p-menth-1-en-8-yl acetate

Endpoint summary

Description of key information

Alpha-Terpinyl Acetate is considered to be not irritating to the skin when using CLP criteria. It is not irritating to eye and is not expected to be a respiratory irritant either.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study is performed in compliance with OECD guideline 404 but reliability 2 is assigned because observations were not followed until the reversibility of the effects (only 7 days)

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

The skin observations should have been extended after 7 days because they were not fully reversible.

GLP compliance:

yes

Species:

rabbit

Strain:

other: albino

Type of coverage:

semiocclusive

Preparation of test site:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume with unit): 0.5 mL

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

Test 1: 3
Test 2&3: 4

Details on study design:

SCORING SYSTEM: Draize scoring system
- tested in a multi-patch study

Irritation parameter:

erythema score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 h

Score:

1.89

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Test 1

Irritation parameter:

edema score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 h

Score:

1.67

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Test 1

Irritation parameter:

erythema score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Test 2

Irritation parameter:

edema score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 h

Score:

2.33

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Test 2

Irritation parameter:

erythema score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 h

Score:

1.75

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Test 3

Irritation parameter:

edema score

Basis:

mean

Remarks:

all animals

Time point:

other: 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: Test 3

Irritant / corrosive response data:

Test 1: One animal showed marked desquamation at the 7 day time interval. The other two animal showed desquamation at the 7 day time interval.
Test 2:One animal showed desquamation at the 7 day time interval. One other animal showed slight desquamation at the 7 day time interval.
Test 3: One animal showed minimal desquamation at the 7 day time interval. The other three other animals showed slight desquamation at the 7 day time interval.

 

Test 1

	1 hr	1 day	2 days	3 days	7 days	Mean (24, 48, 72 hrs)
Animal No. 1
Erythema	0	1	2	2	1	1.67
Oedema	0	1	1	1	1	1
Animal No. 2
Erythema	2	2	2	2	1	2
Oedema	1	2	2	2	1	2
Animal No. 3
Erythema	1	2	2	2	2	2
Oedema	2	2	2	2	1	2

 

Test 2

	1 hr	1 day	2 days	3 days	7 days	Mean (24, 48, 72 hrs)
Animal No. 1
Erythema	2	2	2	2	2	2
Oedema	3	3	3	3	1	3
Animal No. 2
Erythema	2	2	2	2	1	2
Oedema	2	2	2	2	1	2
Animal No. 3
Erythema	2	2	2	2	1	2
Oedema	2	2	2	2	1	2
Animal No. 4
Erythema	1	2	2	2	1	2
Oedema	0	2	2	2	1	2

 

Test 3

	1 hr	1 day	2 days	3 days	7 days	Mean (24, 48, 72 hrs)
Animal No. 1
Erythema	2	2	2	2	1	2
Oedema	1	2	2	2	1	2
Animal No. 2
Erythema	1	2	2	1	1	1.67
Oedema	1	2	1	0	0	1
Animal No. 3
Erythema	1	2	2	2	1	2
Oedema	1	1	1	0	0	0.67
Animal No. 4
Erythema	1	2	1	1	0	1.33
Oedema	0	1	0	0	0	0.33

Interpretation of results:

other: not irritating using CLP criteria

Remarks:

Criteria used for interpretation of results: other: the substance is irritating using DSD criteria

Conclusions:

For substances CLP is leading and therefore the substance is considered to be not irritating.

Executive summary:

In three primary dermal irritation studies performed according to OECD guideline 404 and in compliance with GLP, three or four albino rabbits were dermally exposed to 0.5 mL of undiluted alpha-Terpinyl Acetate under a semi-occlusive patch for 4 hours to intact skin of the flank. Animals were then observed for 7 days. Irritation was scored by the method of Draize at 1, 24, 48 and 72 hours and 7 days after exposure. Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals of the first test were 1.66, 2 and 2 for erythema score and 1, 2 and 2 for edema score. For the second test 2, 2, 2 and 2 for erythema score and 3, 2, 2 and 2 for edema score. For the third test the mean individual scores were 2, 1.67, 2 and 1,33 for erythema score and 2, 1, 0.67 and 0.33 for edema score. Under the test conditions the substance is considered to be irritating to the skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not 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:

31 August 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliant, guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)

GLP compliance:

yes (incl. QA statement)

Remarks:

TNO Triskelion, Utrechtseweg 48, Zeist, the Netherlands

Species:

other: ROSS, spring chicken eyes

Details on test animals or tissues and environmental conditions:

Approximately 7 weeks old, male or female chickens (ROSS, spring chickens), body weight range approximately 1.5 - 2.5 kg, were used as eye-donors. Heads of these animals were obtained from poultry slaughterhouse v. Miert, Breukelen, the Netherlands. Heads of the animals were cut off immediately after sedation of the animals by electric shock and incision of the neck for bleeding, and before they reached the next station on the process line. The heads were placed in small plastic boxes on a bedding of paper tissues moistened with isotonic saline. Next, they were transported to the testing facility. During transportation, the heads were kept at ambient temperature.

Vehicle:

unchanged (no vehicle)

Controls:

other: Control eyes

Amount / concentration applied:

30 µL

Duration of treatment / exposure:

10 seconds

Observation period (in vivo):

The eyes were examined at approximately 0, 30, 75, 120, 180 and 240 minutes after treatment.

Number of animals or in vitro replicates:

Three test eyes per test sample, one negative control eye and three positive control eyes were selected for testing.

Details on study design:

Within 2 hours after kill, eyes were carefully dissected and placed in a superfusion apparatus using the following procedure: First the eye-lids were carefully removed without damaging the cornea and a small drop of Fluorescein sodium BP 2.0% w/v (Minims, Chauvin, England) was applied to the corneal surface for a few seconds and subsequently rinsed off with isotonic saline at ambient temperature. Next, the head with the fluorescein-treated cornea was examined with a slit-lamp microscope (Slit-lamp 900 BP, Haag-Streit AG, Liebefeld-Bern, Switzerland) to ensure that the cornea was not damaged. lf undamaged (e.g., fluorescein retention and corneal opacity scores of ≤ 0.5), the eye was further dissected from the head without damaging the eye or cornea. Care was taken to remove the eye-ball from the orbit without cutting off the optical nerve too short. The enucleated eye was placed in a stainless steel clamp with the cornea positioned vertically and transferred to a chamber of the superfusion apparatus (TNO, Zeist, the Netherlands). The clamp holding the eye was positioned in such a way that the entire cornea was supplied with isotonic saline from a bent, stainless steel tube, at a target rate of 0.10 - 0.15 mL/min (peristaltic pump set at speed 5.00, Watson-Marlow 205C4, Rotterdam, the Netherlands). The chambers of the superfusion apparatus as well as the saline were temperature controlled at approximately 32°C (water pump set at 36.4°C; Lauda 103, Germany). After placing in the superfusion apparatus, the eyes were examined again with the slit-lamp microscope to ensure that they were not damaged. Corneal thickness was measured using the Depth Measuring Attachment No. I for the Haag-streit slit-lamp microscope. Corneal thickness was expressed in instrument units. An accurate measurement was taken at the corneal apex of each eye. Eyes with a corneal thickness deviating more than 10% of the average corneal thickness of the eyes, eyes showing opacity (score higher than 0.5), or were unacceptably stained with fluorescein (score higher than 0.5) indicating the cornea to be permeable, or eyes that showed any other signs of damage, were rejected as test eyes and replaced. Three test eyes per test sample, one negative control eye and three positive control eyes were selected for testing. Each eye provided its own baseline values for corneal swelling, corneal opacity and fluorescein retention. For that purpose, after an equilibration period of 45-60 minutes, the corneal thickness of the eyes was measured again to determine the zero reference value for corneal swelling calculations. The isolated chicken eyes were exposed to a single application of 30 uL of the test sample for 10 seconds followed by a 20 mL saline rinse. After rinsing, each eye in the holder was returned to its chamber. The eyes were examined at approximately 0, 30, 75, 120, 180 and 240 minutes after treatment. Fluorescein retention was only scored at approximately 30 minutes after treatment. All examinations were carried out with the slit-lamp microscope. After the final examination, the test substance treated eyes, the negative and positive control eyes were preserved in a neutral aqueous phosphate-buffered 4% solution of formaldehyde. The corneas were embedded in paraffin wax, sectioned at 5 µM and stained with PAS (Periodic Acid-Schiff). The microscopic slides were filed in the archives and kept available for histopathological examination if considered relevant. ln the case of Terpinyl Acetate Alpha, histopathological examination was considered not relevant, because no borderline severe corneal effects were observed.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 30 min

Score:

0.5

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 75 min

Score:

1

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 120, 180, 240 min

Score:

1.2

Max. score:

4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 30 min

Score:

1

Max. score:

100

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 75, 120 , 180 min

Score:

4

Max. score:

100

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 240

Score:

3

Max. score:

100

Irritation parameter:

other: Fluorescein retention

Basis:

mean

Time point:

other: 30 min

Score:

0.7

Max. score:

3

Irritant / corrosive response data:

Terpinyl Acetate Alpha caused very slight corneal swelling, slight or slight to moderate corneal opacity, and very slight or slight fluorescein retention. The calculated lrritation lndex was 42 (max possible score is 200). The negative control eye did not show any corneal effect and demonstrated that the general conditions during the tests were adequate. The positive control eyes showed severe corneal effects and demonstrated the suitability and sensitivity of the ICE to detect severe eye irritants.

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the UN-GHS and the EU-CLP classification schemes of the lCE, Terpinyl Acetate Alpha is considered to be "Not Classified".

Executive summary:

In a GLP compliant isolated chicken eye test performed according to OECD guideline 438, the eye irritation potential of Terpinyl Acetate Alpha was evaluated. Isolated chicken eyes were exposed to a single application of 30 uL of the test sample for 10 seconds followed by a 20 mL saline rinse. Three main parameters were measured to disclose possible adverse eye effects: corneal thickness (expressed as corneal swelling), corneal opacity and fluorescein retention of damaged epithelial cells. Terpinyl Acetate Alpha caused very slight swelling of the cornea, slight or slight to moderate corneal opacity, and very slight or slight fluorescein retention. The calculated lrritation lndex was 42 (max possible score is 200). The negative control (saline) caused no corneal effects. The positive control BAC 5% caused moderate corneal swelling, severe opacity and severe fluorescein retention. The calculated lrritation lndex was 42 (max possible score is 200). According to the UN-GHS and the EU-CLP classification schemes of the lCE, TerpinylAcetate Alpha is considered to be "Not Classified".

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

For alpha-Terpinyl Acetate, skin irritation information is available. Four in vivo skin irritation tests according to the OECD TG 404 have been performed. Three of these have been reported in ECETOC (1995) and one is performed by the Toxicol Lab. In ECETOC report, three primary dermal irritation tests are presented. These are performed according to OECD guideline 404 and in compliance with GLP, three (test 1) or four (test 2 and 3) albino rabbits were dermally exposed to 0.5 mL of undiluted alpha-Terpinyl Acetate under a semi-occlusive patch for 4 hours to intact skin of the flank (ECETOC 1995). Animals were then observed for 7 days. Irritation was scored by the method of Draize at 1, 24, 48 and 72 hours and 7 days after exposure. Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals of the first test were 1.66, 2 and 2 for erythema score and 1, 2 and 2 for edema score. For the second test the mean individual scores were 2, 2, 2 and 2 for erythema score and 3, 2, 2 and 2 for edema score. For the third test the mean individual scores were 2, 1.67, 2 and 1,33 for erythema score and 2, 1, 0.67 and 0.33 for edema score. At the end of the observation period of 7 days, reversibility of the effects was seen but in some animals slight desquamation was visible.

In the skin irritation study by the Toxicol Lab four albino rabbits were dermally exposed to 0.5 mL of undiluted alpha-Terpinyl Acetate, under a semi-occlusive patch for 4 hours to the clipped skin of the trunk (Hayes 1986). Animals were then observed for 7 days. Erythema and oedema was scored at 1, 24, 48 and 72 hours and 7 days after exposure. The average (24 + 48 + 72 hours) erythema score was 1.8 and the average oedema score was 1.0. Seven days after dosing very slight reaction remained in three animals and desquamation from the surface of the treated skin was apparent in all four rabbits. In this study, alpha-Terpinyl Acetate was found not to be skin irritant when applied topically to the rabbit.

Eye irritation

In a GLP compliant isolated chicken eye test performed according to OECD guideline 438, the eye irritation potential of alpha-Terpinyl Acetate was evaluated (TNO Triskelion 2012). Isolated chicken eyes were exposed to a single application of 30 uL of the test sample for 10 seconds followed by a 20 ml saline rinse. Three main parameters were measured to disclose possible adverse eye effects: corneal thickness (expressed as corneal swelling), corneal opacity and fluorescein retention of damaged epithelial cells. Alpha-Terpinyl Acetate caused very slight swelling of the cornea, slight or slight to moderate corneal opacity, and very slight or slight fluorescein retention. The calculated lrritation lndex was 42 (max possible score is 200). The negative control (saline) caused no corneal effects. The positive control BAC 5% caused moderate corneal swelling, severe opacity and severe fluorescein retention. The calculated lrritation lndex was 42 (max possible score is 200). According to the UN-GHS and the EU-CLP classification schemes of the lCE, alpha-Terpinyl Acetate is considered to be "Not Classified".

Respiratory irritation

For respiratory irritation mostly human data are used for the assessment because no suitable in vitro or in vivo tests are available that can identify respiratory irritation (REACH guidance R.7.2.3). There are no human data such as indicated in R7.2.3 of the ECHA guidance that indicate respiratory reactions of the substance e. g. from consumer experience or occupational exposure. Despite the slight skin irritating potential, the substance is not corrosive or severely irritating which further minimizes the respiratory irritation hazard (REACH guidance: 7.2.1.2).

Justification for selection of skin irritation / corrosion endpoint:
Four skin irritation studies are available performed with alpha-Terpinyl Acetate. Though the substance need to be classified and labelled according to DSD. It is decided that for the risk characterisation the conclusion on CLP is leading because this is the classification scheme for substances under REACH.

Justification for selection of eye irritation endpoint:
One eye irritation study is available, which is performed in accordance with OECD 438 and GLP. This study is adequate for covering this endpoint.

Justification for classification or non-classification

The skin irritation results in all four tests are fairly consistent. The erythema and oedema scores are around 2. The effects are reversible but at the end of the observation period at day 7 slight erythema (circa 1) and some desquamation may be seen. The interpretation for classification and labelling are presented in the table below:

Test 1/3 animals	Test 1	Test 2	Test 3	Test 4
Reference	ECETOC	ECETOC	ECETOC	Toxicol lab
Erythema average	< 2 (1.89)	2	< 2 (1.75)	< 2 (1.67)
Oedema average	< 2 (1.66)	2.3	< 2 (1)	< 2 (0.75)
Number of animals erythema score >=2	2/3	4/4	2/4	2/4
Number of animals oedema score >=2	2/3	4/4	1/4	1/4
Erythema/Oedema and desquamation (de) at the end of the observation period	1-2 + de	Mostly 1+ and de	1 + de	0-1 + de
DSD criteria	Yes based on scores 2 or > in 2/3 animals	Yes based on scores 2 in 4/4 animals	No: based on scores lower than 2 and only 2 and 1 out of 4 animals showing the erythema or oedema, respectively	No based on scores lower than 2 and only 2 and 1 out of 4 animals showing the erythema or oedema, respectively
Number of animals erythema score >=2.3	0	0	0	0
Number of animals oedema score >=2.3	0	1 out of 4 showed oedema 3,1 showed oedema 2.3all others were 2	0/4 showed	0
CLP criteria	Not reached scores remain <2.3 and no animals having scores > 2.	No only two out of 4 animals shows oedema score>2.3	No all scores are <2.3	No all scores are <2.3

The erythema score of ≥2, seen at 2/3 or 10/15 animals, result in the DSD classification and labelling. Since no animals scored erythema >=2.3 and only two animals oedema of >= 2.3, CLP labelling is not needed. The slight irritation effects seen at the end of the observation period of 7 days indicate reversibility and are therefore not considered for classification and labelling because it can be anticipated that these effects are fully reversible within 14 days. These effects and the desquamation seen at the end of the test do not indicate effects needed for classification.

The effects seen result in Xi:R38 according to the EU Directive 67/548 and absence of classification and labelling according to the EU Classification, Labelling and Packaging of Substances (CLP) Regulation (EC) No. 1272/2008.

The in vitro eye irritation study with alpha-Terpinyl Acetate showed minor eye irritation which does not warrant classification according to the EU Directive 67/548 for mixtures and according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

The absence of skin and eye irritation potential shows that the substance is not corrosive or severely irritating which minimizes the respiratory hazard (REACH guidance: 7.2.1.2). Therefore, classification for respiratory irritation is not warranted.