Extract from the seeds of Trigonella foenum-graecum (Fabaceae) obtained by extraction with polar solvents

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 November 2019 - 11 December 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Extract from the seeds of Trigonella foenum-graecum (Fabaceae) obtained by extraction with polar solvents

EC Number:

950-727-0

Molecular formula:

Fenugreek extract

IUPAC Name:

Extract from the seeds of Trigonella foenum-graecum (Fabaceae) obtained by extraction with polar solvents

Specific details on test material used for the study:

Identification/Synonym FENUGREEK EXTRACT
Substance type UVCB substance
EC No. 950-727-0
Recommended storage Ambient (18-20°C) and protected from temperatures below 4°C; dry (< 70% rh);
conditions to be stored under dark conditions

Test animals / tissue source

Species:

chicken

Strain:

other: Ross 308

Details on test animals or tissues and environmental conditions:

Head collection was performed by a slaughter house technician. Heads were removed immediately after sedation of the chickens. The heads were transported at the earliest convenience for use approximately within 2 hours from collection. The ambient temperature was optimal (19.3ºC to 20.2ºC) during the transport. After collection, the heads were inspected for appropriate quality and wrapped with paper moistened with saline, then placed in a plastic box that can be closed (4-5 heads/box).

Test system

Amount / concentration applied:

Used as such, whole surface of eye covered

Duration of treatment / exposure:

10 seconds

Duration of post- treatment incubation (in vitro):

The control and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse.

Number of animals or in vitro replicates:

3 samples
3 positive controls (acetid acid)
1 negative control ( NaCl solution)

Details on study design:

SELECTION AND PREPARATION OF ISOLATED EYES
A Drop of fluorescein solution was applied onto the cornea surface and rinsed off with isotonic saline. Checking of the integrity of the cornea, removing of the eyeball. Unnecessary material was removed from the eye. Eye was fixed with steel clamp in vertical position.

EQUILIBRATION AND BASELINE RECORDINGS
At the end of the acclimatization period, a zero reference measurement was recorded for cornea thickness and opacity to serve as a baseline (t=0) for each individual eye. The cornea thickness of the eyes should not change by more than ±5-7 % within approximately 45 to 60 minutes before the start of application. Changes in thickness were not observed in the eyes. Following the equilibration period, the fluorescein retention was measured. Baseline values were required to evaluate any potential test item related effects after treatment.

NUMBER OF REPLICATES
3 samples

NEGATIVE CONTROL USED
1 negative control ( 30 µL saline solution)

POSITIVE CONTROL USED
3 positive controls (30 µL 10 % w/w acetid acid)

APPLICATION DOSE AND EXPOSURE TIME
undiluted
10 seconds

OBSERVATION PERIOD
30, 75, 120, 180 and 240 minutes

REMOVAL OF TEST SUBSTANCE
Cornea surface was rinsed thoroughly with saline solution at ambient temperature.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

percent corneal swelling

Run / experiment:

75 min

Value:

2

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

percent corneal swelling

Run / experiment:

240 min

Value:

3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Value:

0.5

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

fluorescein retention score

Value:

0.2

Other effects / acceptance of results:

Tables of ICE classification, see tables attached below.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The overall ICE score was 3xI. According to the guideline OECD 438, Test Substance is categorized as “No Category”.