D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-16 (even numbered) and C18 unsaturated acyl] derivs.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-07-21 to 2015-09-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Glucamide CC

IUPAC Name:

Glucamide CC

Constituent 2

Reference substance name:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-16 (even numbered) and C18 unsaturated acyl] derivs.

EC Number:

940-422-0

Cas Number:

1591783-13-9

Molecular formula:

C15H31NO6 (C8 derivative) C17H35NO6 (C10 derivative) C19H39NO6 (C12 derivative) C21H43NO6 (C14 derivative) C23H47NO6 (C16 derivative) C25H51NO6 (C18 derivative) C25H49NO6 (C18 unsatd. derivative)

IUPAC Name:

D-Glucitol, 1-deoxy-1-(methylamino)-, N-[C8-16 (even numbered) and C18 unsaturated acyl] derivs.

Test material form:

other: solid

Details on test material:

Chemical Name: D-Glucitol, 1-deoxy-1-(methylamino)-, N-coco acyl derivs.
CAS No.: 1591783-13-9
Colour: white to slightly yellowish
Storage Conditions: at room temperature
Safety Precautions: The routine hygienic procedures were sufficient to assure personnel health and safety.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test System
Species/strain: healthy New Zealand White Rabbits, Crl: KBL (NZW)
Source: Charles River Deutschland, 97633 Sulzfeld, Germany
Sex: male
Body weight at the beginning of the study:> 2 kg
Age at the beginning of the study: approximately 15 - 16 weeks old
Number of animals: 3
The animals were derived from a controlled full-barrier maintained breeding system (SPF). According to Art. 9.2, No. 7 of the German Act on
Animal Welfare the animals were bred for experimental purposes.

Housing and Feeding Conditions
- Semi barrier in an air-conditioned room
- Temperature: 18 +/- 3 °C (recommendations of TVT, GV-SOLAS)
- Relative humidity: 55 +/- 10%
- Artificial light, sequence being 12 hours light, 12 hours dark
- Air change: at least 10 x / hour
- Free access irradiated hay briquettes and to Altromin 2123 maintenance diet for rabbits (lot no. 0944), rich in crude fibre
- Free access to tap water (drinking water, municipal residue control, microbiological controls at regular intervals)
- Certificates of food, water and bedding are filed for two years at BSL Munich and afterwards archived at Eurofins Munich
- Housed in ABS-plastic or Noryl rabbit cages, floor 4200 cm2
- Adequate acclimatisation period (at least 5 days) under laboratory conditions

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

A dose of 0.1 g of the test item was applied to the test site.

Duration of treatment / exposure:

The test item was applied at a single dose in the conjunctival sac of one eye of each test animal after pulling the lower lid away from the eyeball.
The lids were then gently held together for about 1 second in order to prevent loss of the material. The untreated contralateral eye served as control.
The treated eye was not rinsed after the application.

Observation period (in vivo):

The animals were observed for 72 hours after dosing.
To determine the reversibility of the observed effects in animals no. 2 and no.3, the observation period was extended up to 21 days after dosing.

Number of animals or in vitro replicates:

The in vivo test was performed initially using one animal.
The results of the initial test did not indicate the test item to be corrosive or a severe irritant to the eye using the procedure described.
In order to confirm the response, two additional animals were treated in the same manner.

Details on study design:

Preparation of the Animals
Approximately 24 hours before the test and immediately prior to the application both eyes of each animal were examined. A health inspection was
performed to ensure the good state of health of the animals.
Approximately 24 hours (animal no. 1), 20 hours (animals no. 2 and no. 3) before the application the eyes were also examined with the aid of a
fluorescein solution (Fluoreszein SE Thilo, Alcon Pharma, lot no. H 402, expiry date: 30/06/2016). The eyes were rinsed with physiological saline
0.9% NaCl (AlleMan Pharma, lot no. 050315-1, expiry date: 28/02/2018) after the examination.
None of the animals showed eye irritation, ocular defects, or pre-existing corneal injury.

Application
One hour before the application of the test item, 0.01 mg/kg of buprenorphine (Reckitt Benckiser, lot no. 5448, expiry date: 30 June 2017)
was administered subcutaneously in order to achieve a therapeutic level of systemic analgesia. Approximately 5 minutes prior to the application
of the test item, 1-2 drops of an ocular anaesthetic (proparacaine hydrochloride ophtalmic 0.5% solution, Ursapharm Arzneimittel, lot no. 282974,
expiry date: 30 June 2016) were administered in both the treated and the control eye of each animal.
The test item was applied at a single dose in the conjunctival sac of one eye of each test animal after pulling the lower lid away from the eyeball.
The lids were then gently held together for about 1 second in order to prevent loss of the material. The untreated contralateral eye served as control.
The treated eye was not rinsed after the application.
To prevent pain and distress after the application of the test item, animal no. 2 was treated with the following doses of buprenorphine (see above) and meloxicam (Boehringer Ingelheim, lot no. F20808A, expiry date: 30/06/2017).
Administration of Analgesic Animal No. 2
Time Post Application Analgesic Route and Dose of Administration Frequency on Medication Day
day 3 – day 20 Buprenorphine subcutaneous: 0.01 mg/kg bw once
Meloxicam subcutaneous: 0.5 mg/kg bw once
day 21 no analgesic medication

Clinical Observation
The eyes were examined for signs of irritation throughout the observation period. The eye irritation was scored and recorded according to the
grades in the table below.
For the calculation only the 24, 48 and 72-hour readings were used.
72 hours post-application as well as at the end of the prolonged observation period the treated eyes were examined with the aid of a fluorescein
solution (see above). The eyes were rinsed with physiological saline 0.9% NaCl (B. Braun Melsungen, lot no. 1406785, expiry date: 05/2017) after the
examination.

Evaluation of Results
Individual reactions for each animal were recorded according to the scoring system described below at each time of observation.
For the calculation only the 24, 48 and 72-hour readings were used.
Nature, severity and duration of clinical observations were described.
The body weight changes were summarised in a tabular form.
With few exceptions, data were captured using the validated departmental computerised system
E-Workbook (version 9.4.0, ID Business Solutions Ltd.).

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: reversible effects in 1 out of 3 animals within 72 hours

Time point:

other: 72 hours

Remarks on result:

other: Eye irritation fully reversible within 72 hours

Results 2

Irritation parameter:

other: irreversible effects in 2 out of 3 animals

Time point:

other: 21 days

Reversibility:

not fully reversible within: 21 days

Irritant / corrosive response data:

After the application into the eyes of three male NZW rabbits the test item produced irritant effects in all animals, which were fully reversible within 72 hours in animal no. 1 and not fully reversible until the end of the observation period of 21 days in animals no. 2 and no. 3.
Neither mortalities nor significant clinical signs of toxicity were observed. Upon fluorescein examinations at the end of the observation period of 72 hours no corneal lesions were found in animal no. 1 after test item application. Corneal lesions upon fluorescein examinations were found in the treated eye of animals no. 2 and no. 3 72 hours after test item application and at the end of the prolonged observation period. Conjunctival redness, chemosis and discharge were observed in all animals. Corneal effects were observed only in animals no. 2 and no. 3.

Thus, irreversible effects were observed in animal no. 2 and no. 3 but not in animal no. 1.

Other effects:

The body weight development of all animals was within the expected range.

Any other information on results incl. tables

 

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: other: EU CLP

Conclusions:

Under the conditions of the present study, a single ocular application of the test item Glucamide CC to rabbits at a dose of 0.1 g produced irritant
effects, which were not fully reversible within 21 days post-application in 2 out 3 animals.
Neither mortalities nor significant clinical signs of toxicity were observed.

Executive summary:

Summary Results

The eye irritation potential of Glucamide CC was investigated in rabbits according to OECD TG 405.

Species/strain:                                       New Zealand White Rabbits Crl: KBL (NZW)

Number of animals:                              3

Amount of substance:                           0.1 g per test site

First time of effects:                              animal no. 1: 1 hour post-application redness grade 1, chemosis grade 1 and discharge grade 2

                                                                animals no. 2 and no. 3: 1 hour post-application redness grade 1, chemosis grade 2 and discharge grade 3

Last time of effects:                             animal no. 1: 48 hours post-application redness grade 1

animal no. 2 : 21 days post-application redness grade 1, chemosis grade 1 and corneal opacity grade 1

animal no. 3: 21 days post-application redness grade 1 and corneal opacity grade 1

Reversibility of observed effects:   animal no. 1: the changes were fully reversible within 72 hours post-application

animals no. 2 and no. 3: the changes were not fully reversible within 21 days post-application

Conclusion

Under the conditions of the present study, a single ocular application of the test item Glucamide CC to rabbits at a dose of 0.1 g produced

irritant effects, which were not fully reversible within 21 days post-application in 2 out 3 animals. Neither mortalities nor significant clinical signs of toxicity were observed. Based on the test results, Glucamid CC is considered to be an eye irritant category 1 according to UN GHS / EU CLP.