Distilled acetalization product between glucose and C12-18(even numbered)alcohol

Endpoint summary

Description of key information

The substance is not irritating to the skin in animals and humans. It is slightly irritating to the eye in animals, without classification being required. For skin and eye irritation the conclusion is “Not classified - based on specific, valid data on the substance”. 
Considering these results in high-sensitivity assays, there is no reason to suspect any relevant respiratory irritation, especially as the granulometry of the substance (as the substance is in the form of pearls) leads to negligible exposure of terminal airways. For this route, the conclusion is “Not classified - based on weight-of-evidence analysis”.
Value used for CSA:
Skin irritation/corrosion: not irritating
Eye irritation: not irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2007-07-31 to 2007-08-10

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: read across from a category grouping, GLP study, accepted OECD guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Test animals:
Three male albino New Zealand rabbits were supplied by the Elevage de Gérome (Quartier Labaste –
F40260 Linxe). They were kept during a minimal 5-day acclimatisation period. During the test, the
animals weighed between 2.22 kg and 2.63 kg.

Housing:
Each animal was kept in an individual box installed in conventional air conditioned animal
husbanding; the environmental conditions were:
- temperature : between 19°C and 23°C
- relative humidity : between 47% and 65%
- lighting time: 12 hours daily
Drinking water (tap-water from public distribution system) and foodstuff were supplied freely.
Microbiological and chemical analyses of the water were carried out once every six months by the
Institut Européen de l'Environnement de Bordeaux (I.E.E.B.).

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g

Duration of treatment / exposure:

4 hours

Observation period:

The skin reactions were appreciated 1, 24, 48 and 72 hours after removal of the test item.

Number of animals:

3 animals

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

ca. 0

Max. score:

0

Reversibility:

fully reversible

Irritant / corrosive response data:

No cutaneous reactions (erythema and oedema) were observed whatever the examination time (1, 24, 48 and 72 hours).

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The results obtained, under these experimental conditions, enable to conclude that the test item LCE07050 according to the scales of interpretation dont' need to be classified, according to the criteria for classification, packaging and labelling of dangerous substances and preparations in compliance with the E.E.C. Directives 67/548, 2001/59 and 99/45. No symbol and risk phrase are required.

Executive summary:

The test item LCE07050 was applied, as supplied, at the dose of 0.5 g, under semi-occlusive dressing

during 4 hours on an undamaged skin area of three rabbits. The experimental protocol was established

from the O.E.C.D. guideline n° 404 dated April 24th, 2002 and the test method B.4 of the directive n°

2004/73/EC.

No cutaneous reactions (erythema and oedema) were observed whatever the examination time (1, 24,

48 and 72 hours).

The results obtained, under these experimental conditions, enable to conclude that the test item

LCE07050 need not to be classified, according to the criteria for classification, packaging and

labelling of dangerous substances and preparations in compliance with the E.E.C. Directives 67/548,

2001/59 and 99/45. No symbol and risk phrase are required.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2010-06-08 to 2010-06-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, accepted OECD guideline

Qualifier:

according to guideline

Guideline:

other: OECD GUIDELINE n°437

GLP compliance:

yes (incl. QA statement)

Remarks:

2009-12-09

Vehicle:

physiological saline

Remarks:

NaCl to 0.9%

Amount / concentration applied:

The test item was diluted to 20% (W/W) in NaCl to 0.9%
Quantity applied : 750 uL per cornea

Duration of treatment / exposure:

contact timepoint = 4 hours ± 10 minutes

Number of animals or in vitro replicates:

not applicable

Details on study design:

BIOLOGICAL TEST SYSTEM
Bovine eyes (from cattle less than 12 months) collected at the slaughter-houses of Corbas – France (69960) and carried in a stable medium (Hanks medium buffered with sodium bicarbonate), at room temperature, and prepared 4 hours maximum after killing the animals.
This biological test system is adapted to evaluate the ocular irritancy potential.

METHODOLOGY
Media and reagents preparation
Buffered Hanks medium:
- Hanks medium : 0.98 g
- Sodium bicarbonate (CAS N° 144-55-8) : 0.035 g
- Distilled water : sufficient quantity for 100 mL
- shaking until complete dissolution of the compounds, measurement of the pH and adjustment to 7.4 with a 4M HCl solution.

Transport medium:
buffered Hanks medium, completed with 0.2% of Penicillin-Streptomycin solution (CAS N° 69-57-8 / CAS N° 3810-74-0).

Conservation medium:
- M199 medium : 0.98 g
- Sodium bicarbonate (CAS N° 144-55-8) : 0.22 g
- Dextran (CAS N° 9004-54-0) : 5 g
- Distilled water : sufficient quantity for 100 mL
- shaking until complete dissolution of the compounds, then addition of the following solutions:
- L-glutamine 200 mM (CAS N° 56-85-9) : 0.34%
- Penicillin/Streptomycin : 0.4%
- shaking once again, then measurement of the pH and adjustment to 7.4 with a 4M HCl solution.

Nutritive medium:
- Minimum Essential Medium : 0.94 g
- Sodium bicarbonate(CAS N° 144-55-8) : 0.22 g
- Distilled water : sufficient quantity for 100 mL
- shaking until complete dissolution of the compounds, then addition of the following solutions:
- Foetal Bovine Serum : 1%
- L-glutamine 200 mM(CAS N° 56-85-9) : 1%
- Penicillin/Streptomycin solution : 0.25%
- shaking once again, then measurement of the pH and adjustment to 7.4 with a 4M HCl solution.
Nutritive medium with phenol red (for rinsing): addition of 0.1% of a solution of phenol red to 0.2%.
Fluorescein solution :
- Fluorescein (CAS N° 518-47-8) : 0.5 g
- DPBS : sufficient quantity for 100 mL
- validation of the trial: dilution of this previous solution to 1/500th (20 µL in 10 mL) in the nutritive medium (final fluorescein concentration of 10 µg/mL).

Biological system preparation

Eyes selection:
detection of defects of the cornea (neovascularization, pigmentation, opacity, scratches) by immersing the eyes in a jar containing buffered Hanks medium and putting them under lighting.

Dissection and immersion of corneas:
for each selected eye
- incision on the scleral ring, using a scalpel, then dissection of the cornea, using scissors, leaving 2 or 3 mm of the scleral ring
- immersion of the corneas in buffered Hanks medium, the epithelial side upward
- use of the preserved corneas, preservation for a maximum duration of 24 hours.

Preservation of the corneas:
- 3 successive rinsings of 15 ± 3 minutes with Hanks enriched with antibiotics
- deposit of each cornea in conservation medium
- preservation of the corneas at 5 ± 3 °C for a maximum duration of 24 hours.

Mounting of the holders:
- deposit of each preserved cornea (previously rinsed with buffered Hanks medium), epithelial side upward, on the posterior part of the holders
- placement of the anterior part of the holder above the corneas
- screwing on with an electric screwdriver in order to ensure the waterproofness of the system without any risk for the cornea
- identification of each cornea with the corresponding holder number.

Pre-incubation of the corneas:
- in order to eliminate the air bubbles, filling, excessively, of the anterior compartments (epithelial side) and posterior (endothelial side), with nutritive medium at room temperature
- incubation of the holders in a water-bath at 32 ± 1 °C, for at least one hour, in a horizontal position and immersed in three quarters of their height.

Measurement of the initial opacity of the corneas

- realised using an OP KIT opacitometer, which determines a change in light transmission between an "empty" holder (containing nutritive medium only) and a "treated" holder, and displays a numerical value of opacity.
- calibration of the opacitometer, with specific calibrators, before the measurement of the corneal opacity
- values of opacity of the calibrators:
- calibrator 1: 73 to 77
- calibrator 2: 147 to 153
- calibrator 3: 220 to 240
- if necessary: adjustment with the "calibrator" button
- change of the nutritive medium of the posterior compartment
- wiping of each holder
- measurement of the initial opacity (OPT0) of each cornea versus an "empty" holder (containing only nutritive medium).


Preparation and modalities of application of controls ant test item

Carrying out in the same chronological order of each operation (treatment, rinsing and readings), by series of 3 corneas. One contact timepoint is carried out: it depends on the physical aspect of the test item.
- negative control: Sodium Chloride to 0,9% (W/V) (NaCl – CAS N° 7647-14-5)
- positive control: 20% Imidazole solution (CAS N° 288-32-4) (W/W) in NaCl to 0.9%
- measurement of the pH of each control
- control of the homogeneity and stability of each control in the trial conditions
- test item:
- determination of the modalities of application :
. category: UVCB (non-surfactant) / aspect : solid (pellets)
. to be diluted to 20% / contact timepoint = 4 hours ± 10 minutes
- measurement of the pH of the preparation
- control by I.E.C. of the homogeneity of the test item and confirmation by the Sponsor of the stability in the storage conditions
- extemporaneous preparation of the test item:
- crush the pellets with a mortar so as to obtain a powder
- dilute the powder to 20% (W/W) in NaCl to 0.9%
- control of the homogeneity and stability of the preparation in the trial conditions

- aspiration of the nutritive medium of the anterior compartment using a specific vacuum pump
- remove the nut and the glass disk
- application of 750 µL of the preparation and of each control on the cornea using a micropipette
- starting of the chronometer for each series, after the treatment of the 1st cornea
- incubation in a water-bath at 32 ± 1 °C, in a vertical position (screw upward) for 4 hours ± 10 minutes.

Rinsing (at the end of the contact timepoint)

- aspiration of the test item (or the preparation)
- rinsing at least 3 times with nutritive medium with phenol red at 32 ± 1°C: the corneas are rinsed more than 3 times if the phenol red is still discolored or the test substance is still visible
- elimination of the excess product which had stuck to the walls of the support with a cotton bud
- final rinsing with nutritive medium
- filling of the anterior with nutritive medium at 32 ± 1 °C.

For the 4-hour exposure period: no 2-hour incubation period.


Measurement of the corneal opacification

- replacing of the nutritive medium in the posterior compartments using a vacuum pump
- wiping of each holder
- measurement of the opacity OPT2 (=OPT "2 hours") of each cornea versus an "empty" holder.


Measurement of the corneal permeability

Addition of the fluorescein solution:
- aspiration of the anterior compartment using a vacuum pump
- deposit of 1 mL of the fluorescein solution (0.5%), in the anterior compartment.
- incubation of the holders at 32 ± 1 °C for 90 ± 5 minutes in a vertical position.

Measurement of optical density:
- sample of the medium contained in each posterior compartment
- deposit of each medium in tubes identified by the holder number
- transfer of 360 µL of this medium onto microplates of 96 wells
- measurement of the optical density (O.D.) with a spectrophotometer at 490 nm (blank = nutritive medium).

Determination of the theoretical O.D. :
Determined for each new batch of fluorescein by establishing a standard curve connecting the O.D. to the concentration of fluorescein (1-2-3-4-5-6-7-8-9-10-12-14-16-20-40 µg/mL). The graph being under a linear form, the calculation of the gradient permitted the calculation of theoretical O.D. for the concentration at 10 µg/mL.

It also permitted to determine a value of optical density beyond which the medium had to be diluted (beginning of the plate stage). Each medium of which the O.D. exceeded this value was diluted to 1/4 in the nutritive medium. The O.D. value obtained was then multiplied by 4, during the final calculations.

Corneas examination

- dismantling of each holder
- observation of the cornea condition: epithelium detachment, visible modification of the cornea (oedema, colouring, …).

Irritation parameter:

other: OPT2-OPT0

Basis:

mean

Time point:

other: 4 hours

Score:

ca. 5.3

Max. score:

5.7

Irritation parameter:

other: OD

Basis:

mean

Time point:

other: 4 hours

Score:

ca. 5.9

Max. score:

6.4

Irritant / corrosive response data:

Calculation
- calculation of the means of each parameter (OPT2-OPT0 and O.D.)
- correction of the values by subtracting the negative controls corneas
- calculation of the in vitro irritancy score (IVIS) from the corrected values:
IVIS = (OPT2-OPT0) + (15 x O.D.)

Sore for the test item (4 hours) : 5.9 ± 0.6
Test item not classified corrosive or severe irritant

Interpretation of results:

other: not "classified corrosive or severely irritant"

Remarks:

Criteria used for interpretation of results: other: OECD GUIDELINE 437

Conclusions:

From the results obtained under the experimental conditions adopted, the test item designated as "LCE10045 (batch n° T93611 of 31/08/2009)", applied diluted to 20 % (W/W) in NaCl to 0.9%, is not "classified corrosive or severely irritant" for the isolated bovine cornea, after 4 hours of contact.

Executive summary:

The aim of this study was to assess the corrosive or severe irritant potential of item LCE10045 using the ocular primary irritation B.C.O.P. This study was conduced in occodance to the OECD guideline n°437 and in compliance with the GLP.

Bovine eyes (from cattle less than 12 months) collected at the slaughter-houses of Corbas – France (69960) and carried in a stable medium (Hanks medium buffered with sodium bicarbonate), at room temperature, and prepared 4 hours maximum after killing the animals.

This biological test system is adapted to evaluate the ocular irritancy potential.

RESULTS AND CLASSIFICATION

Category of the test item	Aspect of the provided test item	Concentration	Contact timepoint
UVCB (non-surfactant)	Pellets yellow in colour	Diluted to 20% (W/W) in NaCl to 0.9%	4 hours ± 10 minutes

 

Beginning and end of the experiment	Number of rinsings	Score	Classification
08/06/10	3	5.9 ± 0.6	Not classified corrosive  or severe irritant

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available