2-hydroxy-3-(prop-2-enoyloxy)propyl 2-methyl-2-propylhexanoate

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From June 09, 2015 to June 26, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

yes

Remarks:

typing error

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Specific details on test material used for the study:

Batch no.: JBGJ0045R
Appearance: clear yellowish liquid

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Source: Velaz Prague, Czech Republic
Age: At least 8-12 weeks; female animals were non-pregnant and nulliparous
Acclimatization: at least 5 days
Temperature: 22 ± 2°C, relative humidity : 55 ± 10% and light regimen: 12-hour light /12-hour dark cycle
Diet: laboratory food Altromin (Altromin Spezialfutter GmbH, Germany, ad libitum
Water: tap water for human consumption, ad libitum

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on oral exposure:

The required amount of the test substance (according to the body weight and dose) was mixed with vehicle (carboxymethyl cellulose - 1% solution) shortly before administration. The test substance was administered in a single dose by gavage using a metal stomach tube. Animals were fasted prior to dosing (food but not water were withheld over-night). Following the period of fasting, the animals were weighted and the test substance administered. After the test isubstance had been administered, food was withheld for further 3-4 hours.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 males and 6 females

Control animals:

no

Details on study design:

- Animals were observed individually immediately after the administration of the test substance and then ½, 1, 2, and 4 hours later. Then each animal was inspected daily for the next 14 days. Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Individual weights of animals were determined shortly before the test substance was administered and at weekly thereafter. Weight differences after first and second week after application were calculated and recorded.
- All test animals were subjected to gross necropsy. Full, detailed gross necropsy included careful examination of external surface of the body, all orifices, and cranial, thoracic and abdominal cavities and their contents. All gross pathological changes were recorded for each animal.

Preliminary study:

The starting dose was selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg bw. Available information indicated that the test substance is likely to be nontoxic considering to acute toxicity. A limit dose of 2000 mg/kg bw was used as starting dose. Group of 3 rat’s females were dosed. Test substance-related mortality was not produced during 24 hours; group of 3 rat’s females and group of 3 rat’s males were tested at the same dose.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

>= 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

not determinable due to absence of adverse toxic effects

Mortality:

All 6/6 females and 3/3 males survived the limit dose 2000 mg/kg bw. No further dosing was necessary.

Clinical signs:

other: No mortality was observed during the study. Animals lived through observation period without important visible signs of intoxication. Neither change of health nor negative reactions were registered.

Gross pathology:

All animals (6 females and 3 males) were necropsied. During necropsy, no macroscopically changes were noticed.

Based on the results, the test substance was classified in category 5/Unclassified with the cut off LD50 ≥ 5000 mg/kg bw, after single oral administration to Wistar rats.

Interpretation of results:

other: CLP criteria not met

Remarks:

does not need to be classified

Conclusions:

Under the study conditions, the rat LD50 was determined to be ≥ 2000 mg/kg bw.

Executive summary:

A study was conducted to determine the acute oral toxicity of the test substance according to OECD Guideline 423, in compliance with GLP. The test substance was administered by gavage to 6 female and 3 male rats at a limit dose of 2000 mg/kg bw. All 6/6 females and 3/3 males survived the limit dose of 2000 mg/kg bw. No further dosing was necessary. No body weight losses were recorded one and two weeks after administration of the test substance. No important symptoms were observed at the dosage of 2000 mg/kg bw during first 4 h neither in females nor in males or in 14 day observation period. During necropsy, no macroscopically changes were noticed. Under the study conditions, the rat LD50 was determined to be ≥ 2000 mg/kg bw (Hozova, 2015).

Cross-reference 2

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From June 17, 2015 to July 21, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

yes

Remarks:

but did not affect the validity of the study

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

yes

Remarks:

but did not affect the validity of the study

GLP compliance:

yes (incl. QA statement)

Type of study:

mouse local lymph node assay (LLNA)

Justification for non-LLNA method:

-

Specific details on test material used for the study:

Batch no.: JBGJ0045R
Purity: 100% (UVCB)
Appearance: clear yellowish liquid

Species:

mouse

Strain:

CBA/Ca

Remarks:

CBA/CaOlaHsd

Sex:

female

Details on test animals and environmental conditions:

Source: Harlan Laboratories B.V. Postbus 6174 5960 AD Horst / The Netherlands
Acclimatization: at least 5 days
Body weight: ca. 20g
Temperature: ca. 22±2°C, relative humidity : ca. 45-65% and light regimen: 12-hour light /12-hour dark cycle
Diet: 2018C Teklad Global 18% protein rodent diet, ad libitum
Water: tap water, ad libitum

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

0, 0.5, 1 and 2.5% (w/w)
(Pre-tests: 50, 25, 10 and 5%)

No. of animals per dose:

4

Details on study design:

Three groups each of four female mice were treated with different concentrations (25 µL of 0, 0.5, 1, and 2.5% (w/w) in acetone/olive oil (4+1, v/v)) of the test substance by topical application at the dorsum of each ear once daily each on three consecutive days. The highest concentration tested was the highest concentration that could be achieved whilst avoiding systemic toxicity and excessive local skin irritation as confirmed by three pre-experiments. A control group of four mice was treated with the vehicle only. Five days after the first topical application, the mice were intravenously injected into a tail vein with radio-labelled thymidine (3H-methyl thymidine; 3HTdR). Approximately five hours after intravenous injection, the mice were sacrificed and the draining auricular lymph nodes excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes, which were subsequently washed and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was then determined by the incorporation of 3H-methyl thymidine measured in a β-scintillation counter.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Mean values and standard deviations

Positive control results:

alpha- hexylcinnamaldehyde 25% (w/w) in acetone/olive oil (4+1, v/v): S.I. value of 9.5 and was therefore regarded as a sensitiser in the LLNA test, since the exposure to one or more test concentrations resulted in a 3-fold or greater increase in incorporation of 3HTdR compared with concurrent controls.

Key result

Parameter:

SI

Value:

1

Test group / Remarks:

0.0% (w/w) in acetone/olive oil (4+1, v/v)

Remarks on result:

other: negative control

Key result

Parameter:

SI

Value:

0.84

Test group / Remarks:

0.5% (w/w) in acetone/olive oil (4+1, v/v)

Remarks on result:

other: at 0.5% conc; not a skin sensitiser

Key result

Parameter:

SI

Value:

0.92

Test group / Remarks:

1.0% (w/w) in acetone/olive oil (4+1, v/v)

Remarks on result:

other: at 1% conc; not a skin sensitiser

Key result

Parameter:

SI

Value:

1.22

Test group / Remarks:

2.5% (w/w) in acetone/olive oil (4+1, v/v)

Remarks on result:

other: at 2.5% conc; not a skin sensitiser

Cellular proliferation data / Observations:

The animals did not show any signs of systemic toxicity during the course of the study and no cases of mortality were observed. The body weight of the animals, recorded prior to the first application and prior to treatment with 3HTdR, was within the range commonly recorded for animals of this strain and age. A very slight erythema (score: 1) was observed in the animals of the high dose group on test day 3. In this study, Stimulation Indices (S.I.) of 0.84, 0.92, and 1.22 were determined with the test substance at concentrations of 0.5, 1, and 2.5% (w/w) in acetone/olive oil (4+1, v/v), respectively. The test substance was not a skin sensitiser under the test conditions of this study.
(The estimated concentration of test substance required to produce a S.I. of 3 is referred to as the EC3 value. In this study, the EC3 value could not be calculated, since none of the tested concentrations induced a S.I. greater than the threshold value of 3.)

Interpretation of results:

other: CLP criteria not met

Remarks:

does not need to be classified

Conclusions:

Under the study conditions, the test substance was not a skin sensitiser (LLNA).

Executive summary:

A study was conducted to determine the skin sensitisation potential of the test substance according to OECD Guideline 429 and EU Method B.42, in compliance with GLP. Groups of 4 female CBA/Ca mice each were treated with 0, 0.5, 1, and 2.5% (w/w) preparations of the test substance in acetone/olive oil (4+1, v/v)) or with the vehicle alone. The highest concentration tested was the highest concentration that could be achieved whilst avoiding systemic toxicity and excessive local skin irritation as confirmed by three pre-experiments. The study used 3 test groups and 1 control groups (vehicle). 25 µL of the respective test substance preparation or of the vehicle were applied to the dorsum of both ears of each animal once daily for three consecutive days. Five days after the first topical application, the mice were intravenously injected into a tail vein with radio-labelled thymidine (3H-methyl thymidine; 3HTdR). Approximately five hours after intravenous injection, the mice were sacrificed and the draining auricular lymph nodes excised and pooled per group. Single cell suspensions of lymph node cells were prepared from pooled lymph nodes, which were subsequently washed and incubated with trichloroacetic acid overnight. The proliferative capacity of pooled lymph node cells was then determined by the incorporation of 3H-methyl thymidine measured in a β-scintillation counter. Mortality / viability was reported at least once daily from experimental start to necropsy. Body weights was measured prior to the first application and prior to treatment with 3HTdR. Clinical signs (local / systemic) were recorded at least once daily. The animals did not show any signs of systemic toxicity during the course of the study and no cases of mortality were observed. The body weight of the animals, recorded prior to the first application and prior to treatment with 3HTdR, was within the range commonly recorded for animals of this strain and age. A very slight erythema (score: 1) was observed in the animals of the high dose group on test day 3. In this study, Stimulation Indices (S.I.) of 1.00, 0.84, 0.92, and 1.22 were determined with the test substance at concentrations of 0, 0.5, 1, and 2.5% (w/w) in acetone/olive oil (4+1, v/v), respectively. The EC3 value could not be calculated, since none of the tested concentrations induced a S.I. greater than the threshold value of 3. Under the study conditions, the test substance was not a skin sensitiser (LLNA) (Roth, 2015).

Cross-reference 3

Reason / purpose for cross-reference:

data waiving: supporting information

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 14, 2001 to May 18, 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Batch no.: F016143 05
Appearance: Light-yellow, clear viscous liquid

Species:

rabbit

Strain:

New Zealand White

Remarks:

females

Details on test animals or test system and environmental conditions:

Source: Charles River Wiga, D-97633
Acclimatization: 5 days
Body weight: 2.0-2.4 kg
Temperature: ca. 20.3°C, relative humidity : ca. 58.8% and light regimen: 12-hour light /12-hour dark cycle
Diet: laboratory food Altromin 2123, ad libitum
Water: tap water for human consumption, ad libitum

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

1, 24, 48 and 72 h after patch removal

Number of animals:

3

Details on study design:

- Hair was clipped on the dorsal areas of the trunks one day before the application of the test substance. The test sites were median on the dorsal thoracal region. 0.5 mL of the test substance was applied via a cellulose patch to a site of about 2.5 cm x 2.5 cm of the intact skin of each of 3 rabbits and covered by a semi-occlusive dressing.
- The skins were examinated for local alterations one day before the administration of the test substance (after clipping of the hair) and immediately before the administration. The treated areas and the surrounding untreated skin (control area) of the animals were examinated (using a cold light source KL 1500 electronic) for erythema/eschar and oedema as well as for other local alterations approximately 1, 24, 48 and 72h after patch removal. The animals were analysed once daily.
- Primary irritation index was calculated according to EPA: all scores for erythema and oedema for the 3 animals and the 4 reading times were added and divided by 12.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

>= 0 - <= 0

Max. score:

0

Reversibility:

other: no effects were seen

Remarks:

no effects were seen

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

>= 0 - <= 0

Max. score:

0

Reversibility:

other: no effects were seen

Remarks:

no effects were seen

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

>= 0 - <= 0

Max. score:

0

Reversibility:

other: no effects were seen

Remarks:

no effects were seen

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

>= 0 - <= 0

Max. score:

0

Reversibility:

other: no effects were seen

Remarks:

no effects were seen

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

>= 0 - <= 0

Max. score:

0

Reversibility:

other: no effects were seen

Remarks:

no effects were seen

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

>= 0 - <= 0

Max. score:

0

Reversibility:

other: no effects were seen

Remarks:

no effects were seen

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

All exposed skin sites were normal at each examination term; neither erythema/eschar nor oedema were observed. The primary irritation index was 0.0. The test substance was considered not irritating to the skin.

Other effects:

No general toxic effects of the test substance were observed.

Interpretation of results:

GHS criteria not met

Remarks:

does not need to be classified

Conclusions:

Under the study conditions, the test substance was considered not irritating to rabbit skin.

Executive summary:

An in vivo study was conducted to determine the skin irritation potential of the test substance according to OECD Guideline 404, EU Method B.4 and EPA-Guideline OPPTS 870.2500, in compliance with GLP. Hair of 3 female rabbits (New-Zealand White) was clipped on the dorsal areas of the trunks one day before the application of the test substance. 0.5 mL of the test substance was applied via a cellulose patch to a site of about 2.5 cm x 2.5 cm of the intact skin of each of 3 rabbits and covered by a semi-occlusive dressing. The skins were examined for local alterations one day before the administration of the test substance (after clipping of the hair) and immediately before the administration. The treated areas and the surrounding untreated skin (control area) of the animals were examined for erythema/eschar and oedema as well as for other local alterations approximately 1, 24, 48 and 72h after patch removal. The animals were analysed once daily. Primary irritation index was calculated according to EPA: all scores for erythema and oedema for the 3 animals and the 4 reading times were added and divided by 12. No general toxic effects of the test substance were observed. All exposed skin sites were normal at each examination term; neither erythema/eschar nor oedema was observed. The primary irritation index was 0.0. Under the study conditions, the test substance was considered not irritating to the skin (Wolf, 2001).