Sage, Salvia lavandulifolia, ext.

Endpoint summary

Description of key information

SKIN CORROSION / IRRITATION :

According to an in vitro skin irritation study (OECD 439) the test item SPANISH SAGE OIL (Salvia lavanduliflia) has to be classified in Category 2 “Irritating to skin” or in Category 1 “Corrosive”. The hazard statement  “H315:  Causes  skin irritation” with the signal word “Warning” or “H314: Causes severe skin burns and eye damage” with the signal word “Danger” are required.

According to an in vitro skin corrosion study (OECD 431) the test item SPANISH SAGE OIL (Salvia lavandulifolia) does not have to be classified in Category 1 “Corrosive”.

EYE CORROSION / IRRITATION :

According to an in vitro eye corrosion / irritation study (OECD 438), the results obtained under these experimental conditions lead to the category  “no prediction can be  made”.

According to an in vitro eye corrosion / irritation study (OECD 492) :

The viability of the two treated RHCE tissues are respectively 61.64% and 56.78%. In this case, they do not have the same classification (no classification for tissue with viability >60% and classified for tissue with viability < 60%).

However, in order to maximized the risks, the test item SPANISH SAGE OIL (Salvia lavandulifolia) has to be identified as potentially requiring classification and labeling according to UN GHS Category 2 or Category 1.

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:

25 july to 8 august 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: Reconstructed Human Epidermis (RHE) Test Method)

GLP compliance:

yes

Remarks:

GLP certificate in annex 1 of the final report

Test system:

human skin model

Source species:

other: Reconstructed human epidermis

Cell type:

other:

Control samples:

yes, concurrent negative control

Amount/concentration applied:

after a treatment of 3 min and 1 hour (see annexes 2 and 3 of the final report)

Duration of treatment / exposure:

3 minutes at room temperature and 1 hour at 37°C +/- 1°C

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

dose of 50 µL

Duration of treatment / exposure:

3 minutes and 1 hour

Irritation / corrosion parameter:

% tissue viability

Remarks:

3 min

Value:

102.77

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Remarks:

1 hour

Value:

35.86

Negative controls validity:

valid

Positive controls validity:

valid

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

3 minutes and 1 hour after the test item application, the mean percent viability of the epidermis skins treated with the test item were 102.77% (considered as 100%) and 35.86%, versus 8.19% and 0.66%, respectively, with the positive control item (potassium hydroxide 8N).
In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions enable to conclude that the test item SPANISH SAGE OIL (Salvia lavandulifolia) does not have to be classified in Category 1 “Corrosive”.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

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)

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Remarks:

For SkinEthic RHE models of 0.50cm^2

Source species:

human

Cell type:

non-transformed keratinocytes

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

At dose of 16µL

Duration of treatment / exposure:

Following the exposure to the test item (during 15 minutes for the EPISKIN SM or during 42 minutes for the SkinEthic RHE model).

Duration of post-treatment incubation (if applicable):

Viability measurements are performed after a 42 hours post-treatment incubation period of the rinsed tissues in fresh medium.

Number of replicates:

3 replicates.

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

10

Value:

7.2

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

11

Value:

10.2

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

12

Value:

11

Negative controls validity:

valid

Positive controls validity:

valid

Interpretation of results:

study cannot be used for classification

Conclusions:

In accordance with the Regulation EC No. 1272/2008 and in absence of information on skin corrosion, the test item SPANISH SAGE OIL (Salvia lavanduliflia) has to be classified in Category 2 “Irritating to skin” or in Category 1 “Corrosive”. The hazard statement “H315: Causes skin irritation” with the signal word “Warning” or “H314: Causes severe skin burns and eye damage” with the signal word “Danger” are required.
That is the reason why the test OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method) has been performed.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 july to 3 october 2018

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:

yes

Remarks:

The eyes were incubated between 45 and 64 min instead of 45 and 60 min. As the results obtained with the eyes with the neg control, were conformed to what was expected, the deviation is considered as without impact on the conclusion of the study.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

• Sponsor’s identification: SPANISH SAGE OIL (Salvia lavanduliflia)
• CAS No.: 8016-65-7
• Batch No.: LS104585
• Date received: 28 March 2018
• Container: smoked glass flask (n=3)
• Storage: room temperature, darkness
• Quantity (container + content) : 113.20 g + 113.50 g + 114.20 g
• Form: liquid
• Colour: colorless to slight yellow
• Production date: August 2017
• Expiry date: August 2021

Species:

chicken

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The eyes collected from chickens obtained from a slaughterhouse (Etablissement Brun, 33820 Etauliers, France) where they are killed for human consumption have been used for this assay.
The age and weight of the chickens used 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).
Heads have been removed immediately after sedation of the chickens by electric shock, and incision of the neck for bleeding. The heads have been collected on 23 July 2018 at 8:25 am.
Because eyes were dissected in the laboratory, the intact heads were transported from the slaughterhouse at ambient temperature in plastic boxes humidified with towels moistened with physiological saline.
The eyes were enucleated at Phycher on 23 July 2018 at 8:25 am.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

0.03 mL

Duration of treatment / exposure:

The test item (liquid or solid) is applied for 10 seconds and then rinsed from the eye with physiological saline (approximately 20 mL) at ambient temperature. If the rinse volume exceeds approximately 20 mL, the rinse volume used will be reported.

Observation period (in vivo):

Treated corneas are evaluated prior to treatment and at 30, 75, 120, 180, and 240 minutes (± 5 minutes) after the post-treatment rinse.

Duration of post- treatment incubation (in vitro):

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.

Number of animals or in vitro replicates:

3

Irritation parameter:

cornea opacity score

Run / experiment:

eye no 7

Value:

<= 0

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

cornea opacity score

Run / experiment:

eye no 8

Value:

<= 0

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

cornea opacity score

Run / experiment:

eye no 9

Value:

<= 0

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein retention score

Run / experiment:

eye no 7

Value:

>= 0.5 - <= 3

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein retention score

Run / experiment:

eye no 8

Value:

>= 0.5 - <= 3

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein retention score

Run / experiment:

eye no 9

Value:

>= 0.5 - <= 3

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

percent corneal swelling

Run / experiment:

eye no 7

Value:

>= 0 - <= 2

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

percent corneal swelling

Run / experiment:

eye no 8

Value:

>= 0

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

percent corneal swelling

Run / experiment:

eye no 9

Value:

>= 0 - <= 2

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

Combinantion of the 3 Endpoints : 1 X IV , 2 X I : no prediction can be made.
No morphological effects were noted, whatever the examination time.

The ocular reactions observed in eyes treated with the test item were:

-  maximal mean score of corneal opacity: 0.0, corresponding to ICE class I;

-  mean score of fluorescein retention: 3.0, corresponding to ICE class IV;

-  maximal mean corneal swelling: 1%, corresponding to ICE class I.

The combination of the three endpoints for the test SPANISH SAGE OIL (Salvia lavandulifolia) was 1 x IV, 2 x I.

The combination of the three endpoints for the positive control, 5% Benzalkonium chloride, was 3 x IV. Therefore, the positive control is classified as “Corrosive/Severe Irritant”, as expected.

The combination of the three endpoints for the negative control, physiological saline, was 3 x I. Therefore, the negative control is classified as “No Category”, as expected.

Interpretation of results:

study cannot be used for classification

Conclusions:

In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions lead to the category “no prediction can be made”, as defined by the OECD guideline No.438.
Therefore, the test item SPANISH SAGE OIL (Salvia lavandulifolia) is not predicted as causing serious eye damage (Category 1) or as not classified for eye irritation/serious eye damage (No category) with the Isolated Chicken Eye test method.
Additional testing (in vitro and/or in vivo) is required to establish a definitive classification.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

50µL

Duration of treatment / exposure:

Following the exposure to the test item, viability measurements are performed immediately after a post-treatment incubation period of the rinsed tissues in fresh medium.

Duration of post- treatment incubation (in vitro):

Tissues treated with liquid test items and with control substances tested concurrently to liquid test items are incubated for 30 ± 2 minutes at standard culture conditions

Number of animals or in vitro replicates:

two tissue replicates

Irritation parameter:

other: percent tissue viability (%)

Run / experiment:

sample 5

Value:

61.64

Irritation parameter:

other: percent tissue viability (%)

Run / experiment:

sample 6

Value:

56.78

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The mean percent tissue viability of the RhCE replicates treated with the test item SPANISH SAGE OIL (Salvia lavandulifolia) was 59.21% versus 1.32% in the positive control (Methyl acetate).
The viability of the two treated RHCE tissues are respectively 61.64% and 56.78%. In this case, they do not have the same classification. However, in order to maximized the risks, the test item SPANISH SAGE OIL (Salvia lavandulifolia) has to be identified as potentially requiring classification and labeling according to UN GHS Category 2 or Category 1.

Executive summary:

An in vitro study is conducted on the sage oil iself but it did not permit to establish a definitive classification between category 1 and category 2.

The compounds presents in the sage oil are not classified as a eye corrosive substances according to CLP and GHS criteria but classified as eye irritant cat 2.

Hence, according to table 3.3.3 of CLP regulation, sage oil should be classified as a eye irritant substance (cat 2) according to CLP and GHS criteria.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

.

Justification for classification or non-classification

Furthermore, the sage oil does not contain any component (any molecule) classified and labelled according to UN GHS cat 1 only molecule classified according to UN GHS category 2.

Moreover, the mean percent tissue viability after exposure and post-exposure incubation is close to the threshold of 60% (61.64% and 56.78%) for not requiring classification and labelling according to UN GHS.

In conclusion, it should be determined that sage oil is classified and labelled cat 2 according to UN GHS.