3-cyclohexylaminopropane-1-sulphonic acid

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP OECD 437 guideline study without deviations on the registered substance itself

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 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

OECD Guideline No. 437, edition adopted 26. Jul. 2013: “Bovine Corneal Opacity and Permeability 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.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Version / remarks:

EU Method B. 47: “Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants” adopted 09. Dec. 2010

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesamt für Umwelt, Waserwirtschaft und Gewerbeaufsicht, Kaiser-Friedrich-Straße 7, 55116 Mainz, Germany

Test material

Test material information

Constituent 1

Reference substance name:

3-cyclohexylaminopropane-1-sulphonic acid

EC Number:

214-492-1

EC Name:

3-cyclohexylaminopropane-1-sulphonic acid

Cas Number:

1135-40-6

Molecular formula:

C9H19NO3S

IUPAC Name:

3-cyclohexylaminopropane-1-sulphonic acid

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): 3-(Cyclohexylamino)-propane sulfonic acid, 3-CAPS
- Storage condition of test material: room temperature (20 ± 5 °C) in a closed vessel

Test animals / tissue source

Species:

other: Bos primigenius Taurus (fresh bovine corneas)

Strain:

not specified

Details on test animals or tissues and environmental conditions:

not applicable

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: 0.9% NaCl in demin. water (negative control), 20% Imidazole dissolved in 0.9% sodium chloride solution (positive control)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 250.4 mg, 255.8 mg, or 292.7 mg. The test item was given on the epithelium in such a manner that the cornea was completely covered with test item.

Duration of treatment / exposure:

4 h

Observation period (in vivo):

incubation with fluorescein solution for further 90 min

Number of animals or in vitro replicates:

3 replicate corneas for each treatment group (negative control solution, test item and positive control)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): thorough rinsing with cMEM with phenol red and final rinsing with cMEM without phenol red
- Time after start of exposure: 4 h

SCORING SYSTEM: Opacity value, calculated from the measured absorbance at 570 nm; permeability of the cornea was measured as optical density of the liquid at 490 nm

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

cornea #1 (test item)

Value:

-0.04

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: IVIS ≤ 3 -> No Category (UN GHS, Eye damage)

Results 2

Irritation parameter:

in vitro irritation score

Run / experiment:

cornea #2 (test item)

Value:

-0.38

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: IVIS ≤ 3 -> No Category (UN GHS, Eye damage)

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

cornea #3 (test item)

Value:

0.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: IVIS ≤ 3 -> No Category (UN GHS, Eye damage)

Results 4

Irritation parameter:

in vitro irritation score

Run / experiment:

mean over 3 corneas (test item)

Value:

-0.01

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: IVIS ≤ 3 -> No Category (UN GHS, Eye damage)

Other effects / acceptance of results:

In the negative control, no signs of eye irritation were observed (IVIS = 0.89; mean over 3 corneas).
The positive control induced serious eye damage (IVIS = 77.74; mean over 3 corneas), which would be classified as GHS category 1.
The test item 3-(Cyclohexylamino)-propane sulfonic acid showed no effects on the cornea of the bovine eye. The mean calculated IVIS (in vitro irritancy score) is -0.01.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Testing was performed via a GLP OECD 437 guideline study on the registered substance itself. The present in vitro method is recommended in a tiered testing approach and the validity criteria are met, making the results sufficiently reliable to assess the corrosivity of 3-(Cyclohexylamino)-propane sulfonic acid to the eye. The present in vitro method allows the identification of corrosive chemical substances and mixtures or of substances which do not require classification.
One valid experiment was performed. The test item 3-(Cyclohexylamino)-propane sulfonic acid showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is -0.01. The experiment is considered as sufficient for the classification of the test item, because all three replicates of the test item lead to the same assessment for the test item. This single in vitro test can serve as a stand-alone test as it was consistently shown that the test item does not require classification (eye damage).

Executive summary:

In an OECD 437 (in vitro eye corrosion) study under GLP, one valid experiment was performed.

Bovine corneas were used. They were collected from slaughtered cattle which were between 12 and 60 months old.

The test item 3-(Cyclohexylamino)-propane sulfonic acid was brought onto the cornea of a bovine eye which had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 h and whose opacity had been measured. The test item was incubated on the cornea for4 h at32 ± 1 °C. After removal of the test item, opacity and permeability values were measured.

 

Physiological sodium chloride solution was used as negative control. The negative control showed no irritating effect on the cornea and the calculated IVIS (in vitro irritancy score) is 0.89.

20% Imidazole solution was used as positive control. The positive control induced serious eye damage on the cornea and falls within two standard deviations of the current historical mean. The calculated IVIS (in vitro irritancy score) is 77.74.

 

Under the conditions of this study, the test item 3-(Cyclohexylamino)-propane sulfonic acid showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is – 0.01.

According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS ≤ 3 requires no classification for eye irritation or serious eye damage.