Tar, Pinus Sylvestris L. A complex combination distillate produced from trunks, stumps and roots of Pinus sylvestris L. by a traditional pine tar pit, kiln or retort process.

Endpoint summary

Description of key information

Pine tar / Tar, Pinus sylvestris does not have to be classified in Category 1 “Corrosive”.

In accordance with the Regulation EC No. 1272/2008 and with the classification non-corrosive obtained with in vitro membrane barrier test method for skin corrosion, the test item Pine tar / Tar,Pinus sylvestris has to be classified in Category 2 “Irritating to skin”. The hazard statement “H315: Causes skin irritation” with the signal word “Warning” are required.

In accordance with the Regulation (EC) No. 1272/008, the results obtained under these esperimental conditions enable to conclude that the test item Pine tar / Tar, Pinus sylvestris has to be classified in Category 1 "Irreversible effects on the eye". The signal word "Danger" and hazard statements H318 "Causes serious eye damage" are required.

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:

The experiment was performed between 21 February 2017 and 03 March 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 435 (In Vitro Membrane Barrier Test Method for Skin Corrosion)

Version / remarks:

The experimental protocol was established in accordance with O.E.C.D. Test Guideline No. 435 adopted 28 July 2015.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate attached in the study report

Test system:

artificial membrane barrier model

Source species:

other: CORROSITEX®

Cell type:

other: The test system is composed of two components, a synthetic macromolecular bio-barrier (MEMBRANE DISCS, batch No. CT060115) and a Chemical Detection System (CDS).

Cell source:

other: CORROSITEX® kit

Source strain:

other: The test system is composed of two components, a synthetic macromolecular bio-barrier (MEMBRANE DISCS, batch No. CT060115) and a Chemical Detection System (CDS).

Details on animal used as source of test system:

The in vitro membrane barrier test method was performed using a commercially available kit CORROSITEX® (InVitro International supplied by INT.E.G.RA. Srl, batch No. CT060115) received on 29 September 2016 and stored at 6°C ± 3°C.

Justification for test system used:

The test system is composed of two components, a synthetic macromolecular bio-barrier (MEMBRANE DISCS, batch No. CT060115) and a Chemical Detection System (CDS).
Prior to performing the membrane barrier test, a compatibility test was performed to insure that the test item and the CDS reagent were compatible. The CDS is a measurement system that must respond to the presence of the test item.

Vehicle:

other:

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

The test item Pine tar / Tar,Pinus sylvestris was applied as supplied, at the dose of 500 μL, onto 4 synthetic proteinaceous bio-barriers.

Duration of treatment / exposure:

Observation times points were in between 25 - 245 minutes.

Number of replicates:

4

Irritation / corrosion parameter:

penetration time (in minutes)

Run / experiment:

60 minutes

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

In accordance with the Regulation EC No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item Pine tar / Tar,Pinus sylvestris does not have to be classified in Category 1 “Corrosive”. The hazard statements “H314: causes severe skin bums and eye damage” with the signal word “Danger” are not required.

Executive summary:

The aim of the study was to evaluate the possible effects of the test item Pine tar / Tar, Pinus sylvestris after deposition on a synthetic proteinaceous bio-barrier and monitoring using a Chemical Detection System (CDS).

The test item Pine tar / Tar,Pinus sylvestris was applied as supplied, at the dose of 500 μL, onto 4 synthetic proteinaceous bio-barriers. Timescale Category Test showed that the test item belongs to Corrositex Category 2. The experimental protocol was established in accordance with O.E.C.D. Test Guideline No. 435 adopted 28 July 2015.

The test item did not disrupt the membrane after 60 minutes in the four replicates.

As expected, the negative control (propionic acid 6% (v/v)) did not disrupt the membrane and was not

corrosive. As expected, the positive control (sodium hydroxide) was found to be corrosive (GHS subcategory

1B) and disrupt the membrane after 16 minutes and 29 seconds. In accordance with the Regulation EC No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item Pine tar / Tar,Pinus sylvestris does not have to be classified in Category 1 “Corrosive”. The hazard statements “H314: causes severe skin bums and eye damage” with the signal word “Danger” are not required.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The experiment was performed between 09 March 2017 and 16 March 2017

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)

Version / remarks:

The
experimental protocol was established in accordance with O.E.C.D. Test Guideline No. 439 adopted 28 July 2015 and the Test method B.46 of Council regulation No. 761/2009 dated 23 July 2009 (EU Journal L220) - ATP Council regulation No. 440/2008 of 30 May 2008 (E.U. Journal L142).

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

other: SkinEthic RHE® model

Cell source:

other: The 0.50 cm² reconstructed epidermis (Episkin SA, RHE/S/17 Batch No. 17-RHE-031)

Duration of treatment / exposure:

42 minutes

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

The mean corrected percent viability of the treated tissues was -15.3% (considered as 0%), versus 0.9% in the positive control (5% Sodium Dodecyl Sulfate).

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

In accordance with the Regulation EC No. 1272/2008 and with the classification non-corrosive obtained with in vitro membrane barrier test method for skin corrosion, the test item Pine tar / Tar,Pinus sylvestris has to be classified in Category 2 “Irritating to skin”. The hazard statement “H315: Causes skin irritation” with the signal word “Warning” are required.

Executive summary:

The aim was to evaluate the possible irritating effects of the test item Pine tar / Tar,Pinus sylvestris

after topical application on in vitro human reconstructed epidermis (SkinEthic RHE® model).

The test item Pine tar / Tar,Pinus sylvestris was applied as supplied, at the dose of

16 μL, to 3 living and 2 killed Reconstructed Human epidermis (SkinEthic RHE® model) during

42 minutes, followed by a rinse with 25 mL of DPBS and a 41 hours and 15 minutes post-incubation

period at 37°C, 5% CO2. Cell viability was then measured by enzymatic conversion of the vital dye

MTT into a blue formazan salt that was quantitatively measured after extraction from tissues. The

experimental protocol was established in accordance with O.E.C.D. Test Guideline No. 439 adopted

28 July 2015 and the Test method B.46 of Council regulation No. 761/2009 dated 23 July 2009 (EU

Journal L220) - ATP Council regulation No. 440/2008 of 30 May 2008 (E.U. Journal L142).

A yellow to brown coloration was noted on all Reconstructed Human epidermis after the rinse.

The mean corrected percent viability of the treated tissues was -15.3% (considered as 0%), versus

0.9% in the positive control (5% Sodium Dodecyl Sulfate).

In accordance with the Regulation EC No. 1272/2008, the test item Pine tar / Tar,Pinus sylvestris

has to be classified in Category 2 “Irritating to skin”. The hazard statement “H315: Causes skin

irritation” with the signal word “Warning” are required.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

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:

The experiment was performed on 10 April 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

GLP compliance:

yes (incl. QA statement)

Species:

chicken

Details on test animals or tissues and environmental conditions:

Historically, eyes collected from chickens obtained from a slaughterhouse where they are killed for human consumption have been used for this assay, eliminating the need for laboratory animals. Only the eyes of healthy animals considered suitable for entry into the human food chain are used.

Although a controlled study to evaluate the optimum chicken age has not been conducted, the age and weight of the chickens used historically in this test method are that of spring chickens traditionally processed by a poultry slaughterhouse (i.e., approximately 7 weeks old, 1.5 - 2.5 kg).

Vehicle:

physiological saline

Controls:

yes

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Test item was applied at the dose of 30 µl, to 3 enucleated chicken eyes, during 10 seconds.

Duration of treatment / exposure:

Test item was applied at the dose of 30 µl, to 3 enucleated chicken eyes, during 10 seconds. Treated corneas were evaluated pretreatment and starting at 30, 75, 120, 180, and 240 minutes after the post-treatment rinse.

Observation period (in vivo):

Treated corneas were evaluated pretreatment and starting at 30, 75, 120, 180, and 240 minutes after the post-treatment rinse.

Duration of post- treatment incubation (in vitro):

Treated corneas were evaluated pretreatment and starting at 30, 75, 120, 180, and 240 minutes after the post-treatment rinse.

Number of animals or in vitro replicates:

3

Details on study design:

All observations of the cornea and measurement of corneal thickness are performed using a Haag-Streit BP900 slit-lamp microscope with depth-measuring device no. I. For the measurement of corneal thickness, the slit-width is set at 9½, equalling 0.095 mm.

Treated corneas are evaluated prior to treatment and at 30, 75, 120, 180, and 240 minutes (± 5 minutes) after the post-treatment rinse. These time points provide an adequate number of measurements over the four-hour post-treatment period, while leaving sufficient time between measurements for the requisite observations to be made for all eyes.

The endpoints evaluated are corneal opacity, swelling, fluorescein retention, and morphological effects (e.g., pitting or loosening of the epithelium). All of the endpoints, with the exception of fluorescein retention (which is determined only at prior to treatment and at 30 minutes after test item exposure) are determined at each of the above time points.

Irritation parameter:

cornea opacity score

Value:

ca. 2.7

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Interpretation of results:

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

Conclusions:

In accordance with the Regulation (EC) No. 1272/008, the results obtained under these esperimental conditions enable to conclude that the test item Pine tar / Tar, Pinus sylvestris has to be classified in Category 1 "Irreversible effects on the eye". The signal word "Danger" and hazard statements H318 "Causes serious eye damage" are required.

Executive summary:

The test item Pine tar / Tar, Pinus sylvestris was applied at the dose of 30 µl, to 3 enucleated chicken eyes, during 10 seconds. A positive and a negative control was included. Damages of the test item were assessed by determination of corneal swelling, opacity, and fluorescein retention at 30, 75, 120, 180 and 240 minutes post-dose. In accordance with the Regulation (EC) No. 1272/008, the results obtained under these esperimental conditions enable to conclude that the test item Pine tar / Tar, Pinus sylvestris has to be classified in Category 1 "Irreversible effects on the eye". The signal word "Danger" and hazard statements H318 "Causes serious eye damage" are required.  

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Pine tar / Tar, Pinus sylvestris does not have to be classified in Category 1 “Corrosive”.

In accordance with the Regulation EC No. 1272/2008 and with the classification non-corrosive obtained with in vitro membrane barrier test method for skin corrosion, the test item Pine tar / Tar,Pinus sylvestris has to be classified in Category 2 “Irritating to skin”. The hazard statement “H315: Causes skin irritation” with the signal word “Warning” are required.

In accordance with the Regulation (EC) No. 1272/008, the results obtained under these esperimental conditions enable to conclude that the test item Pine tar / Tar, Pinus sylvestris has to be classified in Category 1 "Irreversible effects on the eye". The signal word "Danger" and hazard statements H318 "Causes serious eye damage" are required.