Allyltributylphosphonium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study initiation dated : 13 Jun 2018 and experimental completion date 28 Jun 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

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 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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Allyltributylphosphonium chloride

EC Number:

216-231-7

EC Name:

Allyltributylphosphonium chloride

Cas Number:

1530-48-9

Molecular formula:

C15H32P.Cl

IUPAC Name:

tributyl(prop-2-en-1-yl)phosphanium chloride

impurity 1

Reference substance name:

Tetrahydrothiophene 1,1-dioxide

EC Number:

204-783-1

EC Name:

Tetrahydrothiophene 1,1-dioxide

Cas Number:

126-33-0

Molecular formula:

C4H8O2S

IUPAC Name:

tetrahydrothiophene 1,1-dioxide

impurity 2

Reference substance name:

Tributyl-1-propen-1-yl-phosphonium chloride

Molecular formula:

C15H32P.Cl

IUPAC Name:

Tributyl-1-propen-1-yl-phosphonium chloride

Test material form:

other: solid : wax like

Details on test material:

Homogeneity : homogeneous if totally melted
Note: For every use the test item was completely melted and the melted test item
was used for the test.

Specific details on test material used for the study:

Designation in Test Facility: 18032901G
Date of Receipt: 29. Mar. 2018
Condition at Receipt Room temperature, in proper conditions
Batch no. INT 18 192
Appearance Beige crystallised mass
Purity 95.0 % (NMR 1H and 31P)
Homogeneity homogeneous if totally melted
Expiry date 17. Oct. 2019
Storage Room temperature (20 ± 5°C)
The following additional information was relevant to the conduct of the study, according to
OECD 437 and provided by the sponsor:
CAS No. 1530-48-9
EINECS-No. 216-231-7
Chemical Class not stated
Volatility < 0.1 hPa at 30 °C
pH-value unknown
Stability H2O: unknown; EtOH: unknown; acetone: unknown;
CH3CN: unknown; DMSO: unknown
Solubility H2O: > 1 g/L; EtOH: unknown; acetone: unknown;
CH3CN: unknown; DMSO: unknown
Surface activity no*

Test animals / tissue source

Species:

other: Bos primigenius Taurus (fresh bovine corneas)

Strain:

other: bovine

Details on test animals or tissues and environmental conditions:

Fresh bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and 60 months old. The eyes were transported to the test facility in Hanks’ Balanced Salt Solution with 1% Penicillin-Streptomycin solution (Penicillin 100 U/mL, Streptomycin 100 μg/mL) in a suitable cooled container within 1 hour.

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

The test item is a solid non-surface-active substance.
In a non-GLP pre-test, the solubility of the test item was determined in Hank’s Balanced Salt Solution (HBSS) 10-fold concentrated, diluted in demin. water (1:10). The test item is soluble in a concentration of 20% in HBSS.
Therefore, the test item was tested as solution with a concentration of 20 % in Hank’s Balanced Salt Solution (HBSS) 10-fold concentrated, diluted in demin. water (1:10). On the day of the test, an aliquot of the test item was heated to 100°C in the water bath and stirred until complete melting of the substance to achieve homogeneity. Then it was diluted in HBSS and used for the assay.

Duration of treatment / exposure:

Incubation time 4 h

Details on study design:

Method Description
After the initial incubation, the medium was completely changed and the baseline opacity for each cornea was recorded. None of the corneas showed tissue damage; therefore, all corneas were used. For each treatment group (negative control solution, test item and positive control), three replicates were used.
After removal of the pre-incubation medium (cMEM without phenol red), 750 μL negative control solution, 750 μL test item solution and 750 μL positive control solution were applied to each replicate to the epithelial side of the cornea.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

123.69

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

According to the guideline, the test is considered as valid if the positive control causes an
IVIS that falls within two standard deviations of the current historical mean.
The mean IVIS of the negative control has to show an IVIS ≤ 3.
In the negative control, no signs of eye irritation were observed.
The positive control induced serious eye damage, which would be classified as GHS category
I.The test item Allyltributylphosphonium chloride induced serious eye damage on the cornea of the bovine eye. The calculated mean IVIS was 123.69.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS > 55 induces serious eye damage, that should be classified as UN GHS Category I.

Executive summary:

Two experiments were performed. The first experiment was not valid, because the IVIS of the negative control was above the acceptable limit. Therefore, the experiment was declared invalid and repeated. The results are not reported, but the raw data are kept in the GLP-archive of the test facility. The second performed experiment was valid. Bovine corneas were used. They were collected from slaughtered cattle that were between 12 and 60 months old.

The test item Allyltributylphosphonium chloride was brought onto the cornea of a bovine eye which had been previously incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been measured.

The test item was incubated on the cornea for 4 hours at 32 ± 1 °C. After removal of the test item, opacity and permeability values were measured.

Hank’s Balanced Salt Solution (HBSS) was used as negative control. The negative control showed no irritating effect on the cornea and the calculated mean IVIS (In Vitro Irritancy Score) was 2.57. 20% imidazole solution was used as positive control. The positive control induced serious eye damage on the cornea and was within two standard deviations of the current historical mean. The calculated mean IVIS was 102.27. Under the conditions of this study, the test item Allyltributylphosphonium chloride induced serious eye damage on the cornea of the bovine eye. The calculated mean IVIS was 123.69. According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS > 55 induces serious eye damage, that should be classified as UN GHS Category I.