Hesperidin

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant guideline study without significant deviations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Hesperidin

EC Number:

208-288-1

EC Name:

Hesperidin

Cas Number:

520-26-3

Molecular formula:

C28H34O15

IUPAC Name:

5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-4-oxo-3,4-dihydro-2H-chromen-7-yl 6-O-(6-deoxy-alpha-L-mannopyranosyl)-beta-D-glucopyranoside

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

orange/brown powder
Batch# 120323
Purity: 91.15% based on dry mass

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.27 or 2.36 kg and were twelve to twenty weeks old. After an acclimatisation period of at least five days each animal was given a number unique within the study which was written with a black indelible marker-pen on the inner surface of the ear and on the cage label.
The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness. The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

A volume of 0.1 ml of the test item, which was found to weigh approximately 75 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball.

Duration of treatment / exposure:

72 hours in total with score reading following 1, 24, 48 and 72 hours after initial exposure.

Observation period (in vivo):

Terminated after 72 hours due to lack of irriation.

Number of animals or in vitro replicates:

2 animals were used in this experiment

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.
Initially, a single rabbit was treated. A volume of 0.1 ml of the test item, which was found to weigh approximately 75 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made.
After consideration of the ocular responses produced in the first treated animal, a second animal was treated. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation (from Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).
Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Any clinical signs of toxicity, if present, were also recorded. Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Due to the very weak response regards irritation, a third test animal was not required and the test was terminated as the substance was shown to be not an eye irritant.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Remarks:

of 2 animals

Time point:

other: 1 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Remarks:

of 2 animals

Time point:

24 h

Score:

0

Max. score:

4

Results 3

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Remarks:

of 2 animals

Time point:

48 h

Score:

0

Max. score:

4

Results 4

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Remarks:

of 2 animals

Time point:

72 h

Score:

0

Max. score:

4

Results 5

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

mean

Remarks:

of 2 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 6

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 2 animals

Time point:

other: 1 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 48 h

Results 7

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 2 animals

Time point:

24 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 24 h

Results 8

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 2 animals

Time point:

48 h

Score:

0

Max. score:

2

Results 9

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 2 animals

Time point:

72 h

Score:

0

Max. score:

2

Results 10

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 2 animals

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible

Results 11

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 2 animals

Time point:

other: 1 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 72 h

Results 12

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 2 animals

Time point:

24 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 48 h

Results 13

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 2 animals

Time point:

48 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 24 h

Results 14

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 2 animals

Time point:

72 h

Score:

0

Max. score:

3

Results 15

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of 2 animals

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible

Results 16

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 2 animals

Time point:

other: 1 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Results 17

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 2 animals

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Results 18

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 2 animals

Time point:

48 h

Score:

0.5

Max. score:

4

Reversibility:

fully reversible within: 24 h

Results 19

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 2 animals

Time point:

72 h

Score:

0

Max. score:

4

Results 20

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 2 animals

Time point:

24/48/72 h

Score:

0.5

Max. score:

4

Reversibility:

fully reversible

Irritant / corrosive response data:

As both rabbits tested showed corneal opacity <1, iritis <1, conjunctival redness < 2 and conjunctival oedema (chemosis) <2, calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and all scores on both animals returned to zero within 72 hours, the substance is not subject to classification according to GHS (Globally Harmonized System of Classification and Labelling of Chemicals) and CLP (Regulation (EC) No 1272/2008). As a result, for animal welfare, a third animal was not used in this test.

Any other information on results incl. tables

No corneal effects were noted during the study. Iridial inflammation was noted in both treated eyes one and 24 hours after treatment. Moderate conjunctival irritation was noted in both treated eyes one hour after treatment. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye at the 24-Hour observation with minimal conjunctival irritation noted in both treated eyes at the 48-Hour observation. Both treated eyes appeared normal at the 72-Hour observation.

Both animals showed expected gain in bodyweight during the study.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

As a result of this test hesperidin is not subject to classification for eye-irriation according to CLP and/or DSD .

Executive summary:

As both rabbits tested showed corneal opacity <1, iritis <1, conjunctival redness < 2 and conjunctival oedema (chemosis) <2, calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and all scores on both animals returned to zero within 72 hours, the substance is not subject to classification according to GHS (Globally Harmonized System of Classification and Labelling of Chemicals) and CLP (Regulation (EC) No 1272/2008). As a result, for animal welfare, a third animal was not used in this test.