Amines, N-(C16-18 and C18-unsatd. alkyl)trimethylenedi-, dioleates

Endpoint summary

Description of key information

An acute oral toxicity study with Tallow-diamine dioleate resulted to an LD50 > 5000 mg/kg, and one acute dermal study with Oleyl-diamine dioleate resulting to an LD50 > 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:

Aug 08, 1984 - Aug 22, 1984

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Reasonably wel reported and performed under GLP.

Qualifier:

according to guideline

Guideline:

EPA OTS 798.1175 (Acute Oral Toxicity)

Deviations:

not specified

GLP compliance:

yes

Remarks:

EPA GLP regulations of 29 Dec 1983

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Ace Animals
- Age at study initiation: male and non-pregnant and nulliparous female Wistar Ablino rats, approximately 8 weeks old
- Weight at study initiation: between 200-300 grams,
- Fasting period before study: 16-20 hours prior to dosing.
- Housing: housed 5/sex/cage in suspended wire mesh cages. Bedding was placed beneath the cages.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not stated
- Humidity (%): not stated
(temperature and humidity controlled)
- Air changes (per hr): not indicated
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: Aug 08, 1984 To: Aug 22, 1984

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

syringe and dosing needle at a dose level of 5.0 g/kg - no vehicle.
Sample preparation: 20 g of test article was heated gently to a melting point of 44°C

Doses:

Single dose 5.0 g/kg

No. of animals per sex per dose:

5 male + 5 female

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 1, 2 and 4 hours post dose and once each morning and afternoon thereafter
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight. Abnormal tissues were preserved in 10% buffered formalin for possible future microscpic examination.

Statistics:

Not required for single dose: The LD 50 was considered to be greater than 5.0 g/kg if there was no compound related mortal ity at the 5.0 g/kg level.

Preliminary study:

No

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: 3/10 animals died at 5000 mg

Mortality:

3 females on observation days 7, 8 and 12 respectively.

Clinical signs:

other: Yes - see table under other information

Gross pathology:

Yes - see table under other information

Clinical signs (duration average number of days)

	Males	Females
		Death	Sarcrifice
Number treated	5	3	2
Diarrhea	1(1)	3(5)	2(4)
Piloerection	1(1)	3(3)
Ptosis	1(1)	2(2)
Emaciation	1(1)	1(1)
Lethargy		2(1)
Anogenital area stained brown	2(6)	3(7)	2(11)
Nose/mouth stained brown		3(6)	2(8)
Body surfaces soiled	1(2)

 

Necropsy observations

	Males	Females
		Death	Sarcrifice
Number necropsied	5	3	2
Normal			2
Cannibalized		2
Anogenital area stained brown		1
Lungs congested		1
Stomach areas red		1
Stomach distended with fluid		1	2(11)
Intestines red		1	2(8)
Intestines distended with fluid		1

 

Body weights:

Animal-sex	Dose vol. cc	Day 0 g	Day 7 g	Day 14 g
1-M	1.2	216	246	330
2-M	1.3	219	242	328
3-M	1.3	218	184	290
4-M	1.4	247	300	384
5-M	1.3	224	242	332
Mean		224.8	242.8	332.8
S.D.		12.8	41.1	33.5
6-F	1.2	209	188	234
7-F	1.5	253	270	322
8-F	1.3	218	202	dead d.12 (144 g)
9-F	1.3	218	164	dead d.8*
10-F	1.2	211	dead d.7*
Mean		221.8	206	278
S.D.		17.9	45.5	62.2

*Termina1 body weight was not recorded since the animals were cannibalized.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Acute oral toxicity was determined in a limit test in 5 males and 5 females at 5 g/kg bw, undilted. LD50 > 5.0 g/kg bw for males and females combined.

Executive summary:

Method: Five healthy male and five healthy non-pregnant and nulliparous female albino rats were dosed orally with Duomeen TDO, Lot #1400407 at 5.0 g/kg of body weight.

The rats were observed 1, 2 and 4 hours post dose and twice daily for 14 days for mortality and toxicity. Body weights were recorded pretest, weekly, at death and at termination in the survivors. All animals were examined for gross pathology. Abnormal tissues were preserved in 10% buffered formalin for possible future microscopic examination.

Although there was compound related mortality, a multi-dose LD50 was not performed as per client request.

 

Results: Seven of ten animals survived the 5.0 g/kg oral dose.

Three females died between Day 7 and Day 12 with pre-death physical signs of diarrhea, lethargy, ptosis, piloerection, emaciation, soiling of body surfaces and brown staining of the nose/mouth and anogenital areas. Two animals which died were not necropsied since they had been cannibalized. The other death had abnormalities of the lungs and gastrointestinal tract, as well as brown staining of the anogenital area. Physical signs of chromorhinorrhea, diarrhea, piloerection, emaciation, ptosis, soiling of body surfaces, and brown staining of the nose/mouth and anogenital areas were noted in survivors and were more prevalent in females.

Body weight increases were normal in 5/7 survivors. One male and one female lost weight during the first week of the study.

Necropsy results were normal in survivors.

Conclusion: The LD 50 is greater than 5.0 g/kg in males, less than 5.0 g/kg in females and greater than 5.0 g/kg for combined sexes.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

5 000 mg/kg bw

Quality of whole database:

Valid GLP study, sufficiently adequate reported.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

01 Nov 2011 to 02 Mar 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed according to current OECD/EC guidelines and according to GLP principles.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

study was conducted according to the acute toxic class method designed for assessment of acute oral toxicity (OECD No. 423): 2x3 females were used instead of 5 animals.

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.

Principles of method if other than guideline:

For treated animals, an external macroscopic examination was conducted next to an internal macroscopic examination.
The study integrity was not adversely affected by the deviation.

GLP compliance:

yes (incl. QA statement)

Remarks:

wih exception of the additional histopathology of the skin.

Test type:

other: 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: approx. 12 weeks old
- Weight at study initiation: Body weight variation did not exceed +/- 20% of the sex mean.
- Housing: Individually in Makrolon cages
- Diet: ad libitum, pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: ad libitum, tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3-21.9
- Humidity (%): 40-59
- Air changes (per hr): 15
- 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 25 cm² for males and 18 cm² for females
- % coverage: appr. 10% of the total body surface
- Type of wrap if used: a surgical gauze patch (Surgy 1D), successively covered with aluminum foil and Coban elastic bandage

REMOVAL OF TEST SUBSTANCE
- Washing: with tap water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount applied: 2.26 mL/kg bw (= 2000 mg/kg bw)

Duration of exposure:

24 hours

Doses:

2000 mg/kg

No. of animals per sex per dose:

6 females (2x3 f were dosed in a stepwise manner)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: mortality was checked twice daily, clinical signs once daily.
- Frequency of weighing: day 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: histopathological examination of treated skin with macroscopic abnormalities.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Hunched posture, lethargy, chromodacryorrhoea, piloerection and/or ptosis were shown by all animals between Days 1 and 10. The treated skin-area of all animals showed general or focal erythema, scales, scabs, a thickened area and/or fissures during the ob

Gross pathology:

Three animals for which an external macroscopic examination was conducted showed scab formation on the dorso-lumbar region of the skin. No other abnormalities were found at macroscopic post mortem examination of the animals.

Other findings:

Treatment-related microscopic skin findings were as follows:
− Ulceration (characterized by full thickness loss of the epithelium through the basement membrane) was present in 5/6 2000 mg/kg treated rats at slight (3/6) or marked (2/6) degree.
− A fibrinous exudate (a focally fibrillar eosinophilic mass of exudative proteinaceous material and cellular debris) was present on the skin surface of all rats at a minimal (1/6), slight (1/6) or moderate (4/6 degree). The most severe exudate was seen in ulcerated areas.
− Acanthosis, thickening/hyperplasia of the epidermis (increase in the number of epithelial cell layers which may extend into the dermis), was present in the non-ulcerated /less severely affected areas of all rats at a slight (5/6) or moderate (1/6) degree. This was likely a reactive response to the test item.
− All rats had lymphogranulocytic inflammation (characterized by a mixture of predominately lymphocytic and some granulocytic inflammatory cells) present in the dermis at minimal (3/6), slight (1/6) or moderate (2/6) degree. Inflammation extended focally into the underlying panniculus muscle which separates the dermis from the underlying subcutaneous tissue.

Interpretation of results:

not classified

Remarks:

Migrated information for dermal acute toxicity Criteria used for interpretation of results: EU

Conclusions:

The dermal toxicity of oleyl diamine was tested according to current OECD/EC guidelines and under GLP principles. The LD50 was determined to be > 2000mg/kg.

Based on these results, Oleyl diamine, dioleate 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 (2007),
- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Executive summary:

Assessment of acute dermal toxicity with Oleyl diamine, dioleate in the rat.

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, B1tris: "Acute Oral Toxicity, Acute Toxic Class Method"

-      EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity"

-      JMAFF guidelines (2011) including the most recent partial revisions.

-      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"

Oleyl diamine, dioleate was administered to two subsequent groups of three female Wistar rats 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 after terminal sacrifice (Day 15). Histopathology was conducted on the treated skin area from all rats.

Results: Hunched posture, lethargy, chromodacryorrhoea, piloerection and/or ptosis were shown by all animals between Days 1 and 10. The treated skin-area of all animals showed general or focal erythema, scales, scabs, a thickened area and/or fissures during the observation period.

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.

Three animals for which an external macroscopic examination was conducted showed scab formation on the dorso-lumbar region of the skin. No other abnormalities were found at macroscopic post mortem examination of the animals.

Treatment-related histopathological findings in the treated skin area consisted of ulceration (complete loss of the epidermis) in most rats with a superficial fibrinous exudate (this correlated with the scab formation seen macroscopically) in all rats. There was hyperplasia of the epidermis (acanthosis) in the non-ulcerated areas of all rats. Furthermore, dermal lymphogranulocytic inflammation was present in all rats.

 

The dermal LD50 value of Oleyl diamine, dioleate in Wistar rats was established to exceed 2000 mg/kg body weight.

 

From the histopathological results it is concluded that Oleyl diamine, dioleate is an irritant and has corrosive potential when applied with an occlusive dressing to the rat skin at 2000 mg/kg for 24 hours.

Based on these results, Oleyl diamine, dioleate does not have to be classified and has no obligatory labeling requirement for acute dermal toxicity according to the:

- Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2007),

- Regulation (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

discriminating dose

Value:

2 000 mg/kg bw

Quality of whole database:

Recent, high quality study; Based on read across from very similar substance Oleyl-diamine dioleate

Additional information

Acute oral toxicity:

The acute oral toxicity of Tallow-diamine dioleate was evaluated in a study performed in compliance to GLP. In this study, five male and five female rats were dosed orally with Tallow-diamine dioleate (pure, without vehicle) at 5.0 g/kg of body weight. The rats were observed 1, 2 and 4 hours post dose and twice daily for 14 days for mortality and toxicity. Body weights were recorded pretest, weekly, at death and at termination in the survivors. All animals were examined for gross pathology and abnormal tissues were preserved for possible future microscopic examination.

Seven of ten animals survived the 5.0 g/kg oral dose. Three females died between Day 7 and Day 12 with pre-death physical signs of diarrhea, lethargy, ptosis, piloerection, emaciation, soiling of body surfaces and brown staining of the nose/mouth and anogenital areas. Two animals which died were not necropsied since they had been cannibalized. The other death had abnormalities of the lungs and gastrointestinal tract, as well as brown staining of the anogenital area. Physical signs of chromorhinorrhea, diarrhea, piloerection, emaciation, ptosis, soiling of body surfaces, and brown staining of the nose/mouth and anogenital areas were noted in survivors and were more prevalent in females.

Body weight increases were normal in 5/7 survivors. One male and one female lost weight during the first week of the study. Necropsy results were normal in survivors.

Conclusion: The LD 50 is greater than 5.0 g/kg in males, less than 5.0 g/kg in females and greater than 5.0 g/kg for combined sexes.

 

 

Acute dermal toxicity:

Testing for acute dermal toxicity was done using Oleyl-diamine dioleate rather than Tallow-diamine dioleate. Cross-reading between the two substances is acceptable on the basis of similarities of structure with same functional groups, and properties leading to common biological activity, and possible common metabolic degradation. Contrary to oleyl do tallow-alkyl chains also contain some C16-chains next to the C18-alkyl chains. If anything, the higher level of unsaturation in oleyl-alkyl chains can be considered a worst case representation compared to tallow. Due to this very great similarity between these two substances, are results obtained from Oleyl-diamine dioleate fully applicable for the evaluation of Tallow-diamine dioleate as well.

 

The acute dermal toxicity of Oleyl-diamine dioleate was evaluated following a single dermal application at 2000 mg/kg body weight for 24 hours in two subsequent groups of three female Wistar rats. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). Histopathology was conducted on the treated skin area from all rats.

All animals showed hunched posture, lethargy, chromodacryorrhoea, piloerection and/or ptosis between Days 1 and 10. The treated skin-area of all animals showed general or focal erythema, scales, scabs, a thickened area and/or fissures during the observation period.

Treatment-related histopathological findings in the treated skin area consisted of ulceration (complete loss of the epidermis) in most rats with superficial fibrinous exudates, correlating with the scab formation seen macroscopically, in all rats.

The dermal LD50 value of Oleyl-diamine dioleate in Wistar rats was established to exceed 2000 mg/kg body weight.

 

Acute inhalation toxicity:

No data available. Due to irritating properties respiratory irritation would be in principle possible. Due to the low vapour pressure (< 0.0015 Pa at 20°C based on C12/14-diamine rather than Tallow-diamine dioleate salt) inhalation of vapours leading to irritation of airways will not occur. The indicated use pattern also does not give rise to a high concern for specific exposures via inhalation of these oleyl-diamine dioleate salts.

 

Tallow-diamine dioleate is a solid at 20°C, and its melting point is 38 °C. No classification for aspiration hazard is therefore warranted.

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

Justification for selection of acute toxicity – inhalation endpoint
No inhalation studies are needed when vp < 0.1 Pa at 20°C

Justification for selection of acute toxicity – dermal endpoint
Only study available.

Justification for classification or non-classification

The evaluation of available data leads to the conclusion that for Tallow-diamine dioleate, both the acute oral and acute dermal LD50 are determined to be above 2000 mg/kg bw, Consequently, Tallow-diamine dioleate does not need to be classified for acute oral or dermal toxicity for EU-CLP.

For inhalation route no data is available. However, in view of low likelihood of exposures related to very low vapour pressure no further testing is needed.

Tallow-diamine dioleate is a solid at 20°C, and its melting point is 38 °C. No classification for aspiration hazard is therefore warranted.