Pine, Pinus pinaster, ext.

Endpoint summary

Description of key information

Acute toxicity, oral: LD50 (rat) > 2000 mg/kg bw (K, Rel.1).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08 January-12 February 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted in compliance with OECD Guideline 420 without any deviation.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

UK GLP Compliance Programme (inspected on 10 July 2012/ signed on 30 November 2012)

Test type:

fixed dose procedure

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Oxon, UK.
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 147-156 g (main study)
- Fasting period before study: Animals were fasted overnight before treatment and food was given approximately 3-4 h after dosing.
- Housing: Animals were housed in groups of up to four in suspended solid-floor polypropylene cages furnished with woodflakes.
- Diet: 2014C Teklad Global Rodent diet (Harlan Laboratories UK Ltd., Oxon, UK), ad libitum
- Water: Mains drinking water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 °C
- Humidity: 30-70 %
- Air changes: 15 changes/h
- Photoperiod (hrs dark / hrs light): 12 h dark / 12 h light

Route of administration:

oral: gavage

Vehicle:

arachis oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 30 mg/mL (for 300 mg/kg bw)
- Justification for choice of vehicle: Arachis oil BP was used because the test item was not dissolved/suspended in distilled water.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw for 300 mg/kg bw dose; 2.31 mL/kg bw for 2000 mg/kg bw dose (specific gravity – 0.869)

DOSAGE PREPARATION (if unusual): Test item at 2000 mg/kg bw dose was used as concentrate. For 300 mg/kg bw dose, test item was freshly prepared as solution in arachis oil BP. The test item was formulated within 2 h of dosing and assumed that the formulation was stable for this duration..

Doses:

- Sighting study: 300 and 2000 mg/kg bw
- Main study: 2000 mg/kg bw

No. of animals per sex per dose:

- Sighting study: 1 female/dose (300 and 2000 mg/kg bw)
- Main study: 4 females (2000 mg/kg bw)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical signs and mortality: Each animal was checked for morbidity and mortality twice daily for 14 days. Clinical observations were made 0.5, 1, 2 and 4 h after dosing and then daily for 14 days.
Body weight: Body weight of each animal was recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: Yes; on completion of the observation period, animals were killed by cervical dislocation and macroscopic examination was performed.

Statistics:

None

Preliminary study:

- No mortality and systemic toxicity were observed in the initial animal treated at 300 and 2000 mg/kg bw dose
- Animal showed expected gains in body weight over the observation period
- No abnormality was noted at necropsy

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

- No mortality was observed

Clinical signs:

other: - Hunched posture and increased salivation were observed in the additional four animals treated at 2000 mg/kg bw and one of these animals showed noisy respiration.

Gross pathology:

- No abnormalities were noted at necropsy.

Other findings:

None

None

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions, the oral LD50 for Pinus Pinaster oil is higher than 2000 mg/kg bw in rats therefore it is not classified according to the Directive 67/548/EEC and the Regulation (EC) N° 1272-2008.

Executive summary:

In an acute oral toxicity study performed according to OECD Guideline 420 and in compliance with GLP, a group of five female Wistar (RccHan™:WIST) rats were given a single oral dose of the test item Pinus Pinaster oil at 2000 mg/kg bw. Animals were then observed for mortality, clinical signs and bodyweights for 14 days and were all sacrificed for macroscopic examination. Before the main test, a sighting test was also conducted on 1 female rat/dose at 300 and 2000 mg/kg bw and animals were observed for 14 days. As no mortality occurred, the test item was administered to four additional animals at the dose level of 2000 mg/kg bw (main test).

No mortality was observed. No sign of systemic toxicity was observed in the initial animal treated at 300 and 2000 mg/kg bw dose level (sighting study). Signs of systemic toxicity observed in animals of the main study treated at a dose level of 2000 mg/kg bw were hunched posture, increased salivation and noisy respiration. All animals showed expected gains in body weight. No abnormalities were noted at necropsy. In this study, the oral LD50 of Pinus Pinaster oil was considered to be higher than 2000 mg/kg bw in female rats.

Under the test conditions, the oral LD50 for Pinus Pinaster oil is higher than 2000 mg/kg bw in rats therefore it is not classified according to the Directive 67/548/EEC and the Regulation (EC) N° 1272-2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

OECD guildeline study in compliance with GLP (Kr. 1)

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute toxicity: via oral route:

A key study was available (Pooles, 2013, Rel. 1). In this acute oral toxicity study conducted according to the OECD guildeline 420 and in compliance with the GLP, 2 female rats were administered a single oral dose of pinus pinaster of 300 and 2000 mg/kg bw, respectively in the sighting study. 4 other females were treated with 2000 mg/kg bw. The animals were observed for mortality for 14 days.

No mortality was observed. Hunched posture and increased salivation were observed in the additional four animals treated at 2000 mg/kg bw and one of these animals showed noisy respiration. Therefore the LD50 of Pinus pinaster in rat was higher than 2000 mg/kg bw.

Justification for selection of acute toxicity – oral endpoint
Only one study available

Justification for classification or non-classification

Harmonized classification:

Pinus pinaster oil has no harmonized classification according to the Regulation (EC) No. 1272/2008.

Self-classification:

Acute toxicity via Oral route:

Based on the available information, Pinus pinaster oil is:

- not classified according to the Regulation (EC) No. 1272/2008 as the LD50 is greater than 2000 mg/kg bw

- not classified according to the Directive 67/548/EEC as the LD50 is greater than 2000 mg/kg bw.

Acute toxicity via Dermal route:This information is not available and is not required for this tonnage band.

Acute toxicity via Inhalation:This information is not available and is not required for this tonnage band.

Specific target organ toxicity: single exposure (Oral):

The classification criteria according to the Annex VI of the Regulation (EC) No. 1272/2008 as specific target organ toxicant (STOT) – single exposure, oral are not met since no reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value (oral) for a Category 1 classification (C ≤ 300 mg/kg bw) and at the guidance value (oral) for a Category 2 classification (2000 mg/kg bw ≥ C > 300 mg/kg bw). Therefore no classification is required.

Specific target organ toxicity: single exposure (Dermal):This information is not available.

Specific target organ toxicity: single exposure (Inhalation):This information is not available.

According to the Regulation (EC) No. 1272/2008, Substances in Category 1for aspiration hasard include but are not limited to certain hydrocarbons, turpentine and pine oil. Based on its composition (> 10% of aspiration toxicants, i.e. pinene), Pinus pinaster oil should be classified for aspiration hazard:

- H304,May be fatal if swallowed and enters airways according to the regulation (EC) No. 1272/2008

- Xn, R65, Harmful: may cause lung damage if swallowed according to the Directive 67/548/EEC.

Source: ECHA disseminated dossiers

-Pinene alpha:

http://apps.echa.europa.eu/registered/data/dossiers/DISS-9d952924-c8ed-4614-e044-00144f67d249/DISS-9d952924-c8ed-4614-e044-00144f67d249_DISS-9d952924-c8ed-4614-e044-00144f67d249.html