conjunctivae score

Remarks:

mean

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Reversibility:

fully reversible within: 72h

Results 8

Irritation parameter:

conjunctivae score

Remarks:

mean

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible within: 48h

Results 9

Irritation parameter:

conjunctivae score

Remarks:

mean

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Reversibility:

fully reversible within: day 7

Results 10

Irritation parameter:

chemosis score

Remarks:

mean

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 11

Irritation parameter:

chemosis score

Remarks:

mean

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 12

Irritation parameter:

chemosis score

Remarks:

mean

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Reversibility:

fully reversible within: 72h

Irritant / corrosive response data:

The ocular reactions observed during the study have been slight to moderate and totally reversible in the three animals (results of individual and mean scores are presented in Table 1 in "any other information on results incl. tables" section):
- at the conjunctivae level: a slight to moderate redness noted 1h after the test item instillation and totally reversible between days 2 and 7, associated with a slight chemosis noted 1h after the test item instillation and totally reversible between days 1 and 3;
- at the cornea level: a slight opacity, noted in one animal 1h after the test item instillation and completely resolved 24 h post exposure.

Other effects:

- Lesions and clinical observations: none
- Ophthalmoscopic findings: none
- Histopathological findings: not performed
- Effects of rinsing or washing: not applicable
- Other observations: none

Any other information on results incl. tables

Table 1.Individual and mean scores for Conjunctivae, Iris and Cornea

Animal #	Time after treatment	Conjunctivae		Iris	Cornea
		Chemosis (A)	Redness (C)	Lesion (D)	Opacity (E)
1	24h	0	1	0	0
	48h	0	1	0	0
	72h	0	0	0	0
Mean		0.0	0.67	0.0	0.0
2	24h	0	1	0	0
	48h	0	0	0	0
	72h	0	0	0	0
Mean		0.0	0.33	0.0	0.0
3	24h	1	3	0	0
	48h	1	2	0	0
	72h	0	1	0	0
	7 days	0	0	0	0
Mean (24-48-72h)		0.67	2.0	0.0	0.0

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions of this study, the test item Saccharomyces cerevisiae cell wall, extracted was not irritating to the rabbit eyes. Thus, the test material is not classified for eye irritation / corrosion according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) criteria and the UN-GSH system.

Executive summary:

This GLP-compliant study was performed to assess the potential of Saccharomyces cerevisiae cell wall, extracted to induce eye irritation in vivo, according to OECD Guideline 405 (Acute Eyeal Irritation/Corrosion) and EU method B.5 (Acute Toxicity: Eyel Irritation / Corrosion).

Material and methods

0.1 g of test item were instilled, as supplied, into the conjunctival sac of one eye of each of the three New Zealand White rabbits. The test item was applied as a single dose. The lids were gently held together for about one second. The other eye remained untreated and served as control. No washing was performed.

The degree of eye irritation/corrosion is evaluated by scoring lesions of conjunctivae, cornea and iris on both right and left eyes 1h, 24, 48 and 72 hours following treatment. If no reaction is observed 72 hours after instillation, the study is terminated. In case of persistent reactions, additional observations can be carried out from day 4 to day 21 in order to determine the reversible character of the lesions observed. Other effects in the eye and adverse systemic effects are also described.

Results

There was no mortality during the observation period.

The ocular reactions observed during the study have been slight to moderate and totally reversible in the three animals:

- at the conjunctivae level: a slight to moderate redness noted 1h after the test item instillation and totally reversible between days 2 and 7, associated with a slight chemosis noted 1h after the test item instillation and totally reversible between days 1 and 3;

- at the cornea level: a slight opacity, noted in one animal 1h after the test item instillation and completely resolved 24 h post exposure.

The animals’ individual mean scores (24, 48 and 72 h) for corneal opacity were 0.00, 0.00 and 0.00 respectively.

The animals’ individual mean scores (24, 48 and 72 h) for iris were 0.00, 0.00 and 0.00 respectively.

The animals’ individual mean scores (24, 48 and 72 h) for conjunctivae were 0.67, 0.33 and 2.00 respectively.

The animals’ individual mean scores (24, 48 and 72 h) for chemosis were 0.00, 0.00 and 0.67 respectively.

Conclusion

Under the experimental conditions of this study, the test item Saccharomyces cerevisiae cell wall, extracted was not irritating to the rabbit eyes. Thus, the test material is not classified for eye irritation / corrosion according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) criteria and the UN-GSH system.