Amines, rosin

Endpoint summary

Description of key information

Acute oral toxicity

In one key acute oral toxicity study, Rosin amine 90 was administered in a single dose to female rats at one or more defined dosages.

The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. At 2000 mg/kg, one animal was sacrificed for humane reasons on Day 8 (more than 20% body weight loss) and one animal was found dead on Day 10. At 300 mg/kg, no mortality occurred.

The oral LD50 value of Rosin Amine 90 in Wistar rats was established to be within the range of 300- 2000 mg/kg body weight.

the LD50 cut-off value was considered to be 2000 mg/kg body weight.

Acute dermal toxicity

In one key acute dermal toxicity study, Rosin amine 90 was administered to 5 female and 5 male Wistar rats via a single dermal application at 2000 mg/kg body weight for 24 hours.

Animals were subjected to daily observations, body weight was determined weekly and macroscopic examinations were performed on the day of death or after terminal sacrifice (Day 15).

2 animals were sacrificed for humane reasons, but no mortality due to systemic toxicity of Rosin Amine 90 was observed. Body weight gain was in the expected range and no abnormalities were seen in the macroscopic post-mortem examination of the animals.

The dermal LD50 was determined to be more than 2000 mg/kg bw.

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:

From 15 September 2015 to 15 October 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conclusive, performed to a valid guideline (OECD TG 423 adopted 17 December 2001) and was conducted under GLP conditions. No deviations from the test methods were noted.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Remarks:

Statement of GLP compliance signed by study director 18 December 2015

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Approximately 9-12 weeks
- Weight at study initiation: 165-203 g
- Fasting period before study: Overnight prior to dosing and until 3-4 hours after administration of the test substances
- Housing: Group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) provided ad libitum outwith fasting period
- Water (e.g. ad libitum): Tap water provided ad libitum
- Acclimation period: At least 5 days before start of treatment under laboratory conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24
- Humidity (%): 40 to 70
- Air changes (per hr): At least 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2.0202 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Limit test as specified by testing guideline

Doses:

2000 mg/kg (2.0202 mL/kg) body weight.
300 mg/kg (0.303 mL/kg) body weight.

No. of animals per sex per dose:

3 (stepwise treatment)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/viability observations made twice daily, body weights recorded Days 1, 8 and 15 and at death
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs observations made at periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15

Statistics:

No statistical analysis was performed.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

>= 300 - <= 2 000 mg/kg bw

Based on:

test mat.

Mortality:

At 2000 mg/kg, one animal was sacrificed for humane reasons on Day 8 (more than 20% body weight loss) and one animal was found dead on Day 10.
At 300 mg/kg, no mortality occurred.

Clinical signs:

other: At 2000 mg/kg, hunched posture, piloerection, hypotonia, uncoordinated movements, lean appearance and salivation were noted for the animals between Days 1 and 13. Additionally, the animals that died during the study showed lethargy, chromodacryorrhoea (sn

Gross pathology:

At 2000 mg/kg, abnormalities of the spleen (discolouration, pale), thymus (reduced in size), esophagus (contents: hemorrhagic/clotted blood), stomach (focus/foci, several, dark red) were found in the animals that died during the study. Additionally, it was noted that the animal sacrificed for humane reasons was emaciated.
Macroscopic post mortem examination of the surviving animals treated at 2000 mg/kg and the animals treated at 300 mg/kg, did not reveal any abnormalities.

Please see the 'Attached background material' section below for all tables of results.

Interpretation of results:

Category 4 based on GHS criteria

Remarks:

Iinformation Criteria used for interpretation of results: EU

Conclusions:

The oral LD50 value of Rosin Amine 90 in Wistar rats was established to be within the range of 300-2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 2000 mg/kg body weight.

Based on these results, according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments), Rosin Amine 90 should be classified as Category 4 and should be labelled as H302: Harmful if swallowed.

Executive summary:

The study was carried out based on the guidelines described in: OECD No.423 (2001) "Acute Oral Toxicity, Acute Toxic Class Method"

Commission Regulation (EC) No 440/2008, B1 tris: "Acute Oral Toxicity, Acute Toxic Class Method" EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity"

JMAFF Guidelines (2000), including the most recent revisions.

Initially, Rosin Amine 90 was administered by oral gavage to three female Wistar rats at 2000 mg/kg body weight. In a stepwise procedure three additional groups of three females were dosed at 2000, 300 and 300 mg/kg body weight. The animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

At 2000 mg/kg, one animal was sacrificed for humane reasons on Day 8 (more than 20% body weight loss) and one animal was found dead on Day 10. At 300 mg/kg, no mortality occurred.

At 2000 mg/kg, hunched posture, piloerection, hypotonia, uncoordinated movements, lean appearance and salivation were noted for the animals between Days 1 and 13. Additionally, the animals that died during the study showed lethargy, chromodacryorrhoea (snout) and red staining of the mouth and nose. At 300 mg/kg, hunched posture was noted for all animals on Day 1.

At 2000 mg/kg, all animals showed body weight loss or reduced body weight gain between Days 1 and 8. The surviving animals showed normal body weight gain on Day 15 compared to Day 1, except for one animal which showed reduced body weight gain. At 300 mg/kg, the body weight gain shown by these animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.

At 2000 mg/kg, abnormalities of the spleen (discolouration, pale), thymus (reduced in size), esophagus (contents: hemorrhagic/clotted blood), stomach (focus/foci, several, dark red) were found in the animals found dead. Additionally, it was noted that the animal sacrificed for humane reasons was emaciated. Macroscopic post mortem examination of the surviving animals treated at 2000 mg/kg and the animals treated at 300 mg/kg, did not reveal any abnormalities.

The oral LD50 value of Rosin Amine 90 in Wistar rats was established to be within the range of 300-2000 mg/kg body weight. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 2000 mg/kg body weight.

Based on these results and according to the UN Globally Harmonized System of Classification and Labelling of Chemicals (GHS) (2011) (including all amendments), Rosin Amine 90 should be classified as: harmful if swallowed (Category 4) for acute toxicity by the oral route; - according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments), Rosin Amine 90 should be classified as Category 4 and should be labeled as H302: Harmful if swallowed.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 October-29 October 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material: 542700
- Expiration date of the lot/batch: 31 March 2016
- Purity: not specified

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: Stable under the storage conditions until the expiry date

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test substance was dosed undiluted as delivered by the sponsor.
- No correction was made for the purity/composition of the test substance.
- An adjustment was made for the specific gravity of the test susbtance.

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany.
- Age at study initiation: Approx. 10 weeks old
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean.
- Fasting period before study: None
- Housing: Individually housed in labeled Makrolon cages (MIII type, height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) provided ad libitum
- Water (e.g. ad libitum): Tap water provided ad libitum
- Acclimation period: At least 5 days before the start of the treatment under laboratory conditions.During the acclimatization period the animals were group housed in Makrolon cages (MIV type, height 18 cm).

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 24
- Humidity (%): 40 to 70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: approximately 5 x 7 cm. On the day before exposure this area on the back of each animal was clipped.
- % coverage: 10 of total body surface (approximately 25 cm^2 for males and 18 cm^2 females)
- Type of wrap if used: surgical gauze patch (Surgy 1D) successively covered with aluminium foil and Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Skin cleaned of residual test substance using tap water.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution): 0.99 g/mL
- Constant volume or concentration used: yes
- For solids, paste formed: not applicable

Duration of exposure:

24 hours after which dressings were removed and the skin was cleaned of residual substance using tap water.

Doses:

2000 mg/kg (2.02 mL/kg) body weight

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed:Yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: body weights observations were performed at Days 1 (pre-administration), 8 and 15 and at death (if sacrificed after Day 1)., clinical signs were monitored at periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

In consultation with a veterinarian, one male and one female animal were sacrificed for humane
reasons on Day 6, due to severe irritation of the treated skin area. No further mortality occurred. For full table of results see attached document.

Clinical signs:

other: Flat posture, hunched posture and/or chromodacryorrhoea were noted for all animals between Days 1 and 3. Additionally, one animal showed hunched posture up to Day 8. General erythema, erythema maculate, scales, scabs, thickened areas, fissures, wounds a

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals. For full table of results see attached document.

Please see the 'Attached background material' section for the tables of results.

Interpretation of results:

not classified

Remarks:

Information Criteria used for interpretation of results: EU

Conclusions:

Based on the results of the study, since there were no mortalities observed as a result of systemic toxicity caused by the test substance, the dermal LD50 value of Rosin Amine 90 in male and female Wistar rats was established to be more than 2000 mg/kg bw. According to GHS and EC Regulation No. 1272/2008, the test substance does not have to be classified and has no obligatory labelling requirements for acute dermal toxicity.

Executive summary:

This study was conducted to assess the systemic toxicity of the test substance, Rosin Amine 90, in a process carried out in accordance with the following guidelines; OECD No.402 (1987) "Acute Dermal Toxicity", Commission Regulation (EC) No 440/2008, B3: "Acute Toxicity (Dermal)", EPA, OPPTS 870.1200 (1998), "Acute Dermal Toxicity" and JMAFF Guidelines (2000), including the most recent revisions. The study was thought to provide a rational basis for risk assessment in humans, since the dermal route is a possible route of exposure during the manufacture, handling or use of the test substance.

Rosin Amine 90 was administered to five Wistar rats of each sex by a single dermal application at 2000 mg/kg body weight for 24 hours. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed on the day of death or after terminal sacrifice (Day 15).

The main clinical signs observed for all animals from Day 1 -3 were flat posture, hunched posture and/or chromodacryorrhoea. Additionally, one animal showed hunched posture up to Day 8. During he observation period, general erythema, erythema maculate, scales, scabs, thickened areas, fissures, wounds and/or brown staining were noted in the treated skin-area of the animals. In addition, one animal had scabs on the left flank during the observation period. Two animals (one male and one female) were sacrificed for human reasons on Day 6, but there were no mortalities as a result of systemic toxicity caused by the test substance. The changes noted in body weight gain in males and females were within the range expected for rats used in this type of study and were therefore considered not indicative of toxicity. There were no abnormalities found during the macroscopic post-mortem examination of the animals.

Since no mortality occurred due to systemic toxicity by the test substance, the dermal LD50 value of Rosin Amine 90 in male and female Wistar rats was established to be more than 2000 mg/kg body weight. Based on these results, Rosin Amine 90 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2011) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Justification for classification or non-classification

Acute oral toxicity

According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 2000 mg/kg body weight.

According to the Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments), Rosin Amine 90 should be classified as Category 4 and should be labeled as H302: Harmful if swallowed

Acute dermal toxicity

Rosin Amine 90 does not have to be classified and has no obligatory labelling requirement for acute dermal toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United

Nations (2011) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures (including all amendments).