Pentaerythritol tetrabenzoate

Endpoint summary

Description of key information

A skin corrosion study in the Reconstructed Human Epidermis (RHE) Test Method following OECD Guideline 431 was conducted in order to determine the skin corrosive properties of Pentaerythritol tetrabenzoate. After 3 minutes treatment with the test item, the mean value of relative tissue viability was increased to 102.3%. This value is above the threshold for corrosion potential (50%). After 1 hour treatment, the mean value of relative tissue viability was reduced to 94.4%. This value is also above the threshold for corrosion potential (15%). Pentaerythritol tetrabenzoate is considered non-corrosive to skin in the Reconstructed Human Epidermis (RHE) Test Method.

An in vitro study following OECD 439 was performed in order to evaluate the potential of Pentaerythritol tetrabenzoate to evoke skin irritation in a reconstructed human epidermis (RhE) test method.

The test item Pentaerythritol tetrabenzoate is considered as non-irritant to skin. After the treatment, the mean value of relative tissue viability was reduced to 96.9%. This value is above the threshold for skin irritation (50%) according to OECD 439. The optical density of the negative control was well within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8.

The test item Pentaerythritol tetrabenzoate showed no effects on the cornea of the bovine eye and can be considered not to be eye damaging. The calculated IVIS is 0.48. According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS ≤ 3 requires no classification for serious eye damage (UN GHS No Category) The negative control (HBSS) and the positive control (20% imidazole solution) have met the validity criteria. No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19.02.2018 - 22.02.2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name: Pentaerythritol tetrabenzoate
Appearance: off-white solid
Composition: Pentaerythritol tetrabenzoate >96%
CAS No.: 4196-86-5
EINECS-No.: 224-079-8
Molecular formula: C33H28O8
Molecular weight: 552.576 g/mol
Purity: >96%
Homogeneity: homogeneous
Expiry date: Nov. 2018
Storage: Room Temperature (20 ± 5°C)

Test system:

human skin model

Source species:

human

Cell type:

other: See details on test system

Cell source:

other: See details on test system

Source strain:

other: See details on test system

Details on animal used as source of test system:

n.a.

Vehicle:

water

Details on test system:

The test system is a commercially available EpiDermTM-Kit, procured by MatTek. The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDermTM tissues are cultured on specially prepared cell cultures inserts.

EpiDermTM tissues were procured from MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia.
Designation of the kit: EPI-200-SCT
Day of delivery: 20. Feb. 2018
Batch: 25882

Control samples:

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

Tissue 1: 26.5 mg (3 minutes), 25.9 mg (1 hour)
Tissue 2: 26.4 mg (3 minutes), 25.6 mg (1 hour)

Duration of treatment / exposure:

3 min and 1 hour

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

3 replicates per group

Irritation / corrosion parameter:

% tissue viability

Value:

>= 94.4 - <= 102.3

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Other effects / acceptance of results:

The mean value of relative tissue viability of the test item was increased to 102.3% after 3 minutes treatment. This value is above the threshold for corrosivity (50%). After 1 hour treatment, the mean value of relative tissue viability of the test item was reduced to 94.4%, lying above the threshold for corrosivity (15%). Therefore, the test item is considered as non-corrosive to skin.

The criterion for optical density of the negative control (≥ 0.8 and ≤ 2.8) was fulfilled: optical density was 1.8 (3 minutes) resp. 1.9 (1 hour). The positive control showed clear corrosive effects. The criterion for the viability of the 1 hour experiment, expressed as % of the negative control (< 15%), was fulfilled, too. The mean value of relative tissue viability was 6.6% Values for negative control and for positive control were within the range of historical data of the test facility. Therefore the experiment is considered valid.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item Pentaerythritol tetrabenzoate is considered non-corrosive to skin. After 3 minutes treatment, the mean value of relative tissue viability of the test item was increased to 102.3%. This value is well above the threshold for corrosivity according to OECD TG 431 (50%). After 1 hour treatment the mean value of relative tissue viability of the test item was reduced to 94.4%. This value is well above the threshold for corrosivity (15%) according to OECD TG 431.

Executive summary:

A skin corrosion study in the Reconstructed Human Epidermis (RHE) Test Method following OECD Guideline 431 was conducted in order to determine the skin corrosive properties of Pentaerythritol tetrabenzoate.

After 3 minutes treatment with the test item, the mean value of relative tissue viability was increased to 102.3%. This value is above the threshold for corrosion potential (50%). After 1 hour treatment, the mean value of relative tissue viability was reduced to 94.4%. This value is also above the threshold for corrosion potential (15%).

Values for negative control and for positive control were within the range of historical data of the test facility confirming the validity of the experiment.

Pentaerythritol tetrabenzoate is considered non-corrosive to skin in the Reconstructed Human Epidermis (RHE) Test Method.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19.02.2018 - 23.02.2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name: Pentaerythritol tetrabenzoate
Appearance: off-white solid
Composition: Pentaerythritol tetrabenzoate >96%
CAS No.: 4196-86-5
EINECS-No.: 224-079-8
Molecular formula: C33H28O8
Molecular weight: 552.576 g/mol
Purity: >96%
Homogeneity: homogeneous
Expiry date: Nov. 2018
Storage: Room Temperature (20 ± 5°C)

Test system:

human skin model

Remarks:

The test system is a commercially available EpiDermTM-Kit, procured by MatTek. The EpiDermTM tissue consists of human-derived epidermal keratinocytes which have been cultured to form a multi-layered, highly differentiated model of the human epidermis.

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: EpiDermTM tissues were procured from MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia.

Vehicle:

other: The tissues were wetted with 25 μL DPBS buffer before applying the test item and spreading it to match the tissue size.

Details on test system:

DPBS-buffer: Solution for the rinsing of the tissues and solvent for MTT concentrate, also used as negative control. A subset was procured by MatTek In Vitro Life Science Laboratories; the other subset was prepared by LAUS GmbH.
Composition of the subset from MatTek In Vitro Life Science Laboratories (batch no.: 092817MGKA):
KCl 0.2 g
KH2PO4 0.2 g
NaCl 8.0 g
Na2HPO4 * 7H2O 2.16 g
H2O ad 1 L

Composition of the subset from LAUS GmbH (batch no.: 20171114):
KCl 0.2 g
KH2PO4 0.2 g
NaCl 8.0 g
Na2HPO4 * 2H2O 1.44 g
H2O ad 1 L

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Tissue 1: 25.5 mg
Tissue 2: 26.0 mg
Tissue 3: 25.3 mg

Duration of treatment / exposure:

Tissues were dosed in 1-minute-intervals. After dosing the last tissue, all plates were transferred into the incubator for 35 minutes at 37 ± 1°C and 5.0 ± 0.5% CO2. hour after the first application, the inserts were removed from the plates using sterile forceps and rinsed immediately in 1-minute-intervals.

Duration of post-treatment incubation (if applicable):

After rinsing, each tissue was blotted with sterile cellulose tissue and then transferred into a new 6-well-plate with fresh assay medium (0.9 mL). Then, the tissues were set in the incubator for 23 hours and 25 minutes at 37 ± 1°C and 5.0 ± 0.5% CO2.

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Tissue 1

Value:

ca. 96.5

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Tissue 2

Value:

ca. 97

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Tissue 3

Value:

ca. 97.1

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

The mean value of relative tissue viability of the test item was reduced to 96.9% after the treatment. This value is above the threshold for skin irritation (50%). Therefore, the test item is considered as non-irritant to skin.

All validity criteria were met. Values for negative control and for positive control were within the range of historical data

of the test facility.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item Pentaerythritol tetrabenzoate is considered as non-irritant to skin. After the treatment, the mean value of relative tissue viability was reduced to 96.9%. This value is above the threshold for skin irritation (50%) according to OECD 439. The optical density of the negative control was well within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8. The positive control has met the acceptance criterion too, for thus ensuring the validity of the test system.
Variation within replicates was within the accepted range for negative control, positive control and test item (required: ≤ 18%). For these reasons, the result of the test is considered valid.

Executive summary:

This in vitro study following OECD 439 was performed in order to evaluate the potential of Pentaerythritol tetrabenzoate to evoke skin irritation in a reconstructed human epidermis (RhE) test method.

The test item Pentaerythritol tetrabenzoate is considered as non-irritant to skin. After the treatment, the mean value of relative tissue viability was reduced to 96.9%. This value is above the threshold for skin irritation (50%) according to OECD 439. The optical density of the negative control was well within the required acceptability criterion of 0.8 ≤ mean OD ≤ 2.8. The positive control has met the acceptance criterion too, for thus ensuring the validity of the test system. Variation within replicates was within the accepted range for negative control, positive control and test item (required: ≤ 18%).

For these reasons, the result of the test is considered valid.

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08.02.2018 - 15-02-2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

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)

Specific details on test material used for the study:

Name: Pentaerythritol tetrabenzoate
Appearance: off-white solid
Composition: Pentaerythritol tetrabenzoate >96%
CAS No.: 4196-86-5
EINECS-No.: 224-079-8
Molecular formula: C33H28O8
Molecular weight: 552.576 g/mol
Purity: >96%
Homogeneity: homogeneous
Expiry date: Nov. 2018
Storage: Room Temperature (20 ± 5°C)

Species:

cattle

Strain:

other: Bos primigenius Taurus (fresh bovine corneas)

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.

Vehicle:

Hank's balanced salt solution

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

20% in Hank’s Balanced Salt Solution (HBSS) 10-fold concentrated, diluted in demin. water (1:10).

Duration of treatment / exposure:

4 h

Duration of post- treatment incubation (in vitro):

90 min

Number of animals or in vitro replicates:

3

Irritation parameter:

in vitro irritation score

Run / experiment:

1

Value:

ca. 0.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

2

Value:

ca. 1.2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

3

Value:

ca. -0.25

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Interpretation of results:

GHS criteria not met

Conclusions:

The test item Pentaerythritol tetrabenzoate was tested as a 20% suspension in HBSS. Under the conditions of this test, the test item Pentaerythritol tetrabenzoate showed no effects on the cornea of the bovine eye and can be considered not to be eye damaging. The calculated IVIS is 0.48. According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS ≤ 3 requires no classification for serious eye damage (UN GHS No Category) The negative control (HBSS) and the positive control (20% imidazole solution) have met the validity criteria. No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid.

Executive summary:

This in vitro BCOP study following OECD 437 was performed to assess corneal damage potential of Pentaerythritol tetrabenzoate by quantitative measurements of changes in opacity and permeability in a bovine cornea. Two experiments were performed. The first experiment was discontinued after the second opacity measurement, because the opacity values of two replicates of the negative control were too high. Therefore the first experiment was declared invalid and was discontinued.

The results and data of the invalid experiment are not included in this final report but will be archived with the raw data in the GLP Archive of the Test Facility. The second experiment was valid and reported. The test item Pentaerythritol tetrabenzoate was brought onto the cornea of a bovine eye which previously had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been determined. 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. The test item was tested as a 20% suspension in HBSS. Under the conditions of this test, the test item Pentaerythritol tetrabenzoate showed no effects on the cornea of the bovine eye and can be considered not to be eye damaging. The calculated IVIS is 0.48. According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS ≤ 3 requires no classification for serious eye damage (UN GHS No Category). The negative control (HBSS) and the positive control (20% imidazole solution) have met the validity criteria. No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid.